Abstract:The traditional sinus lifting uses bone grafts to fulfill extended space in order to promote bone formation. However，it has disadvantages such as donor site deformity，immunologic rejection，high cost-effect etc. Therefore，with the improvements of tissue engineering and biomaterials，some bone substitute materials，eg. blood products and injectable hydrogel，were used in bone augmen－tation procedures. Besides，sinus lifting without any grafts was reported to be successful. Furthermore，in order to avoid the complications and complexity of sinus augmentation，some alternative strategies had been introduced to enable implantation without sinus lifting. There are still debates about the necessity and optimal choices of grafting materials and alternative treatment of sinus lifting proce－dures，so the aim of the study is to make a mini-review about the alternative therapy of sinus augmentation including the osteogenesis mechanism，influencing factors，grafting material selection and alternative surgery.

Abstract:Objective：To investigate the effect of tissue engineering biomaterials based on adipose-derived stem cells（ADSCs） for healing calvarial defects. Methods：ADSCs were isolated，cultured and conducted the three-lineage differentiation of osteogenesis，adipogenesis and chondrogesis. Porous chitosan-based scaffolds were prepared and ADSCs were seeded onto the surface of scaffold. Microstructural changes before and after the implantation of ADSCs were detected by scanning electron microscope（SEM）. Cell viability after ADSCs loaded on scaffolds was determined by cell viability assays. A total of 15 male Sprague Dawley（SD） rats were selected to create cranial defect models，and the defects were left in empty or implanted with chitosan scaffolds which were seeded with low（2.5×105） or high densities（1×106） of ADSCs. After operation of 8 weeks，samples were collected for Micro-CT analysis. And then，those samples were decalcified，paraffin embedded and sectioned；hematoxylin-feosin staining and immunohistochemistry stain－ing were performed to observe the bone formation biomarker of bone sialoprotein. Results：ADSCs were able to induce osteoge－nesis，chondrogenesis and adipogenesis. Data from SEM and cell viability assays showed that chitosan-based scaffold had porous structure，and ADSCs were able to attach to the surface of scaf－folds and proliferate，and proliferation effects on scaffolds sur－face were not significantly different when compared with those on the culture plate. In the rat calvarial defect model，the bone volume ratio（BV/TV） in the high-density ADSCs/chitosan scaf－fold group was increased by 140.8%（P=0.014） and 37.4%（P=0.036） when compared with that in the blank control group and the low-density ADSCs/chitosan group，respectively. Compared with that in the control group，bone density in the low-density ADSCs/chitosan group and high-density ADSCs/chitosan group were in－creased by 29.4%（P=0.123） and 64.1%（P=0.022），respectively. Histological results showed that distribution and quantity of the new bone tissue in the bone defect area were consistent with results of Micro-CT analysis. Expression of bone formation markers of bone sialoprotein had an increasing tendency in the high-density group. Conclusion：High density of ADSCs-loaded chitosan scaffolds can effectively promote the healing of calvarial defect in rats.

Abstract:Objective：To investigate the effect of precursor brain-derived neurotrophic factor（proBDNF） on the migration and invasion of SCC-4 cells in oral squamous cell carcinoma（OSCC），and to explore its underlying mechanism. Methods：After stimulation of SCC-4 cells with 20 ng/mL proBNDF，the cell migration was evaluated by Transwell migration assay and wound healing assay，the cell invasion was evaluated by Transwell invasion assay，and the mRNA and protein expression of p75 neurotrophin receptor （p75NTR） in SCC-4 cells was determined by qRT-PCR and Western blot，respectively. Furthermore，immunofluorescence assay was used to observe the subcellular localization of proBNDF and p75NTR in SCC-4 cells. After knockout of p75NTR，the effects of proB－NDF on cell migration，cell invasion，and expression of genes promoting cell migration and invasion were evaluated by Transwell mi－gration assay and wound healing assay，Transwell invasion assay，and qRT-PCR，respectively. Results：Twenty ng/mL proBDNF signif－icantly promoted the migration and invasion of SCC-4 cells（both P=0.000）. Moreover，pro-BDNF and p75NTR were colo－cated in SCC-4 cells. ProBDNF promoted the mRNA and protein expression of p75NTR in SCC-4 cells（both P=0.000）. Knock－out of p75NTR reversed proBNDF-induced SCC-4 cell migra－tion and invasion and reduced the mRNA expression levels of connective tissue growth factor（CTGF），vimentin（VIM），anex－elekto（AXL），high-mobility group A2（HMGA2），a disintegrin and metalloproteinase 19（ADAM19），fibronectin leucine-rich trans－membrane protein 2（FLRT2），and phosphodiesterase 1C（PDE1C）（all P<0.05）. Conclusion：ProBDNF promotes the migration and invasion of SCC-4 cells by up-regulating p75NTR expression，and its mechanism of action may be related to the up-regulation of the expression of downstream migration and invasion-promoting genes（namely，CTGF，VIM，AXL，HMGA2，ADAM19，FLRT2，and PDE1C） through the binding between proBDNF and p75NTR.

Abstract:Objective：To explore the clinical efficacy of Le Fort I and π osteotomy guided by 3D digital technology in the correction of severe dento-maxillofacial deformities（DENTO）. Methods：seven patients displayed developmental DENTO with irregular maxillary arch were selected. In this study，12 measurement indicators（SNA，SNB，ANB，A-CP，B-CP，PG-CP，OP-FH，MP-FH，N-ANS，ANS-ME，NA-PA，SNPg） were selected to perform 3D head model measurement analysis. The 3D head model measurement data was compared between preoperative 1-week（T0） and postop－erative 3months（T2）. In addition，Lateral cephalograms（T0 and T2） were taken with the patient oriented to the Frankfort Hori－zontal Plane for soft tissue measurements. All data were presented as mean ± SD. Paired t test was used to detect the differences between（T0） and （T2） by SPSS 21.0 software. Equivalence test was used in preoperative simulation（T1） and three months after operation（T2） by SPSS 21.0 software. Results：All patients in this group，T0 and T2 measurements showed that the difference in SNA angle，SNB angle and other major soft and hard tissues were statistically significant（P<0.01）. All measurements of T1 and T2 had no statistical significance. Because both sides of the 90% confidence interval are in the range of （-Δ，Δ）. indicating that under the guidance of 3D digital technology. The pre-surgically designed effects in facial appearance and oral function were obtained after the surgery. Conclusion：there is an effective treatment to correct complex deformities of the maxilla by using the Le Fort I osteotomy combined with π osteotomy under the guidance of 3D digital technology.

Abstract:Objective：To observe the effect of a single intravitreal injection of conbercept in the treatment of neovascular age-related macular degeneration（nAMD），to evaluate the short-term efficacy of conbercept in the treatment of nAMD through investigating the correlation between retinal anatomy and retinal function by spectral-domain optical coherence tomography（SD-OCT） and multifocal electroretinogram（mf-ERG），and to explore the value of mf-ERG in predicting therapeutic effect and optimizing therapeutic regimen. Methods：A retrospective analysis was conducted in 36 patients with nAMD（36 eyes）. All patients were treated with a single intravit－real injection of conbercept 0.05 mL/0.5 mg. The best corrected visual acuity（BCVA），the central retinal macular thickness（CRT），the volume of RPE uplift in the 1，3，and 6 mm circles with the macula lutea as the center（1RV，3RV，6RV），and the amplitude density and latency of P1 wave in mf-ERG R1 ring were recorded before and at 1 month after treatment. Statistical analysis was performed using SPSS 22.0 software. Continuous data were compared between two groups using the paired t-test，and the correlation between macular structure and function was evaluated by Pearson cor－relation analysis，with a test level of 0.05. Results：After treat－ment with conbercept in the 36 eyes of 36 patients，the mean BCVA significantly improved（0.19±0.16 vs. 0.26±0.17），the mean CRT significantly decreased[（472.14±181.53） μm vs. （389.5±124.79） μm]，the mean 1RV significantly decreased[（0.36±0.14） mm3 vs. （0.31±0.11） mm3]，the mean 3RV significantly decreased[（3.04±0.98） mm3 vs. （2.59±0.71） mm3]，and the mean 6RV significantly decreased[（8.17±2.91） mm3 vs. （6.66±2.38） mm3] （all P<0.01）. The amplitude density of P1 wave signifi－cantly increased from （36.47±14.25） nv/deg2 before treatment to （47.21±13.13） nv/deg2 after treatment（P<0.01），while the latency of P1 wave showed no significant change after treatment[（38.13±9.75） ms vs. （39.21±9.62） ms，P>0.05]. BCVA had a significant negative correlation with CRT，1RV，3RV，and 6RV before and after operation，but a significant positive correlation with the amplitude density and latency of P1 wave；CRT，3RV and 6RV were significantly negatively correlated with the amplitude density of P1 wave before and after operation（P<0.05）. Subconjunctival hemorrhage occurred in 3 eyes of 3 patients after injection，and all recovered after treatment. No serious ocular and systemic complications were noticed in any of the patients during follow-up. Conclusion：Conbercept has definite short-term efficacy in the treatment of nAMD，which can improve the visual acuity of the affected eyes，and restore the structure and function of the retina with a high safety. BCVA and SD-OCT combined with mf-ERG are more objective and compre－hensive in evaluating the treatment outcome of nAMD，and mf-ERG is expected to be an objective functional indicator for evalu－ating the efficacy and judging the re-treatment during the treatment of nAMD.

Abstract:Objective：To investigate factors influencing prognosis of early vitrectomy on open globe injuries. Methods：Clinical data of 25 patients with open globe injury who underwent early vitrectomy from July 2016 to June 2018 in our hospital were retrospectively analyzed，including visual acuity after injury，prognostic visual acuity，type of ocular trauma，division of ocular trauma，lens injury and/or dislocation，intraocular foreign body，vitreous hemorrhage，retinal detachment and influences of those factors on prognostic visual acuity. Results：Among 25 patients，10 eyes（40.0%） were injured in area Ⅰ，8 eyes（32.0%） injured in area Ⅱ and 7 eyes（28.0%） injured in area Ⅲ；11 eyes（44.0%） had visual acuity after injury≥0.05 and 14 eyes（56.0%） had visual acuity after injury<0.05；16 eyes（64.0%） had prognostic visual acuity≥0.05 and 9 eyes（56.0%） had prognostic visual acuity<0.05. Visual acuity after injury （P=0.017），trauma zone（P=0.013），vitreous hemorrhage（P=0.033） and retinal detachment（P=0.008） were important factors influenc－ing the prognostic visual acuity. Conclusion：After early vitrectomy，most of patients with open globe injury can achieve functional cure and anatomical cure. The better the visual acuity after injury is，the better the curative effect is. The prognostic visual acuity in area Ⅰ is obviously better than that in area Ⅲ. Vitreous hemorrhage and retinal detachment can significantly influence the prognostic vi－sual acuity.

Abstract:Objective：To explore the application of prechop and conventional stop-and-chop techniques in phacoemulsification and to compare their clinical efficacy and effect on corneal endothelial cells. Methods：Sixty patients（60 eyes） with cataract collected from our hospital from January 2016 to April 2017 were enrolled in the study and were divided into group A（30 eyes，phacoemulsification performed using prechop technique） and group B（30 eyes，phacoemulsification performed using stop-and-chop technique） by surgical approach，with 30 cases in each group. Patients in both groups were observed for surgical effect and changes in corneal endothelial cells. Results：Group A had significantly lower actual ultrasonic energy and significantly shorter effective ultrasonic duration than group B[（50.36±2.47）% vs. （55.42±3.28）%，（56.56±10.20） s vs. （67.79±11.45） s，P<0.05]. A repeated measures analysis of variances showed that there were significant differences in the best corrected visual acuity，endothelial cell density，and proportion of hexagonal cells measured at different time points（P<0.05），with no time-group interaction effects observed in the above three indices in the two groups（P>0.05）. Corneal edema was significantly milder in group A than in group B one day after operation（P<0.05）；the corneal edema resolved seven days after operation with no significant difference observed between the two groups（P>0.05）. Conclusion：Both surgical approaches have a good clinical effect；the prechop technique can effectively reduce ultrasonic energy and shorten ultrasonic duration in a cataract surgery；meanwhile，it can reduce the injury of corneal endothelial cells and is easy to operate. All these make it worthy of clinical application.

Abstract:Objective：To construct a lentiviral vector that is capable of knocking down EPCR in hUC-MSCs and to explore the effect of knocking down EPCR on differentiative capacity of hUC-MSCs into fibroblasts. Methods：Three siRNAs were synthesized according to the mRNA sequence of human EPCR and were transferred into hUC-MSCs. Western blot was conducted to analyze the suppression efficiency of three siRNAs to screen for an optimum siRNA. Then the shRNA targeting the optimum siRNA sequence was synthesized，cloned into lentiviral vector and packaged by lentivirus. HUC-MSCs were infected with the recombinant lentivirus. The effici-ency of knocking down was detected both at mRNA and protein levels. HUC-MSCs were stimulated with TGF-β1. The expression levels of α-SMA and Col Ⅰ were detected by Real-time PCR and Western blot for evaluation of differentiative capacity of hUC-MSCs into fibroblasts. Results：Lentiviral vector that targeting to the optimal siRNA sequence of EPCR and can down-regulate the mRNA（t=28.86，P=0.000） and protein（t=88.60，P=0.000） expression of EPCR was constructed successfully. And knocking down of EPCR in hUC-MSCs decreased the mRNA（P=0.044，P=0.000） and protein（P=0.000，P=0.000） expression of COL1A1 and COL1A2. TGF-β1 could induce up-expression of α-SMA（F=369.713，P=0.000；F=451.060，P=0.000），COL1A1（F=42.051，P=0.000；F=759.041，P=0.000） and COL1A2（F=359.205，P=0.000；F=764.348，P=0.000） in mRNA and protein level and transform hUC-MSCs into fibroblasts. However，knocking down of EPCR in hUC-MSCs significantly decreased the expression of α-SMA（P=0.002，P=0.002） and COL1A1（P=0.002，P=0.000） and COL1A2（P=0.000，P=0.000） induced by TGF-β1 compared with that of control group. Conclusion：Knocking down of EPCR in hUC-MSCs are successfully constructed and could decrease the expression of Col Ⅰ as well as suppress the differentiative capacity of hUC-MSCs into fibroblasts.

Abstract:Objective：To study the effect of andrographolide combined with gentamicin on burn wounds infection. Methods：The burn model infected by Pseudomonas aeruginosa mixed with Staphylococcus aureus was established in 48 BALB/c mice，with four wounds in each. The wounds in each mouse underwent blank intervention（control group），gentamicin intervention（gentamicin group），andro－grapholide intervention（andrographolide group） and andrographolide combined with gentamicin intervention（combined intervention group） respectively. The tissue necrosis was observed under pathological section，and the morphology of mixed biofilm by scanning electron microscopy. The expressions of tumor necrosis factor-α（TNF-α），interleukin-6（IL-6） and interleukin-1β（IL-1β） were de－tected by ELISA. The mixed biofilm model in vitro was established. The expression of exotoxin A（ExoA） was detected by Western blot and the expressions of exoS and exoY by qPCR. Results：Electron microscopy confirmed the successful modeling and biofilm de－struction under different interventions. The pathological scores showed that the prognosis of combined intervention group（4.86±0.38） was better than that of gentamicin intervention group（3.29±0.49） and andrographolide intervention group（4.14±0.69），with P value as 0.000 and 0.02，respectively. ELISA showed that the expres－sions of IL-1β，IL-6 and TNF-α in gentamicin intervention group decreased significantly than those in combined interven－tion group（P<0.05）. Western blot showed that the gray values of ExoA in the supernatant and bacterial cells of the gentamicin intervention group were 43.73±1.07 and 73.53±1.90 respec－tively，which were significantly higher than those in the com－bined intervention group（40.99±0.57 and 53.37±2.31）（P=0.017，P=0.000）. The expressions of exoS and exoY in the gentamicin intervention group were higher than those in the combined intervention group，with P values as 0.003 and 0.011. Conclusion：Andro－grapholide enhances the effect of gentamicin on the repair of burn wounds，which is related to the effects that andrographolide can promote the destruction of biofilm，and the reduction of inflammatory factors and virulence factors.

Abstract:Objective：To establish a novel strategy based on proximity binding-induced entropy-driven amplification circuits for high-sensitivity detection of DNA. Methods：Target DNA was used to trigger proximity binding to form a sandwich complex with two affinity probes and then a proximity trigger，which lit up the entropy-driven amplification circuits by mediating branch migration for DNA detection. Results：Under the optimal experimental conditions，the DNA sensing strategy established in this study showed high sensi－tivity and selectivity，with a wide dynamic range of 0.01-100 nmol/L and a limit of detection as low as 6.7 pmol/L. Conclusion：This rapid，cost-effective，and highly efficient signal amplification strategy provides a simple and sensitive platform for DNA detection.

Abstract:Objective：To construct the lentiviral vector of fibronectin type Ⅲ domain-containing protein 5 （FNDC5） overexpression，and to obtain the THP-1 cell line stably transfected with FNDC5. Methods：FNDC5 gene was amplified by PCR and was then inserted to pLenti-EF1a-EGFP-P2A-Puro lentiviral vector. The recombinant vector was amplified in DH5α competent cells，and after se－quencing and identification，the vectors were transfected into 293T cell with the 4-plasmid packaging system to produce overexpression lentiviral particles. Virus titer was then measured. Western blot was used to measure the expression of FNDC5 in recombinant lentivirus；the recombinant lentivirus was transfected into THP-1 cell line，and a fluorescence microscope was used to calculate trans－fection efficiency. After a stably transfected cell line with FNDC5 overexpression was screened out by puromycin，the cells were divid－ed into blank group，empty group，and FNDC5 group，and Western blot and RT-PCR were used to measure the protein and mRNA expression of FNDC5. Oil red O staining was used to observe the effect of FNDC5 overexpression on lipid accumulation in THP-1 macrophage-derived foam cells. Results：PCR product identi－fication and sequencing results showed that a lentiviral vector of FNDC5 overexpression was successfully constructed. Green flu－orescent protein was detected after transfection of 293T cells，and the Flag-tag expression of FNDC5 was detected by Western blot. The titer of the lentivirus after packaging was 1.32×109 TU/mL. Western blot and RT-PCR were used to analyze the THP-1 cell line transfected with the lentivirus；compared with the empty group，the FNDC5 overexpression group had a significant increase in the protein and mRNA expression of FNDC5（238.0±24.9）% and （228.5±3.4）%，respectively，both P=0.000）. There was a reduction in lipid accumulation in THP-1 macrophage-derived foam cells with FNDC5 overexpression. Conclusion：A lentiviral vector of FNDC5 overexpression is successfully constructed in this study and a THP-1 cell line stably transfected with FNDC5 is established by the lentiviral vector of FNDC5 overexpression，indi－cating that FNDC5 can significantly reduce lipid accumulation in THP-1 macrophage-derived foam cells，which lays a foundation for further research.

Abstract:Objective：To study the pharmacodynamics of multifunctional evodiamine butyryl derivative（EAB） and evodiamine butyryl derivative-loaded lipid nanoparticles（EABLNs） in rats. Methods：EABLNs were prepared by the film ultrasound method；the appear－ance，particle size，potential，and encapsulation efficiency of EABLNs were measured. Thirty-two male Sprague-Dawley rats were ran－domly divided into normal group，model group，EAB treatment group，and EABLNs treatment group；the body weight changes of the rats were monitored daily. After continuous administration by oral gavage for 1 week，followed by administration of hypoxanthine by oral gavage and subcutaneous injection of potassium oxonate，the rat model of hyperuricemia was successfully established. The rats were tested for serum levels of uric acid（UA），creatinine（CR），urea nitrogen（BUN），total cholesterol（TC），triglyceride（TG），low-den－sity lipoprotein（LDL），high-density lipoprotein（HDL），alanine aminotransferase（ALT），and aspartate aminotransferase（AST）. Mean－while，histopathological examinations were performed on the heart，liver，spleen，lung，kidney，and brain of the rats. Results：Electron microscopy showed that EABLNs were successfully prepared. Body weight curves showed that there were no differences（except for the results measured at 7 time points） in body weight between the four different groups. Biochemical analysis showed that，the levels of UA，CR，and BUN were significantly higher in the model group than in the other three groups（F=20.080，F=8.459，F=7.169；P=0.000，P=0.007，P=0.012）；there were no significant differences between the four groups in serum levels of ALT or AST（F=1.701，F=3.528；P=0.244，P=0.068），or serum levels of TC，TG，LDL，or HDL（F=3.069，F=0.398，F=0.191，F=3.291；P=0.091，P=0.758，P=0.899，P=0.079）. Histopathological examinations showed that the integrity of the renal units in the model group was seriously damaged，but the damage of the kidney induced by hyperuricemia was reversed by EABLNs，and EABLNs did not cause damage to the heart，liver，spleen，lung，or brain. Conclusion：It is the first time that EABLNs have been found to be feasible for the treatment of hyperuricemia and its complications.

Abstract:Objective：To investigate the alteration of Lysine-specific demethylase 5C（KDM5C） protein level and the regulatory effect of KDM5C on spindle assembly checkpoint（SAC），related mitotic progression of osteosarcoma cell lines. Methods：The various osteosar－coma cells in prometaphase were collected by synchronization. The alteration of KDM5C protein level and its potential posttransla－tional modification in mitosis were determined by Western blot. The effect of inhibitor CPI-455 on mitosis process was detected using Western blot. Results：In mitosis，the alterations of SAC related markers，cell division cycle protein 27（cdc27） and Cyclin B1，were similar to previous literatures，but KDM5C protein level had no obvious change. The shift and ubiquitination of KDM5C truncations were not observed by Western bolt. When demethylase activity of KDM5C was inhibited by CPI-455，SAC related markers，cdc27 and Cyclin B1，had no obvious change. Conclusion：Mitotic process of osteosarcoma cell and the SAC is unaffected by KDM5C protein level and its demethylase activity.

Abstract:Objective：To investigate the effect of miR-505 on proliferation and metastasis of osteosarcoma cells and its mechanisms. Methods：The miR-505 level in human osteoblast of hFOB1.19 and osteosarcoma of cell MG63 was determined by q-PCR. The miR-505 mimics group and the negative control group（miR-NC） were set. Both miR-505 mimics and miR-NC were transfected into MG63 cells with Lipofectamine TM 2000 and cell proliferation rate and invasive cells were compared between two groups. Proteins and mRNA levels of MMP-9，CDK4，Cyclin-D1，HMGB-1，β-catenin and GSK-3β in MG63 cells transfected with miR-505 was further detected via Western blot and q-PCR technologies. Results：The expression level of miR-505 in osteosarcoma cell of MG63 was 0.57±0.06，which was significantly lower than that of 1.03±0.12 in human osteoblast of hFOB1.19（P<0.01）. After transfecting miR-505，expression level in the miR-505 mimics group was 3.59±0.37，which was significantly higher than that of 1.09±0.12 in the miR-NC group（P<0.01）. After transfecting 48 h and 72 h，A450 nm values in the miR-505 mimics group were 0.84±0.09 and 1.29±0.13，respectively，which were significantly lower than those in the miR-NC group（P<0.05）. Invasive cells in the miR-505 mimics group was （42.84±4.35），which was significantly lower than that of （66.84±6.85） in miR-NC group（P<0.05）. Protein lev－els of MMP-9，CDK4，Cyclin-D1 and HMGB-1 in the miR-505 mimics group were lower than those in the miR-NC group（P<0.05）. The mRNA levels of MMP-9，CDK4，Cyclin-D1 and HMGB-1 in the miR-505 mimics group were lower than those in the miR-NC group（P<0.05）. Meanwhile，protein and gene expressions of β-catenin and GSK-3β in Wnt signaling pathway in the miR-505 mim－ics group were significantly lower than those in the miR-NC group（P<0.05）. Conclusion：The miR-505 can down-regulate the expres－sion of MMP-9，CDK4，Cyclin-D1 and HMGB-1 by inhibiting Wnt signaling pathway，so as to inhibit the proliferation and metastasis of osteosarcoma cells.

Abstract:Objective：To predict the extracellular domain and transmembrane domain of HIV-1 gp41 protein，and to investigate its structure and function. Methods：The gp41 gene and its protein-encoding sequence were obtained from the NCBI GenBank database. ExPASy ProtParam tool，ProtScale，SignalP 4.0 Server，TMHMM Server v.2.0，SWISS-MODEL and Predict Protein software were used to predict the biological properties of gp41 protein. Results：The HIV-1 gp41 gene had an ORF length of 666 bp and encoded 222 amino acids. Its molecular formula was C1164H1815N317O325S6，and the isoelectric point was 8.60. It was a stable and hydrophobic protein. This protein had a transmembrane region and the signal peptide cleavage site was located between the amino acids 20 and 21. Its sec－ondary structure mostly consisted of random coil，β-fold，and α-helix，and thus it was an all-helix protein. It also had an N-terminal signal peptide and guided the extracellular secretion of protein. In addition，gp41 protein had 4 protein-protein binding sites and 1 protein-DNA binding site，which could regulate various biological functions of gp41 protein. Conclusion：This study predicts the structure and function of gp41 protein and lays a foundation for research on the molecular mechanism of gp41 protein in disease de－velopment and progression.

Abstract:Objective：To construct attenuated Salmonella typhimurium carrying the lipoprotein 2（Lcn2） gene and to investigate its effect on the growth of mycobacteria. Methods：RNA was extracted from macrophage RAW264.7 cells and reverse transcribed into cDNA. The Lcn2 gene was amplified by PCR using the cDNA as a template and inserted into the multiple cloning sites of the plasmid pBudCE4.1-orim to construct the recombinant plasmid pBudCE4.1-orim-Lcn2. The recombinant plasmid was trans－fected into RAW264.7 cells. The mRNA expression of Lcn2 was measured by RT-qPCR，with the empty plasmid for a control group. Then the recombinant plasmid was transfected into the attenuated Salmonella typhimurium SL7207 to obtain the recom－binant strain SL7207-pBudCE4.1-orim-Lcn2. RAW264.7 cells were infected with the recombinant strain. The protein expres－sion of the Lcn2 gene was measured by Western blot，and its mRNA expression was measured by RT-qPCR，with the empty strain for a control group. Bacterial colonies were counted on the 7H10 plate to observe the effect of the recombinant strain on the survival of bacille Calmette-Guérin（BCG） in RAW264.7 cells，with the empty strain for a control group. Mice were intragastrically given the re－combinant strain to measure the Lcn2 expression in the spleen and the interferon gamma（IFN-γ） level in serum，with the empty strain and normal saline for control groups. Results：The Lcn2 gene was amplified by PCR，and the recombinant plasmid was identified by double-enzyme digestion and DNA sequencing. The protein expression of Lcn2 in RAW264.7 cells infected with the recombinant strain was significantly increased compared with the control group（P=0.000）. The RT-qPCR showed that the mRNA expression of Lcn2 in the recombinant strain group and the recombinant plasmid group was significantly higher than that in the control groups（F=5.281，P=0.000；F=22.570，P=0.000）. The colony number of BCG on the 7H10 plate in the recombinant strain group was significantly reduced compared with the control group（F=11.41，P=0.000）. The expression of Lcn2 in mouse spleen tissues and the level of IFN-γ in serum were significantly higher in the recombinant strain group than in the control groups（F=4.449，P=0.000；F=15.27，P=0.004）. Conclusion：Recombinant attenuated Salmonella typhimurium carrying the Lcn2 gene was successfully constructed. The expression of Lcn2 increased in RAW264.7 cells infected with the recombinant strain，which inhibited the survival of BCG in the cells. The expres－sion of Lcn2 in mouse spleen tissues and the IFN-γ level in serum were elevated after the mice were intragastrically given the recom－binant strain，so that the immune state of mice was beneficial to anti-tuberculosis outcome. This study lays a foundation for further study of the relationship between lipid carrier proteins and Mycobacterium tuberculosis infection and its molecular mechanism.

Abstract:Objective：To investigate the effects of the phospholipase Cδ1（PLCD1），a potential tumour gene，in pancreatic cancer cells，on the proliferation，migration and invasion of pancreatic cancer cells. Methods：Liposome method was used to transfer the pcDNA3.1-PLCD1 to pancreatic cancer cells and the Vector group with empty vector was set. The proliferation,migratory and invasive abilities of pancreatic cancer cells were detected with CCK-8 assay，clone formation assay，Transwell assay and wound healing assay. In additional，Western blot was used to demonstrate the potential mechanism. Results：PLCD1 was significant up-regulated in CAPAN-1 and BX－PC-3 cells after being transiently transfected with the PLCD1 over-expressing plasmid（P＜0.05）. The ability of proliferation showed significant inhibition with CCK-8 and clone formation assay（P＜0.05）. The motility，invasion and migration were remarkable restrained with Transwell assay and wound healing assay（P＜0.05）. Additionally，Western blot showed PLCD1 induced MAPK/ERK pathway molecule p-ERK1/2（P＜0.05），as well as the mesenchymal markers N-cadherin and Vimentin were decreased（P＜0.05）. And the ex－pression of the epithelial marker E-cadherin was increased（P＜0.05）. Conclusion：PLCD1 may inhibit the proliferation，migration and invasion of pancreatic cancer cells through inhibiting the MAPK/EKR pathway and suppressing the EMT，which lay the foundation for the further study of PLCD1 in pancreatic cancer.

Abstract:Objective：To investigate the regulatory effect of berberine on NLRP3/TLRs signal pathway in mice with gouty arthritis. Methods：C57BL/6 mice were randomly divided into 6 groups：normal control group，model group，colchicine group（positive control，0.65 mg/kg），and berberine groups with high，middle and low doses（150，100，50 mg/kg），respectively. Administration for 7 days after modeled，and the swelling degree of the ankle joint was measured every day to observe therapeutic effects. Meanwhile，serum inter－leukin-2（IL-2），interleukin-6（IL-6），tumor necrosis factor-α（TNF-α） and interferon-α（IFN-α） expression levels were measured by ELISA. And the expressions of key target genes and proteins of NLRP3/TLRs in the mice's ankle synovial tissue were detected by RT-PCR and Western blot assays. Results：Compared with the model group（31.34%），the swelling degrees of the ankle joint of the berberine groups with high，medium and low doses（18.35%，21.35%，25.35%，respectively） and the colchicine group（15.35%） were significantly reduced，and the difference was statistically significant（F=11.240，P=0.005）. After the berberine treatment with different doses，inflammatory cell infiltration and edema in the ankle synovial tissue were significantly alleviated. The ELISA results showed that compared with the model group，the serum levels of IL-2，IL-6，TNF-α and IFN-α in the berberine groups were significantly decreased（IL-2：F=5.690，P=0.005；IL-6：F=10.370，P=0.009；TNF-α：F=15.240，P=0.002；IFN-α：F=4.520，P=0.020）. RT-PCR and Western blot results showed that the expression levels of NLRP3，TLR3，TLR7，MyD88 and NF-?资B p65 genes and proteins were obviously decreased in the mice's synovial tissue of the berberine groups with high，medium and low doses（P<0.05）. Conclusion：Berberine can alleviate gouty arthritis in mice，and its mechanism may be related to down-regulating inflammatory cytokine levels and inhibiting NLRP3/TLRs signal pathway.

Abstract:Objective：To investigate the expression of F-box and WD40 domain-containing7（Fbxw7） in acute T lymphocyte leukemia and its effect on cell growth as well as its mechanism. Methods：The mRNA expression of Fbxw7 in 62 patients with acute T lympho－cyte leukemia was detected by RT-PCR. The lentivirus vector carrying Fbxw7 overexpression was transferred to leukemia Jurkat cells and the effect of transfection was tested by RT-PCR and Western blot. And the proliferation of Jurkat cells was measured by MTT as－say. The cell cycle and apoptosis were examined by flow cytometry，and the expression of c-Myc and Cyclin E protein was checked by Western blot. Results：Expression levels of Fbxw7 mRNA in relapse group and initial group were significantly lower than those in normal healthy group（P<0.05）. After transfection，relative expression levels of Fbxw7 mRNA and protein in the Fbxw7 overexpression group（transfected with Fbxw7 overexpressed lentivirus vector） were significantly higher than those in the control group （no transfected cells） and negative control group（transfected with control lentivirus vector）（P<0.05）. Cell proliferation was not significantly influenced（P>0.05） after transfection of 24 h，while was significantly inhibited after transfection of 48 h and 72 h（P<0.05）. After transfection of 48 h，when compared with control and negative control groups，cell apoptosis and the proportion of cells in G0/G1 phase of Jurka cells were significantly increased in Fbxw 7 group，S phase and G2/M phase were significantly decreased，and relative expression levels of c-Myc and Cyclin E were significantly decreased，all with statistically significant difference（P<0.05）. Conclusion：Fbxw7 is low expression in acute T lymphoblastic leukemia，its overexpression can inhibit the proliferation of Jurkat leukemia cells and promote cell apoptosis，and its mechanism may be related to the down-regulation of c-Myc and Cyclin E protein expression.

Abstract:Objective：To explore the diagnostic value of plasma neutrophil gelatinase-associated lipocalin（NGAL） in patients with asthma-COPD overlap（ACO）. Methods：A total of 30 patients with ACO，35 patients with bronchial asthma（asthma），60 patients with chronic obstructive pulmonary disease（COPD） and 25 healthy nonsmokers（control） were enrolled in this study. Their general data were collected and their pulmonary function was completed. Levels of plasma NGAL were determined by ELISA；univariate analysis of variance was used to evaluate the expressive difference of plasma NGAL levels in the ACO group，the asthma group，the COPD group and the control group；the relationship of NGAL and confounding factors was analyzed；the receiver operating characteristic curve （ROC） was used to assess the diagnostic value of NGAL on differentiating ACO and non-ACO. Results：Plasma NGAL levels in the ACO group and the COPD group were （110.50±22.81） ng/mL and （127.40±27.57） ng/mL，respectively，which were higher than those of （85.79±20.02） ng/mL in the asthma group and （86.54±23.46） ng/mL in the control group. Plasma NGAL levels were sta－tistically different between ACO group and asthma group（P=0.000），as well as between ACO group and COPD group（P=0.015）. Plasma NGAL levels were negatively correlated with forced expiratory volume in one second（FEV1）（r=-0.422，P=0.000），and were positively correlated with smoking（r=0.500，P=0.000）. The area under ROC curve（AUC） was 0.565（P=0.273）. Conclusion：Plasma NGAL may have a certain value to differentiate ACO，asthma，as well as COPD，and may be a potential biomarker for ACO.

Abstract:Objective：To explore the relationship between the innate lymphoid cells（ILCs） and the occurrence，development and prognosis of non-small cell lung cancer（NSCLC）. Methods：70 patients diagnosed as NSCLC from January 2012 to January 2014 in the First Affiliated Hospital of Anhui Medical University were enrolled as NSCLC group. 70 healthy examiners with matching gender ratio and age in the hospital for physical examination at the same time period were selected as control group. Flow cytometry was used to detect the proportion of ILCs subsets in the two groups. The levels of ILCs-related cytokines were detected by enzyme-linked immunosorbent assay（ELISA）. The clinical data and follow-up data were collected，and the relationship between ILC1，ILC2 and ILC3 and clinical features and prognosis of NSCLC was analyzed. Results：Compared with control group，the levels of ILC1 and IFN-γ with （8.61±3.78）% and （27.24±12.85） ng/L in NSCLC group decreased（P<0.05），while the levels of ILC2，ILC3，IL-5 and IL-17 with （1.29±0.64）%，（3.64±1.93）%，（22.70±9.94） ng/L and （8.10±2.09） ng/L increased（P<0.05）. ILC1 was positively cor－related with IFN-γ（r=0.780，P<0.05），ILC2 was positively correlated with IL-5（r=0.704，P<0.05），and ILC3 was positively correlated with IL-17（r=0.654，P<0.05）. The levels of ILC1，ILC2 and ILC3 were associated with lymph node metastasis and TNM staging（P<0.05）. The overall survival（OS） was significantly longer among patients with no lymph node metastasis，low TNM stage，ILC1 level ≥8.98% and ILC2 level <1.37%（P<0.05）. The median survival time was 49 months in patients with ILC1≥8.98%，and 38 months in patients with with ILC1<8.98%，and there were significant differences in the survival curves between the two groups（P<0.05）. The median survival time was 34 months in patients with ILC2≥1.37% and 46 months in patients with ILC2<1.37%，and there were significant differences in the survival curves between the two groups（P<0.05）. Multivariate analysis showed that lymph node metastasis，TNM stage III，IV and ILC2≥1.37% were indepen－dent risk factors for OS（P<0.05），and ILC1≥8.98% was an independent protective factor for OS（P<0.05）. Conclusion：NSCLC patients have an imbalance of ILCs，and ILC2 and ILC3 have obvious advantages. The detection of ILC1 and ILC2 can evaluate the development and prognosis of disease to a certain extent.

Abstract:Objective：To explore the effects of high-volume platelet-lowering therapy on the comprehensive coagulation indices and fibrinogen concentration of patients with thrombocytosis using a continuous-flow cell separator（CFCS）. Methods：Patients with throm－bocytosis underwent a platelet-lowering therapy administered by the ELP/MNC program of a COBE Spectra CFCS using ACD-A as an anticoagulant. A total of 32 platelet-lowering treatments were performed for 21 patients with the treated blood volume for each treatment set as 2.5 to 3.0 times the total blood volume（TBV） and the collected platelet suspension volume as 20% to 25% of TBV. The effect of platelet-lowering therapy and the changes in coagulation indices and fibrinogen concentration after treatment were eval－uated. Results：A single treatment had a run time of （212.53±41.54） minutes with a treated blood volume of （8 812.63±2 087.15） mL and a collected platelet suspension volume of （798.84±190.77） mL. The patients’ coagulation indices in circulating blood before and after treatment were as follows：platelet count：（1426.46±530.23）×109 cells/L vs. （778.83±247.25）×109 cells/L（t=10.808，P=0.000），prothrombin time（PT）：（12.82±1.53） s vs. （13.28±1.51） s（t=3.921，P=0.000），activated partial thromboplastin time（aPTT）：35.75 （29.43，37.55） s vs. 35.40（31.20，38.20） s（Z=3.021，P=0.003），thrombin time（TT）：（17.46±1.30） s vs. （17.88±1.41） s（t=2.783，P=0.009），fibrinogen concentration（Fbg）：（2.96±1.18） g/L vs. （2.81±1.11） g/L（t=3.433，P=0.002）. Conclusion：High-volume platelet-lowering therapy using a CFCS can significantly reduce the platelet load within the body of patients with thrombocytosis，and its effects on PT，aPTT，TT，and Fbg are within the normal range of body compensation.

Abstract:Objective：To investigate the clinical application of the pediatric early warning score（PEWS） in septic shock of leukemia children with neutrophil deficiency. Methods：A total of 67 leukemia patients who suffered from neutrophil deficiency and complicated infection after chemotherapy in Children’s Hospital of Chongqing Medical University from January to December，2016 were included in the study as retrospective observation group. A retrospective study was performed on their clinical data. The PEWS was determined based on their monitoring data to establish the receiver operating characteristic curve and obtain the optimal cut-off value of PEWS for septic shock. A total of 72 leukemia patients who suffered from neutrophil deficiency and complicated infection after chemotherapy from January to December，2017 were included in the study as prospective study group. The PEWS was used for the early evaluation of the patients in the prospective study group. According to different PEWS values，the patients were given corresponding interven－tion measures，including anti-infective therapy，oxygen thera－py，fluid resuscitation，vasoactive agent，component transfusion，and recombinant activated protein C. The incidence rate of septic shock and the mortality rate due to septic shock were compared between the two groups. Results：The PEWS value of 3.5 was the optimal cut-off value for predicting septic shock in leukemia children with neutrophil deficiency，with a sensitivity of 83.3% and a specificity of 83.7%. The prospective study group had significantly lower incidence rate of septic shock and mortality rate due to septic shock than the retrospective observation group（P<0.01）. Conclusion：PEWS can be used for effective evaluation of condition and early identification of signs of septic shock in leukemia children with neutrophil deficiency. Accordingly，taking specific treat－ments and nursing interventions at the appropriate time will improve the prognosis of the disease.

Abstract:Objective：To investigate the risk of hypertension in adults with different insomnia symptoms in Jiangbei District of Chongqing，China. Methods：A total of 2000 residents in Jiangbei District of Chongqing were randomly selected as subjects. The resi－dents with one or more insomnia symptoms for at least one week were selected as insomnia group，and the rest of the residents were included as non-insomnia group. According to insomnia symptoms（sleep latency，number of awakenings，early awakening time，and total sleep time），the insomnia group was divided into four subgroups（sleep latency subgroup，sleep maintenance disorder subgroup，early awakening subgroup，and short total sleep time subgroup）. A logistic regression analysis was performed to identify the risk factors for hypertension. Results：A total of 612（30.6%） individuals who were suffering from insomnia for at least one week were included in the insomnia group. Sleep maintenance disorder（8.0%） was the main insomnia symptom. The insomnia group had significantly higher family income，overweight rate，and drinking rate than the non-insomnia group（P<0.05）. Among the four subgroups of the insomnia group，the sleep maintenance disorder subgroup had the highest prevalence rate of isolated systolic hypertension（7.0%）；the sleep latency subgroup had the highest prevalence rate of classical hypertension（systolic pressure>140 mmHg and diastolic pressure >90 mmHg）（6.4%）；the early morning awakening subgroup had the highest prevalence rate（6.6%）；the short total sleep time sub－group had the highest prevalence rate of classical hypertension（6.9%）. The sleep maintenance disorder subgroup had a significantly higher risk of isolated systolic hypertension than the non-insomnia group（OR=1.603，95%CI=1.042-2.466，P<0.05）. The short total sleep time subgroup had a significantly higher risk of classical hypertension than the non-insomnia group（OR=1.711，95%CI＝1.086-2.696，P<0.05）. Conclusion：Insomnia adults have a higher risk of hypertension than those without insomnia. The risk of hyperten－sion is closely associated with sleep maintenance disorder and short total sleep time.

Abstract:Objective：To investigate the behavioral response and physiological reaction to pain before，during，and after the turning procedure in patients undergoing invasive mechanical ventilation，with the application of the Chinese version of Critical-Care Pain Observation Tool（CPOT）. Methods：A total of 120 patients who underwent invasive mechanical ventilation in the intensive care unit （ICU） in Department of Respiratory Medicine in a tertiary general hospital from January 2015 to January 2017 were enrolled. The Chinese version of CPOT was used to observe pain response at three stages of the turning procedure（at 1 minute before the turning procedure，during the turning procedure，and at 10 minutes after the turning procedure）. The scores of CPOT and each item，related physiological indices（mean arterial pressure，heart rate，respiratory rate，and blood oxygen saturation），and percentage of observations with a score of 1 to 2 points on each item of the CPOT scale were recorded at the three stages of the turning procedure. The scores of CPOT and each item and related physiological indices were compared between stages. Results：The score of CPOT was 0.35±0.99 at 1 minute before the turning procedure，1.96±1.77 during the procedure，and 0.12±0.58 at 10 minutes after the procedure. Compared with baseline levels，facial expression had the greatest increase as high as 72%，followed by body movements as high as 47%，compli－ance with mechanical ventilation as high as 36%，and muscle tension as high as 36%. There were significant changes in CPOT score and physiological indices in different stages of the turning procedure（P<0.05）. Conclusion：Observation of behavioral and physiological changes during the turning procedure helps medi－cal staff to perform an objective evaluation of pain in critically ill patients with impaired verbal communication. The Chinese version of CPOT scale can effectively measure the pain process in pa－tients and thus holds promise for clinical application in patients undergoing invasive mechanical ventilation.

Abstract:Objective：To understand the current status and knowledge base of enhanced recovery after surgery（ERAS） in our country，and to provide a reference for accelerating further development of ERAS in China. Methods：A literature search was performed in Web of Science database to collect the relevant literature on ERAS in China. After analyzing the general information of the literature，CiteSpace was used to visually analyze the literature. Results：A total of 169 SCI articles on ERAS in China were collected. The amount of literature showed an increasing trend in the past 10 years. The main type of research in the literature was experimental study. The research content and knowledge base were mainly related to the research fields of gastrointestinal surgery and hepatopan－creatobiliary surgery. Conclusion：ERAS in China needs operational and repeatable practice protocol. The focus on patient-reported outcomes and patient’s perspectives are needed to help improve and optimize the implementation strategy of ERAS.