Abstract:Objective: 4-nitro-1-naphthyl phosphate (NNPP) as a chromogenic substrate was synthesized for the assay of alkaline phosphatase (ALP). Methods: (4-nitronaphthol, 4-NNP) was reacted with phosphorus oxychloride to yield NNPP. By recording the absorbance of 4-NNP during the hydrolysis of NNPP, initial rates of ALP were estimated. Michaelis-Menten constant (Km) was estimated by Lineweaver-Burk plot. Results: The hydrolysis of NNPP by ALP gave the product with a maximum absorbance peak around 460 nm and an isoabsorbance wavelength around 405 nm. In comparison to the absorptivity of 4-nitrophenol, the absorptivity of 4-NNP is at least three-times at pH from 6.0 to 7.0, and is about twice at pH over 7.0. Km for NNPP is about 12 ?mol/L while that for 4-nitrophenylphosphate is about 35 ?mol/L; the competitive inhibition constant of phosphate against both substrates was about 20 ?mol/L. Activity of ALP on NNPP is about 40% of that on 4-nitrophenylphosphate. Conclusion: NNPP is an effective chromogenic substrate for the assay of ALP and is promising for simultaneous assay of two enzymes in one reaction solution by using a derivative of 4-nitrophenol as the chromogenic substrate of another enzyme.

Abstract:Objective: To investigate the effect of murine gamma herpesvirus 68 (γHV68) infection on lipid accumulation in livers of C57BL/6J mice and to explore potential mechanisms. Methods: C57BL/6J mice fed a high fat diet were randomly assigned to receive either γHV68 or PBS treatment. Serum was analyzed for inflammatory cytokines. Liver was taken for histology and lipid analysis. Quantitative RT-PCR was used to measure expression of hepatic tumor necrosis factor alpha (TNFα), sterol regulatory element binding protein-1 (SREBP1), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC). Results: γHV68 infection promoted lipid accumulation in livers of C57BL/6J mice in association with elevated serum and hepatic inflammatory cytokines levels. The expression of SREBP1, FAS and ACC in livers of C57BL/6J mice were increased in the γHV68-infected mice. Conclusion: γHV68 infection promotes liver lipid accumulation in C57BL/6J mice via increasing triglyceride synthesis.

Abstract:objective: To investigate whether inflammation enhances lipid accumulation via SREBP1 in liver of C57BL/6J mice. Methods: 12 male mice were subcutaneously injected with 0.5ml 10% casein or PBS. The mice were fed a high fat diet for 18 weeks. TG and FFA in liver and serum amyloid A (SAA) and IL-6 in serum were assayed. The lipid accumulation in liver was evaluated by Oil Red O staining. The mRNA and protein expression of TNF-α, MCP1, SREBP1, ACCα, FAS were analyzed by Real Time PCR and Western blotting. Results: Casein injection elevated serum levels of IL-6 and SAA in mice. In the liver of casein-injected mice, mRNA and protein expression of TNF-α, MCP-1 were significantly increased compared with control mice, suggesting that inflammatory stress was successfully induced in C57BL/6J mice. The inflammation up-regulated mRNA and protein expression of sterol regulatory element binding protein 1, fatty acid synthase, and acetyl CoA carboxylase alpha in liver of C57BL/6J mice. Conclusions: Inflammation may exacerbate lipid accumulation via up-regulating SREBP1 expression in liver of C57BL/6J mice.

Abstract:Objective：To study the effects of DNA extraction methods on deoxyribonucleic acid-short tandem repeats（DNA-STR） typ－ing of human tissue after high-temperature disposal. Methods：Human tissue samples after high-temperature disposal were initially handled with different concentrations of TritonX-100. DNA was extracted using the method of Chelex-100 and was amplified by re－combination of DNA-STR. Then，ABI-3130 sequence analyzer was used for electrophoresis separation and Gene mapper ID V3.2 gene analysis software was used for gene typing. Results：1% TritonＸ-100+Chelex-100 was used to process the samples before DNA ex－traction. All loci could be detected after QIA quick PCR purification and the loci could be balancedly amplified. Conclusions：This is a simple method with satisfied results，which is recommended in the field of forensic science，especially for the prosecution case.

Abstract:Objective：To evaluate the replication of virus by extracting virus DNA from low copy number of hepatitis B virus （HBV） serum samples，doing PCR amplification to acquire full-length HBV DNA fragments and transfecting HuH7 cells with full-length HBV DNA fragments after treatment. Methods：Primers for fragment and full-length amplification containing BspQⅠ restrictive en－zyme cutting site were designed and HBV DNA from patients was used as templates. Full-length HBV was acquired by nested PCR combined with sectional amplification. Closed circular DNA was got through restrictive enzyme digestion and T4 DNA ligase ligation. HBV replication level was analyzed by Southern blot after HuH 7 cells being transfected with closed circular HBV DNA. Results：Five complete HBV genomes were successfully obtained using the new PCR method. After digestion，ligation and tansfection experiments，Southern blot results all supported replication expression. Conclusions：This method lays a foundation for investigating the correlation of different HBV replication levels and DNA variation in patients with low copy number of virus.

Abstract:Objective：To investigate the practical value of CT angiography（CTA） in laparoscopic splenectomy（LS） with amputation of secondary structures of the spleen peclicel. Methods：Clinical records of 62 patients with idiopathic thrombocytopenic purpura（ITP） were retrospectively analyzed. Clinical data of patients underwent CTA scan（CTA group） and those did not underwent CTA scan but did LS（control group） were compared. Results：Operative time was （185.58±14.09） min and （215.00±22.01） min in CTA group and control group，Intraoperative blood loss was （165.78±88.42） ml and （262.78±80.70） ml，postoperative hospital stay was （8.46±1.48） d and （10.14±1.51） d，with statistical significances（P<0.05）. Conclusions：Anatomical structure of spleen blood ves－sels and their relationship with pancreas can be mastered before surgery in CTA group，which can guide better dissection of spleen blood vessels in surgery and also contribute to Hem-O-lok ligation and cut off of the blood vessels using harmonic scalpel rather than using Endo-GIA. CTA group displays shorter operative time，less bleeding and shorter hospital stay than control group. Therefore，CTA scan is of clinical valuable for ITP patients who need the treatment of LS.

Abstract:Objective：To compare the short-term efficacy between laparoscopy-assisted D2 radical gastrectomy and conventional open surgery for early distal gastric cancer and to explore the value of laparoscopy-assisted D2 radical gastrectomy. Methods：Sixty patients with distal gastric cancer from December 2008 to March 2012 were enrolled；30 patients received laparoscopy-assisted surgery were taken as observation group while 30 patients underwent conventional open surgery as control group. Length of incision，operation time，intraoperative blood loss，the first anal exhausting time，numbers of dissected lymph node，postoperative complications，postoperative hospital duration were observed and compared. Results：Observation group vs. control group: length of incision（5.98±1.56） cm vs.（13.02 ±2.15） cm（P=0.000）；operation time （189.02±13.78） min vs. （150.58±15.31） min（P=0.000）；intraoperative blood loss （100.89±23.03） ml vs. （156.36 ±30.37） ml （P=0.000）；the first anal exhausting time（3.67±1.56） d vs. （5.04±2.08） d（P=0.005）；numbers of dissected lymph node （18.87±5.03） vs. （17.03±3.67）（P=0.011）；postoperative hospital duration（10.58±3.17） d vs. （13.59±4.36） d（P=0.003）. Differences between groups were all of statistical significances. Conclusions：Laparoscopic-assisted D2 radical gastrectomy has more advantages than coventional open surgery in the treatment of distal gastric cancer.

Abstract:Objective：To compare the clinical efficacy of retroperitoneal laparoscopic radical nephroureterectomy and open nephro-ureterectomy in treating transitional cell carcinoma of the upper urinary tract. Methods：Forty-four cases of upper urinary tract transi－tional cell carcinoma were treated with radical nephroureterectomy and were divided into laparoscopy group（n=15） and open surgery group（n=29）. Patients in laparoscopy group underwent complete retroperitoneal laparoscopic nephroureterectomy while those in open surgery group underwent open nephroureterectomy. Operation time，intra-operative blood loss volume，post-operative painkiller appli－cation dosage and time，recovery time of intestinal function，time of getting out of bed，length of hospital stay and complications were compared and analyzed. All cases were followed up for 6-42 months，survival rates，incidences of bladder carcinoma recurrence and distant metastasis were recorded and compared. Results：Operations of both groups were successfully performed. Blood loss volume and application dosage of painkiller were fewer in laparoscopy group than in open surgery group （116.1±29.4） ml vs. （278.6±84.3） ml，（33.3±24.4） mg vs.（155.2±62.8） mg，P=0.000. Application time of painkiller and hospital stay were shorter in laparoscopy group than in open surgery group （0.7±0.5） d vs. （2.3±0.9） d，（8.7±1.6） d vs. （13.1±3.2） d，P=0.000. Time of getting out of bed was earlier in laparoscopy group than in open surgery group （2.1±0.5） d vs. （4.2±1.0） d，P=0.000. No significant difference was found between two groups in operation time （212.1±19.5） min vs. （251.7±24.2） min and recovery time of intestinal function （1.9±0.8） d vs. （2.9±0.8） d as well as in survival rates，postoperative complications，incidences of bladder carcinoma recurrence and distant metas－tasis during the follow-up period（P >0.05）. Conclusions：With the advantages of minimal invasion，complete retroperitoneal laparo－scopic radical nephroureterectomy conforms to the principles of oncology and should be widely applied as an ideal surgical treatment method for upper urinary tract transitional cell carcinoma.

Abstract:Objective：To investigate the relationship between infection and pediatric acute urticaria and to analyze the clinical charac－teristics of pediatric acute urticaria caused by infection. Methods：Clinical data of 149 patients with acute urticaria were collected from January 2009 to March 20l2 and were analyzed retrospectively. Results：（1）Among 149 cases of pediatric acute urticaria，127 cases（85.2%） was caused by infection，especially respiratory tract infection. Numbers of white blood cell，neutrophile granulocyte and C-reactive protein were increased indicating the bacteria infection. （2）In virus detection，positive rate of coxsackie virus was 24.5%，obviously higher than that of other common viruses. （3）Features of skin lesion：the lesion lasted for 0.5-1.0 d and tended to spread to the whole body；annular or geographic papules and plaques were seen in most patients and a few patients might develop into hemor－rhagic lesions；meanwhile，some lesions faded to leave geographic border or brownish pigmentation；besides，53.5% cases were accom－panied with angioedema；skin manifestations might come with the severity of infection；the duration was about 7.0-11.0 d. （4）64.4% cases were accompanied with digestive symptoms and 43.5% cases were accompanied with increased of creatine kinase-MB；（5）An－ti-allergic agents as well as anti-infective drugs had positive effect. Conclusions：（1）Infection is the main cause of pediatric acute urticaria and the bacteria may dominate the infection. （2）In virus infection，coxsackie virus is the most common. （3）There are some special features in skin lesion of pediatric acute urticaria caused by infection. （4）In fact，quite a few patients are accompanied with di－gestive symptoms and some of which may get myocardial injury. （5）We should attach importance to enough course of anti-infection treatment.

Abstract:Objective：To study advantages and disadvantages of single stapling technique（SST） and double stapling technique（DST） in transanal pull-through rectal resection operations for ultra low rectal cancer. Methods：Totally 1 138 patients with rectal cancer were treated in our department from July 2008 to July 2011.One hundred and thirteen cases underwent transanal pull-through rectal resec－tion operations：49 cases by SST and 64 cases by DST. Operative time，postoperative complication incidences including anastomotic leakage，bleeding and stenosis，pelvic floor recurrence within 1-4 years，hospitalization duration and costs were analyzed and com－pared between the two groups. Advantages and disadvantages of the two stapling techniques were analyzed statistically. Results：Differ－ences in average operative time and postoperative hospitalization duration between the two groups were not statistically significant（op－erative time t=0.172，P=0.864；average hospitalization duration Z=-0.562，P=0.547）. Postoperative complications：differences in anas－tomotic leakage between the two groups were statistically significant（ χ2=6.592，P=0.01）；differences in other complications between the two groups were not statistically significant（ χ2=0.037，P=0.848）. Differences in pelvic floor local recurrence within 1-4 years between the two groups were not statistically significant （ χ2=1.839，P=0.175）. Differences in average hospital cost were statistically significant（Z=-5.782，P=0.00）. Conclusions：Transanal pull-through rectal resection has become a commonly used technique for ultra-low rectal cancer. With the advantages of simpler and safer manipulation，lower cost and recurrences and more complete resection，SST is superior to DST and is worthy of clinical application.

Abstract:objective To investigate the willingness to accept PrEP and its influencing factors among regular sex partners of HIV-infected individuals in Sichuan and Xinjiang province . Methods Face-to-face anonymous questionnaire was conducted to 473 qualified objects by non-randomly sampling method. test was employed to conduct univariate analysis, multivariate analysis was conducted using logistic regression .Results Awareness of PrEP among PAR was not high in Sichuan and Xinjiang province, 85.41% had never heard of PrEP, but reached to 80.34% after being taught on PrEP related knowledges by investigators. Logistic regression showed that the multiple influencing factors of the willingness to accept PrEP among PAR population were nation (Uyghur / Han:OR=2.471,P=0.0093), monthly income levels(<= 1000/1001-3000: OR = 2.046, P =0.0184), whether have sex with spouse within six months (yes / no: OR = 2.263, P = 0.0192), self-awareness of the possibility to be infected with HIV from their spouses (medium possibility/great: OR=0.485,P=0.0651；low possibility/great:OR=0.447,P=0.0273), whether would recommend friends to use PrEP (would / would not: OR = 9.416, P <.0001), the time when the spouse was diagnosed with HIV infection (within three years /one year: OR = 0.478, P = 0.0599; three years ago / within one year: OR = 0.374, P =0 .0094).Conlusion Awareness of PrEP among PAR was not high in Sichuan and Xinjiang province, but with a high rate of willingness to accept, propaganda of PrEP in AIDs intervention should be strengthened in the future thus to improve their awareness of PrEP and their willingness to accept PrEP. In the meanwhile, peer education could be possibly carried out among this population which might be an effective way to promote PrEP.