Objective To investigate the clinical significance of ligating the portasystemic shunt confirmed by preoperative CT evaluation during orthotopic liver transplantation. Methods From January 2007 to August 2008, 35 patients in Tianjin First Central Hospital underwent preoperative three-dimensional CT scan, among them 23 patients had spontaneous major portasystemic shunts, the other 12 patients did not have portasystemic shunts. 16 out of the 23 cases with significant shunts underwent shunt ligation based on portal blood flow volume measured by intraoperative portal vein flowmetry. The shunt of the other 7 patients were left untreated. Results The portal blood flow in the 12 patients without portasystemic shunt as judged by preoperative CT scanning were (1101±70) ml/min. The shunts in 7 patients with portal blood flow greater than 1000 ml/min were not ligated, that of the 16 patients with portal blood flow volume lower than 1000 mL/min were ligated. The portal blood flow volume in those 16 patients before and after ligating the shunt were (657±112) m//min and (1136±161) ml/min, respectively (P<0.05). Postoperatively 2 patients suffered from portal vein thrombosis, among them 1 patient suffered from intermittent disturbance of consciousness, 2 patients died within 3 months, with one dying of respiratory failure from pulmonary aspergillus infection one dying of hepatic failure in 2 months after operation because of graft dysfunction.The other 19 patients with normal blood flow and well-functioning graft were alive. Conclusion The ligation of portasystemic shunt is mandatory in patients when pretransplant CT evaluation showing a major porto-systemic shunts and portal blood flow volume was less than 1000 ml/min.

Objective To observe the effect of traditional Chinese medicine internal and external treatment on chronic osteomyelitis.Method Chronic osteomyelitis experimental animal models were set up with debridement, then it was random divided into two groups ( antibiotics group for the control group, antibiotics and traditional Chinese medicine group for the observation group).After treatment for 10 days, new capillaries in the tissues surrounding sinus crossings were detected by pathological observation and VEGF expression was determined by ELISA.Result VEGF expression and vascular points of tissues surrounding sinus crossings of antibiotics with traditional Chinese medicine group were obviously higher than that of antibiotics group[ (47.48 ±3.22) μg/ml vs (28.26 ±2.61)μg/ml, P ＜0.01;8.03 ±1.73 vs5.17 ±2.89, P＜0.05 ].Conclusion Traditional Chinese medicine internal and external treatment can improve VEGF expression and increase capillary number inside tissues surrounding sinus.crossings , thus it can promote the healing of chronic osteomyelitis.

Objective To observe the effect of small interfering RNA(siRNA) targeting Survivin gene on survivin expression,proliferation and apoptosis of hepatocellular carcinoma cell line MHCC-97H.Methods Survivin sequence specific siRNA was designed and synthesized.siRNA/liposome complex was transfected into hepatocellular carcinoma cell line MHCC-97H.The MHCC-97H cells were divided into Survivin siRNA group(Si-survivin),negative control siRNA group(NC group)and blank group (normal control group).Survivin mRNA and protein expressions were detected by reverse transcription-PCR and Western blot,respectively.The proliferation of MHCC-97H was measured by methythiazolydiphenyl-tetrazolium bromide assay.The Annexin V/PI double labeled flow cytometry was employed to measure the apoptosis at 24 h after transfection in different concentrations of Survivin siRNA(12.5,25.0 and 50.0 nmoL/L,respectively).Results After 48 h of transfection,the Survivin mRNA levels were 0.55 ± 0.16 (Si-survivin group),0.85 ± 0.28 (NC group) and 0.93 ± 0.40 (normal control group),respectively,which were significantly different among 3 groups (F =414,P ＜ 0.01).The level of Survivin mRNA was the lowest in Si-survivin group,which was statistically different with NC group and normal control group (t =-20.56,-28.37,all P ＜ 0.001).The levels of Survivin protein expression in 3 groups were 0.602 ± 0.005 (Si-survivin group),0.835 ± 0.007 (NC group) and 0.993 ± 0.003 (normal control group) at 48 h after transfection,which were statistically different among 3 groups (F =238,P ＜0.01).The lowest level of protein expression was in Si-survivin group,which was statistically different with NC group and normal control group (t =-40.17,-66.03,all P ＜ 0.001).After 72 h and 96 h of transfection,the inhibitory rate of cell growth was significantly higher in Si-survivin group [(19.5 ± 3.6)％,(12.0 ± 0.9)％] compared with that in NC group [(3.6 ± 0.9) ％,(-1.3 ± 6.1) ％] (t--36.18,42.53,all P ＜ 0.05).The apoptosis rates in 12.5,25.0,50.0 nmol/L Survivin siRNA were (22.64 ± 2.54) ％,(35.37 ± 3.28) ％ and (53.28 ± 4.35) ％,respectively.However,in NC group and normal control group,the apoptosis rates were (8.77 ± 1.25) ％ and (9.72 ± 1.37) ％.The rates were statistically different among those 5 groups(F =35.93,P ＜0.01).And in the apoptosis rates of siRNA groups in different concentratiom were statistically different when compared between each two groups (t =-29.73,-38.57,all P ＜ 0.001).Conclusion Survivin specific siRNA can inhibit the proliferation and induce the apoptosis by blocking Survivin gene expression in hepatocellular carcinoma cell line MHCC-97H.

Anterior shoulder instability is a very difficult issue to treat,especially with glenoid bone defect.When the defect is small,there is little influence on shoulder instability.The larger the defect is,the more influence there will be.Most authors agree that glenoid bone reconstruction should be considered when glenoid bone defect is more than 20％-25％.In this condition soft tissue procedures alone are not enough to provide stability to the shoulder.To date,there is still not an ideal typing of glenoid bone defect.There are many methods of assessing the size of bone defect.Pico system is one of the most common methods,as it is easier and more precise.Numerous surgical procedures have been described to address the bone defect.The Bristow procedure,the Latarjet procedure and the Eden-hybinette procedure are effective and most popular around the world.The Latarjet procedure can provide more bone blocking than the Bristow procedure,and is more popular.The Eden-hybinette procedure dose not need coracoid transfer and then has no damage of normal anatomical structure.But it also lack the hanging effect of the conjoint tendon.After all,each procedure has its advantage and disadvantage in treating anterior shoulder instability associated with glenoid bone defect and should be chosen depending on the characteristics of each patient and the preference of each surgeon.Furthermore,more new and effective treatments are still needed.

Objective To develop fast detection techniques for the diagnosis of gonococcal infections.Methods Prokaryotic expression vector for Neisserial surface protein A (NspA) was constructed using the NspA gene cloned by PCR.Mice were immunized with the renatured recombinant NspA (rrNspA)to prepare antibodies against NspA.Western blot and ELISA was used to analyze the binding of NspA antibodies to lysate of gonococcal cells or to intact gonococci.Results NspA antibodies that were prepared by the rrNspA expressed in E.coli could bind to rrNspA,the natural NspA existing in gonococcal cells,or intact gonococci.Conclusion RrNspA and its antibodies have potential value in developing fast diagnostic kits for gonococcal diseases.

Objective:To determine the expression of transglutaminase 2 (TGM2) in peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis (AD), and to analyze its correlation with AD-related inflammatory factors and disease severity.Methods:A total of 29 AD patients and 15 healthy controls were collected from the First Affiliated Hospital of Fujian Medical University from July 2020 to January 2021. Ten milliliters of peripheral blood samples were collected from each subject, so was the clinical information, including age, gender, course of disease, eosinophil counts, basophil counts, total IgE levels, Scoring AD index (SCORAD), etc. PBMCs were isolated by density gradient centrifugation. Fluorescence-based quantitative PCR was performed to determine the mRNA expression of TGM2 and AD-related inflammatory factors (interleukin [IL]-1β, IL-4, IL-6, IL-8, IL-10, IL-13, IL-17, thymic stromal lymphopoietin [TSLP], P2RX7 [purinergic receptor P2X, ligand-gated ion channel, 7], etc.) in PBMCs from 29 AD patients and 15 healthy controls, and flow cytometry to determine TGM2 protein expression on PBMCs. Mann-Whitney U test was used to analyze differences between groups, and Spearman correlation analysis to evaluate the correlation. Results:The relative mRNA expression of TGM2 in PBMCs did not differ between the AD group and control group ( M[ Q1, Q3]: 0.509 [0.325, 0.958] vs. 0.475 [0.328, 1.051], U = 210.50, P = 0.872). Compared with the control group, the AD group showed significantly decreased IL-4 mRNA expression (0.171[0.049, 0.449] vs. 0.824 [0.397, 1.378], P < 0.001), but significantly increased mRNA expression of IL-8 and IL-13 ( P = 0.011, 0.006, respectively). Spearman correlation analysis showed that the mRNA expression level of TGM2 in PBMCs was positively correlated with the mRNA expression levels of IL-4 and P2RX7 in the AD group ( rs = 0.42, 0.40, P = 0.024, 0.034, respectively), while there were no correlations between TGM2 mRNA expression and AD severity-related indicators (all P>0.05), such as age (21[16, 29] years), course of disease (4[1,10] years), eosinophil counts (0.33[0.18, 0.65] × 10 9/L), basophil counts (0.04[0.03, 0.06] × 10 9/L], SCORAD scores (60.5[46.98, 66.13] points), and serum total IgE levels (373 [40, 1 815] IU/ml). The relative protein expression levels of TGM2 on the surface of PBMCs did not differ between the AD group and control group (54.9 [47.6, 62.8] vs. 55.55 [51.5, 60.25], U = 112.00, P = 0.922) ], and no correlations were observed between the protein expression of TGM2 on PBMCs and AD severity-related indicators in the AD group (all P > 0.05) . Conclusion:No significant differences were observed in TGM2 mRNA expression in PBMCs or TGM2 protein expression on the surface of PBMCs between the AD patients and healthy controls, and there were no correlations between the TGM2 mRNA and protein expression and AD severity.

Objective To investigate the biological response of survivin siRNA in A549 human lung cancer cells and Hela S3 human cervical cancer cells as well as K562 human erythroleukemia cells,in order to screen the working sequences of survivin siRNA.Methods Three sequences of survivin-targeted siRNA were designed and synthetized,and human cancer lines of A549,Hela S3,K562 were transfected with Hiperfect liposome entrapped survivin siRNA,respectively.The expression of survivin mRNA was detected by means of real-time polymerase chain reaction (RT-PCR) with SYBR Green Ⅰ.Cell proliferation was detected by way of WST-8 cell count kit at 48 hours and 72 hours after transfection.Results The expression of survivin mRNA in all 3 cancer cells studied was significantly inhibited by all siRNA at 48 hours and 72 hours after transfection,gene expression inhibition ratio were 57.47％-88.53％ after transfection 48 hours and were 69.94％-95.03％ after transfection 72 hours.Cell proliferation was also significantly inhibited 48 hours and 72 hours after transfection,cell multiplication inhibition ratio were 27.88％-47.36％ after transfeotion 48 hours and were 42.59％-57.29％ after transfeciton 72 hours.Sequence 1 had the most inhibition efficacy on survivin gene expression and proli-feration in tumor cells.Inhibition rate of the three tumor cell gene expression of survivin at 48 hours are above 75％,72 h all over 90％,48 hours of cell proliferation inhibition rate are above 40％,72 hours of more than 50％.Conclusions The chemo-synthesized siRNAs can significantly down-regulate survivin mRNA expression in cancer cell studies.Survivin siRNA is capable of inhibiting the proliferation of tumor cell in vitro and it might be a new strategy for tumor-targeted therapy.Sequence 1 is the most efficacious working survivin siR-NA in the study.