[Objective]To study the proliferation inhibition effect of decetaxel combined with cisplatin on osteosarcoma cells,and the effect of the drugs during the course of chemotherapy for osteosarcoma.[Method]The effect of docetaxel and cisplatin alone or combined with osteosarcoma cell line HOS-8603 was examined by cell count,morphologic observation,flow cytometry(FCM).The animal model of human osteosarcoma was established in nude mice.They were given docetaxel or cisplatin alone and combined,and a blank control group was set up.Observations on the growth and pathologic markers of osteosarcoma were carried out and pathological examinations were made.[Result]Docetaxel and cisplatin either alone or combined inhibited significantlythe proliferation of osteosarcoma cells and induced obviously apoptosis,and the effect of the drags combined was more effective than that of the drugs alone,Growth inhibition ratio ≥59.34%(P

0.05). Conclusions: There exists an abnormal reduction of neural viability and function on bilateral hippocampus of patients with first-episode MDD.

AIM: To study the effect of double gene transduction mediated by lentiviral vectors on the characteristics of human embryonic stem cells (hESCs). METHODS: Using the backbone from inducibal dual and excisable transgene vector (iDuet101) , lentiviral vectors overexpressing cytotoxic T lymphocyte - associated molecule - 4 immuno-globulin (iDuet101 - CTLA4Ig) , indoleamine 2, 3 dioxygenase (iDuet101 - IDO) , and ubiquitin - C promoter - lueifer-ase - ires - puromycin ( ULIP) were constructed and packaged according to the standard recombinant techniques. Human embryonic stem cells (hESCs) were first transduced with iDuet101, iDuet101 - CTLA4Ig or iDuet101 - IDO, then, after the selection, were transduced again with ULIP. The expression and function of the exogenous genes were detected. Immu-nohistochemistry, RT - PCR and flow cytometry were applied for detection of embryoid bodies ( EB) formation in vitro and in vivo teratoma formation. RESULTS: Double - transduced hESCs showed typical shape of cell clones and positive staining of tumor rejection antigen -1 - 60 ( Tra -1 - 60 ) and octomer transcription factor - 4 ( OCT - 4 ). The formation of EB was observed, in which a - fetoprotein (AFP), paired box gene 6 ( Pax6) and Musashi 1 ( MSI1) were positively expressed. The cells formed teratomas, and the luciferase signals existed until 28 days after xeno - transplantation. CONCLUSION : Double transduction of non - transcriptional factors mediated by lentiviral vectors does not affect the cell growth rate and their differentiation ability.

ObjectiveTo study the risk factors and pathogenic characteristics of ventilator associated pneumonia(YAP). MethodsRisk factors of VAP,pathogens and drug resistance in 118 patients in ICU were analyzed retrospectively. ResultsThe incidence of VAP was 44.9％, and age, the mechanical ventilation time, state of consciousness,tracheotomy,the antibiotic combination are risk factors of incidence of VAP. 53cases of VAP(68.2％ )were infected by Gram negative bacilli,25.9％ were by Gram positive cocci,and 5.9％ were by fungi. Drug resistance was observed obviously. ConclusionThe occurrence of VAP was related with multiple factors. The gram negative bacteria are the major pathogens of VAP, and the rate of drug resistance was high. The occurrence of VAP could severely affect patients' prognosis.

Objective To investigate apoptosis of polymorphonuclear neutrophil and activity of caspase-3 in the peripheral blood and discuss their correlation with development of multiple organ dysfunc- tion syndrome (MODS) after multiple injury. Methods A total of 55 patients with multiple injury were included in the prospective study, and divided into two groups, ie, MODS group (multiple injury patients who developed MODS,) and non-MODS group (multiple injury patients who were free from MODS). The activity of caspase-3 was detected by flow cytometry and the serum levels of IL-6 and IL-10 were detected by ELISA. Then, we evaluated whether the neutrophil apoptesis was correlated with the ser- um levels of IL-6 and IL-10 in patients with MODS. Results Compared with non-MODS group, neu- trophil apoptesis was significantly reduced and activated caspase-3 decreased significantly in MODS group (P <0.05). In MODS group, serum IL-6 was increased significantly while serum IL-10 was decreased significantly compared with non-MODS group. The apoptosis of neutrophil in multiple injury patients with MODS had a negative correlation with IL-6 levels but a positive correlation with IL-10 levels. Conclu- sions The delayed apoptosis of neutrophil due to decreased activated caspase-3 may play partial roles in the development of MODS after multiple injury. IL-6 and IL-10 may contribute to the apoptotic changes.

[Objective]To compare the clinical outcomes of fresh embryo transfer of the in vitro fertilization/intracytoplasmic sperm injection and embryo transfer(IVF/ICSI-ET)in different age groups as well as in different responders using gonadotropin-re-leasing hormone agonist(GnRH-a)long protocol or GnRH antagonist(GnRH-ant)protocol.[Methods]A retrospective analysis was performed on 737 IVF/ICSI cycles,including 386 cycles of GnRH-a long protocol(group A)and 351 cycles of GnRH-ant protocol (group B),from August 28,2015 to December 31,2016. Then all the cycles were divided into sub-groups by ages and retrieved oo-cyte numbers:group a1(<38 years),group a2(≥38 years);group b1(n≤5),group b2(6≤n≤15),group b3(n>15). The basic information of patients and clinical outcomes were compared.[Results](1)Comparable results were obtained from group A and group B in these following variables such as fertilization rate,normal fertilization rate,biochemical pregnancy rate and miscarriage rage. But the stimulation period,the total gonadotropin(Gn)dosage,estradiol(E2)level and endometrial thickness on the day of human chorionic gonadotropin(hCG)administration,number of oocytes retrieved and mature oocytes,ovarian hyperstimulation syn-drome(OHSS)rate,implantation rate and clinical pregnancy rate were significantly higher in group A than group B(P<0.05),and significantly higher cancellation rate of fresh embryo transfer was observed in group B(P<0.001).(2)When divided by ages,no mat-ter in sub-group a1 or sub-group a2,the implantation rate was slightly lower in GnRH-ant protocol than in GnRH-a long protocol, although they failed to reach significant difference(sub-group a1:32.6%vs 39.8%,P=0.067;sub-group a2:9.7%vs 17.9%,P=0.066). The clinical pregnancy rate was comparable using these two protocols in sub-group a1(54.8%vs 50.4%,P=0.429),but it was significantly lower by using GnRH-ant protocol than GnRH-a long protocol in sub-group a2(19.6%vs 39.1%,P=0.021).(3) When divided by numbers of oocytes retrieved,the implantation rate was significantly lower when using GnRH-ant protocol in sub-group b1(13.1%vs 26.0%,P=0.026),but we failed to observe significant differences in other two sub-groups. The clinical preg-nancy rates were comparable in all sub-groups ,whereas differed considerably in sub-group b1 (36.6% vs 19.3%,P = 0.056).[Conclusion]Overall,the implantation rate and clinical pregnancy rate were higher in GnRH-a long protocol than those in GnRH-ant protocol. Nevertheless,GnRH-ant protocol could reduce the dosage of Gn,shorten the treatment duration,and effectively reduce the occurrence of OHSS. There were similar pregnancy outcomes in two protocols for normal responders and high responders ,while for advanced patients or other poor responders,the implantation rate and clinical pregnancy rate were higher in GnRH-a protocol.

Auxiliary liver transplantation reserves whole or part of recipient' native liver while implanting the graft. It call be classified as auxiliary heterotopic liver transplantation and auxiliary orthotopic liver transplantation according to the location of the graft. On the basis of graft volume. it can be categorized as whole liver auxiliary liver transplantation and partial liver auxiliary liver transplantation. Either cadaveric or living donor is available. Because auxiliarg orthotopic liver transplantation is only the partial liver transplantation, so it is also called aunliry orthotopic partial liver transplantation. The. technique of auxiliary liver transplantation progresses with the devehopment of liver transplantation. Great attention is paid to auxiliary liver transplantation because of its special superiority, in spite of some unresolved problems.

Objective To describe the feasibility of biliary reconstruction methods and to explore the preventing and curing methods for biliary complications. Methods A total of 77 cases of living donor liver transplantation were included in our study for retrospective analysis. The operation types included right lobe contain middle hepatic vein (n=29), right lobe without middle hepatic vein (n=45), left lobe (n=1) and left lateral lobe transplantation (n=1). The biliary reconstruction was performed by duct-to-duct and Roux-en-Y epaticojejunostomy. Results Fifty-four grafts had 1 bile duct and 23 had multiple ones. The duct-to-duct biliary reconstruction was performed for 75 adult cases and T-tube drainage was used in 63 patients. Meanwhile, the conventional Roux-en-Y epaticojejunostomy was utilized for 2 pediatric patients. Total biliary complication incidence was 36.4% (28/77) and the complications included bile leakage (10.4%, 8/77) and biliary stricture (26.0%, 20/77).The rate of biliary complication was remarkably different between single hepatic duct group and multiple group (P<0.05). All the 8 patients suffering from bile leakage were cured by percutaneous bile drainage guided by ultrasound. By percutaneous balloon cholangioplasty and placing stent through the T-tube tract or endoscopic retrograde balloon cholangioplasty, the liver function of biliary stricture cases got better and most cases were cured. Conclusion Protecting the blood supply of bile duct at the cutting surface and harvesting one bile duct stump as much as possible can reduce biliary complication effectively. Radiate intervention treatment such as percutaneous balloon cholangioplasty and endoscope treatment etc. are effective for treatment of biliary complications.

Objective To investigate vitamin K3 (VK3) effect on apoptosis of human liver cancer cells and its mechanism. Methods The SMMC-7721 cells were cultured in the experiment. The inhibitory effects of VK3 on SMMC-7721 cells were tested by CCK-8. Morphological evaluation of apoptosis was performed by Hcechst33342 staining. The distribution of cell cycle and apoptosis, and the expression of Fas were assayed by flow cytometry. Fas mRNA expression were detected by RT-PCR. And the concentration of soluble Fas(sFasL) in the culture supernatant were measured by EIJSA. Results The inhibitory rates ofVK3 (at concentrations of 2,5,10,20,25 umol/L for 48 h) on SMMC-7721 growth were 33.8% ,50.1%,63.9% ,78.5% and 84.7%, respectively. Compared with the control group, the number of cells in the G0/G1 phase increased, while that of S phase decreased. The apoptotic cell rates were 18.75%, 25.80%,38.80% ,29.92% and 26.18% ,respectively. The apoptosis cells were strongly stained by Hcechst33342.On exposure to VK3 at the concentration of (2,5,10 umol/L) after 48 h, the mean fluorescence intensity ofFas on cell surface and the expression of fas mRNA and the concentration of FasL in the culture supematantin SMMC-7721 were increased, but they all decreased at the high concentration of VK3. Conclusion VK3can inhibit the proliferation of SMMC-7721 cells and induce apoptosis via up-regulating expression of Fas and sFasL.

Objective To investigate the effect of different time of hypoxia reoxygenation (HR) on cell autophagy and cell viability.Methods According to the different reoxygenation time,H9C2 cells were divided into five groups:normal control group (group A),hypoxia group (group B),reoxygen 2h group (group C),reoxygen 12h group (group D) and reoxygen 24h group (group E).The expression of LC3 Ⅱ/LC3 Ⅰ in each groups was detected by Western Blot,and the activity of myocardial cells was detected by MlT.Results MlT results showed that the viability of the myocardial cells after hypoxia 2h/reoxygenation 2h was significantly lower than that in the normal control group (P＜0.05).The ratio ofLC3 Ⅱ/LC3 Ⅰ of the cells in group B (hypoxia group),group C (reoxygen 2h group),group D (reoxygen 12h group),group E (reoxygen 24h group) was significantly higher than that of group A (normal control group) (P＜0.05).Among them,the ratio of LC3 Ⅱ/LC3 Ⅰ of the cells in group C and D were significantly higher than that of group B (P＜0.05),paticlularly the group D was the highest (P＜0.05).The activity of myocardial cells in group D was the lowest among all groups.Conclusion Culturing in the three gas incubator can obviously decrease the viability of H9C2cells.Hypoxia can activate autophagy,and autophagy increases significantly after reoxygenation.Autophagy may be activated most obviously at 12 hours and decrease to the level of hypoxia group at 24 hours.Cells can be damaged by hypoxia,and further increase of cellular damage may occur after oxygen inhalation.At 12 hours,the cell viability is the lowest,and the cell viability may be improved at 24h.