Objective To investigate the effect of Fas and FasL abnormal expression on immune escape mechanism of renal cell carcinoma(RCC). Methods The expression of Fas and FasL was detected in 44 cases of RCC and 10 cases of normal kidney tissue by immunohistochemical method. Tumor infiltrating T lymphocyte's apoptosis in 44 cases of RCC was examined by TUNEL assay. FCM was used to detect the expression of Fas and FasL as well as apoptosis in RCC cell line 786-0. In addition, the apoptosis of Jurkat T cell with Fas exprssion was also analysed with FCM. Results Fas expression rate (22.8%) in RCC was significantly lower than that (53.8%) in normal kidney tissue(P

Objective To test the effects of restuscitation with air or oxygen on the blood gas and cerebral superoxide dismutase (SOD) concentration in neonatal rats with experimental intrauterine asphyxia. Method Seventy-seven neonatal rats were randomly (random number) divided into three experimental groups: sham operation group (SHAM), air resuscitation group (AR), and oxygen resuscitation group (OR). In groups AR and OR, animal models of intrauterine asphyxia were established and then resuscitated with air (AR) or oxygen (OR) for 30min. Comparison was made between groups including the mortality 0 hour after resuscitation, and the levels of blood gas and cerebral SOD concentrations 0 h, 6 h and 24 h after resuscitation. Results Mortality of neonatal rats in SHAM group, AR group and OR group were 0 (0/24), 0 (0/26) and 3.7% (1/27), respectively (P ＞0.05). The average levels of blood PaO2 in OR group and AR group 0 h after resuscitation were (69.2 ± 8.2)mmHg and (55.5±10.3) mmHg,respectively (P=0.004). Blood pH and PaCO2 and BE levels in OR group 0 h after resuscitation were not significantly different from those in AR group (P＞0.05). Blood pH, PO2, PCO2and BE levels in OR group were also not significantly different from those in AR group 6 h and 24 hours after resuscitation. The average concentrations of cerebral SOD in OR group 0 h and 6 hours after resucitation were (38.3±9.8) U/mgprot and (8.6±3.6) U/mgprot, and those in AR group were (53.8± 10.6) U/mgprot and (13.0±4.6) U/mgprot, respectively (P = 0.003, 0.04). The cerebral SOD concentration in OR group 24 hours after resuscitation was not significantly different from that in AR group (P＞0.05). The cerebral SOD concentrations in SHAM group 0 h,6 h and 24 hours after resuscitation were much higher than those in OR group and AR group (P＜0.05). Conclusions Resuscitation with air is as good as pure oxygen in neonatal resuscitation, in respect of early mortality and improvement of acidosis in neonatal rats after intrauterine asphyxia. Resuscitation with air will generate less radical oxygen species than pure oxygen in neonatal rats after intrauterine asphyxia.

In recent years,along with social economic development and population aging,more and more fracture patients have received internal fixation and artificial joint replacement.Postoperative infection related to orthopedic implants is a catastrophic complication which imposes serious psychological,physiological and economic burdens on the patients.Therefore,it is necessary to be able to make an early diagnosis of the infection.However,diagnosis of implant-related infection is always a clinical problem for orthopedists.Currently,it is thought to be a valuable way to judge orthopedic implant infection by monitoring inflammatory cytokines.This article will offer an overview on the progress in research of diagnosis values of different inflammatory cytokines in postoperative infection related to orthopedic implants.

Objective To observe the protective effects of AG on the myocardial ultrastructure of diabetic rats.Methods STZ-induced diabetic male SD rats were divided into two groups: aminoguanidine(AG group,50 mg?kg~(-1) body weight by daily gavage) and diabetes mellitus groups(DM group).Age-matched normal male SD rats served as normal control(NC group).After 10 weeks of treatment the level of blood glucose was measured and the rats were killed.Cardiac muscle were observed by transmission electronic microscope.Results The changes of cardiac ultrastructure had no significant difference between AG-group and NC-group and were better in AG group than in DM group as following:(1) Myofibril arranged tidily with intact regular edge;(2) Mitochodria were big and normomorph with crests arranging densely,some mitochondria gathered locally;(3)Base membrane of blood vessels didn′t thicken with lumen not narrowed;(4) The collagenous fiber in stroma reduced but amorphous material didn′t reduced. Conclusions The ultrastructure pictures of diabetic cardiomyopathy is inhibited or delayed by AG,which suggests that AGEs may play some role in the development of diabetic cardiomyopathy.

OBJECTIVE To survey the distribution of the pathogenic microorganisms in biles collected from cholelithiasis patients and their susceptibility to antibiotics,to guide the selection of reasonable antibiotics.METHODS Totally 546 bile samples were cultured and tested for antibiotics susceptibility.RESULTS Bacteria were cultured from 332 samples with a positive rate of 60.81%,the first three were Escherichia coli(32.96%),Enterococcus faecalis(12.74%),and Klebsiella pneumoniae(9.97%).The pathogenic microorganisms were more sensitive to the third and fourth generation cephalosporins,antibiotic/?lactamase inhibitor combination,carbopenems and aminoglycosides antibiotics,while less sensitive to the first and second generation cephalosporins,penicillins,macrolide antibiotics and fluoroquinolones.CONCLUSIONS The pathogenic microorganisms in bile are very various,E.coli,E.faecalis and K.pneumoniae are usually cultured,fungous and anaerobic infections cannot be ignored.Doctors should pay more attention to analyze the bacterial resistance profile and use the antibiotics properly.

Objective To explore the effect of autophagy regulator on the injury of rat hippocampal neurons induced by oxygen-glucose deprivation (OGD).Methods Rat hippocampal neurons were cultivated in primary and subjected to OGD to simulate neuronal hypoxic ischemia injury for 2 hours or 6 hours followed by reperfusion for 12 hours with or without 3-methyladenine (3-MA, 20μmol/L) or rapamycin (0.2μmol/L). The morphology of neurons was observed with optical microscope. The expression of autophagy-related protein (LC3, P62) and apoptosis-related protein (cleaved caspase-3) were assessed by Western Blot analysis. The apoptosis of neurons was detected by flow cytometry, the release rate of lactate dehydrogenase (LDH) was calculated by automatic biochemical analyzer, and the cell activity was determined by methyl thiazolyl tetrazolium (MTT) colorimetric assay.Results Compared with the control group, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.091±0.160, 3.422±0.186 vs. 0.256±0.021), cleaved caspase-3 (gray value: 0.230±0.025, 0.440±0.051 vs. 0.050±0.007), neuronal apoptotic rate, LDH release rate [(38.50±4.15)%, (59.60±5.65)% vs. (12.40±1.32)%] were increased, while the expression of P62 (gray value: 0.290±0.025,0.120±0.026 vs. 0.450±0.040), neuronal activity [(71.40±7.23)%, (42.80±4.12)% vs. (100.30±2.30)%] were decreased at 2 hours or 6 hours after OGD (allP < 0.05). When the time of OGD was 2 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ (gray value: 2.281±0.121), the neuronal activity [(51.10±5.73)%] were decreased, while the expression of P62 and cleaved caspase-3 (gray scale: 0.410±0.037, 0.330±0.027, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (47.30±4.43)%] were increased (allP < 0.05). When the time of OGD was 2 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.689±0.214), the neuronal activity [(85.30±8.56)%] were increased, while the expression of P62 and cleaved caspase-3 (gray value: 0.170±0.040, 0.090±0.096, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (24.30±2.14)%] were decreased (allP < 0.05). On the contrary, when the time of OGD was 6 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.021±0.178, 0.240±0.017), neuronal apoptotic rate, the injury of neurons [LDH release rate: (36.60±3.45)%] were decreased, while the expression of P62 (gray value: 0.350±0.060), the neuronal activity [(59.70±6.13)%] were increased (allP < 0.05). When the time of OGD was 6 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.923±0.201, 0.590±0.062), neuronal apoptotic rate, the injury of neurons [LDH release rate:(71.20±7.81)%] were increased, while the expression of P62 (gray value: 0.070±0.008), the neuronal activity [(27.30±2.12)%] were decreased (allP < 0.05).Conclusion The enhancement of autophagy has protective effect on neurons under the condition of mild OGD, while it can aggravate the injury of neurons induced by a long-time OGD.

OBJECTIVE To investigate the change in antibiotic resistance of Pseudomonas aeruginosa(PAE) in our hospital,for the guidance of rational use of antibacterials. METHODS We reviewed the isolation rates of antimicrobial agents from 2004 to 2007,and its construction ratio among the commonly encountered Gram-negative bacilli,and the resistance rates of P.aeruginosa to 14 kinds. RESULTS We totally isolated 823 strains of PAE and the isolation rates of PAE was 13.3% in 2004 but 18.5% in 2007.In the four years,there was the highest isolation rate of 72.54% in sputum and throat mucus specimens.The lowest resistance rate was to amikacin(from 12.2% to 6.21%,the average was 8.75%),and the next was to cefoperazone/sulbactam(from 6.10% to 12.26%,the average was 8.99%);the total resistance rates to ticarcillin and piperacillin was more than 36.00% and there was a lower resistance rate to ticarcillin/clavulanic acid(32.93%)after adding the enzyme inhibitor,while to piperacillin/tazobactam was 20.16%.Drug-resistance rate to ceftazidime was from 19.51% to 43.17%(the average was 33.05%);to cefepime from 22.34% to 34.16%(the average was 29.40%);to imipenem from 19.51% to 23.29%(the average was 22.60%);to meropenem from 11.17% to 18.18%(the average was 15.43%);to the fluoroquinolones antibiotics ciprofloxacin from 32.61% to 44.59%(the average was 38.52%);and to levofloxacin was 26.40% in average. CONCLUSIONS There is commonly increasing resistance of PAE.Reviewing the resistance situation dynamically and selecting the antibiotics reasonably can produce great significance for clinical therapy and postpon its multidrug-resistance.

Objective To investigate the ability of induction of specific cytotoxic T lymphocytes (CTL) stimulated by dendritic cells (DCs) co-transfected with MUC1 and survivin mRNA of human pancreatic cancer,and to provide the experimental basis for the treatment of human pancreatic cancer with multi-epitope DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) of 6 patients with pancreatic cancer.Human pancreatic cancer cell line MiaPaCa-2 was routinely cultured,after being transcripted and amplified by RT-PCR,MUC1 and survivin mRNA were co-transfected or individually transfected into DCs by electroporation,and they were named as DC-MUC1,DC-survivin,DC-MUC1 + survivin.The expression of MUC1 and survivin mRNA in DCs were detected by real-time PCR.The survival rate of transfected DCs were determined by MTT method.The lymphocyte proliferation ability was evaluated by mixed cell culture method.The Th1 cytokine releasing of antigen-specific CTLs were measured by ELISA assay.Results Mature DCs were obtained,the positive expression rates of surface markers CD40,HLA-DR,CD83 and CD86 were 34.31％,50.21％,89.17％ and 73.62％,respectively.The expression amount of MUC1 mRNA of DC-MUC1 was 36.24 ± 5.17,and the expression amount of survivin mRNA of DC-survivin was 34.53 ± 4.02,while the expression amounts of MUC1,survivin mRNA of DC-MUC1 + surviving were 31.79 ±4.26 and 14.67 ± 2.96,which were significantly lower than that in individual transfection group (P ＜ 0.05).The survival rate of DC-MUC1 + surviving was decreased in a time dependent manner,which was significantly lower than that in individual transfection group (about 50.21％ vs 80％ at 24 h,P ＜0.05).When DC/T cells ratio was 1∶ 10,1∶ 20,the autologous T cell proliferation index of MUC1 and survivin mRNA in co-transfection DC group was significantly higher than that in individual transfection group (P ＜ 0.05) ;when DC/T cells ratio was 1∶ 40,1∶ 80,the difference of proliferation index was not statistically significant.When DC/T cells ratio was 1∶ 10,after 14 d culture,the expressions of IL-2 in DC-MUC1,DC-survivin,DC-MUC1 + surviving were (892.73 ± 32.9),(713.62 ± 56.37),(1884.37 ± 95.21) pg/ml,and the expressions of granzyme B were (501.62 ± 12.30),(203.84 ± 12.55),(1193.15 ± 86.04) pg/ml ; and the expressions of IFN-γ were (981.50 ± 47.82),(696.05 ± 41.66),(2237.94 ± 189.55) pg/mL.The corresponding values in DC-MUC1 + surviving group were significantly higher than those in individual transfection group (P ＜ 0.05) ; while the difference of IL-10 was not statistically significant.Conclusions DCs co-transfected with MUC1 and survivin mRNA have a stronger ability to stimulate specific CTL in vitro than individual antigen loaded DCs.

Objective To discuss the feasibility and clinical outcomes of mini-open reduction through lateral deltoid muscle and locking plate fixation in treatment of proximal humeral fractures.Methods A retrospective study was carried out on 39 patients with proximal humeral fractures treated with mini-open reduction through lateral deltoid muscle and locking plate fixation from September 2012 to September 2015.There were 7 males and 32 females,with a mean age of 66.8 years (range,33-86 years).According to the Neer's classification system,there were 26 patients with two-part fracture and 13 with three-part fracture.visual analogue scale (VAS),Constant-Murley shoulder score,time of bone healing and complications were evaluated for all patients.Results All patients were followed up for 12-48 months (mean,21.3 months).All fractures were proved to be bone healing with duration of (12.0 ± 2.7) weeks.The VAS was improved from preoperative (5.7 ± 1.0) points to (0.9 ± 0.6) points at the final follow-up (P ＜ 0.05).The Constant-Murley shoulder score was increased from preoperative (42.3 ±5.1) points to (89.2± 2.5)points at the final follow-up (P ＜ 0.05).Ranges of shoulder motion were improved with satisfactions.There were no complications like axillary nerve injury,loss of reduction,nonunion or humeral head necrosis.Conclusion For proximal humeral fractures,the miniopen reduction through lateral deltoid muscle and locking plate fixation has advantages of minimal invasion,excellent shoulder function,high rate of bone healing and low incidence of complications.

Objective To investigate the anti-tumor immune response induced by human pancreatic cancer mucin 4 mRNA and human telomerase reverse transcriptase (hTERT) mRNA cotransfected dendritic cells (DC),and to provide the experimental evidences for the treatment of pancreatic cancer with multi-epitope loaded DC vaccine.Methods DC were isolated from peripheral blood mononuclear cells (PBMC) of six patients with HLA-A2+ pancreatic cancer and cultured.Mucin 4 mRNA and hTERT mRNA were transcripted and amplified in vitro,which were transfected into DC separately or in order by eleetroporation.DC were cultured for 48 hours.The expressions of mucin 4 and hTERT in DC were detected by quantitative real-time polymerase chain reaction (PCR) and Western blot.The survival rates of transfected DC were determined by methylthiazolyl tetrazolium (MTT) method.The cytotoxic T lymphocyte (CTL) activation induced by mucin 4 mRNA and hTERT mRNA transfected DC were evaluated by interferon (IFN)-γ release assays (enzyme linked immunosorbent assay) method.The cytotoxicity of CTL induced by mucin 4 mRNA and hTERT mRNA transfected DC in pancreatic cancer cell lines MiaPaCa-2,Capan-2,AsPC-1 and Pane-1 was measured by 51Cr standard cytotoxicity test.Student t test was performed for statistical analysis.Results After in order transfection of mucin 4 mRNA and hTERT mRNA for 48 h,the relative quantity of the expression of mucin 4 and hTERT in DC were 30.09±5.24 and 12.87±3.36,which were lower than the relative quantity of the expression in DC transfected separately (38.54±6.21 and 36.35±5.03,t=3.469,6.721,both P＜0.05).After transfected in order for 96 hours,the survival rate of DC decreased to 52.17％,which was lower than that of DC transfected separately (around 80％).The quantity of IFN-γ releasing of specific CTL induced by mucin 4 mRNA and hTERT mRNA cotransfected DC was (32.57±2.01) U/mL in 24 hours,which was higher than that of CTL induced by DC transfected with mucin 4 mRNA ((23.06±4.74) U/mL) or hTERT mRNA ((16.82±3.67) U/mL) separately (1=5.092,7.141,both P＜0.05).After co-transfected with mucin4 mRNA and hTERT mRNA,DC could effectivly induce HLA-A2+/mucin 4+/hTERT+ specific CTL immune responses,however there was no significant cytotoxicity in HLA-A2+ pancreatic cancer cells.Conclusion The induction of CTL anti-tumor immune response by DC co-transfected with mucin4 mRNA and hTERT mRNA is more significant compared with that by single antigen loaded DC.