Objective To investigate the signal mechanism of protein kinase C alpha(PKC-α)participated in enhanced expression of type Ⅰ inositol 1,4,5-trisphophate receptors (IP3 RI) induced by tumor necrosis factor alpha(TNFα),in order to delineate the mechanisms of decreased glomerular filtration rate (GFR) in hepatorenal syndrome caused by TNFα.Methods The glomerular mesangial cells (GMCs)line from rats was chosen as experimental material.GMCs were divided into control (D),TNFα-2 h,TNFα-4 h,TNFα-8 h,and TNFα-24 h groups.Moreover,another two groups were sanflngol-8h (S),TNFα + Sanfingol-8h(TS)groups.The effect of TNFα on the expression of IP3RI was detected by immunocytochemical staining,Western blotting,teal time-polymerase chain reaction (PCR) assays.Results Immunocytochemical staining demonstrated that IP3 RI was mainly distributed in cytoplasm of GMCs.Enhanced positive staining was determined in all TNFα-treated groups,especially in TNFα-8 h group.Western blotting demonstrated that the expression of IP3RI protein was significantly higher in TNFα-4 h,TNFα-8h and TNFα-24 h groups than control group(4 h:1.82 ± 0.63 ; 8 h:2.95 ± 0.66 ; 24 h:2.48 ± 0.72 ; D:1 ±0.02 ; F =9.24,P ＜ 0.05).The expression of IP3 RI protein was the highest in TNFα-8 h and TNFα-24 h groups(P ＜0.05).No difference was found among S,TS,and control groups(S:1.39 ±0.65; TS:1.35± 0.37 ; P ＞ 0.05).Real time-PCR found the expression of IP3 RImRNA was significantly higher in all TNFα-treated groups than control group(2 h:3.35 ± 1.97; 4 h:3.16 ± 1.35; 8 h:3.70 ± 1.76; 24 h:4.49±1.70; D:1 ±0.01; F =6.167,P ＜0.05).No difference was found among all TNFα-treated groups(P ＞0.05).No difference was found among S,TS,and control groups(S:1.53 ±0.79; TS:1.32 ± 0.38 ; P ＞ 0.05).Conclusions IP3 RI was mainly distributed in cytoplasm of GMCs.TNFa could enhance the expression of IP3 RI protein and IP3 RI mRNA,which could be blocked by sanfingol,a PKCα inhibitor.It might be an important signal in the mechanisms of GFR decrease caused by TNFα in hepatorenal syndrome.

Objective To observe and analyze the correlation between the DNA methyltransferase-3B (DNMT3B) gene single nucleotide polymorphisms (SNP) rs 1569686 and the susceptibility to colorectal cancer (CRC) in Tianjin Han population.Methods A case-control study was performed.89 CRC patients and 94 normal controls were examined,and Sequenom MassArray system was applied to detect the genotype of rs1569686 in DNMT3B gene polymorphisms.The association results were analyzed of the SNP and CRC.Results The frequencies of genotypes TT,GT and GG at SNP rs1569686 were 74.2％,24.7％ and 1.1％ in CRC patients respectively,which there were no significant differences from that in controls(73.4％,23.4％ and 3.2％; P＞0.05).Conclusion It suggests that the SNP rs1569686 of DNMT3B gene may not be related to susceptibility of CRC in Tianjin Han population.

Objective To explore the application of bumetanide to inhibition of tumor cell proliferation.Methods In different cell lines, the expression of natrium,kalium, chloride cotransporter 1 ( NKCC1) was detected by Western blotting while the proliferation of different tumor cells was examined by CCK-8 kit.Results The target protein NKCC1 expression in lung cancer cell line ( A549 ) and colorectal cancer cell line ( HCT116 ) was significantly higher than that in chronic myelogenous leukemia cell line (K562), esophageal cancer cell line (Eca109), cervical carcinoma cell line (HeLa), T lymphocytic leukemia cell line (Jurkat) and breast cancer cell line (MCF7).IC50 Values of bumetanide were significantly lower in A549 and HCT116 than in K562, Eca109,HeLa,Jurkat and MCF7.Furthermore, the inhibiory rate and the target protein expression level were positively correlated.Conclusion Bumetanide can inhibit tumor cell proliferation and NKCC1 can serve as a potential target of anticancer drugs.

OBJECTIVE To investigate the efficacy of rneropenem for the meatment of the patients with serious community-acquired pneumonia(SCAP).METHODS Sixty-four patients with SCAP were randomly divided into two groups.Group A(the treatment group,41 cases) was treated with meropenem(5-1.0 g iv q8-12 h?7 d) and group B(the control group,23 cases) treated with mezlocillin(5-1.0 g iv q8-12 h?7 d) in an open clinical study.RESULTS The efficacy rate of group A was 85.0%,with a bacteria elimination rate of 86.4% wheveas 57.0% and 64.3% in group B.There were significant difference between two groups(P

Objective To study the changes of platelet, endothelial functions and fibrinolytic activity of coronary artery disease (CAD) patients before and after PTCA Methods 26 negative patients of coronary angiocardiography (CAG) serving as negative control and 16 positive ones of CAG but unsuitable for PTCA as positive control, we observed the dynamic changes of plasma P selectin? thrombomodulin and plasminogen activator inhibitor 1 of 24 CAD patients before and after PTCA Results In PTCA group, the levels of plasma Ps and TM were higher 5 min after PTCA than those before PTCA ( P

Objective To study the regulatory T cells (Treg-cell) frequencies in patients with childhood acute leukemia and evaluate its clinical application value by investigating the relationship between the increasing numbers of Treg cells and minimal residual disease of leukemia (MRD), Methods Foxp3-FITC/CD25-PE/CD4-PerCP/CD3-APC four-color staining flow cytometry was established to identify Treg cells. Treg cells frequencies both in 10 healthy controls and in 33 patients with newly diagnosed childhood acute leukemia ( B-ALL 17 cases, T-ALL 9 cases, AML 7 cases) were detected. The possibility of the accumulation of Treg cells being the prognostic marker for acute leukemia was evaluated by comparing the results of Treg cells frequency with that of MRD. Results The percentage of Treg cells in CD+4 CD+3 T cells was M = 8. 09% in normal bone marrows, which was significantly different from the results in the bone marrows of newly diagnosed childhood acute leukemia ( M = 12.77% , U = 3.41, P < 0.01 ), but it showed no significantly differences among B-ALL, T-ALL and AML groups. No association was observed between the expression of Treg cells and clinical-biologic characteristics studied. In addition, Treg cells frequency in MRD positive group was significantly different from that in MRD continuously negative group ( M = 14. 74% vs M=11.3%, t =252.5,P<0.05). Conclusions Consistent with results from solid tumor, the study identifies a significantly increased numbers of Treg cells in patients with childhood acute leukemia. The situation of accumulation of Treg cells is closely associated with MRD results during chnical remission. High level of Treg cells may cause poor prognosis and increase the possibility of relapse.

In this paper, a colorimetric method with high sensitivity, reproducibility and low cost for determining glutamate decarboxylase (GAD) activity was developed based on Berthelot reaction. The optimum substrate system is 0.2 mol/L MacIlvaine buffer pH4.7, containing 0.1 mmol/L pyridoxal phosphate (PLP) and 10 mmol/L L-monosodium glutamate (L-MSG). The reaction mixture consisted of 200 ?L substrate solution and 1～100 ?L enzyme preparation from lactic acid bacteria and was incubated at 30℃. The enzymatic reaction was terminated by immersion in ice-water and addition of 200 ?L 0.2 mol/L sodium borate buffer, pH9.0, then 1 mL 6% phenol solution and 400 ?L sodium hypochloride were added. Color development was carried out in boiling water for 10 min then immediately put in ice-water bath for 20 min. The optical density is read at 630 nm. ?-aminobutyric acid (GABA) produced was calculated using the standard curve. Applications of the method in GAD studies were discussed.

Objective To investigate the effects of rehabilitation training on the microenvironment for axon regeneration and the possible mechanisms promoting axon regeneration,reconstruction and functional compensation after spinal cord injury (SCI) in beagle dogs.Methods Fifteen beagles were divided into a sham operated group,an SCI model group and a rehabilitation training group.Spinal cord hemi-transection injury was carried out.From the 8th day after SCI,the rehabilitation training group was given treadmill training.All dogs were sacrificed 60 days postinjury and samples were harvested.Chondroitin sulphate proteoglycan (CSPG) expression of astroglial cells around the injured area was observed using an immunofluorescence technique.The expression of glial fibrillary acidic protein (GFAP) and CSPG protein in each group were semi-quantified by Western blot analysis.Axon regeneration was observed by silver staining.Motor function was assessed using modified Tarlov scores.Results In the rehabilitation training group,compared with the model group astroglial production of CSPG was reduced remarkably.Both CSPG expression and axon regeneration were enhanced and functional deficits improved.Conclusions Rehabilitation training can inhibit astroglial production of CSPG,improve the microenvironment for injured axon regeneration,and promote reconstruction and functional compensation after traumatic SCI.

Objective To investigate the effects of soybean peptide calcium on apparent calcium absorption rate and bone mineral density of rats.Methods Totally 50 male Wistar rats were divided into 5 groups at comparable body weight:the low calcium control group,calcium carbonate control group and 3 groups with low,medium and high dose of soybean peptide calcium(SPC).Rats of each group were fed with designed forage.At the end of the fourth week of the feeding.calcium metabolic experiment was performed for 3 days and the apparent calcium absorption rate was calculated.Three months later,the left and right femurs of the rats were stripped to assay the bone calcium and bone density.Results Apparent calcium absorption rate of the calcium carbonate control group was 69.9 ％,which was lower than medium(82.6 ％) and high doses(77.4％) of soybean peptide calcium groups(F =31.371,P=0.000,0.001).Compared with the low calcium control group,rats with medium and high doses of SPC had obviously increased levels of weights(F =3.150,P =0.010,0.002),bone mass(F =3.195,P =0.015,0.002),calcium content (F=4.506,P =0.004,0.001)and bone mineral density of midpoint (F=6.185,P =0.006,0.000) and far end of femur (F =6.081,P=0.027,0.000).Despite of the weight index (P＞0.05),the high dose of soybean peptide calcium group also had the same results as the calcium carbonate control group (P=0.046,0.024,0.013,0.001).Conclusions Soybean peptide calcium can promote the apparent calcium absorption rate and increase the bone mineral density.

Objective To assess whether a follow-up endoscopy after endoscopic submucosal dissec-tion(ESD)for gastric lesions is effective for prevention of delayed bleeding. Methods Papers published in Medline,Embase,Cochrane library,Elsevier,Wanfang,VIP and CNKI were searched for the studies eval-uating the efficacy of follow-up endoscopy for reducing delayed postoperative bleeding after ESD for gastric le-sions. Statistic analyses were performed using RevMan 5. 1 software. Results A total of 10 studies including 4 randomized control trials and 6 cohort studies were enrolled. Analysis of these studies showed that the inci-dence of delayed bleeding was not significantly different between the patients receiving a follow-up endoscopy and those not(OR= 1. 38,95% CI:0. 89-2. 13,P= 0. 15). Also,there was no difference in the incidence of delayed bleeding between the prophylactic hemostasis and nonprophylactic hemostasis groups( OR = 1. 45, 95% CI:0. 65-3. 22,P= 0. 36). Conclusion A follow-up endoscopy after gastric ESD may contribute little to the prevention of delayed bleeding,but may be useful for the high-risk patients,which can control the bleeding rate at a low level.