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 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 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 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.

To improve the solubility of quercetin ( QT) , one of flavonoids that can inhibit the proliferation of vari-ous types of cancer cells, the novel amphiphilic polymer N-( 4-methylimidazole)-hydroxyethyl-chitosan ( MHC) , synthetized by chemical derivatization from chitosan, was used as the self-assembly micelles of QT. The formed polymer was characterized by 1 H NMR, elemental analysis and pyrene fluorescence spectrometry. The formulation of MHC micelles loading quercetin was optimized through single factor experiment. Then the optimized formulation was obtained as follows:the concentration of MHC was 0. 67% and the ratio of drug and carrier was 1 ∶10. The micelles particle size was ( 99. 21 ± 1. 71) nm, Zeta potential was +( 20. 01 ± 0. 72) mV and drug loading was ( 5. 42 ± 0. 32 )%. The in vitro release curve was investigated and was found to conform to Higuchi equation of Q=0. 1101 t1/2 -0. 064. The results of in vivo experiment showed that the mean rentention time and bioavail-ability of the MHC-QT micelles were 21. 42 h and 57. 49 μg h/mL, respectively, compared to 0. 30 h and 2. 50 μg h/mL of the free QT solution. These indicated that the MHC micelles could significantly improve the solubility of QT, the drug sustained-release effect and bioavailability, which would used as carrier for the anti-tumor drugs.

Objective To investigate the influential factors for the colonoscopic polyp detection rate (PDR) and adenomade detection rate(ADR) in high-risk population in community colorectal cancer screening.Methods Data of patients who were determined as high-risk population in community colorectal cancer screening tests and received colonoscopy in the Digestive Endoscopic Center of Tongji Hospital from May 2014 to November 2014 were collected.PDR and ADR were calculated by different genders, ages, history of abdominal operation, anesthesia assistance, quality of bowel preparation and experience of operators.Thex2 test and logistic regression analysis were used to compare the PDR and ADR between different groups.Results A total of 417 patients received complete colonoscopies.The total PDR was 35.25％.Univariate analysis revealed that gender (OR =2.298,95％ CI: 1.525-3.463), history of abdominal operation (OR =0.596,95％CI:0.390-0.910), anesthesia assistance (OR =1.864,95％ CI: 1.086-3.200) and quality of bowel preparation were significantly associated with PDR while multivariate analysis revealed that only gender, history of abdominal operation and quality of bowel preparation were independent influential factors for PDR.The total ADR was 30.22％.Like PDR, univariate analysis revealed that gender(OR =2.113,95％CI: 1.382-3.229), history of abdominal operation(OR =0.604,95％ CI:0.387-0.941), anesthesia assistance(OR =2.344,95％ CI: 1.283-4.281) and quality of bowel preparation were significantly associated with ADR.However, multivariate analysis revealed that only gender, history of abdominal operation and quality of bowel preparation were independent influential factors for ADR.Conclusion For patients who were highrisk in community colorectal cancer screening, men have greater risk to develop polyps and adenomas than women.Colonoscopy should be more careful in patients with history of abdominal operation, for quality of bowel preparation is directly related to PDR and ADR.