Objective To investigate the effect of berberine on inhibiting HT-29 human colon cancer cell proliferation induced by deoxycholic acid(DCA).Methods The berberine with concentration of 1,5,10 or 20?mol/L were added into the HT-29 human colon cancer cell culture media containing 200?mol/L DCA.The effects of berberine on cell proliferation were studied by the method of MTT.RT-PCR was applied to measure the expression of cyclooxygenase-2(COX-2)mRNA.Cellular immunochemical stain was applied to label COX- 2 protein expression.Concentration of prostaglandin E2(PGE2)was measured by radioimmunoassay. Results HT-29 cells were incubated with DCA for 6 h,COX2 expression of cells were increased prominent compared to controls(65.5%?5.6% vs.6.2%?1.1%).The level of PGE2 were increased(24.1 ng/L?1.4 ng/L vs.10.6 ng/L?0.8 ng/L).One?mol/L berberine reduced the proliferation rate of HT-29 in- duced by DCA over 6 h,the proliferation rate was 7.4?3.5%.Both COX-2 mRNA expression and the level of PGE2 were inhibited when the concentration of berberine was over 1?mol/L,and in a concentration-time dependent manner.Conclusions Berberine can inhibit the proliferation of HT-29 human colon cancer cell in- duced by DCA.Berberine can also suppress the expression of COX-2,and decrease the production of PGE2. These data provide new insights into the mechanism of its anti-cancer properties.

Objective To investigate the efficacy of using a calibrator with values assigned with the reference method for improving the comparability of serum gamma-glutamyltransferase (GGT) measurements.Methods The IFCC reference method for GGT was established and the performance was verified by testing a certified reference material (CRM).A calibrator was prepared and its value for GGT was assigned with the reference method.Forty serum samples were measured on different (including HITACHI 7600,7060,7170,7180 and BECKMAN LX20,OLYMPUS AU 400) chemistry analyzers with Zhongsheng GGT reagent kits calibrated with the calibrator.The samples were also measured on the same analyzers using a theoretical factor.Biases of results obtained with the calibration and with the theoretical factor based calculation were compared.Results The reference method resuhs on the CRM agreed the certified value within the stated uncertainty.Serum results calculated from the theoretical factor showed various biases and inter-analyzer variations.When the analyzers were calibrated with the calibrator,the number of results with biases less than 10% became significantly higher and those with biases more than 20% significantly lower.The variation of the results on 5 serum samples was reduced from 11.0%～14.0% to less than 5% by using the calibrator.Conclusion Accuracy and comparability of GGT measurements with of ZhongSheng GGT kits can be improved by using a calibrator that has a value assigned with the reference method.

Objective To detect the Yersinia pestis plasmid and molecular weight along the Qinghai-Tibet Railway.Methods Yersinia pestis plasmids molecular weight detected and analyzed using alkaline lysis,phenol-chloroform extraction of Yersinia pestis plasmid by agarose gel electrophoresis.Results The 18 Yersinia pestis strains of Qinghai-Tibet Railway contained 6×106,45×106,52×106,65×106,92×106plasmid,varing in the range of the 52×106-92×106.Conclusions The Yersinia pestis of Qinghai-Tibet Railway has a standardplasmid graphics,with the biggest Yersinia pestis plasmid changing in a certain regular degree,which providessignificance in the study of plague natural foci of the spatial structure and the genetic.characteristics of Yersiniapestis.

Objective To investigate the relationship between myocardial ischemia and slow coronary flow phenomenon with 99Tcm-methoxyisobutylisonitrile (MIBI) adenosine myocardial perfusion SPECT imaging. Methods Forty-four patients were divided to three groups according to the result of coronary angiography(CAG). There were GAG-positive(P-GAG) (n=12),slow coronary flow (CSF) (n =22),and normal coronary flow (NCF) (n = 10). Results of adenosine myocardial perfusion imaging were compared among these three groups. Semi-quantitative visual scoring method was used to evaluate the myocardial perfusion:0 = normal,1 = mild decrease,2 = moderate decrease,3 = severe decrease,4 = defect. Statistical analysis was performed using variance analysis,t-test and x2-test. Results No significance was observed at age ( t =0.27,0. 54 and 0. 59),sex (x2 = 0. 92),hypertension,hyperlipemia and diabetes (x2 = 1.23,all P ＞ 0.05 ) among the three groups. A significantly higher frames of the coronary thrombolysis in myocardial infarction (TIMI) flow was noted in CSF than in NCF groups (33.7 ±5.5 vs 17.6 ±3.9,t = 9. 58,P ＜0. 001 ). The positive adenosine myocardial perfusion imaging rate were significant among these three groups with 100% (12/12) in P-CAG group,77.3% (17/22) in CSF group,and 20% (2/10) in NCF group. When using semi-quantitative visual scoring method,significantly higher average ischemia segments were noted in CSF group than in NCF group ( 1.06 ± 0.77 and 0. 91 ± 0.80,t = - 2. 02,P ＜ 0. 05 ),but was less than that in P-CAG group (2.41 ±0.79,t =4. 54,P ＜0.001 ). The degree of ischemia of CSF group was higher than that in NCF group ( 8.01 ± 6.06,and 2.73 ± 2.60,t = - 2.07,P ＜ 0.05 ) and was less than that in P-CAG group (14. 07 ±12. 77 ,t=1.44,P＞0. 05). Conclusion Slow coronary flow phenomenon can be detected by adenosine myocardial perfusion image to offer the evidence of diagnosis and treatment for the chest pain patients with negative coronary angiography results.

Objective To investigate the accuracy and comparability of ?-glutamyltransferase (?- GT) measurement results on human serum samples and controI materials before and after calibration with a common human serum calibrator.Methods A human serum calibrator was prepared by pooling fresh serum aliquots and assigning a value for ?-GT with the IFCC reference method.The calibrator together with 5 human serum samples and 10 control samples were sent to 15 clinical laboratories and the sermn and control samples were measured with different analytical systems before and after a calibration with the calibrator.The results were analyzed for biases and interlaboratory variations.Results For the serum samples,the calibration resulted in reductions in biases from -9.0%～-14.2% to -0.8%～-7.9%,and in interlaboratory variations from 6.9%～11.6% to 2.8%～4.4%.No improvement was observed on the control samples.Conclusions Accuracy and comparability of serum ?-GT measurements can be improved by using a common human serum calibrator.Some control materials may not be commutable for human serum in ?-GT measurements.

OBJECTIVETo evaluate the in vitro and in vivo function of anti-human bladder tumor human-mouse chimeric antibody ch-BDI and its future clinical application.

METHODSWith ch-BDI in high-expression cell-line medium, affinity chromatography was used for the purification. Labeled with (99m)Tc through reduction method, its immunoreactive fraction and association constant were measured. The constant was injected into nude mice with xenografted human bladder tumor. The biodistribution of the labeled ch-BDI was studied with radioimmunoimaging.

RESULTSch-BDI showed desirable immunoreactive fraction (76%) and association constant (3.56 x 10(9) M(-1)) in vitro and a terrific specific targeting effect in vivo.

CONCLUSIONch-BDI has fairly good function against human bladder tumor both in vitro and in vivo, and is promising in clinical use.

Animals ; Antibodies, Monoclonal ; immunology ; Antibodies, Neoplasm ; immunology ; Antibody Affinity ; Humans ; In Vitro Techniques ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Recombinant Fusion Proteins ; immunology ; Urinary Bladder Neoplasms ; immunology

OBJECTIVETo investigate the effect of stromal interaction molecule 1 (STIM1) silencing on EPCs cell cycle.

METHODSRat bone marrow derived endothelial progenitor cells (EPCs) were isolated and cultured in L-DMEM with 20% FBS. Ad-si/rSTIM1 and Ad-hSTIM1 were then transfected into EPCs and the expression of STIM1 mRNA was detected by RT-PCR. The cell cycle was determined using flow cytometry analysis and intracellular free Ca2+ was measured using LSCM. Co-immunoprecipitation was performed to examine the interaction between STIM1 and TRPC1. Protein levels of inositol 1, 4, 5-trisphosphate were analyzed with ELISA assay.

RESULTSForty-eight hours after transfection, the expression of STIM1 mRNA was significantly downregulated (0.37 +/- 0.02 vs. 1.00 +/- 0.02, P < 0.05) and intracellular free Ca2+ level was significantly reduced (34.07 +/- 4.10 vs. 86.51 +/- 14.12, P < 0.05) in Ad-si/rSTIM1 group compared with control group. The cell cycle was arrested at G1 phase [(90.91 +/- 1.10)% vs. (77.10 +/- 0.56)%, P < 0.05] and the store-operated channel entry was strikingly inhibited in EPCs after treatment with Ad-si/rSTIM1. However, cotransfection of Ad-hSTIM1 with Ad-si/rSTIM1 significantly reversed these responses. Interestingly, co-immunoprecipitation study showed that STIM1 co-precipitated with TRPC1, and IP3 levels measured by ELISA were similar among three groups (P > 0.05).

CONCLUSIONsiRNA-mediated knockdown of STIM1 inhibited EPCs proliferation by reducing intracellular free Ca2+ through TRPC1-SOC signaling pathway.

Adenoviridae ; genetics ; Animals ; Cell Cycle ; Cell Proliferation ; Cells, Cultured ; Endothelial Cells ; cytology ; Gene Silencing ; Genetic Vectors ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; RNA, Small Interfering ; Rats ; Stem Cells ; cytology ; Stromal Interaction Molecule 1 ; Transfection ; Transient Receptor Potential Channels ; metabolism

Objective: To establish an echocardiography parameter scoring system for assessing the risk of 1 year re-admission in patients with left ventricular systolic dysfunction (LVSD). Methods: A total of 412 chronic LVSD patients treated in our hospital from 2007-01 to 2016-01 were studied and the end point event was 1 year re-admission. The data included in 280 patients from 2007-01 to 2014-12 for establishing the scoring system and 132 patients from 2015-01 to 2016-01 for verifying the system. Based on 7 echocardiography parameters, the patients were divided into 7 sets of groups: ① Left ventricular diameter (LVD): Group0, n=290 and Group1, n=122;② Mitrial regurgitation (MR): Group0, n=203, Group1, n=138 and Group2, n=71; ③ Tricuspid regurgitation (TR): Group0, n=302, Group1, n=90 and Group2, n=20; ④ LVEF: Group0, n=272 and Group1, n=140; ⑤ Pulmonary artery systolic pressure: Group0, n=282 and Group1, n=130; ⑥ Hydropericardium: Group0, n=347 and Group1, n=65; ⑦ Hydrothorax:Group 0, n=261, Group1, n=86 and Group2, n=65. The parameters were identified by COX regression analysis, weighted value of scoring system was calculate by hazard ratio (HR), predictive value for1 year re-admission was assess by ROC curve and finally, scoring integration was verified by validation data group. Results: The integration score was calculated as follows: LVD>60mm=1 point; TR: Group1=1 point and Group2=3 points; MR: Group1=2 points and Group2=4 points; Hydrothorax: Group1=2 points and Group2=3 points;Hydropericardium=1 point. COX regression analysis indicated that for 1 year re-admission: HR=1.552 in Group1 vs Group0, HR=3.374 in Group2 vs Group0 and HR=4.562 in Group3 vs Group0, all P<0.05. The AUC of ROC for establishing the data was 70.0% (95% CI 0.640-0.761) and for verifying the data was 70.4% (95% CI 0.616-0.792); the best integration score was 4 points. Conclusion: Echocardiography parameter scoring system may better predict the risk of 1 year re-admission in LVSD patients which is superior to single echocardiography parameter.

Objective:To study changes and clinical significance of serum interleukin-33(IL-33) of patients with adult measles complicated acute liver damage. Methods: Totally,88 adult measles patients were selected and divided into acute liver damage group (n=60) and no liver damage group(n=28). At the same time,20 healthy adults were selected as healthy controls. Serum IL-33 levels were measured by enzyme linked immunosorbent assays. Venous blood samples of IL-33 were drawn on the day of admission and were repeated at 7 and 14 days later. The correlation of IL-33 and biochemical indicators were analyzed. 60 patients with liver damage were divided into protecting liver and without protecting liver group in accordance with the method of random numbers. The level of serum IL-33 was compared at 7 and 14 days between the two groups of patients. Results: Patients with adult measles complicated acute liver damage and no liver damage had elevated serum IL-33 levels than that in the controls [( 205. 20 ± 25. 74 ) pg/ml, ( 168. 70 ± 18. 14)pg/ml,vs(132. 17±12. 41)pg/ml,P＜0. 05 for all],especially in acute liver damage group. The expression of serum IL-33 in no protecting liver group was significantly higher than protecting liver group at 7th day. IL-33 was no significant difference in patients between protecting liver and without protecting liver groups at 14th day(P＞0. 05). The level of IL-33 was a significant decrease in the treatment and there was no significant difference between healthy controls at 14th day(P＞0. 05). The level of IL-33 and liver damage were consistent. Serum IL-33 levels in patients with liver damage showed positive correlation with levels of ALT,GGT and IL-6 ( r=0. 392 1,0. 503 9,0. 724 9,P＜0. 001). Conclusion: IL-33 may have proinflammatory effects in the phase of adult measles complicated acute liver damage and its serum level reflects the severity of liver inflammation.

OBJECTIVETo investigate the biological characteristics of dermal fibroblasts of the diabetic rats with deep partial thickness scald, and to explore its relationship with delayed wound healing due to diabetes.

METHODSSprague-Dawley rats weighing 250 g were randomly divided into control (NM, n=40) and STZ-induced diabetic (DM, n=50) groups, and then deep partial thickness scald involving 10% TBSA were reproduced in the two groups. Skin samples were harvested from the wounds on 0, 3, 7, 14 and 21 post scald day (PSD) for the determination of certain histological characteristics.

RESULTSThe thickness of dermis layer in DM group before injury was obviously thinner than that in NM group (P < 0.01). There was an infiltration of a large amount of chronic inflammatory cells and increased content of cutaneous glucose in the dermal tissue in DM group (2.77 mg/g) compared with 0.85 mg/g in NM group, (P < 0.01). An accumulation of advanced glycation end products (AGEs) was found in the dermal tissue in DM group. After the scalding, the percentage of fibroblasts in S phase and hydroxyproline synthesis in DM group was evidently lower than those in NM group. But the apoptosis rate of fibroblasts was much higher in DM group than that in NM group (P < 0.05 or 0.01).

CONCLUSIONIt is found that the high contents of glucose and AGEs in diabetic skin exert untoward effects on biological characteristics of dermal fibroblast, probably constituting one of the underlying mechanisms of delay wound healing of scald in diabetic rats.

Animals ; Burns ; metabolism ; pathology ; Diabetes Mellitus, Experimental ; Fibroblasts ; cytology ; Glycation End Products, Advanced ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; metabolism ; pathology ; Wound Healing