Hepatocellular carcinoma(HCC)is one of the most common malignancies worldwide.Due to the difficulty of diagnosis in the early stage of HCC, most HCCs are diagnosed in intermediate-advanced stage.Moreover, the high invasion, metastasis and recurrence rate of HCC result in the high mortality of HCC.MicroRNAs(miRNAs) are a class of highly conserved, endogenous, small, non-coding ,single stranded RNA with the length of 22 nucleotides.There are plentiful of miRNAs in liver.MiRNAs not only can regulate the growth and development of liver, but also are closely related to the formation of HCC.In the process of HCC formation, miRNAs could function as oncogenes or tumor suppressor genes to regulate multiple biological processes related to HCC, including cell differentiation,proliferation,tumorigenesis,angiogenesis,invasion,and metastasis.With the intensive study of molecular mechanisms of miRNAs in the process of HCC formation, increasingly studies have revealed that miRNAs could become sensitive biomarkers and effective therapeutic targets for HCC.

AIM To study the effects of Biejiajian Pills (Colla Carapacis Trionycis,Asini Corii Colla,Nidus Vespae,etc.) on NF-κB,p65,p50 and IκB in NF-κB signaling pathway and target gene expression in HSC-T6 cells of rats.METHODS HSC-T6 cells were cultured with Biejiajian Pills drug serum for 24 hours,the expressions of p65,p50,VEGF and TIMP-1 mRNA were determined by qPCR;the expression of p65 was measured by immunofluorescence;the expressions of IκBα,IκBβ and α-SMA were determined by Western blot.RESULTS The Biejiajian Pills middle-,high-dose and positive control groups showed significantly lower expressions of p65,VEGF and TIMP-1 mRNA as compared with the blank control group and negative control group,the expressions of p50 mRNA among various groups showed no significant differences.But immunofluorescence showed that the expression of p65 in cytoplasm was decreased.Meanwhile,Biejiajian Pills showed significantly higher IκBα protein expression and obvious down-regulation of α-SMA expression in a dose-dependent manner,but had no significant influence on the expression of IκBβ.CONCLUSION Biejiajian Pills' therapeutic effects on hepatic fibrosis may be related to influencing NF-κB signaling pathway and inhibiting the expression of down-stream target gene.

BACKGROUND: The study of cell transplantation to repair injured cardiac muscle and improve cardiac function of dilated cardiomyopathy (DCM) has become a hotspot in recent years. However, the effect of cardiac electrophysiology following transplantation is still unknown.OBJECTIVE: To explore the influence of ailogenic implanted mesenchymal stem cells (MSCs) on cardiac function, structure and electrophysiology of rabbits with DCM.DESIGN, TIME AND SETTING: Randomized controlled animal trial was performed at Electrophysiology Laboratory of Institute of Pediatrics, Chongqing Medical University between January 2004 and May 2006.MATERIALS: Forty-three New Zealand whiterabbits, weighing 3.0-3.5 kg, irrespective of gender, were selected. Adriamycin was produced by Haizheng Medical Products Company of Zhejiang Province, China, No. 050307. The Ultrasonograph SSD-5000 came from Aloka, Japan, and RM6240 multiplying channel electrophysiolograph of Chengdu Instrument Company was applied.METHODS: All animals were randomized into normal group (n=12), cell transplantation group (n=13), and DCM model group (n=13). Except the normal group, adriamycin was applied to create rabbit DCM model, I mg/kg, twice a week for successive 8 weeks. Bone marrow solution was harvested from the remaining 5 rabbits, and MSCs were isolated, amplified and purified using attachment method. Three weeks after modeling, the MSCs were transplanted into left ventricle anterior wall at four sites in cell transplantation group, model group was injected with matching DMEM/FI2 medium, while the normal group was not given any treatment.MAIN OUTCOME MEASURES: At four weeks postoperatively, left ventricular end-diastolic dimension, end systolic dimension,left ventricular ejection fraction, and shortening fraction were monitored by ultrasonograph; the value for monophasic action potential amplitude (MAPA) and the maximum velocity in 0 phase (V,,~), the value for 50% monophasic action potential durations (MAPDs0) and 90% monophasic action potential durations (MAPDg0) were detected by electrophysiolograph. In addition, the cardiac tissues harvested from implanted region were observed by light microscope and electron microscope, and also the expression of cardiac troponin T (cTnT) and connexin 43 (Cx43) was detected through immunohistochemistry.RESULTS: Of 43 rabbits, 9 rabbits each of the transplantation group died of the acute or chronic toxic effects of Adriamycin, finally, 25 rabbits were included in final analysis. Compared with model group, the end systolic dimension was diminished, and the left ventricular ejection fraction and shortening fraction in cell transplantation group were significantly increased (P ＜ 0.05). The MAPDs0 and MAPDgo in cell transplantation group prolonged significantly compared with model group (P ＜ 0.05). In model group,cardiac myocytes appeared mitochondria swelling and hyperplasia, and their sarcomere had different lengths, arranged unevenly,accompanied by abnormal contraction bands. In addition, myolysis was found in parts of myocardium under electron microscope.However, the structural abnormalities in the cell implantation group were less than the DCM model group, and the implanted MSCs could express cTnT and Cx43.CONCLUSION: Allogenic MSCs transplantation can improve cardiac function, lessen structural abnormalities and may inhibit the progression of electrophysiology derangement.

Objective To validate the value of Simplified Renal Index Score(SRI) in predicting acute renal injury requiring renal replacement therapy(RRT-AKI) after cardiac valve surgery in Chinese adult patients.Methods An analysis was conducted for all the adult patients who underwent cardiac valve surgery from January 2010 to December 2014 in Changhai Hospital,Shanghai.A total of 3 183 adult patients were included.Based on SRI Score,the patients were divided into 3 risk stages:0 to 1 point,2 to 3 point,and 4 to 8 point.The incidence of RRT-AKI was compared between different stages.And the prediction value of the SRI model was assessed by area under the receiver operating characteristic curve (AU-ROC) and the model calibration was assessed with the Hosmer-Lemeshow (H-L) test.Results After surgery 52 (1.6％) patients developed acute kidney impairment and subsequently underwent renal replacement therapy.Patients with low values of simplified renal index (0-1),medium(2-3) and high values (4 and more) were found to have increasingly higher risk for renal replacement therapy of 0.8％ (95％ CI:0.005-0.012) 、3.8％ (95％ CI:0.026-0.052) 、20％ (95％ CI:0.010-0.720),respectively.TheAU-ROCwas0.68(95％ CI:0.610-0.760,P＜0.01).The H-L test was x2 =2.45,P=0.29.Conclusion SRI model gives a certain clinical significance,suggesting that high-values patients may occur RRT-AKI with a significantly higher risk than low-values patients.However,SRI model cannot give an accurate prediction value for RRT-AKI in Chinese adult patients after cardiac valve surgery.Direct clinical use of the model should be considered cautiously.

Objective To explore the change regulation of blood coagulation function before and after treatment of snake bite pa‐tients .Methods A total of 226 hospitalized patients with clear diagnosis belong to what kind of snakes in the past two years were selected in this study ,39 cases were bit by coral ,76 cases were bit by trimeresurus stejnegeri ,47 cases were bit by cobra ,24 cases bit by king cobra ,40 cases were bit by adder .Five blood coagulation indexes were determined before and after treatment various pe‐riods for these patients .The tested results were made to statistical analysis according to kind of snakes ,periods and disease condi‐tion .Results D‐D level obvious .rise before treatment for the severe cases patients of coral snake bite (P<0 .05) .Plasma prothrom‐bin time (PT) ,activated partial clotting enzyme live time (APTT) ,thrombin time (TT)and D‐D level rise and Fib level reduced be‐fore treatment for the severe cases patients of trimeresurus stejnegeri bite ,and Fib and D‐D level before and after treatment had sta‐tistical significant difference with the mild patients (P<0 .05) .Every index for the cobra and king cobra had no statistical signifi‐cant differences (P>0 .05) before and after treatment ,only D‐D level of the severe cases had statistical significance before treat‐ment in contrast to the mild cases of cobra bite (P< 0 .05) .The changes of 5 item blood coagulation indexes were all very large whether mild or severe for viper bite patients ,a number of comparisons had statistical significance difference (P<0 .05) whether before or after treatment and whether mild or severe for viper bite patients .Conclusion The bite of coral snake ,cobra and king co‐bra affect little for the blood coagulation function .The severe cases patients of trimeresurus stejnegeri bite might lead to more seri‐ous blood coagulation function abnormality but the recovery is faster after treatment .The viper bite might lead to maximum blood coagulation function abnormality and the recovery is slow after treatment .

Objective To validate the value of Cleveland Clinical Score in predicting acute renal injury requiring renal replacement therapy(RRT-AKI) after cardiac valve surgery in Chinese adult patients.Methods An analysis was conducted for all the adult patients who underwent cardiac valve surgery from January 2010 to December 2014 in Changhai Hospital,Shanghai.A total of 3 230 adult patients were included.Based on Cleveland Clinical Score,the patients were divided into 3 risk stages:0 to 2 point,3 to 5 point,and 6 to 8 point.The incidence of RRT-AKI were compared between different stages.And the predictive value of the Cleveland Clinical Score model was assessed by area under the receiver operating characteristic curve(AUC-ROC) and the model calibration was assessed using the Hosmer-Lemeshow test.The patients were also divided into two groups:Non-RRT group and RRT-AKI group.The mortality were compared between these two groups.Results The incidence of RRT-AKI was 1.67％ vs the predicted ratio of RRT-AKI 1.70％ (x2 =0.018,P =0.892).Among the stage 1,2,and 3,the actual incidence of RRT-AKI,was 1.23％,2.66％,and 16.7％ vs the predicted incidence 0.40％,1.80％,and 9.50％,respectively.The AUC-ROC for Cleveland Clinical Score predicting RRT-AKI was 0.64 [95 ％ CI(0.57,0.71),P ＜0.01].Compared with Non-RRT group,the RRT-AKI group got a higher mortality(87.00％ vs 1.50％,x2 =1 330,P ＜0.01).Conclusion The Cleveland Clinical score had no real predictive value for RRT-AKI in Chinese adult patients after cardiac valve surgery.The incidence of RRT-AKI of the whole population and the stage 3 patients could be predicted by the model.And the patients with a high Cleveland score got a higher mortality than that of patients with a low Cleveland score.

Objective To analyze the abnormal detectable rates of different kinds of blood test indexes before and after treatment in the patients with snake bite and to probe into the change condition of these indexes after different snake bite .Methods The inpa‐tients with clearly diagnosed what kind of snake bite in the past two years were selected as the research subjects .The multiple blood test indexes were determined before and after treatment in these patients .The abnormal detectable rates of these indexes were ana‐lyzed and compared among various snake bite patients .Results The different kinds of snake bite all could cause the different de‐grees of changes in some detection indexes among partial patients .Specially ,the detectable rates of WBC ,PT ,APTT ,TT ,D‐D ,CK , CK‐MB ,LDH ,Urea ,Cr and Cys‐C increase and PLT and Fbg decrease caused by viper bite were apparently higher than those caused by other kinds of snake bites (P<0 .05) .The detectable rates of CO2 increase and K+ decrease in the coral snake bite were apparently higher than those in trimeresurus gramineus ,cobra and viper bite (P<0 .05) .Conclusion Because the snake species and toxicities are different in the various snake bites ,so the caused changes and the abnormal detectable rates of blood test indexes also are different .

In order to investigate the expression of endothelin receptor B (ETR-B) in human malignant melanoma (MM) cells A375 and SK-mel-1 and the proliferative effects of endothelin 3 (ET3) on A375 cells, RT-PCR was applied to detect the expression of ETR-B gene in human MM cells A375 and SK-mel-1. MTT method was used to evaluate the growth enhancing effects of ET3 on A375 cell line in vitro. The results showed that ETR-B gene was expressed in both MM A375 and SK-mel-1 cells. ET3 had stronger ability to enhance the proliferation of A375 cells in vitro in a concentration-dependent manner. It was suggested that ET3/ETR-B might play an important proliferative role in MM.
Cell Line, Tumor ; Cell Proliferation/*drug effects ; Endothelin-3/*pharmacology ; Melanoma/*metabolism ; Melanoma/*pathology ; Receptor, Endothelin B/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction

In order to investigate the expression of endothelin receptor B (ETR-B) in human ma-lignant melanoma (MM) cells A375 and SK-mel-1 and the proliferative effects of endothelin 3 (ET3) on A375 cells, RT-PCR was applied to detect the expression of ETR-B gene in human MM cells A375 and SK-mel-1. MTT method was used to evaluate the growth enhancing effects of ET3 on A375 cell line in vitro. The results showed that ETR-B gene was expressed in both MM A375 and SK-mel-1 cells. ET3 had stronger ability to enhance the proliferation of A375 cells in vitro in a con- centration-dependent manner. It was suggested that ET3/ETR-B might play an important proliferative role in MM.