Objective To investigate diagnostic value of miRNAs for esophageal cancer .Methods Blood samples were obtained from patients with esophageal cancer .MiRNAs including miR-30a ,miR-126a ,let-7b were detected and compared with healthy con-trols .After adjusting smoking and insobriety ,Logistic regression was used to judging which was best for clinical guidance .ROC curve was used to analyzed the clinic value of these miRNAs .Results Age of the two groups had no significant difference(P>0 .05) .The levels of smoking ,alcoholism and family history of cancer in the two groups had significant difference (P<0 .05) .Two groups′miR-30a levels in plasma had no significant differences (P>0 .05) .Let-7b and miR-126a levels in plasma in patients with e-sophageal were significantly higher in healthy volunteers (P<0 .05) .After adjustment the factors of smoking and alcohol ,let-7b lev-els were associated with the risk of esophageal cancer (P<0 .05) .Area under the ROC curve was 0 .712 ,which had a certain esoph-ageal cancer diagnostic performance .Conclusion Plasma let-7b could be treated as a biomarker for the diagnosis of esophageal canc-er.

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 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 predict the basic physicochemical properties ,structure and function ,and linear B-cell epitopes of the capsid protein VP1 of coxsackievirus A6(CVA6).Methods: The amino acid sequence of the CVA6 VP1 was analyzed using Bioedit software and various online tools including SubLoc ,TargetP and the others from ExPASy Bioinformatics Resource Portal.Results: The CVA6 VP1 protein was a hydrophilic protein with a relative molecular weight of 33.6 kD and an isoelectric point of 7.92.This protein containsed 24 phosphorylation sites , but no signal peptide , transmembrane domains and possible fatty acylation sites.Its secondary structure was characterized by the richest random coils , and 48.52 percent of its amino acid residues exposed at the solution inter-face.Epitope prediction by Bepipred showed a number of potential B cell epitopes in the protein ,the highest antigenicity index among them located in the region of amino acids residue 155-165.Conclusion:The basic physicochemical properties ,structure and function characteristics ,and potential linear B-cell epitopes of CVA 6 VP1 were successfully predicted , which laid foundations for the further study on the protein and the preparation of vaccines and immunological diagnostic reagents for CVA 6 infection.

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.

To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.
Base Sequence ; Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Genetic Vectors/genetics ; Hep G2 Cells ; K562 Cells ; Molecular Sequence Data ; Receptors, Transferrin/*immunology ; Recombinant Fusion Proteins/biosynthesis ; Recombinant Fusion Proteins/genetics ; Single-Chain Antibodies/*biosynthesis ; Single-Chain Antibodies/genetics

To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complemenary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease Sill, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB 1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively.Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.

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.

AIM: To analyze curative effect of four regulating-intestines prescriptions (Wumei Wan, Baitouweng Tang, Shenling Baizhu San and Tongxie Yao-fang) on the treatment of ulcerative colitis (UC) through summing up the previous animal experimental results.METHODS: We collected the conclusions in the papers related to four regulating-intestines prescriptions for the treatment of UC which were published recently, made comparisons from the aspectsof symptoms, physical sign, pathological change, cytokine and its protein expression, blood adhesion molecule, cell apoptosis and controlling gene and analyzed the pathogenesis of UC and action mechanism of four regulating-intestines prescriptions. Four regulating-intestines prescriptions included Wumei Wan,Baitouweng Tang, Shenling Baizhu San and Tongxie Yaofang. Wumei Wan is from Treatise on Exogenous Febrile Diseases and Synopsis of the Golden Chamber, which consists of dark plum, asarum herb, dried ginger, Chinese goldthread, Chinese angelica root, aconite root, pricklyash peel, cassia twig,ginseng (sun-dried ginseng) and bark of cork tree and has marked effect in treating colic caused by ascariasis and persistent dysentery. Baitouweng Tang, from Treatise on Exogenous Febrile Diseases, consists of medicinal herbs such as pulsatilla root, Chinese goldthread, bark of cork tree and ash bark, which has functions of clearing away the heat-evil, expelling superficial evils and relieving dysentery. In addition, it has marked effect in treating heat-type dysentery. Shenling Baizhu San, from Prescriptions of Peaceful benevolent Dispensary, consists of medicinal herbs such as pulp of lotus seed, coix seed, amomum fruit, balloon flower root, white hyacinth bean, poria, ginseng (sun-dried ginseng), glycyrrhiza, bighead atractylodes and rhizoma dioscoreae, which has the nature of replenishing qi to invigorate the spleen and eliminate wetness to arrest diarrhea and has marked effect on treating diarrhea due to the hypofunction of spleen. Tongxie Yao-fang which is from The Complete Works of Zhang Jingyue consists of four herbals of agehead atractylodes, root of herbaceous peony, dried tangerine peel and ledebouriella root and has the functions of soothing liver and invigorating spleen and stopping diarrhea, and has marked effect on treating liver sthenia and deficient spleen, borborygmus and abdominal pain and diarrhea.RFSULTS: ① Because UC is a chronic protracted dysentery with deficiency of vital energy and existing of evil energy, the vital energy will be harmed if its treatment is specialized in removing and dissolving the stagnation, evil energy will continue to exist and stagnation will continue to accumulate if its treatment is specialized in strengthening vital energy and inducing astringency. Only supporting healthy energy and expelling evil energy is the correct therapy method. This is in accordance with the main treatment of Wumei Wan ②Eliminating dampness and pathogenic heat from the blood to treat diarrhea is the main treatment method of Baitouweng Tang, and this is incompletely suitable for the treatment of UC.③Shenling Baizhu San had the effect on soothing liver and invigorating spleen and stopping diarrhea, and this is also incompletely suitable for the treatment of UC. ④ The prescription of Tongxie Yaofang is used for treating diarrhea caused by deficient spleen and liver sthenia, and spleen controlled by liver, and abnormal ascent and descent. It accords with main pathogenesis of UC, deficient spleen, excessive dampness of deficient spleen and it is weaker in invigorating the spleen in catabasis of UC than Shenling Baizhu San. Therefore, Wumei Wan has the best curative effect,Baitouweng Tang the second, Shengling Baizhu San the third and Tongxie Yaofang a little poor.CONCLUSION: UC belongs to recurrent dysentery, and its pathogenesis is in accordance with the main treatment of Wumei Wan, but not the other three prescriptions, so Wumei Wan is the most efficient prescription in treating ulcerative colitis. Diagnosis and treatment based on an overall analysis of signs and symptoms of TCM is the premise of obtaining the best curative effect. Modeling of animal tests must be consistent with the type of syndrome of the traditional Chinese medicine. Ating ulcerative colitis (UC) from aspects such as symptom, physical sign, pathological changes, adherence factor, cytokine and its protein expression, apoptosis and its controlling gene by means of modeling, which proves their functions and effects are different and their curative effect are also different due to their different ingredient though they all have the functions of treating. Results of this test show Wumei Wan has the best curative effect, Baitouweng Tang the second, Shenling Baizhu San the third and Tongxie Yaofang a little poor[1-11] Mechanism of four regulating-intestines prescription in the treatment of ulcerative colitis is discussed from the viewpoints of traditional Chinese medicine as follows.