OBJECTIVE：Analyzing the epidemic of drug risistance bacteria with extend spectrum β-lactamase(ESBLs)in children's wards to cut off the transmitting pathway and prevent the outbreak of nosocomial infections in time.METHODS：Samples，including sputum，umbilical secretion，pus，pharyngeal swab，were collected from in-hospital child patients in Departments of Newborn，Respiratory Tract Diseases，Cardiology，Nephrology，Hematology，Pediatric Surgery and ICU，then bacterial cultures and drug susceptibility test were carried out.54 strains of ESBLs E.coli and Klebsiella pneumoniae were isolated and their plasmid profiles were analysed by agarose electrophoresis.RESULTS：The percentages of ESBLs in the above-mentioned departments were 46.3，12.9，5.6，12.9，5.6，11.1，5.6 respectively.In analysis，three types of profiles were found and strains with four types of the same profile concentrated in the Newborn Department.The others were dispersed.CONCLUSION：Nosocomial infections possibly occured in the Newborn Department to some extent.

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.

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.

Objective To observe the effect of ambroxol on the structure of Pseudomonas aeruginosa biofilm (BF) in an established biofilm model in vitro. Method Mature BF was formed after 7 days′ culture of Pseudomonas aeruginosa on plate. The BF structure was observed by scanning electron microscope (SEM). The parameters of BF structure were analyzed though pictures from confocal laser scanning microscopy (CLSM) with Image Structure Analyzer (ISA) software. Fluorescence intensity of the viable bacteria in BF treated by ambroxol alone and together with ciprofloxacin were determined with fluorescence microscopy. Results After the treatment of ambroxol, the BF was destroyed and the matrix outsides became thin and abnormal compared with the control by SEM, and small amount of bacteria dispersed. The results of ISA showed that with the treatment of 2 mg/ml ambroxol, BF was decreased in thickness, average diffusion distance (ADD) and textual entropy (TE) were decreased in comparison with the group without ambroxol treatment (P

Objective To explore whether the neurons induced from mesenchymal stem cells(MSCs) have synapse function or not.Methods Passage 4-5 MSCs in good shape was induced by Salvia miltiorrhiza with optimized protocol for several times,and then the observation under inverted phase contrast microscope,immunofluorocytochemistry and the measurement of neurophysiological function were carried out.Ca2+ influx and synapse function were detected with laser-scanning confocal microscopy(LSCM),taking 50 mmol/L KCl as stimuli to evoke action potential.Results The cells that had been induced for 5 h at 4th time from MSCs,looked like neurons and displayed that the processes stretched out to form complex net.Immunofluorocytochemistry presented that the rate of TUJ-1 expression was(96.7?2.8)% and that of synaptophysin was(96.2?2.1)%.When the neuron-like cells were stimulated by high concentration of KCl,intracellular Ca2+ influx enhanced quickly.When the neuron-like cells were stimulated by high KCl solution for the first time,SynaptoRed-C2 anchored onto the membrane,and after the second excitation,the fluorescence intensity decreased quickly.Conclusion The neuron-like cells derived from MSCs that are induced by salvia miltiorrhiza with optimized protocol have synapse function.

Objective To explore the distribution and differentiation of bone mesenchymal stem cells (MSCs) in the brain of neonatal rats with hypoxic ischemic encephalopathy (HIE). Methods MSCs were isolated and purified by adhering to the culture glassware wall and prelabeled with bromodeoxyuridine (BrdU) for 72 h before transplantation. The model of HIE was established. At 24 h after hypoxic ischemia, approximate 4?10 6 cells were injected into the brain of neonatal rats with HIE through the right side bregma. The Nestin, neuron specific enolase (NSE) and glial fibrillary acid protein (GFAP) were detected by immunofluorochemistry at 4 weeks after injection. Results Majority of MSCs were distributed in the cortex, hippocampus of the lesioned hemisphere. The number of MSCs was (2781?254) in the left hemisphere, but (4708?281) in the right hemisphere. There was significant difference (t=18.70, P0.05). The expression ratio of NSE was (3.79?0.95)% in the left hemisphere, but (5.69?1.48)% in the right hemisphere (t=3.404, P0.05). Conclusion MSCs are mainly distributed in the lesioned hemisphere and can differentiate into neuronal-like cells, express the mark of neural stem cells, neurons and neuroglial cells at 28 d after intracortical transplantation.

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 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.

In order to further investigate the effect of annexin II (Ann-II) on tissue plasminogen activator (t-PA)-dependent plasminogen (PLG) activation and its interactive mechanism, recombinant native Ann-II bound t-PA, PLG and plasmin with high affinity was examined. The flow cytometric assay showed that the ann-II expression rate was higher in the human umbilical vein endothelial cell (HUVEC) (87.65%) than in the HL-60 cells as controls (35.79%). Two irrelevant proteins, bovine serum albumin (BSA) and equine IgG (EIG) had no effect on the production of plasmin. Ann-II-mediated enhancement of t-PA-dependent PLG activation was inhibited by epsilon-aminocaproic acid or by pretreatment of Ann-II with carboxypeptidase B with the inhibitive rate being 77.8% and 77.0%, respectively. It was revealed that the effect of Ann-II on PLG activation was specific for t-PA. Urokinase didn't bind to Ann-II, demonstrating the role of receptor-related lysine residues on activation of PLG, showing that the Ann-II-PLG interaction was dependent upon carboxyl-terminal lysine residues. These findings suggest that annexin II-mediated co-assembly of t-PA and PLG may promote plasmin generation and play a key role in modulating fibrinolysis on the endothelial surface.
Annexin A2/*pharmacology ; Cells, Cultured ; Endothelium, Vascular/cytology ; Endothelium, Vascular/*metabolism ; Fibrinolysis ; Plasminogen/*metabolism ; Recombinant Proteins/pharmacology ; Tissue Plasminogen Activator/*metabolism ; Umbilical Veins/cytology