Objective To detect the expression and the biological activity of a recombinant adenovirus expression vector carrying human neurotrophin-3 (NT-3) receptor TrkC gene (Adeno-TrkC) in neural stem cells. Methods The expression of TrkC mRNA in 293 cells infected with Adeno-TrkC was detected by RT-PCR, and the expression of TrkC protein in neural stem cells infected by Adeno-TrkC was detected with immunocytochemistry and Western blotting. The effects of human neurotrophin_3 (NT-3) on the neural stem cells infected by Adeno-TrkC differentiating into neuron-like cells and astrocyte-like cells in vitro were observed. Results The transcription of TrkC mRNA and the expression of TrkC protein was detected in 293 cells and neural stem cells infected by Adeno-TrkC. This kind of TrkC was able to make more neural stem cells differenting into neuron-like cells in vitro with its ligand NT-3 and the percentage of neuron-like cell’s differentiation was 55^2% which was higher than that of other control groups.Conclusion TrkC protein is expressed in neural stem cells transfected with Adeno-TrkC and is able to make neural stem cells differenting into neuron-like cells. The present study may provide the basis on gene therapy of central nervous injury using further NT-3 and TrkC.

This paper will address the development of disaster psychology and its important contribution regarding its theoretical and practical frameworks as it applies to the psychological counseling for survivors suffering from the consequences of natural disasters.This paper will also analyze how the approach of disaster psychology can be applied to work with survivors suffering from natural disasters and to alleviate psychological challenges and its influential factors which affect these individuals.Several strategies have been discussed and proposed.In addition,relevant theoretical and integrated approaches for psychological intervention support and guidance,and rehabilitation have been established.

Objective To investigate correlation between the results of drug susceptibility and the extent of drug-resistances in Mycobacterium tuberculosis clinical isolates. Methods Liquid culture and MTT test were used. Twelve anti-TB drug MICs and drug susceptibility testing of the 163 MTB strains from random clinical isolates were detected, which including RFP, INH, SM, EBM, OFLX, LVFX, MOX, AMK,CPM, PTA, CLA and PAIN. Results There are 67% (42/62) Mycobacterium tuberculosis strains resistant to SM, 63% (51/81) Mycobacterium tuberculosis strains resistant to INH, 77% (50/65) Mycobacterium tuberculosis strains resistant to RFP, 41% ( 15/37 ) Mycobacterium tuberculosis strains resistant to AMK,41% (12/29) Mycobacterium tuberculosis strains resistant to CPM, 20% (12/60) Mycobacterium tuberculosis strains resistant to EMB and 43% (25/58) Mycobacterium tuberculosis strains resistant to OFLX which MICs were equal to or more than 16 μg/ml, 8 μg/ml, 8 μg/ml, 16 μg/ml and 4 μg/ml, 4 μg/ml and 8 μg/ml,respectively. There were significant differences in the MICs of OFLX, LVFX and MOX in OFLX resistant strains (2-128, 1-32 and 0.0625-1 μg/ml, respectively) by ANOVA ( F = 16.874, P ＜ 0.001 ). The MICs of SM, INH, RFP, EMB, OFLX, AMK and CPM in isolates resistant to six or seven drugs (0.5-128,2-64,0.25-128,1-32,1-64,0.5-128 and 1-128 μg/ml,respectively) were higher than those (0.25-128,0.0625-64,0.25-32,0.25-2,0.125-2,0.5-4 and 1-4 μg/ml,respectively) in isolates resistant to one or two drugs (F=20.066, 40.499, 47. 197, 70.373, 91.432, 41.840 and 21.547, respectively, P ＜0.05). The MICs of SM, INH, RFP and EMB in isolates resistant to four drugs (1-128,2-64,0.25-128 and 1-32 μg/ml,respectively ) were higher than those ( 0.25-128,0.0625-64, 0.25-64 and 0.25-2 μg/ml,respectively) in isolates resistant to one or two drugs (F = 26.242, 23.563, 31.541 and 64.469,respectively, P ＜0.05).The MICs of RFP in MDR isolates (2-64 μg/ml) were higher than those (0. 25 μg/ml) in other resistant isolate except M DR isolates (F = 5.613, P <0.05). Conclusions The study shows that there are associations between the results of routine drug susceptibility testing and the resistant extent of anti-TB drugs. This could help doctors select more effective anti-TB regimen for TB patients according to the correlations.

Objective To establish a fluorescent polymerase chain reaction (PCR) method for rapid, sensitive and specific determination of -88/-123 polymorphisms in Myxovirus resistance protein A (MxA) gene promoter so as to provide molecular biology tool for optimized interferon-a treatment in chronic hepatitis B patients. Methods Hepatitis B virus (HBV) genotyping,serum HBV DNA level,and- 88/- 123 polymorphisms in MxA gene promoter of patients who had been treated with interferon-α were detected. The statistical analysis was done by using SPSS software to understand the relationship between MxA gene polymorphisms and interferon-α treatment. Afterwards, an optimal fluorescent PCR system was established to determine -88/-123 polymorphisms in MxA gene promoter. The sensitivity and the specificity of this system were confirmed by DNA sequencing. P-value of chi square test, odds ratios of regression analysis and 95% confidence intervals were employed. Results Patients with- 88 G/T and - 123 C/A in the interferon-stimulated response element in MxA gene promoter were interferon-α sensitive, while patients with - 88 GIG and - 123 C/C were not interferon-α sensitive. The coincidence rate of this system was 99.65% in comparison with DNA sequencing.Conclusion MxA gene polymorphisms could be rapidly and sensitively determined by this fluorescent PCR system.

Background Laser peripheral iridoplasty (LPI) is widely used in the treatment of glaucoma by flattening the iris and widening angle of anterior chamber (AA).However,no evidence suggests the optimal site of LPI in iris.Objective This study was to compare the therapeutic effects of LPI at different sites of iris for glaucoma.Methods Glaucoma models were established in the right eyes of 40 healthy adult male pigment rabbits by intrachamber injection of 0.1 ml compound carbomer solution with 0.3％ carbomer and 0.025％ dexamethasone.The models were randomly divided into model control group,corneoscleral limbus group,one spot from corneoscleral limbus group and two spots from corneoscleral limbus group.LPI was performed at corresponding site of iris by 532 nm argon laser with the spot diameter 500 μm,energy 300 mW,exposure time 0.3 seconds and laser number 24 spots,and the rabbits in the model control group did not receive LPI.Intraocular pressure (IOP),coefficient of outflow facility (C value) were measured and calculated with Schi(o)tz tonometer before LPI and 2,4,7,14 and 30 days after LPI,and anterior chamber depth (ACD),AA,anterior chamber angle opening distance within 500 μm radius from scleral spur (AOD500) were measured with ultrasound biomicroscope (UBM).The eyeballs were extracted 30 days after LPI,and the chamber angle were observed under the optical microscope after hematoxylin and eosin staining.The use and care of the animals complied with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.Results UBM showed that compared with the model control group,the anterior chamber angle was evidently widened in all the LPI groups,with the best effectiveness in the one spot from corneoscleral limbus group and the worst one in the two spots from corneoscleral limbus group.Compared with the model control group,the IOP was evidently reduced,and C values,AA and AOD500 were significantly increased in the corneoscleral limbus group,one spot from corneoscleral limbus group and two spots from corneoscleral limbus group after LPI,showing significant differences among the four groups (IOP:Fgroup =16.848,P ＜ 0.01;C value:Fgroup =9.629,P ＜ 0.01;AA:Fgroup =62.336,P＜0.01;AOD500:Fgroup =77.779,P ＜ 0.01).IOP was reduced and C value,AA and AOD500 were increased in 2,4,7,14 and 30 days after LPI as compared with before LPI,with significant differences over time (IOP:Ftime =3.041,P =0.011;C value:Ftime =4.311,P＜0.01;AA:Ftime =14.627,P＜0.01;AOD500:Ftime =20.378,P＜0.01).Compared with the model control group,the ACD was significantly increased in the corneoscleral limbus group and one spot from corneoscleral limbus group,and that in the two spots from corneoscleral limbus group was significantly reduced,and the ACD was insignificantly increased over time after LPI (Fgroup =18.017,P＜0.01;Ftime =0.022,P =1.000).Hematoxylin and eosin staining showed that the trabecular meshwork and adhesion of tissure were reopened and the anterior chamber angle was widened after LPI.Conclusions LPI can widen anterior chamber angle and lower the IOP.The best therapeutic outcome for glaucoma is displayed when LPI is performed at the iris site corresponding to one spot from the corneoscleral limbus.

Objective:To evaluate the effect of orexin A on morphine-induced gastrointestinal dysfunction in mice.Methods:Forty SPF C57B/6 mice, aged 6-8 weeks, half male and half female, weighing 18-22 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), morphine group (group M) and morphine + different doses of orexin A groups (MOH, MOM and MOL groups). Normal saline 8 ml/kg was subcutaneously injected daily in group C, morphine 6 mg/kg was subcutaneously injected daily in the other four groups, and orexin A 75, 50 and 25 μg/kg were subcutaneously injected daily for 10 days at the same time in MOH, MOM and MOL groups.The fetal water content was calculated and averaged daily.After the last administration, the mice were gavaged with black nutrient paste, and the gastric emptying rate and small intestinal propulsion rate were detected 30 min later.Blood samples were collected from the orbit, and the concentration of serum gastrin (GAS) was detected by enzyme-linked immunosorbent assay.The mice were then sacrificed, and colon tissues were removed for determination of c-kit positive cell area (by immunohistochemistry) and expression of c-kit, substance P (SP) and neural nitric oxide synthase (nNOS) in colon tissues (by Western blot). Results:Compared with group C, the rate of fecal water content, gastric emptying rate, small intestinal propulsion rate and serum GAS concentration were significantly decreased, the area of c-kit positive cells was decreased, and the expression of c-kit and SP was down-regulated, and the expression of nNOS was up-regulated in group M ( P<0.05). Compared with group M, the small intestinal propulsive rate and serum GAS concentration were significantly increased, and the area of c-kit positive cells was increased, and the expression of c-kit was up-regulated in group MOH, the rate of fecal water content, gastric emptying rate, small intestinal propulsion rate and serum GAS concentration were significantly increased, the area of c-kit positive cells was increased, and the expression of c-kit and SP was up-regulated, and the expression of nNOS was down-regulated in group MOM, and the serum GAS concentration and c-kit positive cell area were significantly increased in group MOL ( P<0.05). Conclusions:Orexin A 50 μg/kg can effectively alleviate the gastrointestinal dysfunction induced by morphine in mice, and the mechanism may be related to promotion of GAS secretion, interstitial cells of Cajal growth and SP release and inhibition of NO release.

This article reported a successful cesarean section performed on a woman with 37 +6 weeks of gestation after heart-lung transplantation (HLTx) at the Second Xiangya Hospital of Central South University. The patient was a 29-year-old woman with singleton pregnancy who underwent HLTx seven years ago. During pregnancy, she was treated with azathioprine and tacrolimus to suppress immune rejection. The patient was in New York Heart Association (NYHA) class Ⅰ and had no fever or shortness of breath. Prenatal examinations indicated that the mother and the fetus were in good condition. A baby girl was delivered successfully at full term by a transverse lower-segment cesarean section under spinal anesthesia. Both the mother and the child were healthy at follow-up of 42 d after discharge.

OBJECTIVETo induce Mycobacterium tuberculosis (MTB) resistance with ofloxacin (Ofx) of stepwise increasing concentration in vitro, investigate stability to fluoroquinolone (FQs) antibiotic of MTB, and analyze the molecular mechanism and mutation specialty of drug resistance preliminarily.

METHODSMTB Standard strain H37RV and 24 clinical isolates susceptible to Ofx were selected and experimentally serially subcultured in liquid culture medium containing increasing concentration of Ofx and induced the drug resistance to Ofx. Variety of Minimal Inhibitory Concentrations (MICs) to FQs drugs were detected by microwell-MIC-test method. Mutations of quinolone resistance determining region (QRDR) of gyrA gene were sequenced and identified. Relationship of different mutation sites and drug resistant degree were analyzed. A total of 6 MTB clinical isolates resistant to Ofx and induced drug resistant isolates in vitro were serially subcultured in liquid culture medium without drug. Variety of drug resistant stability, including MIC and mutation of gyrA gene were detected.

RESULTSMIC values of 21 Ofx susceptible isolates after induction were eight times higher than before, which were induced to drug resistant strains successfully and also resistant to Lfx and Mfx. Hot mutations of QRDR of gyrA gene were detected by sequencing, except one strain. Mutation of codon 94 occurred in 60% (12/20) of the strains with mutations and corresponding value of 50% Minimal Inhibitory Concentrations(MIC50) was ≥ 8 µg/ml. In all, 4 of 6 MTB clinical isolates resistant to Ofx harbored mutation of codon 90 (67%) , but the corresponding value of MIC50 was 2 µg/ml. After 21 serially subcultured in liquid culture medium without drug, MIC values of 6 clinical isolates resistant to Ofx were not changed obviously and mutations were also not changed. After 11 times serially subcultured in culture medium without drug, MIC values of induced drug resistant strains were also not changed obviously, but new mutations were detected in QRDR of 3 isolates.

CONCLUSIONMTB strains resistant to three kinds of FQs antibiotic were obtained by induction in vitro with Ofx. Codons 88, 94 mutations of QRDR of gyrA gene were related to the high level FQs drug resistance of MTB. Drug resistant stability of MTB to FQs was strong, and it is difficult for MTB to resume susceptibility.

Antitubercular Agents ; pharmacology ; DNA Gyrase ; genetics ; Drug Resistance, Bacterial ; genetics ; Microbial Sensitivity Tests ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Ofloxacin ; pharmacology

OBJECTIVETo investigate the new biomarkers of acute kidney injury, as well as to confirm the values of response gene to complement 32 (RGC-32) for early diagnosis of acute kidney injury by comparing the values of serum creatinine (Scr) and cystatin C (CysC) in children who had undergone cardiopulmonary bypass (CPB).

METHODSixty-seven patients who had accepted CPB were recruited from the cardiac surgery intensive care unit, Children's Hospital Affiliated to Shanghai Jiao Tong University from March to June 2013 and assigned to acute kidney injury group (group AKI) or non-acute kidney injury group (group non-AKI), on the basis of the definition by the pediatric RIFLE (pRIFLE) criteria. Also 30 healthy control children were recruited. Serum samples were taken regularly from each patient after CPB at 30 min, 2 h, 4 h, 24 h, 48 h and 72 h for RGC-32. Serum samples were tested by enzyme linked immunosorbent assay (ELISA) which was employed to determine the levels of serum RGC-32. Scr and CysC were analyzed by HITACHI 7180 automatic biochemical analyzer. All the data were analyzed by receiver operator characteristic curve (ROC) and area under curve (AUC).

RESULTThe incidence of AKI was 34% (23/67), including 15 cases with risk stage AKI, 4 cases with injury stage AKI, 3 cases with failure stage AKI, 1 cases with loss stage AKI. Three out of four subjects with Failure stage AKI and the one case with Loss stage all accepted renal replacement therapy. CPB group had a higher level of serum RGC-32 than that of pre-operation after CPB 30 minute [(2.88 ± 0.68) µg/L vs. (1.39 ± 0.31) µg/L, P < 0.05]. At the same time, comparing with the non-AKI group, the levels of serum RGC-32 were higher than that of controls 30 min, 2 h, 4 h, 24 h and 48 h after CPB (t = 2.560, 2.180, 2.818, 2.226, 3.017; P < 0.05). The values for the AUC were determined for RGC-32 as 0.770, 0.707, 0.768, 0.728,0.723 and 0.770 after CPB 30 min, 2 h, 4 h, 24 h, 48 h and 72 h. The values for sensitivity of serum RGC-32 30 min, 2 h and 4 h after CPB was 0.914, 0.824, 0.824 and the values for specificity of serum RGC-32 was 0.619, 0.667, 0.810, respectively. But the values for sensitivity of CysC was 0.625, 0.813, 0.813, and specificity 0.571, 0.619, 0.571, respectively. The values for sensitivity of Scr was 0.625, 0.625, 0.813 and specificity was 0.571, 0.571, 0.524, respectively.

CONCLUSIONThe sensitivity of serum RGC-32 for detecting AKI was much higher than that of Scr and serum CysC in children who had accepted CPB, and that RGC-32 may be a new biomarker for early detection of AKI. However, the conclusion needs to be further elucidated.

Acute Kidney Injury ; blood ; diagnosis ; etiology ; Area Under Curve ; Biomarkers ; blood ; Cardiopulmonary Bypass ; adverse effects ; Case-Control Studies ; Cell Cycle Proteins ; blood ; Creatinine ; blood ; Cystatin C ; blood ; Female ; Heart Defects, Congenital ; surgery ; Humans ; Infant ; Intensive Care Units, Pediatric ; Male ; Muscle Proteins ; blood ; Nerve Tissue Proteins ; blood ; Postoperative Complications ; Predictive Value of Tests ; Prospective Studies ; ROC Curve ; Sensitivity and Specificity

OBJECTIVETo obtain the C7C peptide ligands of Mycobacterium tuberculosis by affinity screening based on the phage-displayed random C7C peptide library, and preliminarily identify the binding capacity of the peptide to Mycobacterium.

METHODSInactive Mycobacterium tuberculosis reference strain H37Rv was used as the target molecule to screen the Ph. D.-C7C peptide library, and Mycobacterium bovis, BCG was used for reverse screening. After 4 rounds of affinity screening, single phages eluted by H37Rv and BCG were selected for DNA sequencing. ELISA was used to detect the binding affinities of different single phage clones. The cyclic peptides displayed by the phage clones showing the highest appetency were synthesized in vitro with fluorescent markers. Fluorescence microscopy and flow cytometer was used to detect the binding affinities of synthesized cyclic peptides, comparing with linear binding peptides obtained before.

RESULTSAfter 4 rounds of biopanning, phages that could bind with target molecules were remarkable enriched. 16 common sequences were obtained by sequencing analysis of single phages. With ELISA, phage SB1, SB5, SB8 and SB26 all showed higher affinity with H37Rv and BCG, the ratio to negative control of which were ≥ 2.1, but could not bind to the 3 nonmycobacteria, which were identified as the positive clones. Based on the results of flow cytometer detection, the affinities to H37Rv of 4 cyclic peptides SB1, SB5, SB24, SB26 were (73.2 ± 6.3)%, (63.2 ± 5.3)%, (32.9 ± 3.1)%, (89.4 ± 7.0)%, and to BCG were (65.6 ± 6.1)%, (48.6 ± 4.5)%, (10.3 ± 1.8)%, (86.6 ± 7.9)%, separately, which were all higher than H8 ((4.0 ± 1.0)%, (5.5 ± 1.2)%) . From the results of fluorescence microscopy observation, all of the fluorescent labeled cyclic peptides SB1, SB5, SB24, SB26 could bind to H37Rv and showed higher fluorescence intensities, which also had certain affinities to other 18 mycobacteria, but the fluorescence intensities were lower than H37Rv, and didn't bind to 3 non-mycobacteria.

CONCLUSIONBased on the replacement of linear 7 peptide library with C7C peptide library, new ligands of Mycobacterium tuberculosis were achieved successfully, which showed significantly higher binding affinities to mycobacteria.

Base Sequence ; Enzyme-Linked Immunosorbent Assay ; Ligands ; Mycobacterium tuberculosis ; Peptide Library ; Peptides