Objective To evaluate the effects of nerve growth factor-beta (NGF-β) pretreatment on cell apoptosis induced by endoplasmic reticulum stress during ischemia-reperfusion in isolated rat hearts.Methods Male Sprague-Dawley rats,weighing 180-220 g,were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.The hearts were excised and perfused in a Langendorff apparatus with K-H solution aerated with 95％ O2 and 5％ CO2 at 37 ℃.Thirty-two isolated rat hearts were randomly divided into 4 groups (n =8 each) using a random number table:control group (group C),I/R group,NGF-β pretreatment group (group N) and NGF-β combined with K252a (trkA receptor antagonist) pretreatment group (group N + K).In group C,the hearts were continuously perfused with K-H solution for 195 min.The hearts were perfused with K-H solution for 45 min in group I/R.In N and N + K groups,the hearts were perfused with K-H solution for 15 min,and then with K-H solution containing 0.1 μg/ml NGF-β and 0.1 μg/ml NGF-β mixed with 100 nmol/L K252a,respectively,for 30 min.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in I/R,N and N + K groups.HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 15 min equilibration (baseline) and at 5,30,60 and 120 min of reperfusion.Myocardial specimens were obtained at 120 min of reperfusion for detection of myocardial apoptosis (by TUNEL) and expression of glucose-related protein 78 (GRP78),CCAAT/enhancer-binding protein homologous protein (CHOP),and caspase-12 (by Western blot analysis).Apoptosis index was calculated.Results Compared with group C,HR,LVDP and + dp/dtmax were significantly decreased,and LVEDP,apoptosis index and expression of GRP78 and CHOP were increased in I/R and N groups,and the expression of caspase-12 was upregulated in I/R group.Compared with group I/R,HR,LVDP,and + dp/dtmax were significantly increased,and LVEDP,apoptosis index and expression of GRP78,CHOP and caspase-12 were decreased in group N,and the expression of GRP78 was down-regulated in group N + K.There was no significant difference in cardiac function indexes between group I/R and N + K.Compared with group N,HR,LVDP and + dp/dtmax were significantly decreased,and LVEDP,apoptosis index,and expression of GRP78,CHOP and caspase-12 were increased in group N + K.Conclusion NGF-β pretreatment can protect the isolated rat hearts against ischemia-reperfusion injury,and inhibition of the endoplasmic reticulum stress-triggered cell apoptosis after activating trkA receptors is involved in the mechanism.

Objective To evaluate the role of 1-phosphatidylinositol 3-kinase (PI3K) signaling pathway in nerve growth factor-beta (NGF-β)-produced mitigation of cell apoptosis induced by endoplasmic reticulum stress during hypoxia-reoxygenation (H/R) in rat cardiomyocytes.Methods H9c2 cells were seeded in 96-well plates at a density of 5× 105 cells/ml (100 μl/well).The wells were randomly divided into 5 groups (n =15 each) using a random number table: control group (group C);group H/R;NGF-β group (group N);NGF-β+NGF receptor trkA antagonist K252a group (group N+K);NGF-β+ PI3K inhibitor LY294002 group (group N+L).The cells were exposed to 95％ N2-5％ CO2 for 4 h in an anaerobic incubator, followed by reoxygenation in a standard incubator for 4 h in H/R, N, N+K and N+L groups.In addition, the cells in N, N + K and N + L groups were incubated in a standard incubator containing NGF-β, the mixture of NGF-β and K252a and the mixture of NGF-β and LY294002, respectively, during reoxygenation, and the final concentrations of NGF-β , K252a and LY294002 were 50 ng/ml, 100 nmol/L and 50 μmol/L, respectively.The cell viability was detected by using CCK-8 assay, and the cell survival rate was calculated.The cell apoptosis was examined by flow cytometry, and apoptosis rate was calculated.The expression of glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), caspase-12, Akt and phosphorylated Akt (p-Akt) was detected by Western blot.The ratio of p-Akt to Akt was calculated.Results Compared with group C, the cell survival rate was significantly decreased, and the apoptosis rate was increased in H/R and N groups, and the expression of GRP78, CHOP and caspase-12 was significantly up-regulated in group H/R, and p-Akt/Akt was significantly increased in group N (P＜0.05).Compared with group H/R, the cell survival rate was significantly increased, the apoptosis rate was decreased, the expression of GRP78, CHOP and caspase-12 was up-regulated, and p-Akt/Akt was increased in group N (P＜0.05), and no significant change was found in the parameters mentioned above in N+K and N+L groups (P＞0.05).Compared with group N, the cell survival rate was significantly decreased, the apoptosis rate was increased, the expression of GRP78, CHOP and caspase-12 was up-regulated, and p-Akt/Akt was decreased in N+K and N+L groups (P＜0.05).Conclusion NGF-β can mitigate the cell apoptosis induced by endoplasmic reticulum stress during H/R, and activation of PI3K signaling pathway is involved in the mechanism.

Hydrophidae, one of the precious traditional Chinese medicines, is generally drily preserved to prevent corruption, but it is hard to identify the species of Hydrophidae through the appearance because of the change due to the drying process. The identification through analysis on gene barcode, a new technique in species identification, can avoid the problem. The gene barcodes of the 6 species of Hydrophidae like Lapemis hardwickii were aquired through DNA extraction and gene sequencing. These barcodes were then in sequence alignment and test the identification efficency by BLAST. Our results revealed that the barcode sequences performed high identification efficiency, and had obvious difference between intra- and inter-species. These all indicated that Cyt b DNA barcoding can confirm the Hydrophidae identification.
Animals ; Base Sequence ; China ; Cytochromes b ; genetics ; DNA Barcoding, Taxonomic ; Elapidae ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Reptilian Proteins ; genetics ; Sequence Analysis, DNA

Acute Coronary Syndrome ; complications ; drug therapy ; Adrenergic beta-Antagonists ; therapeutic use ; Aged ; Heart Failure ; complications ; drug therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome

Objective To assess the diagnostic yields of clinical chromosomal microarray ( CMA) testing for patients with neurodevelopmetal disorders ( NDD) , and to characterize the spectrum of pathogenic copy number variation(CNV) in NDD.Methods The study was a cross-sectional study.NDD patients from Shanghai Children′s Medical Center ( SCMC ) from April 2014 to April 2015 were recruited.DNA samples from SCMC cohort were tested on Affymetrix Cytoscan Dx microarray platform.The diagnostic yields of CMA testing were further assessed for the whole NDD cohort and each subgroup.Results A genome-wide genotype-phenotype analysis on a total of 107 NDD cases with CMA testing was conducted.Based on the SCMC clinical cohort, the overall diagnostic yield of CMA testing for NDD patients was 20.6%(22/107). Excluding one case with chromosomal aneuploid, the frequency of non-polymorphic CNVs of the rest NDD cases were 25.5%(27/106).The diagnostic yield for developmental delay/intellectual disorder(DD/ID) and autism spectrum disorder(ASD)were 26.3% (15/57) and 10.2%(4/39) respectively.DD/ID was more likely to be associated with CNV than ASD and attention-deficit/hyperactivity disorder(ADHD).Five recurrent genomic loci were significantly enriched in patients including 1q21.1-q21.2, 15q11.2-q13.1, 22q11.2, 7q11.23 and 17q11.2.Conclusion CNV is an important pathogenesis in NDD.

Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.

Objective To investigate the clinical characteristics,the diagnosis and therapy of synchronous multiple primary esophageal carcinoma.Methods Thirty-two cases of synchronous multiple primary esophageal carcinoma were collected from January 1980 to December 2010 and their clinic data were retrospectively analysised.Results In the whole group,there were totally 66 lesions in 30 cases of double primary lesion and 2 cases of three primary lesion.The length of the lesions were 1 cm to 6.5 cm,and there were 22 lesions in cervical esophagus,10 in upper thoracic esophagus,19 in midthoracic esophagus and 15 in lower thoracic esophagus.Within the 66 lesions,65 lesions were squamous cell carcinoma and 1 was adenocarcinoma.The mucosa between the lesions were normal,with a distance of 4 cm to 9.5 cm,average 7.1 cm.Thirty-two patients with synchronous multiple primary esophageal carcinoma received surgery.Among 32 cases,26 of them were given definite diagnosis before operation.Four of them had exploratory operation,one patient underwent palliative resection,27 patients underwent radical resection.Two cases of the pathologic results of esophageal stump showed carcinoma after operation.The complications occurred in 8 patients.Twenty-eight cases were followed up after operation,the 1,3 and 5-survival rate were 76.9％,43.3 and 14.8％,respectively.Conclusion General pre-operation examination can significant helpful for the definite diagnosis of synchronous multiple primary esophageal carcinoma and surgical treatment is better choice for this disease.

AIM: To evaluate visual outcome and wearing times in eyes fitted with a rigid gas permeable contact lenses and to assess the role of these lenses in therapeutic contact lens practice.METHODS: Patients fitted with RGP from May 2004 to December 2005 were retrospectively reviewed. Ocular diagnosis,flat and steep curvatures by corneal topography. The data included initial base curve, power, and the number of changes made in parameters during the follow-up. The outcome data included visual acuity and the duration of follow-up and lens wear. Complications and complaints were also noted.RESULTS: Forty-Three eyes of 25 patients had keratoconus were reviewed. The mean age of patients was 19.88 ±3.11years. 26 (60.5%)eyes achieved 20/20 visual acuity;15(34.9%) eyes achieved 20/25 to 20/40; and 1 (2.3%) eyes achieved 20/50 to 20/70. 37 (86.0%) eyes achieved visual improvement. No infection or neovascularization was noted; a corneal abrasion occurred in one eye.CONCLUSION: RGP may be a better alternative in selected patients with keratoconus rather than penetrating keratoplasty.

Adult ; Chondrosarcoma ; metabolism ; pathology ; Collagen Type II ; metabolism ; Diagnosis, Differential ; Femoral Neoplasms ; metabolism ; pathology ; Giant Cells ; pathology ; Humans ; Male ; Osteoclasts ; pathology ; Osteosarcoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Vimentin ; metabolism

Objective To investigate the effects of pulsed electromagnetic fields(PEMFs)on proliferation and differentiation of endothelial progenitor cells(EPCs).Methods EPCs were isolated from rat bone marrow by density gradient centrifugation.EPCs were exposed to PEMFs from the 5th day to the end of culture.MTT was used to measure the proliferation of EPCs.The expression ofⅧ-related antigen and NOS_3 was evaluated by flow cytometry. Results Compared with the control,the proliferating ability of EPCs exposed to PEMFs was stronger;the number ofⅧ-related antigen and NOS_3 positive cells increased significantly in EPCs exposed in PEMFs.Conclusion PEMFs promotes the proliferation and differentiation of rat bone marrow EPCs.