AIM:To investigate the inhibition of breviscapine on apoptosis of cultured myocardial cell of neonatal rat induced by hypoxia/reoxygenation. METHODS:Myocardial cell hypoxia/reoxygenation model was established by culturing primary myocardial cells of neonatal rats in vitro. Cultured myocardial cells were divided into 5 groups:control group,hypoxia/reoxygenation group and 3 groups pretreated with breviscapine of final concentration 25,50 and 100 mg/L,respectively. The cell viability was measured with MTT; apoptotic rates were determined by AnnexinV-FITC/PI; the expression of Bcl-2 was detected by immunohistochemical method. Expressions of Cytochrome C (CytC) and Caspase-3 were detected by Western blot. RESULTS:Compared with the control group,the viability of myocardial cell decreased and apoptosis rate elevated after hypoxia/reoxygenation. However after pretreatment with 25,50 and 100 mg/L breviscapine,respectively. Cell viabilities increased and apoptotic rates lowered,and the protective effect on myocardial cell had concentration-dependent. In addition,Expression of Bcl-2 decreased but Caspase-3 activity and CytC release increased in myocardial cells induced hypoxia/reoxygenation. Pretreated with breviscapine,expression of Bcl-2 elevated but Caspase-3 activity and CytC release reduced obviously. CONCLUSION:It is associated with the increase in Bcl-2 expression,inhibition of CytC release and Casepase-3 activity that breviscapine could significantly protect myocardial cell against apoptosis induced by hypoxia/reoxygenation.

Objective To investigate the apoptosis in neural epithelium induced by passive smoking. Methods By using the nuclear fast red-crystal violet staining method and situ TUNEL technology, the quantitative changes of apoptosis in neural epithelium in gold hamster were analyzed. Results Excessive apoptosis and less cell division were observed in neural epithelium compared with the control group. On 9th days of pregnancy, there were most apoptosis cells, the apoptosis index was 9.82 ?3.35 which is statically significant compared with control group (P

Objective To establish a new multiplex-PCR assay to improve the detection rate of mutations in the DMD gene in Chinese patients. Methods A retrospective review of DMD deletion spectrum of 355 DMD patients with deletions all over the gene was performed. All deletions were confirmed by " one-step approach" diagnostic procedure and MLPA analysis. The exons with high frequency of mutations were identified to constitute the amplification system and the PCR conditions were optimized. Results Two new multiplex-PCR assays were established. Assay one was used to detect 10 exons including exon 5, 8, 17, 44, 45, 47, 49, 50, 51 and 52 of DMD gene, in two PCR sets. The theoretical detection rate would be 92% (326/355). Assay two was used to detect 5 exons including exon 12, 19, 35, 43 and 54, which could be used to screen additional 5% (17/355) deletion cases. The method was validated in other 22 DMD patients. Multiplex-PCR results were completely identical to the MLPA results in all 22 DMD patients. Conclusions The two multiplex-PCR assays were established based on the analysis of 355 Chinese DMD patients with gene deletions. It is believed that the new approach would be more applicable for deletion detection on the Chinese DMD patients since the DMD cases involved were from the whole country.

Objective To provide an experimental evidence for the clinical applications of glyceryl trinatrate(GTN)and isosorbide dinitrate(ISDN)against Coxsackievirus B(CVB)-related myocarditis.Methods Coxsackievirus B3 was propagated in HeLa cells.Virus yields were determinded by 50% tissue culture infective dosage(TCID50).BALB/c mice were attacked with 5000 TCID50 of CVB3,meanwhile,the mice were administrated with GTN and ISDN.All mice were killed at the 14th day.The myocardial tissues were harvested for histologic evaluation.Results The infection plaques in the myocardial tissues obtained from CVB3-infected BALB/c mice treated with GTN were siginificantly reduced(0.89?0.18 in GTN group and 1.25?0.22 in ISDN group)compared with that of the untreated CVB3-infected mice(P

Objective To observe the effect of Xiaoyingqiangji decoction on mitochondria in skeletal muscle of hyperthyroid myopathy. Methods Healthy Wistar rats were randomly divided into 4 groups: the group of hyperthyroid myopathy were given L-thyroxine intraperitoneal injection for 42 d; Chinese medicine treated group were treated with Xiaoyingqiangji decoction for 30 d after L-thyroxine injection; the saline control group were given saline solution for 30 d after L-thyroxine injection; normal control group were given saline solution. Then the skeletal muscles were obtained and the succinate dehydrogenase (SDH) were measured. The ultrastructural changes were observed with TEM. Results The activity and content of SDH in the group of hyperthyroid myopathy decreased, and the cristae of mitochondria ruptured and even disappeared. After given Chinese medicine, the activity and content of SDH increased, while the mitochondria showed normal ultrastructural appearance. Conclusion Xiaoyingqiangji decoction can improve the function of skeletal muscles in hyperthyroid myopathy by recovering enzyme activity and mitochondria structure.

Objective To observe the effect of Xiaoyingqiangjitang on the ultrastructure of motor end-plate of skeletal muscle in rats with hyperthyroid myopathy, and changes of acetylcholin esterase (AchE).Methods The 36 healthy female Wistar rats were randomly divided into the normal control group (n=6), and model group, Chinese medicine treated group and saline control group (all n=10). All animals in the model group, Chinese medicine treated group and saline control group were treated with group given L-thyroxine intraperitoneal injection for 42 days to establish the animal model of hyperthyroid myopathy. The animals of the Chinese medicine treated group were treated with Xiaoyngqiangjitang for 30 days after L-thyroxine intraperitoneal injection. The enzyme activity of AchE and ultrastructure of motor end-plate were observed by light and electron microscopes.Results The enzyme activity and content in the group of hyperthyroid myopathy was decreased, the reaction products of AchE located in synapse became weaker and distributed heterogeneously, the junction infolding of motor end-plate became smaller and shorter, the mitochondria in axon terminal became vacuolated. After given Chinese medicine, the enzyme activity and content increased, the junction folding, the mitochondria showed normal ultrastructural appearance.Conclusion Xiaoyingqiangjitang can improve the function of skeletal muscle in hyperthyroid myopathy by recovering the enzyme activity of AchE and ultrastructures of motor end-plate.

Objective To set up a new diagnostic platform based on microarray exon-capture and next-generation sequencing for detecting small mutations in dystrophin gene.The sensitivity and specificity of the method were assessed in clinical settings and the distribution of small mutations in Chinese Duchenne muscular dystrophy/Becker muscular dystrophy (DMD/BMD) patients were also analyzed.Methods Forty-one DMD/BMD patients diagnosed by the clinical criteria without large deletion or duplication (≥ 1exon) were recruited from Peking Union Medical College Hospital consecutively.Genomic DNA was extracted from blood samples.The libraries were prepared.Then exon and intron-exon flanking sequences of DMD gene were captured by custom microarray.Targeted next-generation sequencing and Sanger Sequencing were conducted.The patients who were not detected any disease-causing mutation were performed muscle biopsy.Results Thirty-eight subjects were detected small mutations in DMD gene.All single nucleotide variants (SNVs) and insertion & deletions (INDELs) were validated by Sanger sequencing.Twenty-one novel mutations were reported.The distribution of SNVs and INDELs was similar to other international DMD databases.Upon immunohistochemistry staining of dystrophin protein,1 of 3 mutation-undetected patients was diagnosed as DMD,2 of them were excluded.The specificity of the method was 100％,while the sensitivity was 97.4％.Conclusions Our microarray-captured next-generation sequencing assay could detect SNVs and INDELs with high sensitivity and specificity.Its advantages are economic,time-saving and stable.The platform is suitable for clinical gene diagnosis.

Auxotrophic strains of N1-37 (Phe-) and N2-27 (His-), screened from mutations of Paenibacillus polymyxa JSa-9 previously, were used as the parent strains to screen high-producing LI-F antibacterial lipopeptide fusion strain through protoplast fusion with polyethylene glycol as a promote agent. Fusion strain F5-15 was obtained. Then the product of LI-F antibacterial lipopeptide was quantified by HPLC, and the difference of expression of the key genes of lipopeptide synthase between wild strain JSa-9 and the fusion strain was analyzed by real-time PCR. LI-F antibacterial lipopeptide yield of the fusion strain F5-15 was 3.1-fold of the original strain JSa9's, and the expression levels of the target genes were 10.48, 2.48, 2.1 and 11.8 fold of the initial strain JSa-9, respectively.
Anti-Bacterial Agents ; biosynthesis ; Chromatography, High Pressure Liquid ; Lipopeptides ; biosynthesis ; Paenibacillus ; metabolism ; Protoplasts ; metabolism ; Real-Time Polymerase Chain Reaction

Aim To investigate the effect of astragalus injection on the expression of JNK3(c-jun N terminal kinase)protein and JNK3 mRNA interrelated by apoptosis after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats.Methods The hippocampal neurons cultured for eight days were divided into four groups:normal control group,hypoxia/hypoglycemia and reoxygenation group,astragalus injection group and astragalus solution group.Hypoxia/hypoglycemia and reoxygenation group,astragalus injection group and astragalus solution group were treated with hypoglycemia and reoxygenation after being deprived of oxygen and glucose for 30 minutes.Methods of Western blot,ELISA and RT-PCR were used respectively to measure the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation 0,0.5,2,6,24,72,120 h.Results Compared with normal control group,the mean optic density(MOD)of expression of JNK3 protein and activation of JNK3 protein in hippocampal neurons of rats every time points increased obviously in hypoxia/hypoglycemia and reoxygenation group except 120 h(P<0.05);compared with hypoxia/ hypoglycemia and reoxygenation group,MOD of expression of JNK3 mRNA and activation of JNK3 protein in hippocampal neurons of rats every time points decreased obviously except 120 h in astragalus injection group (P<0.05);compared with hypoxia/hypoglycemia and reoxygenation group,there was no difference in astragalus solution group.Compared with normal control group,MOD of expression of JNK3 mRNA in hippocampal neurons of rats every time points increased obviously in hypoxia/hypoglycemia and reoxygenation group(P<0.05);compared with hypoxia/ hypoglycemia and reoxygenation group,MOD of expression of JNK3 mRNA in hippocampal neurons of rats every time points decreased obviously in astragalus injection group except 120 h(P<0.05);compared with hypoxia/hypoglycemia and reoxygenation group,there was no difference in astragalus solution group.Conclusion Astragalus injection can inhibit the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation,moreover,it can inhibit the expression of JNK3 protein and decrease the activation of JNK3 protein,accordingly it inhibits hippocampal neuronal apoptosis.

Objective To investigate the relationship between serum follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and clinicopathological features and prognosis of serous ovarian cancer retrospectively. Methods A total of 73 patients with serous ovarian cancer treated in the Department of Obstetrics and Gynecology of Tianjin Medical University General Hospital from January 2000 to December 2015 were included in this study. The relationship between serum FSH, LH, PRL and clinicopathological features was analyzed by Mann-Whitney U method. Kaplan-Meier (K-M) method was used to analyze survival rates of patients with different clinical features. Multivariate Cox proportional hazards regression analysis was used to analyze prognostic factors of serous ovarian cancer patients. Results The mean concentrations of serum FSH and LH were significantly higher in the>50 year-old group than those in the<50 year-old group (P<0.05). The mean concentrations of FSH and LH were significantly higher in menopause group than those in non-menopause group (P<0.05). There were no significant differences in serum levels of FSH and LH in patients with other different clinicopathological features (P>0.05). There was no significant correlation between serum PRL concentration and clinicopathological features (P>0.05). Analysis results showed that poor prognosis of patients was related with high serum levels of FSH (>40.13 IU/L), PRL (>14.96 μg/L) and FIGO stage (Ⅲ+Ⅳ) (P<0.05). There was no significant correlation between serum LH concentration and prognosis (P>0.05). COX regression analysis showed that the serum PRL>14.96 μg/L was risk factor for prognosis of serous ovarian cancer [HR(95%CI): 3.530(1.180-10.557),P=0.024]. Conclusion The serum levels of FSH and LH are significantly increased in postmenopausal women than those in menopause women. The serum level of PRL is correlated with the prognosis of serous ovarian cancer.