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

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 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 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 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.

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.

BACKGROUND:Previous studies discovered that pRST98, original y isolated from Salmonel a enterica serovar typhimurium (S.typhimurium) could promote bacterial biofilm formation. In addition, bacterial harboring pRST98 can promote the secretion and expression of interleukin-10 after infection in cells and animals.
OBJECTIVE:In vitro studies have discovered the effects of interleukin-10 at varying concentrations and conjugative plasmid pRST98 on the biofilm formation of S.typhimurium.
METHODS:S.typhimurium wild-type strainχ3306, virulence plasmid-deletion S.typhimurium strainχ3337 and pRST98-transconjugant S.typhimuriumχ3337/pRST98 were established in vitro and cultured for biofilm formation. 1, 10, 100 μg/L interleukin-10 were added during the biofilm formation. 0 μg/L interleukin-10 was set as a control. Crystal violet staining method, semi-quantitative method, confocal laser scanning microscopy and scanning electron microscopy were used to determine the effects of interleukin-10 on the biofilm formation and compare the effects of S.typhimurium with or without pRST98.
RESULTS AND CONCLUSION:Intra-group comparison showed that, compared with the control group, S.typhimurium gathered together and formed thicker biofilm in concentration of 1 and 10 μg/L of interleukin-10. The promotion effects of S.typhimurium on biofilm formation were greatly improved in 10 μg/L. Interleukin-10 in 100 μg/L inhibited S.typhimurium biofilm formation. Inter-group comparison showed that, A570 inχ3337/pRST98 was greatly higher than that inχ3306 andχ3337 under the same concentration of interleukin-10. The results indicate that both 1 and 10 μg/L of interleukin-10 promote biofilm formation, especial y bacteria harboring pRST98.

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.

Objective To determine the clinical features and the factors affecting the recurrence of ovarian borderline epithelial tumors. Methods A retrospective data of 112 cases with ovarian borderline epithelial tumors admitted in General Hospital of Tianjin Medical University from 2000 to 2015 were analyzed. Results The average age was (50.59±16.90) years in 112 patients with FIGO stageⅠof 102 (91.07%) patients, stageⅡof 4 (3.57%) and stageⅢof 6 (5.36%). The serum tumor marker (CA125) was examined in 102 patients, and 27 cases with the elevated indicator (26.47%). Surgical treatment was performed in 112 patients. Younger patients were more likely to choose conservative surgery. Ninety-seven patients were followed up, and 5 of them relapsed. Non fertility preserving surgery was performed in patients with recurrence. The recurrence rates of patients with different clinical pathological factors were compared. The recurrence rate was higher in patients with micro infiltration than that of patients without micro infiltration [37.50%(3/8) vs. 2.25%(2/89), P=0.004]. And the recurrence rate was higher in patients with microemulsion type borderline serous tumor than that of patients with non-papillary tumors [40.00%(2/5)vs. 0(0/41),P=0.019]. Seven patients underwent conservative surgery had normal spontaneous pregnancy. Conclusion The fertility-sparing surgery can be used as the treatment procedures for young patients, which is safe and effective. It is necessary to be on alert of recurrence for the cases with micropapillary pattern, and microinvasive tumor.