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 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 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 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 explore the role of matrix metalloproteinase 12 (MMP12) in airway macrophages and pulmonary neu-rogenic substance P ( SP ) in the pathogenesis of asthma by analyzing their relationship in different categories of asthmatic patients.Methods:Twenty patients of asthma remission phase ( remission asthma group ) , twenty ones of mild acute exacerbation asthma (mild asthma group) and twenty healthy adults (normal control group) were included,respectively.After lung function was measured,the numbers of macrophage in induced sputum were counted.The expression levels of MMP12 mRNA and protein in sputum macrophages were detected by quantitative reverse transcription polymerase chain reaction and Western blot.The concentration of sputum SP was assayed by enzyme immunometric assay.Results: ( 1 ) Compared with the subjects in normal control group, forced expiratory volume in 1 second%predicted ( FEV1 ) and forced expiratory flow rates at 50% of the forced vital capacity % predicted ( FEF50 ) were much lower and the numbers of sputum macrophages were much higher in the patients in different asthmatic groups.Compared with the patients in remission asthma group,FEV1 and FEF50 were much lower in the ones in mild asthma group.(2) MMP12 expressions in the macrophages and the concentrations of SP in sputum were significantly increased in the patients in different asthmatic groups compared with those in normal control group;Furthermore,MMP12 and SP in mild asthma group were much higher than in remission asthma.(3) In all patients from different asthmatic groups,mRNA expressions of MMP12 in the macrophages were positively correlated with the levels of sputum SP or the numbers of sputum macrophages,whereas negative correlations between mRNA expressions of MMP 12 and FEV 1 or FEF50 were observed.Conclusion: The regulatory imbalance of macrophages′MMP12 and pulmonary neurogic SP may participate in the pathogenesis of asthma and become the potential targets for asthma therapy.

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 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.

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.