去卵巢骨质疏松性骨折大鼠胰岛素样生长因子1表达与葛根素的干预效应

目的:观察葛根素对去卵巢骨质疏松性骨折大鼠胰岛素样生长因子1表达的影响。方法:实验于2005-02/2006-01在解放军第四五四医院骨科和南京医学院完成。实验材料:葛根素购自中国生物药品鉴定所。SD雌性大鼠40只,8月龄,体质量(280±19)g,由南京医学院实验动物中心提供。实验方法:将大鼠用4%戊巴比妥钠40mg/kg腹腔内麻醉,在无菌条件下行双侧卵巢摘除术,术后继续饲养2个月,借助骨折器建立股骨闭合性骨折模型。按随机排列表法分为对照组和治疗组,每组20只。治疗组术后给予灌胃20mg/(kg·d)剂量的葛根素,对照组给予2mL生理盐水处理。于骨折后第2周利用Western-blot和RT-PCR方法检测胰岛素样生长因子1蛋白与胰岛素生长因子1mRNA表达,观察葛根素对胰岛素生长因子1表达的影响。结果:40只大鼠全部进入结果分析。①Western-blot检测胰岛素生长因子1蛋白:与对照组相比,治疗组胰岛素生长因子1蛋白表达明显升高(1.40±0.09,3.2±0.31,P<0.05)。②RT-PCR分析胰岛素生长因子1mRNA表达:与对照组相比,治疗组胰岛素生长因子1mRNA表达明显升高(12.3±2.09,23.1±1.32,P<0.05)。结论:葛根素可以通过调节胰岛素生长因子1的表达,从而可能加快去卵巢大鼠骨质疏松性骨折的愈合。     

人骨髓间充质干细胞的贴壁分离与体外培养

目的:验证贴壁方式分离人骨髓间充质干细胞,并进行体外扩增培养的可行性。方法:实验于2006-02/12在解放军济南军区总医院脊髓修复科完成。①实验材料:骨髓来源于解放军济南军区总医院脊髓修复科收治的脊髓完全性损伤患者,对本实验知情同意。基础培养液由含体积分数为0.15胎牛血清和低糖α-MEM配置。②实验方法:无菌条件下髂后上棘穿刺抽取骨髓组织6mL,进行细胞培养,观察细胞生长情况,待细胞融合成片、长满培养瓶底部后,用质量浓度为2.5g/L的胰蛋白酶流过所有细胞表面。倒置显微镜下观察细胞变圆、部分脱壁后,立即加入有血清培养液终止消化。③实验评估:取第3代生长状态良好的细胞,胰蛋白酶消化制成细胞悬液,接种,以细胞数为纵坐标,时间为横坐标,绘制细胞生长曲线。同时每隔2h进行细胞贴壁率检测。结果:①骨髓间充质干细胞的形态学观察:倒置显微镜下,骨髓间充质干细胞接种1d即贴壁,去除悬浮细胞后继续培养3d贴壁细胞开始增殖,伸展为椭圆型、短梭型、多角型及不规则型等。至14d细胞密集在集落中心,基本铺满瓶底,第3～5代细胞呈均匀一致的长梭型,排列成旋涡状或放射状。②骨髓间充质干细胞的生长曲线:细胞传代后3d内处于潜伏期,3d后进入生长期,7d后进入平台期。③骨髓间充质干细胞的贴壁率:随着培养时间的延长,骨髓间充质干细胞贴壁率逐渐升高。传代后2,4,6,8,10,12,14,16,18,20h细胞贴壁率分别为(20.20±0.25)%,(33.00±0.29)%,(46.50±0.32)%,(69.20±0.30)%,(76.60±0.34)%,(86.50±0.27)%,(90.30±0.20)%,(96.10±0.28)%,(98.50±0.12)%,(99.00±0.07)%。结论:贴壁法分离骨髓间充质干细胞操作简便,经体外扩增培养后细胞增殖活性强,传代周期为7d,是比较理想的骨髓间充质干细胞培养方法。     

613例新涂阳肺结核病例的队列分析

目的探讨新登记涂阳肺结核病例的转归,评价肺结核传染源控制效果。方法对613例新发涂阳肺结核病例的资料进行队列分析。结果新登记涂阳肺结核,以青壮年多发,以待业、家务和务农为主,集中于农猎区和林区,营养状况偏低。治愈率呈逐年上升趋势,严格执行全程督导化疗。结论严格执行DOTS的技术规范是控制肺结核传染源的主要措施。     

Effects of Ficus asperifolia on normal rat estrus cyclicity

Objective

To evaluate Ficus asperifolia (Moraceae) (F. asperifolia) effecting on regular estrus cycle of Wistar rats.

Methods

Air-dried fruits of F. asperifolia were extracted using water. Prior to the test, vaginal smear was monitored daily for a 3-week period to select females with normal (regular) estrous cycle. Those with regular estrus cycle weighing between 150–170 g were randomized into three sets of 15 animals each. Each set was then divided into three groups: Group 1 (control) was orally administered with distilled water (10 mL/kg body weight) once a day for 1 week starting from the proestrus stage. Groups 2 and 3 were respectively treated with 100 and 500 mg/kg body weight of the plant aqueous extract. The two other sets of 15 animals each were similarly treated as the first set for 3 weeks and 6 weeks respectively. Estrus cycle pattern was monitored before and during plant extract application whereas lipid profile, ovary, uterus and liver growth indices were determined at the end of each treatment.

Results

F. asperifolia did not disrupt (0%) the order of appearance of normal estrus cycle stages, namely, proestrus, estrus, metestrus and diestrus. Short-term treatment (1 week duration) exhibited high frequency of appearance of proestrus and estrus stages while mid- (3 weeks) and long-term (6 weeks) treatments revealed constancy in the frequency of all stages irrespective to animal groups. The plasma and organ lipid profile, as well as ovary, uterus and liver growth remained unchanged when compared to distilled water-treated animals. Following long-term administration of plant extract (6 weeks), no adverse effect was noticed.

Conclusions

Our data partially support the use of F. asperifolia in common medicine.     

Cardiovascular Effects of the n-Butanol Extract from Kalanchoe Crenata Leaves

The present work examined the effects of the n-butanol extract from the leaves of Kalanchoe crenata (Andrews) Haworth (Crassulaceae) on rat blood pressure and guinea pig papillary muscle contraction and action potential. When administered intravenously at doses of 2, 5, and 10 mg/kg, the n-butanol extract of K. crenata leaves induced a significant transient fall in blood pressure and significantly reduced the cardiac rate for about 10 min. Concomitantly, the extract significantly increased the PR, QRS, and QT intervals of the electrocardiogram (ECG). At concentrations of 300 and 1,000 μ g/ml, the n-butanol extract was found to increase the amplitude of electrical contraction of papillary muscle by 137.6 and 223.0%, respectively. When tested on the ventricular myocardiac cell action potential, the extract (100 μ g/ml) significantly and time dependently delayed the repolarization without affecting the amplitude. Indeed, the 10%, 60%, and 90% repolarization times were respectively increased by 131.6%, 78.6%, and 52.7%. The bradycardic effect of the n-butanol extract may result from the increase of the PR, QRS, and QT intervals which are in accord with the delay in action potential repolarization observed in in vitro studies. The data obtained in the in vitro studies suggest that K. crenata possess potassium channel blockade properties that may account for its cardiac properties.     

Acute and subchronic oral toxicity assessment of the aqueous extract from the stem bark of Erythrina senegalensis DC (Fabaceae) in rodents

Aim of the study

The decoction of the stem bark of Erythrina senegalensis (EAES) is traditionally used in the Western region of Cameroon against liver disorders. The present study evaluated the potential toxicity of this decoction after acute and sub-chronic administration in rodents.

Materials and methods

In acute study, a single administration of EAES was given orally to mice at doses ranging from 1.25 to 12.5 g/kg. General behaviour, adverse effects and mortality were determined for up to 14 days post treatment. In the sub-chronic study, EAES was given orally as a single administration to Wistar rats at doses of 300, 600 and 1200 mg/kg/day for 28 days. Animal body weight was observed throughout the experimental period while haematological and biochemical parameters of blood and urine, as well as kidney and liver tissues histology were evaluated at the end of the experiment.

Results

In the acute study in mice, none of the doses used induced mortality or significant behavioural changes. In the sub-chronic study in rats, daily oral administration of EAES at the dose of 600 mg/kg resulted in a significant increase in the relative body weight at the last week of treatment. The relative weights of organs were not affected by the treatment. No haematological changes were observed a part of a significant increase in WBC count at the dose of 600 mg/kg. Serum AST, ALT, ALP, total protein, total cholesterol and triglycerides decreased significantly while total and conjugated bilirubin significantly increased. Renal function indices assay in blood showed significant modification in all the treated groups compared to control while, in urine, only urea excretion markedly reduced at a dose of 1200 mg/kg. Histological analysis did no show any liver or kidney alteration.

Conclusion

These results demonstrated that there is a wide margin of safety for the therapeutic use of EAES and further corroborated the traditional use of this extract as a hepatoprotective agent.     

Identities and frequencies of mutations of the otoferlin gene (OTOF) causing DFNB9 deafness in Pakistan

Mutations in OTOF , encoding otoferlin, cause non-syndromic recessive hearing loss. The goal of our study was to define the identities and frequencies of OTOF mutations in a model population. We screened a cohort of 557 large consanguineous Pakistani families segregating recessive, severe-to-profound, prelingual-onset deafness for linkage to DFNB9 . There were 13 families segregating deafness consistent with linkage to markers for DFNB9 . We analyzed the genomic nucleotide sequence of OTOF and detected probable pathogenic sequence variants among all 13 families. These include the previously reported nonsense mutation p.R708X and 10 novel variants: 3 nonsense mutations (p.R425X, p.W536X, and p.Y1603X), 1 frameshift (c.1103_1104delinsC), 1 single amino acid deletion (p.E766del) and 5 missense substitutions of conserved residues (p.L573R, p.A1090E, p.E1733K, p.R1856Q and p.R1939W). OTOF mutations thus account for deafness in 13 (2.3%) of 557 Pakistani families. This overall prevalence is similar, but the mutation spectrum is different from those for Western populations. In addition, we demonstrate the existence of an alternative splice isoform of OTOF expressed in the human cochlea. This isoform must be required for human hearing because it encodes a unique alternative C-terminus affected by some DFNB9 mutations.     

Transmission permeability of tuberculosis involving immigrants, revealed by a multicentre analysis of clusters

In recent years, the number of cases of tuberculosis (TB) among immigrants in Spain has increased markedly, and led to this analysis of the recent transmission patterns of TB in the immigrant population in Madrid. The countries from which the highest number of immigrant cases have been reported were Ecuador (21%), Romania (16%), Morocco (12%), Peru (11%) and Bolivia (9%). Fifty-one per cent of the cases were from South America. In a multicentre study (2004–2006), IS 6110 restriction fragment length polymorphism and spoligotyping were used to genotype the Mycobacterium tuberculosis isolates from 632 immigrant cases from 47 countries. A total of 183 cases (29%) were grouped into 59 clusters, which are markers of potential transmission events. Most of the clusters (81%) included patients living in different healthcare districts, and 54% of the clusters were multinational. When a sample of 478 autochthonous cases was included, 53% of the clusters involving immigrants also included autochthonous cases. This study revealed marked transmission permeability among nationalities and between the immigrant and the autochthonous populations.     

2006年陕西公众结核病防治知识核心信息知晓情况调查

目的了解陕西公众结核病知识核心信息知晓情况,为更有效地开展结核病防治健康教育提供依据。方法采用分层系统随机抽样方法,在全省抽取7个县区共8119名城乡居民。使用统一设计的调查问卷对调查对象进行入户面对面询问式调查。结果调查对象的核心信息总知晓率为49.5%,其中男性总知晓率为52.0%,女性为47.0%,城区居民总知晓率为62.7%,农村居民为47.5%。自述周围有人患有肺结核病的人群,其核心信息总知晓率为60.9%。12~19岁组、小学及以下文化程度者、学生和农民知晓率较低。调查对象对肺结核病的传染性、传播途径的知晓率较高,而一半或超过一半的调查对象不知道当地有检查和治疗结核病的专门机构、结核病人应到专门机构检查治疗、国家有免费检查治疗政策、不应该歧视结核病人,尤其是对＂咳嗽咳痰3周以上或有咯血、血痰症状者应怀疑得了肺结核病＂这条核心信息的知晓率只有20.2%。结论被调查人员结核病核心信息知晓率较低,不同特征居民间知晓率有差别,应根据当地居民结核防治知识的知晓情况制定相关政策。