Quantifying anti-HBV effect of targeted ribonuclease by real-time fluorescent PCR

AIM: To quantify the inhibition of HBV replication by targeted ribonuclease by using real-time fluorescent PCR. METHODS: Targeted ribonuclease was introduced into 2.2.15 cells by liposome-mediated transfection or HIV-TAT mediated protein transduction. Forty-eight hours after the transfection and 24 h after the transduction, supernatants of 2.2.15 cells were collected and HBV DNA in the supernatants was quantified by real-time fluorescent PCR with a commercial kit. RESULTS: HBV DNA concentrations in the supernatants of 2.2.15 cells transfected or transducted with targeted ribonuclease were 4.9+/-2.4 x 10(8) copies/L and 8.3+/-4.0 x 10(8) copies/L, respectively. Compared with controls, transfection or transduction of targeted ribonuclease reduced HBV DNA concentration in the supernatants of 2.2.15 cells by 90.4% and 90.1%, respectively (P<0.05). CONCLUSION: Targeted ribonuclease can inhibit HBV replication in 2.2.15 cells.     

Inhibitory Effects of Anti-sense PTTG on Malignant Phenotype of Human Ovarian Carcinoma Cell Line SK-OV-3

To construct eukaryotic expression vector expressing full length anti-sense pituitary tumor transforming gene (PTTG) mRNA and observe its blocking effect on the potential invasion of human ovarian carcinoma cell line SK-OV-3. PCR primers containing designed enzyme cut sites were used for cloning full-length PTTG gene fragment, and the resulting PCR product was inserted into the eukaryotic vector pcDNA3. 1 in the antisense direction. The recombinant vector was then transfected into SK-OV-3 by Lipofectamine. The positive cell clone was screened by G418, PTTG and bFGF at protein level expression were detected by Western blot. The biological behavior change of transfection positive cells was observed by colony formation in soft agar assay. Our results showed that SK-OV-3 clones stably expressing full-length recombinant pcDNA3. 1-PTTGas were obtained. The expressions of PTTG and bFGF protein in transfected cells were decreased by 61.5% and 52.3%, respectively as compared with non-transfected ones. The number of colony formation was reduced significantly in transfected cells as compared with empty vector transfected and non-transfected cells. It is concluded that the recombinant vector pcDNA3. 1-PTTGas is a novel tool and provides an alternative anti-sense gene therapy targeted at PTTG in human carcinoma.     

Inhibitory effect of retroviral vector containing anti-sense Smad4 gene on Ito cell line, LI90

Background Transforming growth factor-β1 (TGF-β1) exerts strong fibrogenic potential in culture-activated HSCs. Smad4 is a key intracellular mediator for the transforming growth factor-β (TGF-β) superfamily of growth factors. The aim of this study was to assess the effects of the antisense Smad4 gene on Ito cell line, LI90.Methods The recombinant retroviral vector pLXSN-Smad4 was constructed by cloning the rat antisense Smad4 cDNA into the retroviral vector pLXSN. Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad4 and pLXSN vectors into PA317 cells. Human hepatic stellate cells (HSCs) LI90 were infected with these retroviruses followed by selection with G418. The expression of Smad4 was detected by Northern and Western blots. Cell biological characteristics, including cell growth curve, 3H-TdR and 3H-proline uptake by HSCs and the production of extracellular matrix were assessed.Results mRNA and protein expressions of Smad4 in LI90 cells transfected with retrovirus containing the antisense Smad4 gene were much lower than those in LI90 cells transfected with empty vector or parental LI90 cells. Cells hypoexpressing the Smad4 gene exhibited a slower rate of growth, a lower uptake of 3H-TdR and 3H-proline (P&lt;0.01), and smaller production of th extracellular matrix, compared with parental LI90 cells and cells transfected with empty retrovirus.Conclusions The antisense Smad4 gene can suppress the expression of the Smad4 gene, reduce endogenous production of Smad4 mRNA and protein, block TGF-β1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix （ECM）. Our results may provide a basis for the development of antifibrotic gene therapy.     

Anti-adjuvant arthritis of recombinant human endostatin in rats via inhibition of angiogenesis and proinflammatory factors

AIM: To investigate the profile of endostatin on adjuvant arthritis (AA) and angiogenesis blockade in synovitis. METHODS: The model of rat AA was induced by injection of intradermal complete Freund's adjuvant (CFA). Hind paw volume of rat was measured by volume meter and the activities of interleukin-1 (IL-1) and IL-2 were measured by the assay of thymocytes proliferation. IL-1beta and tumor necrosis factor-alpha (TNF-alpha) produced by synoviocytes was estimated with radioimmunoassay. The number of new blood vessels in knee joint synovium was counted under microscope by hematoxylin and eosin (HE) staining. RESULTS: The secondary inflammation of AA rats appeared on the 10th day after injection of CFA. The therapeutic administration of endostatin (0.1, 0.5, and 2.5 mg/kg/d, sc, plus 7 d) was given from that time (d 10). It was found that endostatin significantly inhibited the secondary paw swelling and the number of new blood vessels in the synovium of AA rats. Endostatin significantly decreased the production of IL-1 derived from both peritoneal macrophages and synoviocytes and IL-2 from splenocytes, especially at the dose of 2.5 mg/kg. This effect of endostatin also was seen on TNF-alpha produced by synoviocytes. CONCLUSION: The recombinant human endostatin had an inhibitory effect on rat AA, which was related to its anti-angiogenesis and inhibition of proinflammatory cytokines.     

正视问题,勇于改进——评《避免绩效考核的反作用》

在企业绩效管理中,绩效衡量扮演着举足轻重的角色。有效地使用绩效衡量方法可以及时反馈企业的运营状况,为企业的改进方向提供切实的依据。但同时,绩效考核使用不当也有可能打击组织士气、降低团队效率、妨碍质量改进等,这些反作用经常会被管理者们忽视。就笔者所在门店而言,绩效考核的不足之处主要表现在两个方面:一是压根不运用,考评完了就完了,好     

避免绩效考核的反作用

一家有5名店员的社区药店,如果总部下发的任务是发放500份DM单,那么员工会去尽心尽力地投放,但是如果任务是2000份,那可能单子会被直接扔进垃圾桶. 上有政策,下有对策.绩效考核指标如果制定得不合理,那么店长和店员可能会一起想“对策”来应付总部考核,甚至为了指标的完成,员工会采取浪费企业资源或者损失企业利益的做法,这在各行各业都不乏案例.     

影响取决于政策

从整体来说,医疗行业在经济体系中的地位在国内并不高,它的地位如何,与人们的基本素质、国家的富裕程度以及国家对该行业的重视程度有关。医疗行业对推动经济发展的作用也非常有限,若作为健康产业对待,还能起到一定的作用;若像过去那样作为卫生事业,耶对经济发展来讲,则纯粹就是附属保障的作用。可以这么说,经济发展能推动医疗行业的发展,但医疗行业反作用于经济发展则非常有限。     

谈谈中药和针灸的双向调节作用

在中医文献中或临床上常常遇到这样的现象,即同一个方或同一种药,或同针一个穴位而具有两相反的作用,这相反作用,就叫中医“双象调节”作用,又叫“双相性”。一常见到的“双向调节”现象1、具有双向调节作用的方剂:如肾气丸既能治疗肾阳虚引起的浮肿,尿少,服药能使小便次数增多;又能治疗肾阳虚引起的多尿或夜尿多,使小便次数减少。     

慢性胃、十二指肠疾病中西医结合临床合理用药

胃、十二指肠疾病是临床消化系统常见病、多发病，即可局限于本脏器，也可累及本系统，甚至累及全身或其它脏器，俗话说：“十人九胃”便是这个道理。目前，临床可见到消化系统药物非常多，致使临床用药杂乱无章，无法可依，如果不能合理用药，不仅不能达到预期治疗目的，甚至起到反作用。本人通过多年临床观察、结合本类疾病的发病机理、以及临床常用中西药物的药理学等综合分析、探讨，对该类疾病的治疗往往收到满意效果，特作一总结。     

音频电碘离子透入治疗注射后硬结疗效比较

注射后硬结多与注射器械的粗糙，如打钩、毛刺以及注射操作规程的执行错误、注射药液的刺激性等有关，常见于小儿及老人。若硬结不大，多会引起局部疼痛；若硬结过大，有些甚至能压迫坐骨神经，引起臀部肌肉，如臀大肌、臀中肌等的挛缩，从而致下肢跛行。目前中西药对之尚无特效疗法，手术切除的副作用大，还会产生诸如术后疤痕粘连等反作用。笔在康复科接诊202例注射后硬结病人，分别予音频电碘离子透入，音频电 碘离子透入，超声波治疗三种方法进行疗效比较，总结如下。