Active Components Formulation Developed from Fuzheng Huayu Recipe for Anti-Liver Fibrosis / 中国结合医学杂志

OBJECTIVE@#To screen the active components from Fuzheng Huayu Recipe (FZHY) and redesign a new recipe composed of the active components, and validate the effect of active components formulation from FZHY against liver fibrosis.@*METHODS@#Thirty-two components from FZHY were evaluated for their activities against liver fibrosis respectively, with 6 kinds of cell models in vitro, including oxidative stressed hepatocyte in L-02, hypoxia injured/proliferative hepatic sinusoidal endothelial cells in SK-HEP-1 and human hepatic sinusoidal endothelial cells (HHSEC), and activated hepatic stellate cell in LX-2. The comprehensive activity of each component against liver fibrosis was scored according to the role of original herbs in FZHY and cell functions in fibrogenesis. Totally 7 active components were selected and combined with equal proportion to form a novel active components formulation (ACF). The efficacy of ACF on liver fibrosis were evaluated on activation of LX-2 and proliferation of HHSEC in vitro and in liver fibrosis model mice induced by dimethylnitrosamine (DMN). Totally 72 mice were divided into 6 groups using a random number table, including normal, high-dose ACF control (20 µ mol/L × 7 components/kg body weight), model, low-, medium-, high-dose ACF groups (5, 10, 20 µ mol/L × 7 components/kg body weight, respectively). Hematoxylin eosin and Sirius red stainings were used to observe inflammation and fibrosis change of liver tissue; scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were utilized to observe the effect of ACF on ultrastructure of hepatic sinusoids.@*RESULTS@#Fifteen components from FZHY showed higher scores for their activity on against liver fibrosis. Among them, 7 components including tanshinone II A, salvianolic acid B, cordycepin, amygdalin, quercetin, protopanaxatriol, and schizandrin B were recombined with equal proportions to form ACF. ACF at 1,2, 4 µ mol/L showed strong inhibitory effects on activation of LX-2 and proliferation of HHSEC in vitro (all P<0.01). Compared with the model group, ACF attenuated liver collagen deposition, improved sinusoidal capillarization in a dose-dependent manner (all P<0.05).@*CONCLUSION@#ACF exerts a satisfactory effect against experimental liver fibrosis and attenuates sinusoidal capillarization, which warrant a further research and development for herbal components formulation on liver fibrosis.

Treatment of HBV Cirrhosis with Fuzheng Huayu Tablet () and Entecavir: Design of a Randomized, Double-Blind, Parallel and Multicenter Clinical Trial / 中国结合医学杂志

BACKGROUND@#Antiviral therapy can lead to regression of fibrosis in chronic hepatitis B (CHB), but it has a limited effect on cirrhosis. Chinese medicines (CMs), particularly Fuzheng Huayu Tablet (, FZHY), have an antifibrotic effect in patients with CHB.@*OBJECTIVE@#To observe the safety and efficacy of adjunctive FZHY in patients with hepatitis B virus (HBV) cirrhosis, this study was designed as a randomized, placebo-controlled, double-blind, parallel assignment, multicenter trial at 20 centers in China. The total 700 naive patients will be enrolled with compensate cirrhosis due to HBV, and randomly assigned into 2 groups, receiving entecavir (0.5 mg, daily) and FZHY placebo (1.6 g, 3 times a day), or entecavir (0.5 mg, daily) and FZHY (1.6 g, 3 times a day), respectively. The primary endpoint was histological improvement at week 48. The secondary outcome is the decline values of liver fibrosis using the noninvasive methods from baseline to week 48 in each arm of the study. Adverse events such as stomach upset, headache, fatigue, dizziness, nausea will be strictly recorded.@*DISCUSSION@#Through this study, we hope to generate a solid evidence for the therapeutic strategy of HBV cirrhosis with a combination of anti-viral such as ETV and anti-fibrotic herbal product such as FZHY. Protocol version: Version 1.3, Date: 2014.12.4.@*TRIAL REGISTRATION NUMBER@#NCT02241590.

The application of RNA-seq in neurodegenerative diseases / 医学研究生学报

As a noval approach,RNA sequencing(RNA-seq)has been used for studying many diseases in recent years,es-pecially with the introduction of next generation sequencing(NGS),much more and more attention has been gathered on the utilization of RNA-seq for research purpose of central nervous system(CNS). This review focuses on applying of RNA-seq in central nervous sys-tem,taking neurodegenerative diseases for example,including Alzheimer's disease,Parkinson's disease and Huntington's disease,to expound the application of RNA-seq in the studying of central nervous system diseases and the prospective side of finding potential tar-gets in drug treatment of neurodegenerative diseases.

Effect of schisantherin A inhibit liver sinusoid endothelial cell function and action against liver fibrosis relating to angiogenesis / 中国中药杂志

To investigate the effect of schisantherin A on liver sinusoid endothelial cell function and angiogenesis. Different dosages (0-40 μmol•L⁻¹) of schisantherin A were incubated 24 h with SK-HEP-1 cells, and the toxicity of SK-HEP-1 cells was assayed by MTT method. The proliferation of SK-HEP-1 cells were induced by the vascular endothelial growth factor (VEGF), with receptor tyrosine kinase inhibitor sorafenib as the control, at the same time, set up the control group, 2, 20 μmol•L⁻¹ schisantherin A were incubated with SK-HEP-1 cells, cell proliferation was analyzed by EdU DNA cell proliferation kit. Fluorescence probe method was used to assay the intracellular NO levels and NOS activity. Tube formation was observed using cell migration and a matrigel tube formation assay. Rat aortic ring assay was performed to observe the sprouting vessels from aortic ring. The fluorescence vessels, the number of functional blood vessels, and intersegmental vessel changes of transgenic zebrafish were also observed. Compared with control group, the proliferation of SK-HEP-1 cells induced by VEGF increased and and the level of NO and NOS activity induced; compared with model group, 2, 20 μmol•L⁻¹ schisantherin A and sorafenib inhibited the proliferation of SK-Hep-1 cells induced by VEGF, and reduced the level of NO and NOS activity. At the dosage of 20 μmol•L⁻¹, schisantherin A attenuated the migration and tube formation of SK-HEP-1 cells induced by VEGF, and also inhibition the formation of rat aortic rings and intersegmental vessel changes of transgenic zebrafish, and significantly reduce the number of vessels in zebrafish. Schisantherin A has potential effects on function of endothelial cell proliferation and angiogenesis.

Evaluation of cryptotanshinone inhibition of angiogenesis in human hepatic sinusoidal endothelial cells / 药学学报

To investigate the effects of cryptotanshinone (an active ingredient of Salvia Miltiorrhiza) inhibition of angiogenesis, the toxicity of cryptotanshinone was assayed in human hepatic sinusoidal endothelial cells (HHSEC) by CCK8 method. Max dose without toxicity is 10 μmol·L-1. The proliferation of HHSEC were induced by the endothelial cell growth supplement (ECGS), with 2.5 μmol·L-1 sorafenib as the positive control. Cell proliferation was analyzed by EdU assay. Cell viability was analyzed by CCK8 method. The expression of vWF was analyzed by immunofluorescence method. Fluorescence probe method was used to detect the intracellular nitric oxide (NO) levels. Tube formation of HHSEC and transgenic zebrafish were also observed to evaluate the effects of cryptotanshinone against angiogenesis. Compared with normal control, there is a proliferation of HHSEC induced by ECGS. The expression of vWF and the NO levels increased significantly. Cryptotanshinone inhibited the proliferation, down regulated the expression of vWF and the NO levels. Further, cryptotanshinone inhibited the tube formation of HHSEC and reduced the number of fu nctional vessels in transgenic zebrafish. The results suggest that cryptotanshinone could inhibit angiogenesis by regulating the HHSEC cell function.

Epidemiological investigation results of plague in Shanxi province / 中国地方病学杂志

Objective To analyze the data collected from surveillance sites in Shanxi province in epidemiological investigations during 2004 - 2008 and understand the influence of such factors as rat density and vectorial quantities on plague epidemic intensity, thus provide some scientific basis for plague control and prevention. Methods To collect the data of surveillance during 2004 - 2008 in the 12 counties of plague surveillance by retrospective method. Sort and analyze the collected data according to the requirement of operational scheme for plague surveillance-calculate the rat density of spermophilus dauricus, meriones meridianus and the small rodents, and the flea index and the flea infection ratio of body flea, burrow track, cave. The results of serological tests were analyzed at the same time. Results As showed in the results, the mean density of spermophilus dauricus was 1.12 merely in each hectare, and that of meriones meridianus and small rodents were 7.42%(2130/28 700) and 8.03%(3886/48 400), respectively. The flea infection rate of spermophilus dauricus and meriones meridianas were 25.78% ( 1933/7497 ) and 44.19% (2028/4598), respectively;the flea index of spermophilus dauricus and meriones meridianus were 0.59 and 1.84, respectively. The number of nests excavated with flea for spermophilus dauricus was 70, and the numbers of nests and flea were 362 and 333, respectively. The flea infection rate was 19.34 % (70/362), and the flea index was 0.92. The number of nests excavated with flea for meriones meridianus was 34, and the numbers of nests and flea were 177 and 124, respectively. The flea infection rate was 19.21%(34/177), and the flea index was 0.70. The number of caves explored for spermophilus dauricus was 7171,and the number of burrows with flea was 62, so the flea infection rate was 0.86%(62/7171 ), and the flea index was 0.01. Conclusions With a relatively high density and large quantity and considering the epidemic features of animal plague in periphery provinces in recent years, plague may still happen and spread in Shanxi province diespite the negative results of all those serological tests, by means of importing from other provinces.

Differentially expressed genes between a fertile patient and an infertile patient in a large Chinese androgen insensitivity syndrome pedigree / 中国医学科学院学报

<p><b>OBJECTIVE</b>To screen and identify differentially expressed genes between a fertile patient and another infertile patient who belonged to a large Chinese pedigree affected with androgen insensitivity syndrome (AIS).</p><p><b>METHODS</b>We constructed the forward and reversed subtracted libraries using genital skin fibroblasts (GSF), which were obtained from the fertile patient MJ and infertile patient ZGJ, as tester respectively. Candidate clones were screened with colony in situ hybridization, dot blot, and Southern blot analysis step by step and conformed with Northern blot analysis. The potential positive clones were sequenced and the homology of the sequences was analyzed.</p><p><b>RESULTS</b>The forward and reversed subtracted libraries containing differentially expressed pattern of two GSF cell lines were constructed. Two positive clones identified by Northern blot were obtained in the reversed subtracted library. Eleven candidate clones from the two libraries that failed to hybridize with both RNA populations were obtained simultaneously, which might represent differentially expressed low abundance transcripts. Sequencing results and homology analysis demonstrated that the two positive clones were significantly homologous with the genes of autotaxin-t and calcium binding protein calcyclin (S100A6), respectively.</p><p><b>CONCLUSIONS</b>Two positive clones and eleven clones showing no hybridization signals may represent differentially expressed genes between the two GSFs. This finding may be useful to elucidate the molecular mechanisms leading to phenotypic variation and preserved fertility of the AIS pedigree.</p>