Effects of Aqueous Extract of Plumbago zeylanica L. in the Reversal of DMN-induced Hepatic Fibrosis in Rat

[ Objectives ] This study was conducted to evaluate the action of the aqueous extract of a medicinal plant Plumbago zeylanica L. in the reversal of dime- thylnitrosamine （DMN） -induced hepatic fibrosis in rats, and to provide a scientific basis for the utilization of P. zeylanica in treatment of hepatic fibrosis. [Meth- ods ] Hepatic fibrosis in rats was induced by intraperitoneal injection of DMN. The rats were then given the aqueous extract of P. zeylanica at high, medium or low concentration by garage for five weeks. Serum level of alanine aminotransferase （ALT） was determined by lactate dehydrogenase （LDH）-release assay, and serum level of aspartate transaminase （AST） was measured by UV-malate dehydrogenase （MDH） assay. Serum levels of total bilirubin （TBIL）, direct bilirubin （DBIL） and indirect bilirubin （1BIL） were measured by vanadate oxidation assay. Four indices of hepatic fibrosis （hyaluronic acid, laminin, procollagen type III and colla- gen type IV） were determined by radioimmunoassay （RIA） assay. Morphological damage of liver tissue was observed by hematoxylin-eosin staining （H＆E stai- ning）. Immunohistochemical staining was performed to determine the location and area of deposited collagen type I, collagen type III and ct-smeoth muscle actin （a-SMA） in liver tissue. [ Results] Compared with the negative control （rats with diseased fiver and untreated with P. zeylanica aqueous extract）, the serum lev- els of ALT, AST, TBIL, DBIL and IBIL were significantly decreased by P. zeylanica aqueous extract; the levels of the four serum indices of hepatic fibrosis were also obviously reduced. H＆E staining showed that hepatic fibrosis in rats was obviously inhibited or even reversed by P. zeylanica aqueous extract. Immunohisto- chemical staining proved that the aqueous extract of P. zeylanica significantly reduced the area and coloration of collagen type I, collagen type III and ct-SMA in rat liver. [ Conclusions] The aqueous extract of P. zeylanica has a definite effect in reversal of DMN-indueed hepatic fibrosis in rat by promoting the recovery of liver function, reversal of histopathological changes and reducing fibrotic collagen.

Effects of Guangxi Hepu Pearl Hydrolysate on Proliferation Activity and Apoptosis of Human Hepatic Stellate Cells

[Objective] The paper was to observe the effects of Guangxi Hepu pearl hydrolysate on proliferation and apoptosis of human hepatic stellate cells. [Method] The inhibition rate against the proliferation of human hepatic stellate cells was determined by MTT colorimetry after co-incubation with pearl hydrolysate for 48 h. The percentage of early and late apoptotic cells was determined by flow cytometry(Annexin V-FITC/PI staining). [Result] Low, medium and high concentrations of pearl hydrolysate could inhibit the proliferation of human hepatic stellate cells, and the inhibition rates at 48 h were 12.4%, 27.4% and 37.8%, respectively. The low, medium and high concentrations of pearl hydrolysate could induce the apoptosis of cells. The percentages of total apoptotic cells(early apoptotic + late apoptotic) at 48 h were 8.97%, 16.09% and 26.98%, respec-tively. [Conclusion] Pearl hydrolysate can inhibit the proliferation of human hepatic stellate cells and induce the apoptosis of these cells, which may be the mechanism of anti-hepatic fibrosis effect of pearl hydrolysate. These effects are dose-dependent. It is also found that the above cellular effects of high dose pearl hydrolysate are stronger than those of colchicine and compound Biejiaruangan tablets.

Exploration on the Reform of PBL Mode of Course:Comprehensive Experiment of Basic Medicine for Nursing Undergraduates in Colleges and Universities of Traditional Chinese Medicine

The undergraduate education of nursing science produces higher practical operational talents to the society,and the importance of experimental teaching cannot be ignored in the training of nursing students.The basic experimental teaching method of undergraduate nursing has been explored and reformed by the College of Basic Medicine of Guangxi University of Chinese Medicine.After the reform,the experimental teaching adopts the teaching method of PBL mode,and the experimental course is composed of several specific and important medical problems.The specific tasks involved in each problem include relevant morphological knowledge,functional knowledge,diagnostic and laboratory knowledge,and clinical medical knowledge.It makes the teaching content of experimental course more comprehensive,and leads to the transition from basic medicine to clinical medicine.This has gradually and effectively improved the practical ability,scientific research thinking,clinical comprehensive ability and learning interest of undergraduate nursing students.

Effects of Soy Isoflavones on the Sex Hormone Level and Pathological Changes in the Perimenopausal Syndrome Animal Model

[Objectives] To observe the effect of soy isoflavones on the perimenopausal syndrome model of female rats,and to find its mechanism,in order to provide experimental evidence for the clinical treatment of menopausal syndrome. [Methods]50 female SD rats were randomly divided into five groups,with 10 rats in each group,respectively,for the normal control group,sham operation group,model group,diethylstilbestrol group,soy isoflavone treated group,except the normal control group and sham operation group,the other groups were ovariectomized rats,the formation model of perimenopausal rats treated with normal saline,diethylstilbestrol,soy isoflavone by gavage,normal control group and sham operation group were given the same dose of normal saline and soy isoflavone by gavage. After 6 weeks of continuous treatment,abdominal aorta was taken blood,the serum was isolated,and radioimmunoassay method to measure E_2( Eestradiol),P( Progesterone),T( Testosterone). And removal of the pituitary gland,uterine and adrenal prepared pathological section,experimental effect was observed by light microscopy,using immunohistochemical method to observe the brain pituitary FSH( Follicle-Stimulating Hormone) and LH( Luteinizing Hormone) levels. [Results] Radiation immune analysis results showed that soy isoflavone could significantly improve serum E_2 level in ovariectomized rats,compared with the model group,the statistical analysis showed significant difference( P < 0. 05),but the serum P and T level had little effect. The pathological results showed that( i) uterus: the uterine wall and endometrial thickness of soy isoflavone group was between that of model group and sham operation group,the surface epithelium was columnar,glands increased;( ii) pituitary gland: soy isoflavone ovariectomized group cells were between model group and normal group;( iii) adrenal gland: soy isoflavone group adrenal tissue did not change significantly. Immunohistochemical results showed that soy isoflavone group pituitary tissues of FSH and LH cells compared with the model group decreased significantly. [Conclusions] The soy isoflavones could make the uterus wall and endometria of perimenopausal syndrome animal model thicken,improve the serum level of E_2 in ovariectomized rats,and decrease the level of FSH and LH,but had little effect on the adrenal tissues,level of T and P.

Advances in In-vitro Culture Technology of Human Fetal Hepatic Stellate Cells

Liver is an important organ for human metabolism and biological conversion. Medical research on hepatic disease clinic, drug metabolism and drug effi- cacy evaluation all needs an in-vitro model of liver as a research platform. Hepatic stellate cells are core cells for occurrence and development of liver fibrosis. Stud- ies at home and abroad deemed that human fetal hepatic stellate cells are an ideal material for the construction of an in-vitro research model for liver fibrosis. With clinical and basic research of liver going deeper, the requirements to quantity and quality of in-vitro models of fetal hepatic stellate cells become higher and higher. The advances in isolation, culture and cryopreservation technique of human fetal hepatic stellate cells were reviewed in this paper.

Genuine Medicinal Materials and Resources Development in Xinjiang

First of all,this paper introduces the concept,traditional and modern evaluation and conditions of genuine medicinal materials.It focuses on the current situation of genuine medicinal materials in Xinjiang,and suggests that we should strengthen the development,utilization and protection of genuine medicinal resources in Xinjiang from a professional point of view.

Research Progress on the Regulation of Hepatic Sinusoidal Endothelial Cells on Hepatic Fibrosis

Hepatic sinusoidal endothelial cell is a highly differentiated cell in hepatic sinusoid,and plays an important role in the occurrence and development of hepatic fibrosis.The dysfunction of hepatic sinusoidal endothelial cells is considered to be the key cause of a variety of liver diseases.At present,the researches on hepatic fibrosis at home and abroad are mainly focused on inhibiting the activation of hepatic stellate cells and accelerating the hydrolysis of extracellular matrix.However,there are few studies on the important role of the structure and function of hepatic sinusoidal endothelial cells in hepatic fibrosis.This paper reviews the research progress on the effect of hepatic sinusoidal endothelial cells on hepatic fibrosis and its regulatory mechanism in recent years.This paper summarizes the results of the research on the structural characteristics of hepatic sinusoidal endothelial cells,secretion of fibrosis-related cytokines and regulation of hepatic stellate cells activation in the development of hepatic fibrosis.

Growth and Reproduction Characteristics of TLR4 Knockout Mice Used for Liver Fibrosis Experiments

[Objectives]This study was conducted to investigate the similarity and differences between TLR4 knockout mice and C57 BL/6 mice used in liver fibrosis research in terms of growth rate and reproduction ability.[Methods]Twenty TLR4 knockout mice and C57 BL/6 mice,half male and half female,were selected to compare the growth rates of body weight and body length of mice from the 4th to 12th weeks;and 20 pairs of male and female mice of the same strain were compared for the number of baby mice of the second litter.[Results]The growth rates of body weight and body length of the TLR4 knockout mice were significantly lower than those of C57 BL/6 mice(P<0.05)(except for the 4th and 5th weeks when there was no significant difference in body length);and in terms of reproductive ability,the TLR4 knockout mice were significantly lower than the C57 BL/6 mice(the ratio of the total number of baby mice in the second litter of the two strains,72∶147).[Conclusions]Knockout of the TLR4 gene has a significant impact on the growth and reproduction of mice.

Discussion on Serum Technique and Cryopreservation Technique in the Culture of Liver Sinusoidal Endothelial Cells

Hepatic sinusoidal endothelial cells are a kind of highly differentiated cells in hepatic sinusoids, which play an important role in the occurrence and development of liver fibrosis. Hepatic sinusoidal endothelial cells are often used in the study of liver fibrosis. In the process of culturing liver sinusoidal endothelial cells, the treatment of serum for culture and cryopreservation of cells are relatively complicated and error-prone. If the technical details of serum treatment and cell cryopreservation are not handled properly, it will have a more obvious effect on the effect of hepatic sinusoidal endothelial cell culture. We have gained experience in the theoretical study of liver fibrosis and the operation of cell culture experiments, and this paper summarized the details of liver sinusoidal endothelial cell culture such as the problems of serum preservation, thawing, precipitates and heat inactivation, as well as methods and precautions for cell cryopreservation.