Objective To compare the hepatoprotective effects of different extracts in root of Gardenia jasminoides including ethanol extract, petroleum ether fraction, ethyl acetate, chloroform extract, n-butanol extract, and surplus water extract on the jaundice hepatitis mice by the partial least squares (PLS) method and multi index comprehensive evaluation method. Methods Setting up two groups of high and low doses (9 and 3 g/kg) in different extraction sites in root of G. jasminoides respectively, and administered to the stomach for 7 d. Copying a icteric hepatitis model of mice by alpha naphthylisothiocyanate (ANIT) in 5th d. Then, the alanine aminotransferase (ALT), aspartic transaminase (AST), alkaline phosphatase (AKP), γ-glutamyl transpeptidase (γ-GT), total bile acid (TBA), total bilirubin (TBIL), superoxide dismutase (SOD), malonic dialdehyde (MDA), glutathione (GSH) activity were determinated in serum and liver, and pathological changes were observed in mouse liver by HE stain, total hepatoprotective effect of the different extracts in root of G. jasminoides was compared by PLS and multi index comprehensive evaluation method. Results In addition to the high dose group of petroleum ether and low dose group of chloroform, the different extracts in root of G. jasminoides were improved on some indexes or multiple indicators to an extent and compared with the model group, the pathological damage of liver tissue was alleviated obviously, multi index comprehensive evaluation showed that the high dose group of ethyl acetatein root of G. jasminoides had the best hepatoprotective effect. Conclusion The ethyl acetate extract and n-butanol extract in root of G. jasminoides have better inhibitory effect on icteric hepatitis, and the high dose group of ethyl acetate has the best effect, which may be the active site of liver protection, and mechanism may be related to improving the ability of eliminating oxygen free radical, inhibiting lipid peroxidation and enhancing bile bilirubin metabolism and secretion.

The aim of this study was to investigate the effects of tyrosine kinase inhibitor PTK787 on cell proliferation, cell cycle and the expression of fak mRNA of human chronic myeloid leukemia (CML) cell line K562, and to explore the mechanism of PTK787 against acute myeloid leukemia. The MTT method was used to detect the effects of PTK787 in various concentrations and at different time points on proliferation of K562 cells; the flow cytometry was used to determine the effects of PTK787 in different concentrations on cell cycle of K562 cells; the RT-PCR was used to assay the expression of fak mRNA in K562 cells treated with PTK787 for 48 hours. The results showed that along with increasing of the concentration and prolonging of time, the inhibitory rate of PTK787 on K562 proliferation was gradually enhanced. The comparison between various concentration groups at same time or comparison between various time groups in same concentration showed significant differences (p < 0.05), in which the effect of 320 micromol/L PTK787 on cells was strongest, while the continuous increase of PTK787 concentration or prolong of action time did not enhance the inhibitory rate on K562 proliferation. With increasing of drug concentration, the cell proportion in G(1) phase gradually increased, the cell proportion in S phase gradually decreased, the comparison between various groups revealed significant differences (p < 0.05), however the continuous increase of drug concentration from 160 micromol/L did not obviously change the cell proportion in phases of cell cycle. With increasing of drug concentration, the expression of fak mRNA in K562 cells gradually reduced with significant differences between various groups (p < 0.05), but with continuous increase of drug concentration from 160 micromol/L, the effect of PTK787 on the expression of fak mRNA in K562 cells also did not obviously change. It is concluded that the PTK787 shows effect of anti-leukemia cells through inhibiting transformation of the K562 cells from G(1) phase into S phase and decreasing the expression of fak mRNA in cells.
Cell Cycle ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 1 ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Phthalazines ; pharmacology ; Pyridines ; pharmacology ; RNA, Messenger ; genetics

OBJECTIVETo study the morphological, ultramicrostructural and pathological changes of tissues from a patient with severe acute respiratory syndrome (SARS).

METHODSOne autopsy case of diagnosed SARS was investigated. Lung puncture was performed immediately after the patient died, and the autopsy was done after 12 h. The specimens from lymph nodes, spleen, small intestine, colon and bone marrow were studied by immunohistochemical technique. The antibodies used included CD20, CD45RO (UCHL-1), CD4, CD8, CD68 and CD34.

RESULTSThe principal lesions of the SARS case consisted of acute lobular intrastitial pneumonia, hyaloid membranes of pulmonic alveoli and hyperplasia and shedding of alveolar epithelium of. Virus-like inclusions occasionally contained cytoplasm of the alveolar epithelium, which were positive by histochemical staining. The adjacent blood-vessels were changed by hyperplasia and enlargement. The structures of lymph nodes and spleen were damaged with lymph follicles depletion and splenic nodules atrophy. The specific changes included reduction of lymphocytes and hyperplasia of histiocytes, depletion of the follicles of small intestine and colon wall, decreased hyperplasia of the bone marrow and increased number of the megakaryocyte. Meanwhile, in the immunohistochemical study, CD(20)(+) B cells were fully expressed in lymph nodes and spleen, and the CD45RO (UCHL-1)(+) T cells were scatteredly expressed. The number of CD4(+) help T cell was markedly decreased, while the number of CD8+ poisonal T cells increased, and the ratio of the former and latter was no more than 0.5. Under the electronic microscopy observation, virus-like particles with 80 - 160 nm diameter and halo or garland envelope were found in mononuclear macrophage and cytoplasm of alveolar epithelium.

CONCLUSIONThe specific lesions of SARS consist of lobular intrastitial pneumonia with the formation of hyaline membranes of lung, haemorrhage, necrosis, inflammation of blood vessels and the damages of extralung lymphohemopioetic system. The damages were very similar to the pathological features of tissues infected by human immunodeficiency virus, in which numbers of T cells decreased and CD(4)(+) T cell/CD(8)(+) T cell ratio was no more than 0.5. According to the virus-like particles found in lung of the SARS case, it is considered that these virus-like particles may be a new kind of coronavirus which caused the "atypical pneumonia".

Humans ; Immunohistochemistry ; Lung ; pathology ; Lymph Nodes ; pathology ; Male ; Microscopy, Electron ; Middle Aged ; Myocardium ; pathology ; Severe Acute Respiratory Syndrome ; pathology

OBJECTIVETo evaluate the progression in morphologic changes of lungs in SARS patients.

METHODSFour cases of SARS with lung tissue samples available (including one for ultrastructural examination) were enrolled into the study. Histochemical study for VG, Masson, reticulin, orcein, PAS, sirius red stains and immunohistochemical study for vimentin, desmin, smooth muscle actin, HHF-35, CD34, F8, collagen types I and III were also performed.

RESULTSAccording to the morphologic changes, lung lesions in SARS were subcategorized into 3 phases: acute exudative inflammation, fibrous proliferation and the final fibrotic stage. Two cases belonged to the acute exudative phase, in which the course was less than 20 days. The principal lesions consisted of acute alveolar exudative inflammation, hyperplasia of alveolar epithelium, necrosis, alveolar hyaline membrane formation, alveolar desquamation and focal fibroplasia. The acute exudative protein was PAS-positive. There was an increase in reticulin fiber formation. The reactive fibroblasts were highlighted by desmin and vimentin. One case belonged to the fibroproliferative stage, in which the course was around 25 days. Major lesions included proliferative interstitial pneumonia with early pulmonary fibrosis. There was also evidence of organizing pneumonia, with an increase in reticulin fiber formation, which had a glomeruloid appearance on special stain. The mesenchymal cells showed either myofibroblastic (which expressed desmin, HHF-35, smooth muscle actin and vimentin) or fibroblastic (which expressed vimentin only) differentiation. Fibroelastosis and fibroplasia was also noted. The remaining case belonged to the fibrotic stage, in which the course was around 75 days. The main features included diffuse fibrosis and honeycomb change, which were highlighted by sirius red stain. Immunohistochemistry showed mainly types I and IV collagen fibers. In all lesions, there was also an increase of number of CD68-positive macrophages.

CONCLUSIONSThe morphologic progression in lungs of SARS patients is characterized by the development of increased fibrosis. The primitive mesenchymal cells, hyperplastic alveolar epithelial cells and macrophages play an important role in the pathogenesis.

Actins ; metabolism ; Adult ; Collagen Type I ; metabolism ; Desmin ; metabolism ; Humans ; Lung ; metabolism ; pathology ; Male ; Middle Aged ; Pulmonary Fibrosis ; pathology ; Severe Acute Respiratory Syndrome ; metabolism ; pathology ; Vimentin ; metabolism

Seven compounds(deacetylasperulasidic acid methyl ester, gardenoside, chlorogenic acid, geniposide, crocin-Ⅰ, crocin-Ⅱ, chikusetsu saponin Ⅳa)were determined simultaneously by multiple wavelength HPLC with diode array detector(DAD) in different parts of Gardenia jasminoides. The results showed that these components in different parts of G. jasminoides had a different distribution, and there was a large difference in content of each component. Geniposide was mainly distributed in fruits and leaves; chikusetsu saponin Ⅳa was mainly distributed in roots and stems; crocus glycosides existed mainly in fruits; chlorogenic acid had a higher distribution in leaves and stems; gardenoside had a higher distribution in leaves and roots, while ceacetylasperulasidic acid methyl ester had a higher distribution in roots and stems. Based on the analysis of the chemical composition and content difference in different parts of G. jasminoides, the basis for the comprehensive utilization and quality evaluation of resources of G. jasminoides was provided.

Collecting different commodity grade Gardenia jasminoides of wild and cultivated varieties all over the country, obtaining color information from each batch of G. jasminoides by the standard D65 light source and image acquisition system, quantifing the gardenia plumpness information by the digital display vernier caliper, determinating 6 kinds of effective components of G. jasminoides by HPLC, classifing from ten indicators by two step clustering analysis and correspondence analysis method of statistics, clearing the importance of the traditional identification indexes, establishing multiple corresponding relation between the skin color and commercial specification of G. jasminoides,exploring the correlation of the skin color and chemical composition, to provide the reference for the reasonable division of commercial specifications and grades of G. jasminoides. Medicine is divided into two classes and has obvious distinguish meaning, The importance of the skin color is greater than the plumpness in traditional identification characteristics, it can accurately distinguish the specifications of G. jasminoides. We improve and rebuild the standard of commodity specifications and grades of Gardenia jasminoides Ellis and establish the rapid evaluation method by the study, it provide a new way and idea for the comprehensive evaluation of G. jasminoides quality.