ObjectiveProteoglycan TPG-1 isolated from Trametes robiniophila（Huaier） has proved to have anti-hepatoma activity， and this paper aims to explore the molecular mechanism. MethodHuman hepatoma SK-HEP-1 cells were treated with TPG-1 （0， 0.05， 0.1， 0.25， 0.5， 1 g·L^-1）. Then cell survival was detected by methyl thiazolyl tetrazolium （MTT） and apoptosis by flow cytometry. In addition， expression of genes in SK-HEP-1 cells treated with or without TPG-1 was examined by DNA microarray to preliminarily explore the anti-hepatoma molecular mechanism of TPG-1. ResultTPG-1 inhibited the proliferation of SK-HEP-1 cells as compared with the blank group （P<0.01）. After treatment with 1 g·L^-1 TPG-1 for 48 h， the apoptosis rate of SK-HEP-1 cells increased （P<0.01）， and TPG-1 promoted the cleavage of cysteinyl aspartate specific proteinase （Caspase）-3 and Caspase-7， the key mediators of apoptosis （P<0.01）. Additionally， TPG-1 （1 g·L^-1） suppressed the migration of SK-HEP-1 cells （P<0.05）. A total of 971 differentially expressed genes （DEGs） were identified in SK-HEP-1 cells after treatment with TPG-1， with 486 up-regulated and 485 down-regulated. These DEGs were mainly involved in the Gene Ontology （GO） terms of interleukin-6 （IL-6） biosynthesis， antigen processing and presentation， superoxide dismutase activity， positive regulation of mitogen-activated protein kinase kinase kinase （MAPKKK） cascade， nature killer （NK） cell chemotaxis， and chemokine biosynthesis， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathways of nucleotide-binding oligomerization domain （NOD）-like receptor signaling pathway， apoptosis， Toll-like receptor signaling pathway， retinoic acid-inducible gene-Ⅰ （RIG-Ⅰ）-like receptor signaling pathway， T-cell receptor signaling pathway， and chemokine signaling pathway. Western blot results showed that TPG-1 （1 g·L^-1） activated mitogen-activated protein kinase （MAPK） signaling pathway in SK-HEP-1 cells （P<0.01）. ConclusionProteoglycan TPG-1 inhibited the proliferation and migration， and induced apoptosis of human hepatoma SK-HEP-1 cells. Up-regulation of MAPK signaling pathway may be responsible for the growth inhibition of human hepatoma SK-HEP-1 cells by TPG-1.

Background::Estrogen is involved in the pathophysiological process of benign prostatic hyperplasia (BPH), in which epithelial-mesenchymal transition (EMT) plays an important role. Upregulation of aquaporin (AQP) 5, which is directly activated by estrogen, has been reported to promote EMT in multiple cells. This study aimed to examine the effects of AQP5 on estrogen-induced EMT in the prostate.Methods::Normal prostate (NP) tissue samples without any histopathological changes and BPH tissue samples with pathologically confirmed hyperplasia were obtained. An EMT cell model was subsequently established by adding estradiol (E2) to RWPE-1 cells, after which AQP5 knockdown was performed. Tissue morphological and immunohistochemical features were examined using hematoxylin-eosin and immunohistochemical staining. Western blot analysis was performed to determine the expression of AQPs, estrogen receptors, and EMT-related proteins. Cell proliferation was assessed and supernatants were collected for enzyme-linked immunosorbent assay to determine transforming growth factor-β1 (TGF-β1) concentrations. Immunofluorescence staining was performed to assess protein expressions in RWPE-1 cells. Results::BPH tissues exhibited greater EMT (TGF-β1: 1.362 ± 0.196 vs. 0.107 ± 0.067, P = 0.003; vimentin: 1.581 ± 0.508 vs. 0.221 ± 0.047, P < 0.001; E-cadherin: 0.197 ± 0.188 vs. 1.344 ± 0.088, P < 0.001), higher AQP5 (1.268 ± 0.136 vs. 0.227 ± 0.055, P < 0.001) and estrogen receptor (ER) α (1.250 ± 0.117 vs. 0.329 ± 0.134, P < 0.001) expression but lower ERβ (0.271 ± 0.184 vs. 1.564 ± 0.130, P < 0.001) expression than NP tissues. E2-stimulated cells had higher AQP5 expression (1.298 ± 0.058 vs. 1.085 ± 0.104, P = 0.049), increased cell proliferation (1.510 ± 0.089 vs.1.000 ± 0.038, P < 0.001), and EMT (TGF-β1 concentration: 0.352 ± 0.021 ng/mL vs. 0.125 ± 0.014 ng/mL, P < 0.001; vimentin: 1.641 ± 0.120 vs. 0.188 ± 0.020, P = 0.002; E-cadherin: 0.075 ± 0.030 vs. 0.843 ± 0.046, P < 0.001) than controls. E2-stimulated cells with AQP5 knockdown exhibited decreased EMT (TGF-β1 concentration: 0.223 ± 0.041 ng/mL vs. 0.352 ± 0.021 ng/mL, P= 0.016; vimentin: 0.675 ± 0.056 vs. 1.641 ± 0.120, P = 0.001; E-cadherin: 0.159 ± 0.037 vs. 0.075 ± 0.030, P = 0.040) than E2-stimulated cells with non-related small interfering RNA (siRNA). Conclusion::Our findings suggest that estrogen induces BPH possibly by promoting AQP5 expression. Hence, AQP5 might be a novel target for modulating EMT in prostate epithelial cells.

AlM:To discuss the protective effect ofα-mangostin on retinal light damage in mice.METHODS:Totally 30 Balb/c mice, aged 6~8wk, were randomly divided into the control group, light-exposure group and α-mangostin group. Every group contained 10 mice. Mice of α-mangostin group were treated with alpha-mangostin at the dose of 30mg/( kg · d ) body weight by intragastric administration daily for 7d, and then exposed to white light at the 5th d. The light-exposure group and α-mangostin group were exposed to 5 000 ± 200lx white light-emmiting diodes (LEDs) for continuously 1h to establish the mice model of retinal light damage. Flash -electroretinograme was recorded 72h after light exposure. The changes in retinal morphology of mice were observed by light microscopy. Retinas were extracted to detect the malondialdhyde ( MDA ) content change of the retinal homogenate.RESULTS: Flash-electroretinogram ( F-ERG ) showed that retinal dysfunction was less severe in α-mangostin group than in light-exposure group ( P<0. 05 ). Light microscopy test showed that retina structural damage was less severe in α-mangostin group than in light-exposure group (P<0. 05). The level of MDA in retinal tissue of α-mangostin group was significantly lower when compared with light-exposure group (P<0. 05).CONCLUSlON: α-mangostin inhibits lipid peroxidation induced by light damage and protect retina against light damage.

OBJECTIVE Toinvestigatetheantidepressantandantianxietyeffectofproanthocyani-dins(OPC)inchronicallystressedratsanditsunderlyingmechanism.METHODS Onemethodwas selected from 8 different stress methods each day,and the rats were treated with OPC (25,50 and 1 00 mg·kg -1 )1 h before the stress method.The chronically stressed model was established.After 21 d stress experi ment,the i mmobility ti me in force swi mming test,sucrose consu mption and the nu mber of marbles buried in the marble burying test were observed respectively each day.OPC (25,50 and 1 00 mg·kg -1 )was given 1 h before each test.In addition,Western blotting was used to analyze the expression of brain derived neurotrophic factor (BDNF) and phosphorylated cyclic AMP response element-bindingprotein(p-CREB)inthehippocampusandfrontalcortex.RESULTS Comparedwith the control group,the chronically stressed group showed obvious depressive-like and anxiety-like behav-ior,while the immobility time decreased from (90.57 ±4.27)s in chronically stressed group to (78.25 ± 2.53)s (P<0.05),(72.12 ±3.21 )s(P<0.05)and (60.77 ±3.41 )s (P<0.05)when ig given OPC 25,50 and 100 mg·kg -1 respectively,the ratio of sucrose preference increased from (42.80 ±4.92)%to (67.54 ±4.32)%(P<0.05)and (72.21 ±7.99)%(P<0.05)when ig given OPC 50 and 1 00 mg· kg -1 respectively,the number of buried marbles decreased from 1 .57 ±0.21 in chronically stressed group to 0.63 ±0.26 (P<0.05)and 0.44 ±0.1 8 (P<0.05)when ig given OPC 50 and 1 00 mg·kg -1 respectively.The expression of p-CREB in the hippocampus and frontal cortex distinctively increased in OPC group (25,50 and 100 mg·kg -1 )(P<0.05),so did the expression of BDNF in the hippocampus andfrontalcortexinOPCgroup(50and100mg·kg-1)(P<0.05).CONCLUSION OPCcanreverse the depressive-like and anxiety-like behavior in chronically stressed rats,which may be related to the cAMP-CREB-BDNF signal transduction cascades.

To study the constituents of the Prunella vulgaris L, the constituents were isolated by various column chromatography and the structures were identified on the basis of chemical and spectral analysis. One saponin compound (I) and one flavone glycoside compound (II) were obtained from Prunella vulgaris L. Their structures were elucidated as 16-oxo-17-demethyl-3beta,24-dihydroxylolean-12-en-3-O-beta-D-glucuronoside (I), and acacetin-7-O-beta-D-glucopyranoside (II). Compound I is a novel triterpenoid saponin and named as prunelloside A. Compound II was obtained for the first time from the Prunella genus.
Flavonoids ; isolation & purification ; Glucosides ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Prunella ; chemistry ; Saponins ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

Objective To explore the repair of the open compound wounds in lower extremities caused by multiple factors. Methotis Transplantation of cutaneous.musculo-cutaneous or greater omentum flaps were applied to 155 patients of open compound lower extremity wounds. Results The wound healing rate following first operation was 50% and that following two operations was 14.8%.While the wounds were healed in 7.7% of patients after three operations. Conclusion Transplantations of cutaneous,musculo-cutaneous or greater omentum flaps ale effective to repair and reconstruct the open compound lower extremity wounds.

OBJECTIVETo establish the processing method of Cistanche tubulosa decoction pieces.

METHODThe orthogonal test of four factors and three levels was used to optimize the main factors in the process of fresh C. tubulosa decoction pieces processing, including the thickness, temperature, and the time for inactivation of the enzyme in the plant. The result showed that the optimized condition was that fresh C. tubulosa was cut into 4 mm thickness, and heated at 70 degrees C for inactivation the enzyme in the plant for 6 min. Moreover, the optimized method was compared with the method of insolation and traditional dried method.

RESULTThe content of echinacoside in the C. tubulosa decoction piece by the optimized method was 7.3 times of that dried by insolation, and 12.8 times of that by traditional dry method; the content of verbascoside was 6. 5 and 14. 9 times of that dried by insolation and by traditional dry method, respectively; the content of galactitol was 7.1 and 13.2 times of that dried by insolation and by traditional dry method, respectively.

CONCLUSIONThe quality of C. tubulosa decoction pieces could be improved by this method, and its crud drug could be saved, which would protect the source of the mild Herba Cistanche, and produced the better economic and ecological benefits.

Cistanche ; chemistry ; Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Galactitol ; analysis ; Glucosides ; analysis ; Glycosides ; analysis ; Hot Temperature ; Phenols ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the frequencies of human leuckocyte antigens (HLA) -A, B and DRB1 alleles in Chinese patients with primary biliary cirrhosis (PBC) using polymerase chain reaction-based techniques, and to assess the correlation of HLA molecules with other clinical and laboratory profiles.

METHODSGenotyping of HLA-A, B, and DRB1 were performed in 65 well-characterized patients with primary biliary cirrhosis and 431 healthy controls with PCR amplification with sequence-specific primers (PCR-SSP).

RESULTSThe frequency of DRB1*0701 was increased to 29.2% compared with 13.9% in the controls (PC < 0.05, OR = 2.55, 95% CI: 1.4 approximately 4.6). No association was found with HLA-DRB1*08 which had been constantly reported. The A*2 allele (53.8%) was more frequent in the PBC patient group but without a significant statistical difference. The frequencies for the other A, B and DRB1 alleles were similar between patients and healthy controls. There was no difference between patients with or without DRB1*0701 in some clinical and laboratory profiles.

CONCLUSIONSusceptibility to primary biliary cirrhosis in Chinese is associated with DRB1*0701 allele and differs from people in North America, South America, North Europe and even in Japan, but the association is not restricted to any particular subgroup of patients. Valine at position 78 of HLA DRbeta1 may play an important role in the pathogenesis of primary biliary cirrhosis.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Female ; HLA Antigens ; genetics ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DR Antigens ; genetics ; Humans ; Liver Cirrhosis, Biliary ; genetics ; immunology ; Male ; Middle Aged ; Polymorphism, Genetic

OBJECTIVEA study on the value of antimitochondrial antibody (AMA) and its subtypes anti-M2, anti-M4, and anti-M9 in diagnosing primary biliary cirrhosis (PBC).

METHODSAntimitochondrial antibody was detected by indirect immunofluorescence and anti-M2, anti-M4 and anti-M9 by Western blotting. AMA and anti-M2 of 78 PBC patients, of 35 non-PBC hepato-biliary disease patients and 20 healthy controls were studied and anti-M2, anti-M4 and anti-M9 were studied in 30 of the 78 PBC patients.

RESULTS96.2% (75/78) of PBC patients were AMA positive and 94.9% (74/78) of PBC patients were anti-M2 positive. Only three among the 35 non-PBC patients were positive for AMA (one with very low titre). None of the 35 non-PBC patients was anti-M2 positive. AMA and anti-M2 were negative in all the healthy controls. Among the 30 anti-M2 positive patients, 16 patients were anti-M4 positive (16/30, 53.3%) and 4 patients were anti-M9 positive (4/30, 13.3%).

CONCLUSIONAMA and its subtypes (special anti-M2) are important sero-immunological markers for the diagnosis of PBC.

Autoantibodies ; blood ; classification ; Female ; Humans ; Liver Cirrhosis, Biliary ; diagnosis ; immunology ; Male ; Mitochondria, Liver ; immunology

OBJECTIVETo identify the protective effects of hypovolemic hypotension preconditioning on cardiopulmonary function after myocardial ischemia/reperfusion injury and to explore the possible mechanism.

METHODSTwenty-four male white rabbits were randomly assigned to two groups. In the control group, ischemia/reperfusion animals(Group I/R, n=10) were subjected to thirty-minute occlusion of left anterior descending coronary artery and two-hour reperfusion. Animals in hypovolemic hypotension preconditioning group (Group HHP, n=14) experienced brief systemic ischemia preconditioning through blood withdrawal to lower blood pressure to 40%-50% of the baseline before myocardial ischemia/reperfusion. Hemodynamic parameters were recorded. Blood sample was taken to measure superoxide dismutase (SOD), malondialdehyde (MDA) and nitrogen monoxide (NO) changes with blood gas analysis. Myocardium specimens were sampled to examine apoptosis-related gene interleukin-1 beta converting enzyme (ICE) mRNA.

RESULTSCardiac mechanical function and lung gas exchange remained stable in Group HHP with a significant increase in NO level; while in Group I/R without preconditioning, cardiopulmonary dysfunction was present after 2 h reperfusion associated with a significant reduction in NO formation and an increase in MDA (P<0.001). There was negative expression of ICE mRNA in the two groups.

CONCLUSIONSHypovolemic hypotension preconditioning significantly improves cardiopulmonary function and increases NO formation and the protective benefit associated with hypovolemic hypotension preconditioning of the heart may be regulated through NO mediated mechanism.

Animals ; Blood Pressure ; Blood Volume ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Malondialdehyde ; blood ; Myocardial Reperfusion Injury ; physiopathology ; prevention & control ; Nitric Oxide ; blood ; Rabbits ; Superoxide Dismutase ; blood