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Patients with corona virus disease 2019(COVID-19) mainly have fever, cough and other respiratory symptoms, but it is necessary to be aware that some patients are complicated by digestive symptoms, such as diarrhea, nausea and vomiting, and even a few patients show atypical symptoms such as diarrhea as the first presentation. This article reviews the mechanism of SARS-CoV-2 infection with digestive tract symptoms,change of intestinal microbiota, therapeutic drugs, psychosocial factors, cytokine storm and aggravation of primary underlying diseases, etc., so as to sharpen clinicians vigilance against COVID-19 patients with digestive system symptoms, and provide theoretical basis for corresponding treatment.

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Relation recommendation based on common neighbors' property is a hot research branch of link prediction in social network analysis. This paper proposesd a new measure of relation recommendation by introducing a hidden naive Bayesians (HNB) classification model, which model the task by analyzing the dependency among properties and incorporates this idea to measure the influence and contribution among common neighbors. Then built a ranking model to learn the highest similarity associated with each candidate pair by maximizing the likelihood of relationship building and extended the model to CN, AA and RA similarity-based recommendation algorithms. Experimental evaluation by AUC on real social networks proved that the proposed model can achieve a better result than some baseline and LNB. Finally, it also discovered that attributes with different network topologies recommended precision linear effects. [ABSTRACT FROM AUTHOR]

Objective: To compare the clinical efficiencies of three anesthetic techniques used in the transvaginal ultrasound-guided oocyte retrieval. Methods: From December 2011 to March 2012 in our Center, 462 patients undergoing IVF -ET were divided into three groups according to anesthetic techniques during oocyte retrieval, group A (227 cases) used Dolantin (i. m.) only, group B (54 cases) used acupuncture anesthesia, and group C (181 cases) used Propofol (i. v.). The anesthesia effects and adverse effects were evaluated. Results: There were higher oocyte retrieval rate in group C than the other groups (P<0.05), but there were no significant differences in clinical pregnancy rate and miscarriage rate among three groups (P>0.05). The analgesic effects in three groups were similar, but the adverse effects in the group B and C, such as nausea and sweating, were lower (P<0.05) than those in group A. Conclusions: Acupuncture anesthesia and Propofol (i. v.) can reduce pain intensity and adverse effects in patients undergoing transvaginal oocyte retrieval, although treatments do not affect the clinical pregnancy rate and the abortion rate in IVF-ET. [ABSTRACT FROM AUTHOR]

This study investigated the effect of trametenolic acid(TA) on the migration and invasion of human hepatocellular carcinoma HepG2.2.15 cells by using Ras homolog gene family member C(RhoC) as the target and probed into the mechanism, aiming to provide a basis for the utilization of TA. The methyl thiazolyl tetrazolium(MTT) assay was employed to examine the proliferation of HepG2.2.15 cells exposed to TA, and scratch and Transwell assays to examine the cell migration and invasion. The pull down assay was employed to determine the impact of TA on RhoC GTPase activity. Western blot was employed to measure the effect of TA on the transport of RhoC from cytoplasm to cell membrane and the expression of RhoC/Rho-associated kinase 1(ROCK1)/myosin light chain(MLC)/matrix metalloprotease 2(MMP2)/MMP9 pathway-related proteins. RhoC was over-expressed by transient transfection of pcDNA3.1-RhoC. The changes of F-actin in the cytoskeleton were detected by Laser confocal microscopy. In addition, the changes of cell migration and invasion, expression of proteins in the RhoC/ROCK1/MLC/MMP2/MMP9 pathway, and RhoC GTPase activity were detected. The subcutaneously transplanted tumor model of BALB/c nude mice and the low-, medium-, and high-dose(40, 80, and 120 mg·kg~(-1), respectively) TA groups were established and sorafenib(20 mg·kg~(-1)) was used as the positive control. The tumor volume and weight in each group were measured, and the expression of related proteins in the tumor tissue was determined by Western blot. The results showed that TA inhibited the proliferation of HepG2.2.15 cells in a concentration-dependent manner, with the IC&#95;(50) of 66.65 and 23.09 μmol·L~(-1) at the time points of 24 and 48 h, respectively. The drug administration groups had small tumors with low mass. The tumor inhibition rates of sorafenib and low-, medium-and high-dose TA were 62.23%, 26.48%, 55.45%, and 62.36%, respectively. TA reduced migrating and invading cells and inhibited RhoC protein expression and RhoC GTPase activity in a concentration-dependent manner, dramatically reducing RhoC and membrane-bound RhoC GTPase. The expression of ROCK1, MLC, p-MLC, MMP2, and MMP9 downstream of RhoC can be significantly inhibited by TA, as confirmed in both in vitro and in vivo experiments. After HepG2.2.15 cells were transfected with pcDNA3.1-RhoC to overexpress RhoC, TA down-regulated the protein levels of RhoC, ROCK1, MLC, p-MLC, MMP2, and MMP9 and decreased the activity of RhoC GTPase, with the inhibition level comparable to that before overexpression. In summary, TA can inhibit the migration and invasion of HepG2.2.15 cells. It can inhibit the RhoC/ROCK1/MLC/MMP2/MMP9 signaling pathway by suppressing RhoC GTPase activity and down-regulating RhoC expression. This study provides a new idea for the development of autophagy modulators targeting HSP90α to block the proliferation and inhibit the invasion and migration of hepatocellular carcinoma cells via multiple targets of active components in traditional Chinese medicines.

Protein-like dissolved organic matter (pDOM), which is ubiquitous in natural waters, is a critical precursor of nitrogenous disinfection byproducts. Recently, the control and elimination of pDOM have been a growing concern during drinking water treatment processes. In this study, a high-performance size exclusion chromatography system coupled with photo-diode array, fluorescence detector, and online organic carbon detector (HPSEC-PDA/FLD/OCD) was used to determine the removal behaviors of different-sized pDOM from two full-scale drinking water treatment plants (DWTPs). Coagulation and activated carbon adsorption were selected for bench-scale experiments to further assess the removal behavior of pDOM during conventional water treatment processes. The results showed that different-sized pDOM fractions exhibited different removal characteristics. Pre-oxidation can effectively remove some tyrosine-like and tryptophan-like components with high MW, and as the oxidization effect was enhanced, more high MW fractions decomposed into low MW ones. Conversely, some aliphatic pDOM fractions in high MW (e.g., aliphatic proteins) were not subject to pre-oxidation removal. The coagulation-sedimentation unit was efficient in removing high MW fractions, specifically tryptophan-like fractions. Additionally, some pDOM components may be released during coagulation. pDOM with low MW and high hydrophobicity were easily removed during activated carbon filtration. However, long-term operation of the activated carbon filter may breed microorganisms, resulting in the partial release of pDOM fractions. Moreover, UV disinfection processes promoted the degradation of low MW pDOM components. Due to the complex water quality and uncontrollable microbial activities, the aforementioned water treatment units did not exhibit a synergistic effect on pDOM removal. In comparison with humic-like substances, pDOM was susceptible to water quality changes, and its removal was limited in the surveyed DWTPs. Therefore, DWTPs must strengthen pDOM monitoring in influent and effluent and adjust the operating parameters of different treatment units in a timely manner. Moreover, the combination of advanced water treatment processes, such as ozone-biological activated carbon process and nanofiltration, should also be considered to strictly control pDOM component removal.

Objective To verify the expressions of genes associated with colorectal cancer with hyperglycemia and evaluate their diagnostic values.Methods Tumor tissues,distal normal intestinal mucosa,and peripheral blood samples were harvested from 109 colorectal cancer patients and peripheral blood samples from 30 diabetes patients and 30 healthy volunteers. The mRNA expressions of glucose regulated protein 78 (GRP78),NADPH oxidase-1 (NOX1),carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5),heat shock protein 60 (HSP60),and histone deacetylase 1(HDAC1) were detected by real-time quantitative polymerase chain reaction. The correlation between the gene expressions and clinicopathological parameters in colorectal cancer patients were analyzed using Pearson's correlation analysis. Diagnostic test accuracy evaluation was used to calculate the sensitivity,specificity,accuracy,predictability,Youden index,and likelihood ratio of serum gene expressions in colorectal cancer patients,and the receiver operating characteristic (ROC) curves were drawn. The area under the ROC curve was calculated to evaluate the diagnostic efficiency of the combined detection of multiple genes.Results The mRNA levels of GRP78 (P=0.001),NOX1 (P=0.022),CEACAM5 (P=0.000),HSP60 (P=0.044),and HDAC1 (P=0.047) were positively correlated with the fasting blood glucose level. The mRNA expressions of NOX1 (P=0.000,P=0.008) and HDAC1 (P=0.000,P=0.037) in tissues and serum were significantly higher in colorectal cancer patients than in patients with normal blood glucose levels. The NOX1 mRNA expression was positively correlated with the diameter of colorectal cancer (P=0.013),and the HDAC1 mRNA expression was significantly correlated with the tumor site (P=0.049),depth of primary tumor invasion (P=0.025),and TNM stage (P=0.042). The areas under the ROC curves of NOX1,CEACAM5,and HDAC1 were 0.931,0.852,and 0.860 respectively (all P=0.000). The specificity,accuracy,and negative predictive value of NOX1,HDAC1 mRNA expression in colorectal cancer patients with hyperglycemia were all above 90%. The diagnostic sensitivity and specificity of the combined detection of NOX1,CEACAM5,and HDAC1 were 98.82% and 99.93%,respectively.Conclusion Combined detection of genes associated with colorectal cancer accompanied by hyperglycemia can improve the diagnostic efficiency of early screening.

Samples collected from 12 rivers with typical vegetation types in the permafrost regions on the Qinghai-Tibetan Plateau were incubated in the laboratory, and the relationships among the vegetation types, river discharges, the compositions of dissolved organic carbon (DOC), permafrost areas, riverine DOC concentration, biodegradability of dissolved organic carbon (BDOC), and the biodegradation kinetics were examined. The results showed that the DOC concentrations of typical vegetation types in the basin, such as alpine meadow (AM), alpine swamp meadow-alpine meadow (ASM-AM), alpine meadow-alpine steppe (AM-AS), and alpine meadow-alpine steppe-bare soil (AM-AS-BL), were (5.17±0.21), (5.02±0.50), (3.55±0.25), and (2.79±0.41) mg ·L -1 , respectively. The values for the bioavailability of river DOC of different vegetation types were (23.54±2.62)%, (23.66±3.31)%, (18.17±5.26)%, and (11.72±15.56)%, respectively. Correspondingly, the riverine DOC aromaticity increased along with the vegetation cover, while the biodegradation and degradation rates decreased gradually. During the incubation, the reaction of BDOC was in accordance with the first-order kinetics equation. Furthermore, the BDOC in continuous permafrost regions of the rivers was greater than that in the non-continuous permafrost regions. The BDOC in higher discharges were lower than those with lower discharges. Taken together, the results suggested that the vegetation types were the main controlling factors for the BDOC, and BDOC was also related to the discharge and permafrost.

Objective: Using an adenoviral vector, the wild-type PTEN gene was transduced into activated hepatic stellate cell (HSC) cultured in vitro and cell cycle markers and were detect. Thereby, the potential mechanisms of inhibitory effect of the wild-type PTEN overexpression on the proliferation in activated HSC was investigated., Methods: The wild type PTEN gene was transduced into activated HSC (HSC-T6 ) cultured in vitro mediated by adenoviral vector. PTEN expression in HSC was measured by Western blot and Real-time fluorescent quantitation PCR. Flow cytometry (FCM) was then used to detect cell cycle phase of activated HSC. And the expressions of cyclinD1 and cyclin dependent kinase 4 (CDK4) in HSC were determined by Western blot., Results: The data showed that exogenous wild type PTEN gene was successfully transduced and expressed in activated HSC cultured in vitro. The over-expression of wild type PTEN resulted in the increased number of HSC at G0/G1 phase ( P less than 0.01), and the number of HSC at S phase and G2/M phase were decreased significantly, P less than 0.01. Furthermore, there were decreased cyclinD1 and CDK4 expression in HSC infected with Ad-PTEN, P less than 0.01., Conclusion: The over-expression of wild type PTEN inhibit transition of activated HSC in vitro from G1 to S phase and arrest cell cycle of them at G0/G1 phase via the down-regulated expressions of cyclinD1 and CDK4, and then inhibit HSC proliferation.

Objective: To explore the effect of Salvia miltiorrhiza monomer IH764-3 on apoptosis in hydrogen peroxide (H2O2)-stimulated hepatic stellate cells (HSCs)., Methods: HSCs were cultured in medium with different IH764-3 doses (10 mg/L, 20 mg/L, 30 mg/L, 40 mg/L) and without IH764-3. Direct cell count, 3H-thymidine incorporation, Annexin-V/Propidium Iodide double-labeled flow cytometry, TUNEL and transmission electron microscopy were employed to estimate the influence of IH764-3 on proliferation and apoptosis of HSCs. The expression of extracellular signal-regulated kinase 1 (ERK1) mRNA and protein in HSCs were detected using RT-PCR and Western blot respectively., Results: It was showed that H2O2 could promote HSC proliferation. In contrast, IH764-3 at concentrations of 10 mg/L, 20 mg/L, 30 mg/L and 40 mg/L inhibited its proliferation. The inhibition rates were 7.13%, 28.36%, 53.80% and 73.10% (P < 0.01). And the inhibition rates of IH764-3 at concentrations of 30 mg/L at 12 h, 24 h and 48 h were 22.24%, 40.51% and 61.65%. Furthermore, IH764-3 could also induce the HSC apoptosis in dose-dependent an dtime-dependent manners (P < 0.01). In addition, after exposed of HSCs to IH764-3 for 24 h, ERK production decreased and ERK1 mRNA was down-regulated earlier about 2 h after exposure to IH764-3., Conclusion: IH764-3 may inhibit the proliferation and induce apoptosis of HSCs in both dose-dependent and time-dependent manners, which may be related to down-regulation of ERK expression.

Objective: To investigate the mechanisms of Aldosterone stimulating hepatic stellate cells(HSCs) contraction via Ca2+-independent pathways., Methods: HSC-T6 cell line was pre-disposed with Aldo 10mumol/L. The cell contraction was detected by silicone-rubber-membrane cultivation directly. The concentration variation of intracellular free calcium in rat HSC was observed by laser confocal microscopy. Besides, HSC-T6 cell line was under pre-disposal treatment with the blocking agents of Aldo receptor -antisterone, protein kinase C (PKC) special blocking agent-Stauro, Rho kinase blocking agent-Y27632 and MLCK special blocking agent-ML-7 respectively prior to stimulation with aldosterone. RT-PCR was used to detect the expression of Rock2, RhoAGTP and RhoGEF in Ca2+- independent pathways mediated by Rho-kinase., Results: Aldo could induce HSCs contraction. The concentration of intracellular free calcium in rat HSCs had no change after pre-disposal treatment with Aldo. The mRNA expressions of Rock2, RhoAGTP and RhoGEF increased significantly after treatment with Aldo (0.770+/-0.049, 0.960+/-0.096, 0.180+/-0.006, P is less than 0.01).When inhibited with antisterone, the mRNA expressions of the three elements were (0.440+/-0.166, 0.370+/-0.180 and 0.050+/-0.001, P is less than 0.01), lower than that of Aldo group, but higher in ML-7+Stauro + Aldo groups (0.940+/-0.066, 1.330+/-0.192 and 0.160+/-0.007, P is less than 0.05) as compared to the control group (0.140+/-0.023, 0.540+/-0.111 and 0.110+/-0.012). In the Y27632 + ML-7 + Stauro+Aldo group, the mRNA expression of RhoGEF (0.290+/-0.004, P is less than 0.01)was higher than that of the ML-7 + Stauro + Aldo group (0.160+/-0.007)., Conclusion: Aldo could induce HSCs contraction via Ca2+-independent pathways and Rho-Rock pathway involved in the process.

Erythrocyte tropomodulin (E-Tmod) is first isolated from human erythrocyte membrane as a TM-binding protein. Its N-terminus contains two TM-binding sites and one TM-dependent actin capping domain and C-terminus contains 5 leucine-rich repeats and a TM-independent actin capping domain. As the unique capping protein at the slow-growing end of F-actin, E-Tmod binds to N-terminus of TM and actin and decreases the TM-coated F-actin depolymerization. E-Tmod encoding gene is highly conserved and E-Tmod is distributed widely in erythrocytes and cardiomyocytes, etc. E-Tmod plays a pivotal role in organizing F-actin and cytoskeleton and maintaining the mechanical properties of the cells.

Objective: To investigate the clinical value of neoadjuvant chemotherapy in the treatment of operable breast cancer., Methods: A total of 120 patients with pathologically proven operable breast cancer were randomized into two groups to receive 2-4 cycles of neoadjuvant chemotherapy with docetaxel combined with pirarubicin (TThp) or docetaxel combined with epirubicin (TE). Operations were performed two weeks after the neoadjuvant chemotherapy. The short-term therapeutic effect, toxic reaction and the impact of neoadjuvant chemotherapy on the choice of surgical approaches were evaluated., Results: The total effective rate for the primary sites was 87.2% in these patients, and the rate of breast conservation significantly increased from 12.7% to 41.8% (P<0.05), with a tumor resection rate of 97.2%. The major adverse effects of the therapy included leukopenia, nausea, vomiting and alopecia., Conclusion: Neoadjuvant chemotherapy can enhance the breast conservation rate, lower the clinical staging of the tumors and minimize the surgery area to improve the postoperative quality of life of the patients.

Objective: To investigate the dynamic changes of a-AR, b1-AR and b2-AR expression in hepatic fibrosis., Methods: Rat hepatic fibrosis model was established by bile duct ligation (BDL). HE and Masson staining were used to determine hepatic fibrosis levels. Immunohistochemistry was applied to detect alpha -smooth muscle actin (alpha -SMA), a marker of hepatic stellate cell (HSC) activation; Western blot and real-time RT-PCR were used to measure the dynamic changes of alpha -AR, beta(1)-AR, beta(2)-AR expression on protein and mRNA levels, respectively, during the development of hepatic fibrosis., Results: (1) HE and Masson trichrome staining showed that the liver fibrosis models were established successfully. (2) At 1, 2, 3, 4 wk after BDL, alpha -SMA positive area density of the model group (10.58% +/- 1.75%, 24.14% +/- 2.02%, 29.74% +/- 2.59%, 34.28% +/- 2.01%) was significantly higher than that of the sham operation group (4.12% +/- 1.51%), P less than 0.01. (3) The expression of alpha -AR, beta(1)-AR, beta(2)-AR protein and mRNA was increased with the development of the hepatic fibrosis (P less than 0.05). (4) alpha -SMA expression was positively associated with alpha -AR, beta(1)-AR, beta(2)-AR, r values were 0.564, 0.753 and 0.606, respectively., Conclusion: The expression of alpha -SMA is increased dramatically during the fibrosis, and is positively associated with the expression of alpha -AR, beta(1)-AR and beta(2)-AR.

Objective: To investigate the relationship between the expression of glucose transporter protein 1 and the apoptosis of neuron during hypoxic-ischemia brain damage in neonatal rats., Methods: Total 120 10-day old SD rats were divided into normal group, sham control group and hypoxic-ischemia (HI) group. In HI group, hypoxic-ischemia brain damage (HIBD) were generated according to Rice-Vannucci method, brain tissues were harvested at 2, 8, 24, 48 and 72 h after HI. The brain samples were also collected at the same time points in normal group and sham control group. The pathological changes was observed by hematoxylin-eosin (HE) staining, the mRNA expression of glucose transporter 1 (GLUT1) was detected by reverse transcriptase-polymerase chain reaction (RT-PCR), the protein expressions of GLUT1 and cleaved caspase-3 (CC3) were detected by immunohistochemistry, the apoptosis of neuron was measured by TdT-mediated dUTP-biotin nick end labeling (TUNEL) staining., Results: HE staining showed that the degree of brain cell damage increased with time after HI, the loss of neuronal cells peaked at 48 h, while the cells in control group apperanted in an orderly and normal morphology. The mRNA and protein expressions of GLUT1 were increased after HI, which began to increase at 2 h, and reach the peak at 24 h. and the expression levels at each time points were statistically higher (P< 0.01) than those in control group. CC3 protein expression also began to increase at 2 h, peaked at 48 h after HI, which was higher than that of control group (P<0.01). The number of positive cells was significantly increased after HI,with a peak at 48 h., Conclusion: The mRNA and protein expression of GLUT1 in brain tissue increased significantly after hypoxic-ischemia, and the peak time was earlier than that of CC3 protein and cellular apoptosis. This suggests that GLUT1 expression upregulation may be a certain degree of inhibition on neuronal apoptosis.

Objective: To detect the changes in intestinal mucosal permeation in rats with methotrexate-induced small intestinal damage and investigate the protective effects of Changyanqing decoction., Methods: Rat enteritis model was established by methotrexate (MTX) and sodium chloride. The rats were randomly divided into normal control group, model group, N-acetylcysteine (NAC) group and Changyanqing decoction group, and Changyanqing decoction (100 mg/kg) or saline was administered daily in the corresponding groups by gastric irrigation for 6 days. The disease activity index (DAI), colonic mucosal damage index (CMDI) and histological score (HS) of the rats were observed and evaluated. The levels of mRNA expressions of TNF-alpha and IL-1beta were detected by semi-quantitative RT-PCR. The expression of IL-10 was detected by enzyme linked immunosorbent assay, and IkappaB expression was determined with Western blotting., Results: Compared with the normal control group, the model group showed significantly increased DAI, CMDI and HS. The DAI, CMDI, and HS in rats treated with Changyanqing decoction were significantly decreased in comparison with those in the model group (P<0.01). The expressions of TNF-alpha and IL-1beta were significantly higher in MTX-treated group than in the control group. The expression of TNF-alpha and IL-1beta mRNA in the Changyanqing group and NAC group were significantly lower, but IL-10 significantly higher than those of the MTX group. In MTX group, obvious NF-kappaB activation was observed, whose expression was significantly stronger in the cell nuclei, and the IkappaB in the cytoplasm was markedly degraded., Conclusion: Changyanqing decoction offers protection on intestinal mucosa by inhibiting NF-kappaB activation to reduce TNF-alpha and IL-1beta mRNA expressions and increase IL-10 expression.

Objective: To investigate the role of focal adhesion kinase (FAK) in adhesion and migration of hepatic stellate cells (HSC)., Methods: Two recombinant plasmids expressing short hairpin RNAs (shRNAs) targeting FAK were constructed and one plasmid substantially suppressing FAK expression in HSC was selected. Real-time PCR and Western blot were used to detect the knockdown effects of FAK gene. After 48-hour treatment with FAK shRNA, toluidine blue colorimetric assay was used to detect the cell adhesion. Wound-healing assay and improved Boyden double-chamber were used to detect the cell migration induced by FN., Results: The recombinant plasmid expressing FAK shRNA was successfully constructed and transfected into HSC. Compared with the controls, the expression of FAK mRNA and protein in HSC treated with FAK shRNA was markedly down-regulated by 76.82% and 72.53%, respectively. The expression of p-FAK (Tyr397) protein was also decreased by 62.71% 48 h posttransfection. The adhesion of HSC was inhibited by 58.69% at 48 h after shRNA transfection. FAK gene silencing could also dramatically inhibit FN-stimulated HSC migration, and the cell migration distance and the cell number of crossing membrane were decreased by 58.27% and 83.70%, respectively., Conclusions: FAK gene silencing suppresses adhesion and migration of HSC, and FAK may be a potential target for novel anti-fibrosis therapies.

The characteristics of occurrence and distribution of 8 synthetic musks in 15 sewage sludges in Shanghai were studied. The results indicated that HHCB and AHTN were the two main components in all sludges, with the mean concentration of 2.92 mg x kg(-1) (0.81-6.39 mg x kg(-1)) and 1.96 mg x kg(-1) (0.35-3.11 mg x kg(-1)), respectively; and the concentrations of ADBI, AHMI, MK were less. The distributions detected in sludges are in accordance with the usage patterns in China. Total concentrations of 8 synthetic musks were ranged from 1.16-9.57 mg x kg(-1), which were lower than the results in previous studies. Concentrations in sludges are influenced by ratio of domestic wastewater in influent, sewage load and sewage treatment methods. The good linear relationships among HHCB, AHTN and ADBI indicate that these components have the same source: domestic wastewater. The consumption rates of HHCB and AHTN connected to per inhabitant in Shanghai region were estimated, which are low compared with those found in European. The potential impacts on agricultural soil were also assessed.

Aim: To observe the effect of FAK-related non-Kinase (FRNK) plasmid on hepatic stellate cell (HSC) proliferation stimulated by fibronectin (FN)., Methods: FRNK plasmid was transfected into HSC with transient liposomal transfection. The proteins of FRNK, FAK and p-FAK(Tyr397) were assayed by Western blotting analysis. The proliferation of HSC was evaluated by improved MTT assay, and cell cycle pattern was determined by flow cytometry (FCM)., Results: (1) The expression of FRNK protein increased after FRNK transfected HSC, and it was at 48 h that the expression of FRNK protein was the highest (P < 0.01). The protein level of FAK was no significant difference between before FRNK plasmid transfection and after transfection (P > 0.05). The expression of p-FAK(Tyr397) protein was down-regulated after FRNK had been transfected in HSC, (P < 0.01). (2) The HSC proliferation inhibition rates at 12 h, 24 h and 48 h after FRNK transfection were 20.07%, 26.16%, 29.77%, respectively (P < 0.01). (3) Compared with the non-FRNK plasmid group, the FRNK-transfected HSCs almost arrested in G0/G1 phase (71.4 +/- 2.81 vs 48.9 +/- 1.66, P < 0.01)., Conclusion: After FRNK were transfected successfully in HSCs using lipofectamine, the phosphorylation of FAK was inhibited. The HSC proliferation was restrained in a time-dependent manner and the HSC was arrested in G0/G1 phase.

Objectives: To investigate the effects of FAK-related non-kinase (FRNK) on the apoptosis of hepatic stellate cells (HSC) in vitro and on the extracellular signal-regulated kinase (ERK) signal transduction pathway., Methods: HSC were stimulated by fibronectin (FN), and then they were transfected with FAK-related non-kinase (FRNK) plasmids mediated by cationic liposome. The apoptosis of FRNK-induced HSC was examined by annexin-V/propidium iodide double-labeled flow cytometry (FCM), gel electrophoresis and transmission electron microscopy. Levels of FRNK, FAK, p-FAK (Tyr397), ERK1 and p-ERK in HSC were assayed by Western blot on the protein level, and by RT-PCR on the mRNA level., Results: The expression of FRNK was enhanced after FRNK plasmids were transiently transfected into the HSC. The apoptotic rate of the HSC exposed to FRNK plasmids for 48 h was higher than that in the non-FRNK plasmid group (25.37%+/-1.92% vs 9.28%+/-1.05%, P less than 0.01), and was accompanied by a significantly higher activity of caspase-3 both in the protein and in the mRNA levels [(264.17+/-12.60 vs 185.82+/-9.69), P less than 0.01; (4.19+/-0.48 vs 1.07+/-0.27), P less than 0.01]. After exposure of HSC to FRNK plasmids, compared with the non-FRNK plasmid group, the expressions of p-FAK, ERK1 and p-ERK in protein and mRNA levels were lower; on the contrary, compared with the control group, the expressions of p-FAK, ERK1 and p-ERK in the FN group were higher., Conclusion: The expression of FRNK was enhanced and the phosphorylation of FAK was inhibited after FRNK was transiently transfected into HSC in vitro. FRNK induces apoptosis of HSC. FAK-ERK signal transduction pathway perhaps is involved in the process.

Objective: To investigate the effect of Changyanqing decoction, a traditional Chinese medicinal preparation, on the expressions of interleukin-10 (IL-10) and intercellular adhesion molecule-1 (ICAM-1) in the colon mucosa of rats with ulcerative colitis., Methods: The rats with ulcerative colitis induced by trinitrobenzene sulphonic acid and ethanol enema were randomly divided into 3 groups, namely the model group, sulfasalazine (SASP) group, and Changyanqing decoction group. Daily treatment with intragastric administration and enema of normal saline, SASP (100 mg/kg), and Changyanqing decoction (39.75 mg/kg), respectively, were administered 24 h after the establishment of colitis till the end of the experiment. Another group of rats was used as the normal control group. The disease activity index (DAI) and colon mucosa damage index (CMDI) of the rats were calculated. The activity of myeloperoxidase (MPO) was measured by biochemical method, and the expressions of IL-10 and ICAM-1 protein were measured by ELISA and immunohistochemistry, respectively., Results: Compared with the normal group, the model group showed significantly increased DAI, CMDI, HS score and MPO activity in the colon tissues (P < 0.01), with also significantly increased expression of ICAM-1 (P < 0.01) and decreased expression of IL-10 in the rat colon mucosa (P < 0.01). Treatment with Changyanqing decoction resulted in a significant reduction in DAI, CMDI, HS score and MPO activity (P < 0.01), and decreased the expression of ICAM-1 (P < 0.01) and increased the expression of IL-10 (P < 0.01) in the colon mucosa. The expression of ICAM-1 in the colon mucosa was positively correlated to that of IL-10 (r = 0.927, P < 0.01) and the activity of MPO (r = 0.621, P < 0.01)., Conclusions: Changyanqing decoction has protective effect against rat ulcerative colitis, mediated probably by enhancement of IL-10 expression and reduction in ICAM-1 expression and neutrophil infiltration.

Objectives: To investigate the effects of FAK-related non-kinase (FRNK) on expressions of type I collagen and matrix metalloproteinase-2 (MMP-2) mRNA and tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA in rat hepatic stellate cells (HSC)., Methods: Using in vitro cell culture technique, FRNK plasmids were transfected into HSC mediated by cationic liposome. Type I collagen synthesis capability in HSC was examined by 3H-Pro incorporation assay. The levels of FRNK in HSC were assayed by Western blot, and the expressions of MMP-2 and TIMP-2 were assayed by RT-PCR on mRNA levels., Results: The exposure of HSC to FRNK caused the expression of FRNK protein to be up-regulated, and the FRNK protein contents reached the highest point at 48 h after the transfection, P less than 0.05. The expressions of MMP-2 mRNA were up-regulated by FRNK; the expressions of TIMP-2 mRNA were down-regulated by FRNK; the ratios of MMP-2 mRNA/TIMP-2 mRNA were enhanced by FRNK., Conclusion: After FRNK was transfected, the capability of type I collagen synthesis in HSC was inhibited, which may be related to the up-regulation of MMP-2 mRNA/TIMP-2 mRNA.

Objective: To investigate the dynamic expression of PTEN in fibrogenic liver tissue of rats and its effect on the activation and proliferation of hepatic stellate cells (HSC)., Methods: A rat model of hepatic fibrosis was established by common bile duct ligation (BDL). The expressions of PTEN in the rat liver tissues were detected by immunohistochemical staining, Western blot and real-time PCR assay. The expressions of PTEN in activated HSC in the rat liver tissues were detected by immunofluorescence double labeling confocal laser scanning microscopy. The alpha-SMA in the rat liver tissues was determined by immunohistochemical staining., Results: The immunohistochemical staining indicated that there was extensive expression of PTEN in the liver tissues of normal rats, it was expressed mainly in the cytoplasm of the HSC. With the aggravation of hepatic fibrosis, the expression of PTEN in the hepatic tissues decreased gradually (P less than 0.01), while the alpha-SMA positive cells in the hepatic tissues increased significantly (P less than 0.01). The expressions of PTEN protein and mRNA in the rat liver tissues at week 1, 2, 3 and 4 after BDL were all lower than those in the sham operation group (P less than 0.01), and the expressions gradually decreased with the development of hepatic fibrosis (P less than 0.01). Immunofluorescence double labeling confocal laser scanning microscopy showed that PTEN were expressed extensively in activated HSC, especially in the cytoplasm, and with the development of hepatic fibrosis, the PTEN-expressing activated HSC accounted for an increasingly smaller percentage of total activated HSC., Conclusion: The expressions of PTEN mRNA and protein in rat fibrogenic liver tissues were downregulated, and their expressions in HSC in vivo also decreased. The dynamic expressions of PTEN in liver tissues had a significant negative correlation with the activation and proliferation of HSC.

Objective: To investigate the mechanism of Ca(2+)-independent pathways mediated by Rho-kinase in contraction of hepatic stellate cells (HSCs) induced by angiotonin II (Ang II)., Methods: Human HSCs of the line HSC-T6 were cultured and randomly divided into 6 groups: negative control group, Ang II group treated by Ang II10 micromol/L for 15 min, Ang II + irbesartan (Ang II receptor inhibitor) group, exposed to irbesartan for 60 min prior to Ang II treatment, Ang II + Y27632 (Rho kinase specific inhibitor) exposed to Y27632 for 60 min prior to Ang II treatment, Ang II + ML-7 (myosin light chain kinase specific inhibitor) + saturo (protein kinase C specific inhibitor) group exposed to stauro for 60 min prior to Ang II treatment, and Ang II + Y27632 + ML-7 + stauro group, exposed to Y27632 and stauro for 60 min prior to Ang II treatment. The cell contraction was detected by silicone-rubber-membrane cultivation directly. The protein levels of MLC and phosphorylated MLC were detected by Western blotting 5, 15, 30, 60, and 120 min after Ang II treatment. RT-PCR was used to detect the expression of Rock2, RhoAGTP, and RhoGEF in Ca(2+)-independent pathways mediated by Rho-kinase., Results: The silicone-rubber-membrane covered by Ang II treated HSCs showed obvious wrinkles indicating the contraction of HSCs. The ratios of phosphorylated MLC protein at the time pints 5, 15, 30, 60, and 120 min of the Ang II group to the control group (0 min) were 11.7 +/- 0.1, 26.9 +/- 0.1, 11.2 +/- 0.1, 4.1 +/- 0.1, and 1.0 +/- 0.1, showing that Ang II increased the phosphorylated MLC protein level time-dependently with the peak level at the time point of 15 minutes. The levels of phosphorylated MLC protein of the Ang II + irbesartan and Ang II + Y27632 groups were (1.12 +/- 0.09)and (1.22 +/- 0.10) respectively, both significantly lower than that of the Ang II group (1.33 +/- 0.06, both P < 0.01). The level of phosphorylated MLC protein of the Ang I + ML-7 + stauro group was (1.43 +/- 0.09), significantly higher than that of the Ang II + Y27632 group (0.64 +/- 0.04, P < 0. 01). The level of phosphorylated MLC protein of the Ang II + Y27632 + ML-7 + stauro group was (0.64 +/- 0.04), significantly lower than that of the Ang II group (P = 0. 003). The mRNA expression levels of Rock2, RhoAGTP, and RhoGEF of the Ang II group were (0.36 +/- 0.01), (0.80 +/- 0.01), and(0.65 +/- 0.11)respectively, all significantly higher than those of the control group [(0.12 +/- 0.01), (0.40 +/- 0.02), and (0.33 +/- 0.09) respectively, all P = 0. 000], and the mRNA expression levels of Rock2, RhoAGTP, and RhoGEF of the Ang II + irbesartan + group were (0.21 +/- 0.02), (0.62 +/- 0.02), and (0.41 +/- 0.10) respectively, all significantly lower than those of the control group. The mRNA expression levels of Rock2 (0.15 +/- 0.01) and RhoGEF (0.28 +/- 0.08) were lower, but The mRNA expression level of RhoAGTP (1.14 +/- 0.02) was higher in the Ang II + Y27632 group. The mRNA expression levels of Rock2, RhoAGTP, and RhoGEF of the Ang II + ML-7 + stauro group were (0.22 +/- 0.01), (0.55 +/- 0.03), and (0.44 +/- 0.10) respectively, all significantly higher than those of the control group. The mRNA expression levels of Rock2, RhoAGTP, and RhoGEF of the Ang II + Y27632 + ML-7 + stauro group were (0.23 +/- 0.01), (0.83 +/- 0.02), and (0.69 +/- 0.08) respectively, all significantly higher than those of the Ang II + ML-7 + stauro group., Conclusion: Ang II induces HSCs contraction in Ca(2+)-independent pathways mediated by Rho-kinase.

Objective: To investigate the mechanisms of angiotonin II (AngII)-induced Ca(2+)-independent pathways mediated by Rho kinase in hepatic stellate cells (HSCs)., Methods: HSC-T6 cells were treated with 1 micromol/L of AngII, and the subsequent cell contraction was directly observed with silicone rubber membrane culture method. The cells with 10 micromol/L AngII treatment were examined for myosin light chain (MLC) phosphorylation level using Western blotting, and the effects of irbesartan (a specific inhibitor of AngII 1- receptor) and Y27632 (a Rho kinase inhibitor) on AngII-induced MLC phosphorylation were evaluated. RT-PCR was used to detect the expression of Rock2 in Ca(2+)- independent pathways mediated by Rho kinase., Results: AngII induced HSC contraction and time-dependent MLC phosphorylation changes, which peaked 15 min after the treatment followed by gradual reduction. Irbesartan or Y27632 treatment significantly lowered MLC phosphorylation level in AngII-induced cells (P<0.01). The mRNA expression of Rock2 increased significantly after AngII treatment (P<0.01), but decreased following subsequent irbesartan or Y27632 treatment., Conclusion: AngII induces HSC contraction through Ca(2+)-independent pathways mediated by Rho kinase.

Objective: To investigate the signal transduction mechanism underlying the effects of angiotensin II (AngII) on extracellular signal-regulated kinase 1/2 (ERK1/2), early growth response-1 (EGR-1) and platelet-derived growth factor-B (PDGF-B) in hepatic stellate cells (HSCs)., Methods: HSC-T6 cells treated with AngII for 10 or 30 min were examined for phospho-P42/44 protein expression using Western blotting. In another experiment, the cells were preincubated for 1 h in the presence of U0126 (an inhibitor of the MAPK/ERK kinase), irbesartan (an AT-1 receptor blocker), or antioxidant-N-acetylcysteine (NAC) prior to AngII exposure, and the protein expression of phospho-P42/44 and PDGF-B were measured with Western blotting. The DNA binding activity of EGR-1 was analyzed using electrophoretic gel mobility shift assay (EMSA), and the expression of PDGF-B was detected immunohistochemically., Results: AngII induced phospho-P42/44 expression in HSC-T6, which was abrogated by U0126 or irbesartan. NAC did not inhibit phospho-P42/44 expression. EMSA showed that AngII exposure of the HSC cells markedly increased EGR-1 DNA binding activity, reaching the maximum after 60 min of exposure followed by progressive declination; irbesartan and U0126 significantly suppressed AngII-induced EGR-1 activity enhancement. ACEI at 1 micromol/L and 10 nmol/L inhibited EGR-1 activity, but ACEI at the concentration of 0.1 nmol/L resulted in enhanced EGR-1 activity. NAC showed no obvious effect in suppressing EGR-1 activity. AngII increased PDGF-B protein level in the HSCs, the effect of which was inhibited by irbesartan. U0126, NAC and ACEI did not attenuate PDGF-BB protein level in the HSCs., Conclusion: Stimulation of the HSCs with AngII results in EGR-1 activation via the ERK1/2 pathway, leading to up-regulation of PDGF-B expression.

For the odour from fermentation processing of municipal solid waste biological mechanical treatment, a compost biofilter was established to investigate its performance and the characteristics change of compost. The component and TVOC concentration in odors, removal efficiency of biofilter, and total carbon (TC), total nitrogen (TN), total phosphorus (TP), organic matter (OM), pH value and bacterial colony in compost media were measured respectively. SPME-GC-MS analysis showed that the main components of odors were BTEX, terpenes and alkane. The range of TVOC concentrations in odors during a fermentation process period and removal efficiencies of biofilter were 68.3 x 10(-6) -3.3 x 10(-6) (volume fraction), 31.5%-84.8% respectively. After about 4 months operation, TN, TC, TP and OM in compost were kept almost stably, but the dissolved N experienced an increase of 53.7%, and the dissolved P decreased 19.6%. The pH value experienced an increase in the former period and kept stable finally at about 7.8, which excursion was from 7.49 to 7.86. Analytical results for bacterial colony in packing material indicated that bacteria and mould colony counts increased, but yeasts and actinomyces decreased along with biofilter operation, which were 3.1, 3.4, 0.04, 0.07 times of their initial values respectively.

Objective: In order to investigate Hantavirus (HV) infection of captured rodents and to understand the genotypes and the molecular characteristic of Hantaviruses in Shenzhen., Methods: The captured rodents were classified and the density of distribution was calculated. A total of 472 animals were captured, among which Rattus norvegicus was the dominant group. The total viral RNA was extracted from the lung tissues positive with HV antigens by immunofluorescent assay and gene sequence of M fragment was amplified with RT-nested-PCR by using the Hantavirus genotype specific primers. The amplified genes were then sequenced, and subjected to genotyping and homology analysis., Results: The results of genotype analysis showed that the Hantaviruses taken from twenty-one lung specimens in Rattus norvegicus in Shenzhen city belonged to the Hantavirus type II (SEOV). Results in homology analysis suggested that the homology among twenty-one samples should be rather high with 95.4% of nucleotide sequence identity and they belonged to the same subtype. Phylogenetic tree analysis showed that they were branched into at least six different lineages, and were highly homologized with SZ2083. We also found that these virus strains had not shown more highly homology of nucleotide sequence in nearest district, whereas revealed consistency in farther district., Conclusion: The major hosts of Hantaviruses in Shenzhen city were Rattus norvegicus and the epidemic strains were genotyped as SEO-type. Nucleotide sequence and deduced amino acid sequence from different rodents were highly homologous, while nucleotide mutation had also been observed. Further studies are required to explore the possible viruses' sequence mutation.

Objective: Isolation and expansion tumor spheres from colorectal cancer cell line Colo205 cultured in serum-free medium(SFM) supplemented with human recombinant EGF and bFGF., Methods: Colo205 cells were cultivated in SFM,while cells cultivated in serum-supplemented medium(SSM) served as the control. Cells morphology were observed by optical microscope, and expression of intestinal stem cells marker Musashi-1 was detected by immunocytochemical. To induce cell differentiation, tumour spheres were cultivated without EGF and bFGF in the presence of 10% serum. Then we analysed expressions of stem cell surface markers CD133 and CD44 among undifferentiated cell, post-differentiated cells and routine Colo205 cells under serum-supplemented culture condition by flow cytometry. At last we compared cell cycle and spectral karyotype between two groups., Results: In SFM consisting of EGF and bFGF, a minority of Colo205 cells could survive, proliferate and form the suspended tumor spheres. We detected high Musashi-1 expression in these cells. Compared with the SSM group and the post-differentiation SFM group, the expressions of CD133 and CD44 were significantly increased in the undifferentiated SFM group (P<0.05). There was no statistical difference in the expression of CD133 and CD44 between the post-differentiation SFM group and the SSM group (P>0.05). Cell cycle analysis indicated that tumor spheres were of a high proliferation state.We could not find any noticeable difference in the number of chromatosomes between the SFM group and the SSM group., Conclusion: Tumor spheres in which enriched cancer stem cells can be generated under serum-free culture condition with EGF and bFGF.

Aim: To investigate the effect of IH764-3 on the expression of MMP-13 and TIMP-1 by H2O2-stimulated hepatic stellate cell and the alteration of FAK during this process., Methods: The expression of MMP-13 and FAK mRNA was examined by RT-PCR. TIMP-1 mRNA was analyzed by in-situ hybridization. FAK and TIMP-1 were evaluated at protein level through Western blotting method., Results: Being incubated for 2 h, compared with control group, MMP-13 mRNA was upregulated by IH764-3, but TIMP-1 transcription was reduced in a dose-dependent manner, accompanied with the decrease of FAK mRNA. The expression of TIMP-1 and FAK protein in HSC also decreased after being exposed by IH764-3 for 24 h., Conclusion: IH764-3 can induce the expression of MMP-13 and inhibit the expression of TIMP-1. Down-regulating the expression of FAK mRNA may be one of its mechanisms.

Objectives: To investigate the effects of norepinephrine (NE) on the proliferation and apoptosis of hepatic stellate cells (HSCs)., Methods: Cultured HSCs were used in 6 groups: (1) a control group; (2) a NE group; (3) a phentolamine plus propranolol group; (4) a phentolamine (an alpha-AR antagonist) group; (5) a CGP20712A (a beta1-AR antagonist) group; and (6) a ICI118551(a beta2-AR antagonist) group. After NE and the antagonists of adrenoceptor subtypes were administered to the cultured HSCs, MTT assay was used to evaluate the cell proliferation at 24 h, 48 h, and 72 h. Terminal deoxyribonucleotidyltransferase-mediated dUTP nick end labelling (TUNEL) assay and flow cytometry were used to detect cell apoptosis. An inverted microscope was used to observe the morphological changes of HSCs., Results: (1) MTT assay indicated that NE significantly induced HSCs proliferation in a time-dependent manner, which were reduced by antagonist of alpha-AR, beta1-AR and beta2-AR. (2) At 24 h after HSCs exposure to NE, apoptosis rates decreased significantly compared with that of the control group (6.60%+/-3.05% vs 12.60%+/-4.76%). In the antagonists of adrenoceptor subtypes groups, especially of a and beta2 adrenoceptor subtypes, the apoptosis was less. (3) Apoptosis rate of the NE group was significantly lower than that of the control group (2.29%+/-0.22% vs 3.06%+/-0.57%). In the antagonists of alpha and b2 adrenoceptor groups the apoptosis was less. (4) No obvious morphological changes of HSCs were found after administration of NE., Conclusions: Sympathetic neurotransmitter NE can induce proliferation and inhibit apoptosis of the cultured HSCs.

Aim: To study the effect of Arg-Gly-Asp-Ser (RGDS) tetrapeptide on the expression of matrix metalloproteinase-13 (MMP-13) and the tissue inhibitor of metalloproteinase-1(TIMP-1) in hepatic stellate cell(HSC) stimulated by fibronectin (FN)., Methods: The expression of MMP-13 was evaluated by RT-PCR. TIMP-1 was analyzed by in-situ hybridization at mRNA level and by Western blot at protein level., Results: Treated by RGDS tetrapeptide for 2 h, the mRNA expression of MMP-13 was upregulated but the expression of TIMP-1 mRNA and its protein in HSC was inhibited., Conclusion: The expression of MMP-13 can be induced by RGDS tetrapeptide and the expression of TIMP-1 can be inhibited by RGDS tetrapeptide, which is one of the mechanisms of its anti-fibrogenesis.

Objective: For clarifying the situation of the natural infection of rodents having hemorrhagic fever with renal syndrome (HFRS) virus and to type Hantavirus (HV) using molecular technique in Shenzhen city in 2005, and offering guidance for prevention and control of HFRS., Methods: Data on the host animals was collected from the city of Shenzhen. ELISA and indirect immunofluorscent antibody(IFA) test were applied to the specific antibodies against HV in the sera of captured rats. Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues of the HV infected rats were inoculated into Meriones unguiculata to isolate HV. The whole viral RNA was extracted from the lung tissues of the HV infected rats and amplified the partial M fragments with RT-nested-PCR, using the HV genotype specific primers. The amplified genes were then sequenced, and subjected to genotyping and homology analysis., Results: 472 rodents were captured from Shenzhen in 2005. Surveillance on rats demonstrated 9.96% rats carrying HV (with a density of 8.25%) and the main host was Rattus norvegicus. In the blood samples of rats, anti-HV IgG antibodies were detectable in 56 cases by IFA, and proved to be positive in 76 cases by ELISA. We successfully isolated a HV strain designated as SZ2083 from Rattus norvegicus for the first time in Shenzhen and was identified to SEO type by RT-nested-PCR. Compared with the coding region of the M gene of HV L99 virus strain, the homologies of nucleotide among them were 97%, but the homology was 76% of the SZ2083 with HTN 76-118 virus strain., Conclusion: Results showed the existence of natural epidemic areas of HFRS in Shenzhen city. Based on the results of sequencing, it is possible that the Seoul strain of HV might be the predominant serotype of virus harbored.

This paper aims at the reutilization and stabilization of municipal solid waste incineration (MSWI) fly ash to produce a high value added product. The effects of anionic chelating surfactants on the surface modification and fixing capacity of MSWI fly ash were primarily explored. Based on the indexes of active ratio and analysis of IR-spectrometer, the active ratio of ACS1 modified fly ash can be found higher than 95% under the condition of surfactant dosage at 7.0mL/100g fly ash, modification time at 15min and temperature at 75 degrees C . Moreover, Anionic chelating surfactant shows a strong fixing capacity for heavy metals by the United States Environmental Protection Agency's Toxicity Characteristic Leaching Procedure (TCLP).

To investigate the expression patterns of peroxisome proliferator activated receptor2 (PPARgamma2) gene in the differentiation of mouse embryonic stem (ES) cells into adipocytes, mouse ES cells were transfected with the vector of pPPARgamma2-promoter-luciferase, and PPARgamma2 expressions were analyzed by detecting luciferase activities and by detecting the protein expressions using western blotting. The results showed that the gene PPARgamma2 did not express in undifferentiated mouse ES cells and in embryoid bodies (EBs) within the first 2 days of differentiation induction after EB formation, and began to express from the third day of differentiation induction after EB formation to the finish of the differentiation. The gene's expression in differentiated adipocytes was much stronger than that in differentiating preadipocytes. In Conclusion our results reported for the first time the five-step expression patterns of the gene PPARgamma2 during the whole differentiation procedures from mouse ES cells into adipocytes via preadipocytes, and supported the previous studies that PPARgamma2 is a fat-specific gene that expresses only in developed and developing adipose tissues.

Aim: To observe the dynamic expression of ERK1 in fibrotic rat liver., Methods: The rat hepatic fibrosis was induced by bile duct ligation (BDL). Histopathological changes were evaluated by hematoxylin and eosin staining, and by Masson's trichrome method. ERK1 mRNA in liver was determined by reverse transcription-polymerase chain reaction (RT-PCR), while the distribution of ERK1 was assessed by immunohistochemistry. ERK1 protein was detected by using Western blotting analysis., Results: With the development of hepatic fibrosis, the positive cells of ERK1 increased a lot, they were mainly distributed at portal ducts, fiber septa and around the bile ducts, vascular endothelial cells and perisinusoidal cells. Western blotting analysis results displayed that the expression of ERK1 protein were up-regulated with model course, and its levels were the highest at week 4 after operation, achieving to 3.9-fold of that in normal rat liver. ERK1 mRNA expressed in normal rat livers as well, they were up-regulated at day 2 after BDL and its level was the highest at week 4 after BDL., Conclusion: These data suggest that the expression of ERK1 and its mRNA can be increase greatly in fibrotic rat liver.

Objectives: To investigate the effects of Arg-Gly-Asp-Ser (RGDS) tetrapeptide on integrin signaling and apoptosis in fibronectin (FN) -stimulated hepatic stellate cells (HSCs)., Methods: 3H-thymidine incorporation, annexin-V/propidium iodide double-labeled flow cytometry (FCM) and transmission electron microscopy were employed to estimate the influence of RGDS on the proliferation and apoptosis of HSCs. And the adhesion rates were observed by toluidine blue colorimetric assay. The expression of focal adhesion kinase (FAK) mRNA and protein in HSCs was detected using RT-PCR and western blotting analysis, respectively., Results: RGDS tetrapeptide at the concentrations of 25 microg/ml, 50 microg/ml and 100 microg/ml inhibited the proliferation of HSCs and induced HSCs apoptosis in dose-dependent and time-dependent manners, with the apoptotic rates of 9.49%, 27.67%, 31.59%, and the necrotic rates of 3.47%, 5.38%, 9.10%, respectively. Both the rates were higher than those in FN group (apoptotic rate: 3.44%; necrotic rate: 2.39%), F=8.02, P<0.05. After adding RGDS tetrapeptide to HSCs for 2 hours, the adhesive inhibition rates were 8.82%, 29.41% and 45.59%, respectively, which were higher than that in FN group (F=20.58, P<0.01). After exposure of HSCs to RGDS tetrapeptide for 24 hours, FAK protein decreased, and FAK mRNA was down-regulated earlier, about 2 hours after exposure to RGDS tetrapeptide., Conclusion: These results suggest that RGDS tetrapeptide may induce apoptosis of HSC in both dose-dependent and time-dependent manners in vitro, which may be related to the disruption of cell matrix adhesion and down-regulation of FAK expression.

Aim: To investigate the inhibiting role of Salvia miltiorrhiza monomer IH764-3 in proliferation and collagen synthesis and the effect on the expression of focal adhesion kinase in hepatic stellate cells (HSCs) stimulated by H2O2 in vitro, so as to provide evidences for preventing hepatic fibrosis., Methods: The effects of IH764-3 on the proliferation and collagen synthesis of HSCs were examined by 3H-TdR and 3H-proline incorporation assay, respectively. The expression of FAK mRNA was examined by RT-PCR in lysates of cultured HSCs., Results: The proliferation and collagen synthesis were inhibited in dose-dependent and time-dependent manners. The expressions of FAK mRNA in the IH764-3 (10 microg/ml, 20 microg/ml, 30 microg/ml, 40 microg/ml) groups were significantly lower than that in the control group., Conclusion: The present results demonstrate that IH764-3 can inhibit HSC proliferation and collagen synthesis. Meanwhile the mRNA transcription of FAK is reduced significantly, FAK as a signal molecule, can be involved in the effect of IH764-3. This may be one of the anti-fibrotic mechanisms of Salvia Miltiorrhiza.