Background Previous studies showed that after herpes simplex virus-1 (HSV-1) infection of the cornea,matrix metallo proteinase-2 (MMP-2) (produced by corneal cells and corneal epithelial cells) plays an important role in the development of HSK.Focal adhesion kinase (FAK)plays an important role on the expression,release and activation of MMPs.This study explored the expressions of MMP-2 and FAK,which induced by HSV-1 infected human corneal epithelial cells.Objective To investigate MMP-2 and FAK expression in HSV-1 infected human corneal epithelial cells.Methods Human corneal epithelial cells were infected with HSV-1 in vitro to establish cell model of viral infection.The expression of MMP-2 and FAK were detected by reverse transcription PCR (RT-PCR),Western blot,immunohistochemical method and immunofluorescence method at 2 hours,20 hours and 40 hours after infection.Results At 2 hours,20 hours and 40 hours after infection,the expressionis of MMP-2 mRNA and FAK mRNA were significantly increased in comparison with uninfected cells (MMP-2 mRNA:Ftime =0.968,P=0.436 ; Fgroup =47.649,P =0.000 ; Fi ion =0.757,P =0.536.FAK mRNA:Ftime =0.658,P =0.631 ; Fgroup =35.182,P=0.000;Finteraction =1.386,P=0.137).Western blot assays showed that there were no significant differences in p-FAK,FAK or MMP-2 expressions between infected cells and control cells after 2 hours (P＞0.05),but the expression levels of infected cells were significantly increased at 20 hours and 40 hours (both at P ＜ 0.05).Immunohistochemistry results showed that longer infection time was associated with an increased number of cells staining for MMP-2,FAK and p-FAK.Conclusions At the initial stage of HSV-1 infected,p-FAK plays an important role in the process of virus invading and MMP-2 activation.

Antiviral Agents ; pharmacology ; therapeutic use ; DNA, Viral ; blood ; Hepatitis B ; blood ; drug therapy ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Interferon-alpha ; pharmacology ; therapeutic use ; Kinetics ; Lamivudine ; pharmacology ; therapeutic use ; Models, Statistical ; Viral Load

To investigate the effect of recombinant human growth hormone (rhGH) on JAK2-STAT3 pathway and the growth of gastric cancer cell lines at different GHR expression status, the eukaryotic expression vector targeting human GHR (pGPU6/GFP/Neo-shGHR and pGPU6/GFP/Neo-scramble) was constructed and transfected into MGC803 cells by Lipofectamine 2000. Stable expressive cell lines were obtained by G418 screening. The expression of GHR was analyzed by Western blotting. After being stimulated with rhGH, cell growth was detected by MTT assay. Cell cycle and apoptosis were examined by flow cytometry. The components of JAK2/STAT3 signaling pathway were detected by Western blotting. There is no significant difference of GHR expression between MGC803 and pGPU6/GFP/Neo-scramble-transfected cells (named as MGC803-NC) (P > 0.05). Compared with MGC803, the GHR expression in pGPU6/GFP/Neo-shGHR-transfected cells (named as MGC803-shGHR) decreased significantly (protein decreased 50%). The cells were treated with rhGH at 0, 150 and 300 ng x mL(-1), the growth rate of MGC803 and MGC803-NC increased significantly, PI and the number of G2/M phase cells all increased significantly, and apoptosis decreased significantly. Western blotting revealed that the expression of pJAK2 and pSTAT3 was up-regulated after being treated with rhGH in MGC803 and MGC803-NC cells. In contrast, similar change was not observed in MGC803-shGHR cells. Knockdown of GHR gene may decrease the sensitivity of gastric cancer cells to rhGH, and down-regulating of components of the expression of JAK2/STAT3 signaling pathway may be the potential mechanisms.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Human Growth Hormone ; genetics ; pharmacology ; Humans ; Janus Kinase 2 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Somatotropin ; genetics ; metabolism ; Recombinant Proteins ; genetics ; pharmacology ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

OBJECTIVETo investigate whether WASP/Verprolin homologous protein 1 (WAVE1) plays a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).

METHODSWAVE1 mRNA and protein expression in bone marrow mononuclear cells (BMMCs) was measured by RT-PCR and Western blotting respectively in 4 children with ALL relapse, 15 children with ALL in complete remission (CR) and 40 children with newly diagnosed ALL. Ten normal bone marrow samples were used as controls. Jurkat cells were treated with different concentrations of adriamycin (ADM). The cell proliferation was detected with MTT. The apoptosis rate was measured by flow cytometry. WAVE1 mRNA and protein expression of Jurkat cells treated with ADM was detected by RT-PCR and Western blotting respectively.

RESULTSWAVE1 was not expressed or weakly expressed in BMMCs from normal controls and patients with ALL in CR. Higher WAVE1 mRNA and protein expression was found in BMMCs from patients with newly diagnosed ALL and patients with relapse ALL when compared with the controls and the patients in CR (P<0.01). ADM significantly inhibited the proliferation of the Jurkat cells and the inhibitory effect was dose-and time-dependent (P<0.05). After ADM treatment for 24 hrs, the percentage of apoptosis cells increased significantly and WAVE1 mRNA and protein expression of Jurkat cells decreased significantly when compared with the untreated controls (P<0.05).

CONCLUSIONSThe WAVE1 expression increased in children with ALL. WAVE1 may be related to the development of ALL and may be severed as a marker for the evaluation of the severity of ALL in children.

Adolescent ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Cell Proliferation ; drug effects ; Child ; Child, Preschool ; Doxorubicin ; pharmacology ; Female ; Humans ; Infant ; Jurkat Cells ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; etiology ; metabolism ; RNA, Messenger ; analysis ; Wiskott-Aldrich Syndrome Protein Family ; analysis ; genetics ; physiology

By studying the literatures on the adverse reaction of Yuxingcao injection, the clinical features of ADR were summed up, and the causes of ADR were analyzed through raw material, technology, chemical composition, compatibility and the clinical usage. The causes of ADR induced by Yuxingcao injection are complicated, maybe both due to the medicine itself and the incorrect clinical usage. Multi-measures including manufacture, techniques, clinical usage and some other cases should be taken to prevent the occurrence of adverse reaction of Yuxingcao injection.
Dosage Forms ; Drug Administration Schedule ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Humans ; Injections

OBJECTIVETo compare the sensitivity of Brown Norway rats (BN) with Guinea pigs (GP) as allergen assessment animal models.

METHODBN rats and GP were randomly assigned to 1 control group, 2 Bovine serum albumin group (BSA), respectively. Animals in BSA groups of BN rats and GPs were sensitized by intraperitoneal injection of 0.6% BSA 1 ml on day 1, 3, 5, respectively, and irritated by intravenous injection of 2.4% BSA 1 ml on day 7 and day 14 after the last sensitization, while the same volume of normal saline was given to control group on each time point mentioned above. The allergic reactions were scored within 1 h after each irritation treatment, and the sera of both BN rats and GPs were collected to detect IgE concentration by using ELISA. The sera were also applied for passive cutaneous anaphylaxis test (PCA test) in SD rats.

RESULTNo obvious allergic reactions were observed in BSA group of GPs after each irritation treat, however, the score of allergic response in BSA group of BN rats was evidently higher than that in control group after first irritation. PCA test by using sera from BSA group of BN rats after both irritations showed the strong positive result characterized as large amount of subcutaneous effusions of Evans blue in SD rats, however, the sera from BSA group of GP were negative in PCA test. Serum IgE concentration did not increase after each irritation in BSA group of both BN rats and GP.

CONCLUSIONBN rats were more sensitive than GPs on initiative systemic anaphylaxis test and passive cutaneous anaphylaxis test. Meanwhile, BN rats has an advantage in experimental treatment compared with Guinea pigs.

Allergens ; administration & dosage ; toxicity ; Anaphylaxis ; chemically induced ; Animals ; Guinea Pigs ; Hypersensitivity ; etiology ; Male ; Models, Animal ; Rats ; Rats, Inbred BN ; Rats, Sprague-Dawley

Physiotherapy Evidence Database (PEDro) is a free, web-based database containing randomized controlled trial reports, systematic reviews and evidence-based clinical practice guidelines for physical therapy, which has been searched millions of times a year by users around the world, and may improve the development of evidence-based physiotherapy in China.

measured by OCT 1 and 3mo after surgery. Chi-square test was used to compare preoperative and postoperative BCVA. Pearsion’s correlation was used to evaluate relationship between postoperative BCVA and central foveal thickness.RESULTS:The ratio of BCVA<0. 05 was 30%,14%, 11%and 7% respectively for per-operation, 1wk, 1 and 3mo post - operation. After surgery, the central foveal thickness was significantly increased in group with BCVA<0. 3 comparing to group with BCVA≥0. 3. Three month post-operation, central foveal thickness was significantly decreased in both groups comparing to that in 1mo post-operation (P<0. 01). There has significant correlation between 3mo postoperative BCVA and central foveal thickness (r=-0.716, P<0.05).CONCLUSlON: ln this study, BCVA is improved after 3mo follow up. There has significant correlation between postoperative BCVA and central foveal thickness.

ObjectiveTo obtain HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect by transfecting HSC-T6 cells with CRISPR/Cas9 lentiviral vector, and to provide a good method for further functional research and new strategies for the clinical treatment of liver fibrosis. MethodsThe COX-2 gene-specific sgRNAs (COX-2-sgRNA-1, COX-2-sgRNA-2, COX-2-sgRNA-3) were designed, synthesized, and connected to the GV371 vector, and the recombinant plasmid and the packaging plasmid were transfected into 293T cells to form lentivirus particles; the fluorescence method was used to measure virus titer. The most appropriate amount of the virus was calculated based on MOI. Lenti-Cas9-puro was transfected into HSC-T6 cells, and HSC-T6-Cas9 cells were screened out by puromycin; Lenti-COX-2-sgRNA-EGFP was transfected into HSC-T6-Cas9 cells to obtain HSC-T6-COX-2-/- cells. Cruiser enzyme digestion and Western blot were used to verify gene knockout at the gene and protein levels. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsSequencing verified that the COX-2-sgRNA expression vector was constructed successfully. Recombinant expression plasmids and packaging plasmids were transfected into 293T cells to form lentivirus particles, and the fluorescence method showed a virus titer of ＞1×108. HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect were successfully constructed. The HSC-T6-Cas9 group had significantly higher relative mRNA expression of LV-Cas9-Puro than the CON group (541.93±105.76 vs 1.00±0.02, t=12.995, P＜0.01). Cruiser enzyme digestion and Western blot showed that the CRISPR/Cas9 lentivirus expression vectors played a role in the target, among which COX-2-sgRNA-2 knockout had the most significant effect, and this group had a significant reduction in the protein expression level of COX-2 compared with the CON group and the NC group (both P＜0.05), suggesting that COX-2-sgRNA was active. ConclusionA CRISPR/Cas9 lentivirus vector is successfully constructed for COX-2 target gene, and HSC-T6-COX-2-/- cells with stable COX-2 gene knockout are obtained.

ObjectiveTo investigate the effect and significance of cyclooxygenase-2 (COX-2) inhibitors on the expression of the Acsl gene family in the ileum of rats with nonalcoholic fatty liver disease (NAFLD). MethodsA total of 45 Sprague-Dawley rats were randomly divided into normal control group (15 rats given normal diet), NAFLD model group (15 rats given high-fat diet), and nimesulide group (15 rats given high-fat diet and nimesulide). All rats were sacrificed after 12 weeks of feeding, and then blood samples were collected from the inferior vena cava to measure total cholesterol (TC) and triglyceride (TG). HE staining and oil red O staining were performed for the liver to evaluate the degree of hepatic steatosis in each group, and quantitative real-time PCR was used to measure the mRNA expression of the Acsl family genes in the ileum. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in serum TC and TG and marked hepatic steatosis (all P＜0.05); compared with the NAFLD model group, the nimesulide group had significant reductions in serum TC and TG and degree of hepatic steatosis (all P＜0.05). Compared with the normal control group, the NAFLD model group had a significant increase in the expression of COX-2 in the ileum (P＜0.05), and compared with the NAFLD model group, the nimesulide group had a significant reduction in the expression of COX-2 in the ileum (P＜005). Compared with the normal control group, the NAFLD model group had significant increases in the mRNA expression of Acsl3 and Acsl5 in the ileum (both P＜0.05), and compared with the NAFLD model group, the nimesulide group had significant reductions in the mRNA expression of Acsl3 and Acsl5 (both P＜0.05). ConclusionThe COX-2 inhibitor nimesulide can regulate the expression of the Acsl gene family in the ileum of rats with NAFLD, suggesting that COX-2 inhibitors may inhibit the progression of NAFLD through the Acsl gene.