Methyl 3, 4-dihydroxyphenylacetate prevents enterovirus 71 proliferation in rhabdomyosarcoma cells / 药学学报

This article presented the inhibitory activity of methyl 3, 4-dihydroxyphenylacetate on the enterovirus 71 (EV71) infection. The EV71 VP1 capsid protein expression levels were analyzed with Western blotting. Results revealed that the compound is able to inhibit EV71 replication in rhabdomyosarcoma (RD) cells. After being incubated with the compound at a concentration of 0.01 microg x microL(-1) for 48 h, the level of EV71 vp1 mRNA in RD cells decreased by (76.83 +/- 2.47)%. The cytotoxic activity of the compound was evaluated against RD cells by a MTT assay. The results showed that the compound had low toxicity with a CC50 of 0.072 6 microg x microL(-1). These findings suggest that methyl 3, 4-dihydroxyphenylacetate is a novel compound for antiviral therapies against EV71, which merited further investigation.

Effects of multiple interference on borderline hypertension with metabolic syndrome / 中国医师进修杂志

ObjectiveTo investigate the effects of different therapeutic methods on borderline hypertension with metabolic syndrome patients. MethodsNinety borderline hypertension with metabolic syndrome patients were divided into three groups by random digits table with 30 cases: control group,conventional therapy group and intensive therapy group. The control group was given regular observation, the conventional therapy group took drug according to the disease situation; and the intensive therapy group not only formulated the aim of therapy, but also received diet control, sport therapy, healthy education and drug therapy. After 1 year's follow-up, the patients' changes were compared. ResultsAfter 1 year's follow-up,the levels of FPG, 2 h PG, 24 h mAlb and IMT were significantly increased(P ＜ 0.05 ), and the levels of other index had no significant changes (P＞ 0.05) in control group. The levels of FPG,2 h PG,TC and TG were significantly decreased and IMT was significantly increased(P ＜0.05), the levels of other index had no significant changes(P ＞ 0.05 ) in conventional therapy group. The levels of SBP, DBP, PP, FPG, 2 h PG, TC,TG,hs-CRP,24 h mAlb and HOMA-IR were significantly decreased and HDL-C, ABI were significantly increased (P ＜ 0.01 or ＜ 0.05 ) in intensive therapy group. After treatment, the levels of ABI and H DL-C were significantly higher and SBP, DBP, PP,TG, hs-CRP, 24 h mAlb, HOMA-IR, IMT were significantly lower in intensive therapy group than those in conventional therapy group (P ＜ 0.01 or ＜ 0.05 ). ConclusionsDrug therapy is efficient in borderline hypertension with metabolic syndrome patients, and intensive therapy can obviously improve the insulin resistance, to control the developing of hypertension can delay the vascular

Effect of the compound of poly lactic-co-glycolic acid and bone marrow stromal cells modified by osteoprotegerin gene on the periodontal regeneration in Beagle dog periodontal defects / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of the osteoprotegerin (OPG) gene-modified autologous bone marrow stromal cells (BMSCs) on regeneration of periodontal defects, and to provide new experimental evidence to explore the gene therapy for periodontal disease.</p><p><b>METHODS</b>pSecTag2/B-opg was transduced into BMSCs by lipofectamine 2000. The expression of OPG protein in the BMSCs was detected by immunocytochemistry and Western blot. Inverted phase contrast microscope and scanning electron microscopy (SEM) were used to observe the morphology and proliferation of the BMSCs(OPG) on on the surface of the poly lactic-co-glycolic (PLGA). Horizontal alveolar bone defect (4 mmx4 mmx 3 mm) were surgically created in the buccal aspect of the mandibular premolar, and were randomly assigned to receive BMSCs(OPG)-PLGA (cells/material/OPG), BMSCs-PLGA (cells/material), PLGA (material), or root planning only (blank control). The animals were euthanized at 6 weeks post surgery for histological analysis. The height of new alveolar bone and cementum and the formation of new connective tissue were analyzed and compared. All data were statistically analyzed using the q test.</p><p><b>RESULTS</b>The BMSCs transfected by human OPG gene can highly express OPG protein. SEM observations demonstrated that BMSCs(OPG) were able to proliferate and massively colonize on the scaffolds structure. After 6 weeks, the height of new alveolar bone and cementum and the formation of new connective tissue were significantly greater in the experimental group than in the control groups (P < 0.05).</p><p><b>CONCLUSION</b>BMSCs(OPG)-PLGA can significantly promote the regeneration of dog's periodontal bone defects. Gene therapy utilizing OPG may offer the potential for periodontal tissue engineering applications.</p>

Establishment and identification of transiently expression system of bone marrow stromal cells modified by osteoprotegerin gene / 华西口腔医学杂志

<p><b>OBJECTIVE</b>In oder to treat periodontitis by using tissue engineering and gene engineering technology, the article established an transient expression system of bone marrow stromal cells (BMSC) modified by osteoprotegerin (OPG) gene and detected its expression using eukaryotic secreted expression pSecTag2/B-OPG plasmid.</p><p><b>METHODS</b>By solation and culture of BMSC in vitro, the identified recombined plasmid was transiently transfected into BMSC by Lipofectamine 2000 and OPG expression in BMSC was determined by RT-PCR and Western blot in 6 weeks.</p><p><b>RESULTS</b>The fragments of the recombinant plasmid digested with Hind III, EcoR I and BamH I and examined by 10 g/L agarose electrophoresis, were consistent with predicted size. The sequence of OPG was identical to the sequence provided by GeneBank [gi:33878056]. OPG transcribing in BMSC was confirmed by RT-PCR and OPG sustainable expressing in BMSC was detected by Western blot in 39 days.</p><p><b>CONCLUSION</b>The transiently expression system of BMSC modified by OPG gene was successfully established.</p>

Role of caveolin-1 down-regulation by iRNA in human hepatocyte proliferation / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of caveolin-1 down-regulation in human hepatocyte proliferation in vitro.</p><p><b>METHODS</b>The expression vector psiRNA-CAV1 was constructed and transfected into Chang liver cells (CHL). The caveolin-1 down-regulated cell clones were selected by the antibiotic zeocin. The proliferation of the cell strain CAV7 was examined by MTT, in which untransfected CHL and HepG2 cells were set as controls. Expression of caveolin-1, Akt, Erk1/2, p-Akt and p-Erk1/2 in the transfected and control cells was detected by Western blot.</p><p><b>RESULTS</b>After caveolin-1 expression was down-regulated by RNAi, CHL increased faster at first (24 h and 72 h, P<0.05; 96 h, P<0.01), but slower later. P-Akt and p-Erk1/2 expressions were down-regulated, indicating that the growth and proliferation related Akt and Erk1/2 pathways were inhibited after caveolin-1 down-regulation.</p><p><b>CONCLUSION</b>Caveolin-1 may play an important role in hepatocyte proliferation.</p>

Immunological characteristics of primary biliary cirrhosis patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the clinical significance of the immunological characteristics in patients with primary biliary cirrhosis (PBC).</p><p><b>METHODS</b>3000 patients with abnormal liver functions were examined for anti-nuclear antibodies (ANA), anti-mitochondrial antibodies (AMA), anti-smooth muscle antibodies (SMA) and anti-liver kidney microsomal antibody (LKM) using immunofluorescent assays (IFA). LKM-1, liver cytosolic-1 (LC-1), soluble liver antigen (SLA)/liver- pancreas antigen (LP) and subtype of AMA (M2, M4, M9) as well as ANA profile were detected by an immune blotting assay and an enzyme-linked immune absorbent assay (ELISA), respectively. Cytokines were tested by flow cytometry.</p><p><b>RESULTS</b>Of the 3000 patients with liver diseases, 52 (1.7%) were diagnosed with PBC. All the PBC cases were positive for AMA and M2. 94% of them showed high titer of AMA (> or = 1:320), and in 79% of them M2 was >200 RU/L, and 78% of them were ANA positive. Three main fluorescent patterns of ANA seen were nuclear membrane, nuclear dots and centromere patterns. Sjogren's Syndrome A/B (SS-A/B), homogenous, nucleolar or nuclear granular patterns were seen in only a few patients. IgM, ALP and GGT in PBC patients were significantly higher than those in hepatitis B related liver cirrhosis patients. The levels of IL-6, IL-10, TNF-alpha and IFN-gamma in PBC patients were higher than in the normal controls. Among the 52 PBC patients, 5 had autoimmune liver disease overlap syndromes. Two of them were SLA/LP positive, indicated as AIH type III and PBC overlapping, and 1 was LKM-1 positive showing AIH type II overlapping PBC, and 2 had ANA positive and were identified as AIH and PBC by liver biopsy.</p><p><b>CONCLUSION</b>The percentage of PBC in Chinese liver disease patients is about 1% to 2%. Most of the PBC patients have high levels of AMA and AMA-M2, IgM, ALP, GGT and several cytokines, indicating that abnormality of humeral and cellular immunity may be associated with the pathogenesis of PBC.</p>

RANTES gene single nucleotide polymorphisms and expression in patients with chronic hepatitis B virus infection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Regulated on activation, normal T-cell expressed and secreted (RANTES) plays a critical role in T-lymphocyte activation and proliferation. The process is involved in both acute and chronic phases of inflammation. The present study was to ascertain the possible correlations between chronic hepatitis B virus (HBV) infection and the RANTES gene polymorphisms and their expression.</p><p><b>METHODS</b>The study included 130 HBV negative healthy donors and 152 patients with chronic hepatitis B (CHB) virus infection. The polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLPs) were used to detect RANTES gene single nucleotide polymorphisms (SNPs). RANTES levels in the platelet depleted plasma were detected by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>RANTES alleles -403G, -28C and In1.1T were the predominant alleles in the subjects studied. No significant correlation was found between CHB infection and the RANTES alleles, while a significant correlation was found between CHB infection and increased RANTES expression in platelet depleted plasma (P < 0.05).</p><p><b>CONCLUSIONS</b>SNPs in RANTES gene do not affect chronic HBV infection or the outcome of interferon-alpha treatment in patients positive for HBV "e" antigen (HBeAg+). However, patients with CHB infection express the higher levels of plasma RANTES, which is thus associated with CHB infection.</p>

Effects of octreotide on expression of L-type voltage-operated calcium channels and on intracellular Ca2+ in activated hepatic stellate cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The contractility of hepatic stellate cells (HSCs) may play an important role in the pathogenesis of cirrhosis with portal hypertension. The aim of this study was to research the effects of octreotide, an analogue of somatostatin, on intracellular Ca2+ and on the expression of L-type voltage-operated calcium channels (L-VOCCs) in activated HSCs, and to try to survey the use of octreotide in treatment and prevention of cirrhosis with portal hypertension complications.</p><p><b>METHODS</b>HSC-T6, an activated HSCs line, was plated on small glass coverslips in 35-mm culture dishes at a density of 1 x 10(5)/ml, and incubated in DMEM media for 24 hours. After the cells were loaded with Fluo-3/AM, intracellular Ca2+ was measured by Laser Scanning Confocal Microscopy (LSCM). The dynamic changes in activated HSCs of intracellular Ca2+, stimulated by octreotide, endothelin-1, and KCl, respectively, were also determined by LSCM. Each experiment was repeated six times. L-VOCC expression in HSCs was estimated by immunocytochemistry.</p><p><b>RESULTS</b>After octreotide stimulation, a significant decrease in the intracellular Ca2+ of activated HSCs was observed. However, octreotide did not inhibit the increases in intracellular Ca2+ after stimulation by KCl and endothelin-1. Moreover, octreotide did not significantly affect L-VOCC expression. These results suggest that neither L-VOCC nor endothelin-1 receptors in activated HSCs are inhibited by octreotide.</p><p><b>CONCLUSIONS</b>Octreotide may decrease portal hypertension and intrahepatic vascular tension by inhibiting activated HSCs contractility through decreases in intracellular Ca2+. The somatostatin receptors in activated HSCs may be inhibited by octreotide.</p>

Clinical features of severe acute respiratory syndrome in forty-one confirmed health care workers / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the clinical features of severe acute respiratory syndrome (SARS).</p><p><b>METHODS</b>Forty-one medical care workers (aged 23 - 55 years, with a average of 32 years; men/women = 8/32) who were admitted to our hospital and diagnosed with SARS during March and April, 2003 were retrospectively analyzed.</p><p><b>RESULTS</b>Thirteen of all the patients were physicians and the rest were nurses. The disease was mainly transmitted through air droplet in a short distance, and overwork induced tiredness was involved in disease stimulation. Seventy-three percent of the patients presented fever as their first symptom. Ten patients complained inertia and myalgia. One patient showed no clinical symptoms, and bilateral infiltrates was found in his chest X-ray. Among the 41 cases, 6 (15%) were diagnosed as severe type. At the first week, the counts of white blood cells (WBCs), lymphocyte and platelets were (4.4 +/- 1.5) x 10(9)/L, 0.22 +/- 0.12 and (143 +/- 37) x 10(9)/L, which were significantly lower when compared with those at the 2nd to 4th week. Abnormal liver function was found in 27 cases (mostly with elevated serum ALT), with 70% occurred at the 3rd or 4th week. In terms of CT, 30 patients (73%) showed pathological changes in lungs, and bilateral lung involvement was found in 35.59%. Of 36 cases treated with steroids, 86% received middle or low dosage (80 - 240 mg/d). Artificial ventilation was used for twenty-seven patients, and air pipe mechanical ventilation was used for 1 case. Mortality in this study was 5%.</p><p><b>CONCLUSIONS</b>Inertia and myalgia may be the earlier symptoms of health care workers with SARS include, which are parallel to CT manifestations. There is no objective index for the assessment of the severity of the disease at early stage. The medicine associated toxicities may be the main reason of liver lesions. damages. Middle or low dosage of steroid was reasonable to be used as early as possible.</p>

Relationships between periapical lesion and IL-1, TNF-alpha gene expression in rat / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To understand the effects of the IL-1, TNF-alpha cytokines on the pathogenesis of periapical lesions by investigating its gene expression in rat.</p><p><b>METHODS</b>The model was established by surgically exposing mandibular molar teeth and left open to permit infection from the oral environment. The SD rats were killed at 1, 2, 3, 4 weeks and mandibular molar teeth X-ray were taken. In situ hybridization was used to test IL-1alpha, IL-1beta, TNF-alpha mRNA expression in periapical area and the kinds of positive cells were identified. Using image analysis system analyzed gene expression semi-quantified.</p><p><b>RESULTS</b>IL-1alpha, IL-1beta and TNF-alpha mRNA were all expressed beginning at 1 week, peaked at 3 weeks, and declined somewhat at 4 weeks, but IL-1beta mRNA was expressed at much lower levels with the same kinetics (P<0.01). Most of the staining occurred in areas that had heavy inflammatory infiltrate, fibroblasts, or endothelial cells. There was a statistically significant correlations between the area of periapical lesion and the number of positively stained cells for IL-1alpha and for TNF-alpha (IL-1alpha: r=0.875, P<0.001; TNF-alpha: r=0.858, P<0.001), and between the number of positive cells for IL-1alpha and that of for TNF-alpha (r=0.969, P<0.001).</p><p><b>CONCLUSIONS</b>IL-1alpha and TNF-alpha genes are highly expressed in developing periapical lesions in rat, which supports the hypothesis that these two cytokines play a key role in pulpal and periapical pathogenesis, including the concomitant bone destruction.</p>

Detection of SARS-CoV RNA in stool samples of SARS patients by nest RT-PCR and its clinical value / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate of severe acute respiratory syndrome (SARS) convalescent stool shedding by RT-PCR.</p><p><b>METHODS</b>One hundred and three stool samples from 46 SARS patients were collected on May 16th, 20th, and 23rd, 2003. For each sample, RNA was extracted using commercial kit and 7 Nest RT-PCR using a 14-pair different SARS-associated coronavirus (SARS-CoV) special primers were carried out simultaneously.</p><p><b>RESULTS</b>Among these 46 SARS patients, 17 cases (37.0%) were stool SARS-CoV RT-PCR negative, and 29 cases (63.0%) were SARS-CoV RT-PCR positive. The duration of positive cases lasted (31.76 +/- 10.78) d (12-64 d). The longest stool shedding case in this study lasted 64 days. Two serial stool samples and for each sample 2 RT-PCR tests using different primers were positive in this case.</p><p><b>CONCLUSIONS</b>Our study observed longest stool shedding of SARS patients to be 64 days after initial onset of SARS. The average stool shedding was 32 days. Hence it is important to think highly of SARS convalescent patient stool sterilization.</p>