[ABSTRACT]AIM:Toinvestigatetheroleofcysteine-rich61(Cyr61/CNN1)inproliferationandmigrationof bone marrow mesenchymal stem cells ( BMSCs ) .METHODS: The lentiviral vector carrying CCN 1 ( Lenti-GFP-CCN1 ) was constructed and then transfected into the rat BMSCs .The cells were divided into non-transfection group , transfection group ( transfected with Lenti-GFP-CCN1 ) and negative control group ( Lenti-GFP ) .The fluorescence intensity of the transfected BMSCs was observed under inverted fluorescence microscope .The effects of CCN1 on the proliferation and mi-gration of BMSCs were detected by MTT assay and scratch wound healing assay .RESULTS:The proliferation of BMSCs transfected with Lenti-GFP CCN1 had no significant difference compared with negative control group and control group .The width/thickness ratio of migrated BMSCs in wound healing was significantly higher in Lenti-GFP-CCN1 group than that in negative control group and control group (P<0.05).CONCLUSION:Exogenous CCN1 promotes the migration of BMSCs.

Objective To investigate the expressions of apoptosis associated protein 3 (APR3) and nuclear factor 3 of activated T-cell (NFAT3) in the tissue of epithelial ovarian tumors and its correlation with the clinicopathological features.Methods 92 patients with epithelial ovarian tumor were collected,23 cases with malignant tumor,24 cases with borderline tumor,45 cases with benign tumor.The expressions of APR3 and NFAT3 were detected by immunohistochemical methods,and the differences of different types of epithelial ovarian tumor were compared.The correlation of the expressions of APR3 and NFAT3 with the clinicopathological features of epithelial ovarian tumor was analyzed.The correlation of the expressions of APR3 with the expressions of NFAT3 in epithelial ovarian tumor was analyzed.Results The positive expression rate of APR3 in patients with malignant epithelial ovarian tumors (78.26％) was significantly higher than borderline tumors (41.67 ％) and benign tumors (22.22 ％),the differences were statistically significant (x2 =5.864,7.632,all P ＜ 0.05).The expression of APR3 in patients with malignant epithelial ovarian tumors was significantly correlated with differentiation,clinical stage,lymph node and abdominal organs metastasis and ascites (x2 =7.425,7.262,8.421,5.031,all P ＜ 0.05).The positive expression rate of NFAT3 in patients with malignant epithelial ovarian tumors (56.52％) was significantly higher than borderline tumors (29.17％) and benign tumors(17.78％),the differences were statistically significant (x2 =6.829,7.547,all P ＜0.05).The expression of NFAT3 in patients with malignant epithelial ovarian tumors was significantly correlated with differentiation,clinical stage,lymph node and abdominal organs metastasis (x2 =5.253,6.367,8.021,all P ＜ 0.05).The expressions of APR3 and NFAT3 in patients with malignant epithelial ovarian tumors were positively correlated (r =0.032,P ＜ 0.05).Conclusion The expressions of APR3 and NFAT3 in the tissue of malignant epithelial ovarian tumor obviously increase,are significantly correlated with differentiation,clinical stage,lymph node and abdominal organs metastasis and are positively correlated,and it may be correlated with the development and progression of malignant epithelial ovarian tumor.

Objective To study the relationship of MR delayed enhancement with cardiac troponin I in hypertrophic cardiomyopathy(HCM)and to evaluate their values on assessing HCM condition and prognosis.Methods Thirty-five HCM patients who were diagnosed by echocardiography were enrolled.All patients were performed MR scan and cTn Ⅰ test of blood.The relationships of MR delayed enhancement, myocardial hypertrophy and cTn Ⅰ were analyzed.Results(1)DE was found in 25 of total 35 HCM patients(71.4%).19 of 35 HCM patients(54.3%)had abnormal increased eTn Ⅰ value.The medians of cTn Ⅰ in patients with DE and without DE(110,5 ?g/ml,respectively)had statistics significance (P

OBJECTIVETo know the genotype and subtype of hantavirus (HV) which infected persons in Hebei province.

METHODSAccording to G2 coding region of 76-118 and R22 strains, specific type primers were designed to detect and identity the types of HV in HFRS patients' sera with RT-nested PCR. Nucleotides were assayed from partial products after purification and reclaim. Then, gene analysis was done with DNAStar package.

RESULTS17 out of 69 positive serum specimens were successfully detected by RT-PCR and the detection rate was 24.64%, among which, or= 14 days were 0. 17 positive specimens were all belonged to SEO. The nucleotide homology of 9 typical specimens was 92.0%-100%. Between HeB7 and other 8 specimens was 92%-95%, and they belonged to different subtypes. When HeB7 compared with R22 strain, it was 97.7%. HeB7 and R22 belonged to S1 subtype. The 8 specimens except HeB7 was 95.7%-100% and they all belonged to S3 subtype. When compared with 76-118 strain, 9 specimens' nucleotide homology was only 70.3%-72.7%, belonged to different type.

CONCLUSIONSEO was the major type of HV from HFRS patients in Hebei province, S3 was the major subtype and S1 was also existed. In a certain area, the HV which belonged to the same type was correspondingly conservative, and had the characteristic of regional stability.

China ; Genotype ; Hantavirus ; classification ; genetics ; Hemorrhagic Fever with Renal Syndrome ; diagnosis ; prevention & control ; therapy ; virology ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Envelope Proteins ; genetics

OBJECTIVETo evaluate the safety and efficacy of composite sophora colon-soluble capsule (CSCC) in treating ulcerative colitis (UC) of the damp-heat accumulation syndrome pattern (DHAS) and to prepare a basis for a phase III clinical trial.

METHODSA multi-center, randomized, single-blind, and positive drug parallel-controlled design was adopted. There were 126 patients of UC-DHAS stratified and assigned equally to three groups. Patients in two CSCC treated groups, Groups T1 and T2, were treated orally with high (six capsules, thrice a day) and low (four capsules, thrice a day) doses CSCC, and patients in the control group were treated orally with Mesalazine Enteric-coated Tablets (four tablets, thrice a day), respectively, all for eight weeks. The clinical efficacy and safety of treatments were evaluated through clinical symptom observations and colonoscopic examinations.

RESULTS(1) Full analysis set (FAS) and per-protocol set (PPS) analyses showed the comprehensive curative effect in Groups T1, T2, and the control group, obtaining the values of 85.7%, 92.9%, and 71.4% (P=0.330), and 89.5%, 92.7%, and 73.2% (P=0.552), respectively, demonstrating no statistical significance among the three groups. (2) FAS and PPS analysis showed the efficacy on membranous lesions in Groups T1, T2, and the control group, obtaining the values of 83.3%, 92.9%, and 73.8% (P=0.063), and 86.8%, 92.7%, and 75.6% (P=0.070), respectively, showing statistical insignificance among the three groups. (3) FAS analysis showed an efficacy tendency on improving tenesmus (P=0.056). No changes were found in improving the other symptoms, and statistical significance was not shown among the three groups (P>0.05). PPS analysis showed the efficacy on single item symptom in Groups T1, T2, and the control group was not statistically significant among the three groups (P=0.082).

CONCLUSIONSThe comprehensive effect of CSCC in treating UC is basically equivalent to that of Mesalazine enteric-coated tablet; however, the tendency was shown to improve symptoms. Its efficacy could not be raised by increasing the dosage used. Therefore, the recommended dosage of CSCC is four capsules, three times a day.

Adult ; Capsules ; Colitis, Ulcerative ; drug therapy ; Colon ; drug effects ; pathology ; Female ; Humans ; Intestinal Mucosa ; drug effects ; pathology ; Male ; Medicine, Chinese Traditional ; Plant Extracts ; adverse effects ; pharmacology ; therapeutic use ; Single-Blind Method ; Sophora ; chemistry ; Treatment Outcome

BACKGROUNDControl of hypersecretion of certain hormones is one of the key targets in the treatment of pituitary adenomas. RNA interference has been shown to inhibit protein expression, and thus it may represent a promising method for the treatment of pituitary adenomas. In the present study, transfection efficiency of small interfering RNA (siRNA) was optimized in human prolactinoma cells.

METHODSFirst, a method was optimized to extract highly purified human prolactinoma cells in vitro. The extracted cells were verified to retain the physiological features of prolactin (PRL) secretion. Second, three conditions for siRNA transfection were tested by the evaluation of transfection efficiency and cell viability. The proper transfection condition was verified for human prolactinoma cells. Third, the siRNA for prolactin was transfected into the human prolactinoma cells, and the suppression of PRL mRNA was evaluated by quantitative real-time reverse transcription-PCR.

RESULTSThe siRNA of 100 pmol with Lipofectamine 2000 of 5 µl for 1 × 10(6) cells was proved preferable, with transfection efficiency being 53.3% and cell viability being 69.7%. In the preliminary experiment the siRNA against PRL decreased the mRNA of PRL by 34.0%.

CONCLUSIONIt is possible to inhibit hormone hypersecretion by RNA interference, that may eventually enable therapeutic siRNA drugs developed.

Adolescent ; Adult ; Cell Line, Tumor ; Cell Separation ; Female ; Humans ; Male ; Middle Aged ; Pituitary Neoplasms ; pathology ; therapy ; Prolactinoma ; pathology ; therapy ; RNA, Small Interfering ; genetics ; Transfection

Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

Objective To know the genotype and subtype of hantavims (HV) carried by host animals in HFRS natural epidemic area of Hebei province. Methods According to the conservative sequence of 76-118 and R22 strains, specific primers were designed. RT-nested PCR was used to amplify partial M segments from the positive rat lungs screened by IFA. Agarose gel electrophoresis was used to identify the types. Nucleotides were assayed from partial products after purification and reclaim. Gene analysis was carried on with DNAStar package.Results 32 specimens, which were positive screened by IFA, were amplified the specific segment (418bp) and all belonged to type SEO. Sequencing results of 10 partial segments indicated that G2 segment had little variability and nucleotide homology reached to 98.0%-100.0%. Comparing with the R22 and 76-118 strains, homology was 93.3%-94.3% and 67.7%-69.0% respectively. Conclusion According to G2 segment, SEO was the major type in Hebei HFRS natural epidemic area and S3 was the major subtype. HV which belonged to the same subtype had high homology and genetic materials were correspondingly stable. Different rats could carry the same subtype of HV.

OBJECTIVETo explore the factors affecting the long-term survival of patients with curative resection of gastric cardia cancer.

METHODSThe data of 108 patients who underwent radical resection of gastric cardia cancer from Jul. 1994 to Dec. 2003 in our hospital were investigated retrospectively. The Kaplan-Meier method and long-rank test were used for bivariate comparisons of survival. Multivariate analysis was done by the Cox regression model (Backward Wald).

RESULTSSurvival status of the 108 patients was ascertained in Dec. 2004. Among them, 68 were Siewert type II and 40 were Siewert type III. Seventy-four patients had lymph node metastases (68.5%). The mean follow-up time was 37 months (95% CI: 29.3-44.7 months) and the middle follow-up time was 26.6 months (95% CI: 25.8-34.2 months). The 1-,3- and 5-year cumulative survival rates were 77.2%, 33.6% and 21.8%, respectively. According to the Kaplan-Meier and log-rank methods, splenectomy, lesion size, depth of tumor invasion and regional lymph node status were prognostic factors. Multivariate regression analysis indicated that only depth of tumor invasion (P=0.009) and lymph node metastases (P=0.001) were independent prognostic factors.

CONCLUSIONDepth of tumor invasion and lymph node metastases have negative effects on the survival of patients with gastric cardia cancer undergoing curative resection. Splenectomy may only be appropriate for patients with direct tumor invasion to the spleen and the extent of gastric resection does not influence survival in patients with curative gastric cardia cancer.

Adult ; Aged ; Aged, 80 and over ; Cardia ; pathology ; Female ; Follow-Up Studies ; Gastrectomy ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Staging ; Prognosis ; Regression Analysis ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery

OBJECTIVETo study the mechanism of electrophysiologic changes caused by different type of sodium salicylate injection.

METHODSDecapitated three group rats ( acute injected, chronic injected and normal rats ) separately, dissected the temporal bones to collect cochlea, and the otic capsules were removed. Then the cochlear materials from each groups were pooled and homogenized respectively, extracted the total RNA, obtained cDNA from purified total RNA by reversed transcription, cDNA were transcripted to cRNA probes in vitro. Hybridized the cRNA probes with tester chip to evaluate the quality of probes, if good, hybridized the probes with real chip. Obtained three gene expression profiles of different groups of cochlea Analyzed the differentially expressed genes among three groups by SOM. Analogized the SOM result to electrophysiologic changes. Then analyzed the genes in clusters of analog results by Gene Ontology. Then the genes in clusters of analog results were analyzed by Gene Ontology. Hsp27 was chosen to validate the result of gene chip using real time quantitative reverse transcription PCR ( RTQ RT-PCR).

RESULTSThe probes was good, and the chip hybridization results was credible. We obtained 6 clusters genes by SOM analysis, in which we choose cluster 3 and cluster 4 as candidate cluster. There were 46 genes in cluster 3 and 30 genes in cluster 4 employing GO analysis, which involved in cell communication, cell motility, metabolism, immune response and nerve ensheathment, et al. The result of RTQ RT-PCR showed high concordance with that of gene chip.

CONCLUSIONIt's a new method to study the mechanism of electrophysiologic changes caused by sodium salicylate by gene chip and SOM analysis.

Animals ; Cochlea ; drug effects ; metabolism ; Gene Expression Profiling ; Injections ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Sodium Salicylate ; administration & dosage ; pharmacology