Segmented cloning and expression of the pigment epithelium derived factor and its effect on the proliferation of human SCL-1 cells / 医学研究生学报

Objective Studies are rarely reported on the effect of short peptides of the pigment epithelium derived factor (PEDF) on the proliferation of human cutaneous squamous (SCL-1) cells. The purpose of this study is to investigate segmented cloning and expression of the PEDF protein and observe its effect on the proliferation of human SCL-1 cells. Methods The target genes of PEDF1, PEDF2 and PEDF3 were amplified by PCR and the recovered fragments subjected to double digestion of NheⅠ and Hind Ⅲ and inserted into the pET28a(+) plasmid. The product was transformed into human E coli BL21 and induced to express, followed by isolation and purification of the fusion protein. CCK-8 assay was used to detect the proliferation of the SCL-1 cells with PEDF1, PEDF2 and PEDF3 at 100, 400, 800 and 1000 nmol/L at 24, 48 and 72 hours. Results The prokaryotic expression vectors of PEDF1, PEDF2 and PEDF3 were successfully constructed, and their fusion proteins prepared, with the molecular weight of 18 000, 17 000 and 13 000, respectively. The proliferation of the SCL-1 cells was significantly decreased in the 800 and 1000 nmol/L PEDF3 groups compared with that in the 0 nmol/L PEDF3 group at 24 hours (0.16 ± 0.03 and 0.78 ± 0.07 vs 1.00 ± 0.00, P < 0.05), inhibited in a concentration-dependent manner in the 400, 800 and 1000 nmol/L PEDF3 groups at 48 hours (P < 0.05), markedly lower in the 800 and 1000 nmol/L PEDF3 groups at 72 hours (0.53 ± 0.05 and 0.51 ± 0.05) than in the in the 400 and 0 nmol/L PEDF3 groups (0.60 ± 0.05 and 1.00 ± 0.00) (P < 0.05). Conclusion The PEDF fusion proteins were successfully segmentally cloned and expressed and PEDF3 inhibited the proliferation of SCL-1 cells, which has paved the ground for further screening of active functional short peptides of PEDF.

Implementation effect analysis of Global Fund for Tuberculosis Control Project on designated hospital in Wenzhou City of Zhejiang Province / 上海预防医学

Objective] To analyze the implementation effect of the Global Fund for tuberculosis ( TB) control project on designated hospital , and provide basis and recommendation for TB control in Wenzhou City of Zhejiang Province . [ Methods] According to the data from the National TB Manage-ment Information System,quarterly reports of project were collected and analyzed . [Results] The X-ray check rate and sputum test rate of newly diagnosed patients both reached more than 97%.The registration rate of active pulmonary TB and smear-positive TB were 71.45 per 100 000 and 26.41 per 100 000.A total of 1 393 new smear-positive pulmonary TB cases were detected .The completion of case finding target for new smear-positive pulmonary TB was 119.67%.The cure rate and treatment success rate for new smear-positive cases were 88.08%and 90.52%.And 1 657 cases have accepted psychological support .The psychological support rates were 36 .05%.A total of 39 361 person times for TB cases received free liver function test , and the test rate was 100%. [ Conclusion ] This project was implemented well in Wenzhou , and obtained good achievement .The discovery of new smear-positive patients , treatment outcome , psychological support and free liver function test for TB patients have reached the project requirements .

Gastrointestinal stromal tumor with synchronous isolated parenchymal splenic metastasis of ovarian cancer / 中华医学杂志(英文版)

Gastrointestinal stromal tumor (GIST) represents the most common intramural mesenchymal tumor of the gastrointestinal tract, but the synchronous occurrence of GIST in the stomach and gynecological cancer is rare. We present a unique case of a 56-year-old female patient who was diagnosed with the synchronous development of GIST and an isolated parenchymal splenic metastasis of ovarian cancer. She underwent a wide local excision of gastric lesions with splenectomy. A morphological (histological and immunohistochemical) study established a spindle-cell type of gastrointestinal tumor that expressed CD117, and a parenchymal recurrence of ovarian papillary serous adenocarcinoma. The patient has remained alive and disease-free for 30 months since the last operation. A small GIST concomitant with an isolated parenchymal splenic metastasis of ovarian cancer is rarely encountered. The coexistence of GIST with other malignancies constitutes an intriguing oncologic model. Surgeons are advised to be alert against possible primary GIST accompanying other neoplasms.

Prognostic factors in adult granulosa cell tumor of the ovary

To determine the clinicopathologic prognostic factors in adult granulosa cell tumors [GCTs] of the ovary. This retrospective study was carried out over a period of 10 years [1995-2005] in the Gynecology Department of Shengjing Hospital, China Medical University, Shenyang, China. Forty-six patients with GCT were enrolled in this study. Demographic data, pathologic findings, treatments, and survival time were reviewed and analyzed for prognostic significance. It was found that International Federation of Gynecology and Obstetrics [FIGO] stage [p=0.0003], presence of nuclear atypia [p=0.036], and increased mitoses [p=0.002] were the 3 factors that impacted significantly on survival. Age, residual tumor disease, parity, and size of the tumor had no significant effect on survival. The only factor associated with risk of recurrence was rupture of the tumor [p=0.038]. Patients who received chemotherapy had a better median disease-free survival than those who did not [105 versus 78 months], however, this did not reach statistical significance [p=0.080]. The FIGO stage, nuclear atypia, and increased mitoses are the statistically significant prognostic factors, and may be used for selecting patients for adjuvant therapy. A prolonged follow-up is necessary due to risk of recurrences, late, and exceptional for the adult ovarian GCT, especially when the tumor ruptured before, or at operation

The comparison of the three anti-dsDNA antibody detecting test / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To compare the three Anti-dsDNA antibody detecting test (IIF, ELISA, Farr) with 200 serum samples to evaluate which one has higher sensitivity and specificity.</p><p><b>METHODS</b>200 serum samples including 120 serum samples of SLE, 20 serum samples of rheumatoid arthritis, 20 serum samples of MCTD, 20 serum samples of SS, 20 serum samples of PSS and 50 serum samples of healthy measured by IIF, Farr and ELISA.</p><p><b>RESULTS</b>Detection the Anti-dsDNA antibody of the serum sample with the methods of IIF, ELISA and Farr. The positive percentage of Anti-dsDNA in SLE is 25%, 32% and 32%, while in RA is 0, 0 and 0; in PSS is 0, 0 and 5%; in SS is 0, 0 and 0; in healthy is 0, 0 and 0.</p><p><b>CONCLUSION</b>Detection the Anti-dsDNA antibody with two method in the same time, especially with IIF and ELISA, will heighten the positive rate than with single method and will be helpful for the diagnosis of SLE.</p>

Effect of Lewis y antigen on regulating gene expression of partial drug resistance associated proteins in human ovarian cancer cell line RMG-I-H / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the influence of Lewis y antigen on the gene expression of partial drug resistance associated proteins in human ovarian cancer cell line RMG-I-H.</p><p><b>METHODS</b>RT-PCR was used to determine the gene expressions of partial drug resistance associated proteins in RMG-I-H cell line transfected with alpha1, 2-fucosyltransferases gene and RMG-I cell line, as well as in RMG-I-H treated with or without anti-Lewis y monoclonal antibody at the concentration of 10 micro/g/ml. The immunocytochemical method was used to detect the expression of P-glycoprotein (P-gp) in RMG-I and RMG-I-H cell lines. RMG-I and RMG-I-H cells were transplanted into nude mice and the expression of P-gp in the tissues was measured by immunohistochemistry.</p><p><b>RESULTS</b>The mRNA expressions of protein kinase C-alpha (PKC-alpha), topoismerase I ( Topo I ), multidrug resistance-associated protein-1 (MRP-1), and MRP-2 were significantly higher in RMG-I-H cells than those in RMG-I cells (0.46 +/- 0.02 vs. 0.27 +/- 0.05, 0.82 +/- 0.08 vs. 0.52 +/- 0.04, 0.66 +/- 0.07 vs. 0.34 +/- 0.12, and 0.44 +/- 0.08 vs. 0.23 +/- 0.05; all P < 0.05). However, the mRNA expression of multi-drug resistance 1 (MDR-1) was significantly lower in RMG-I-H cells than that in RMG-I cells (0.26 +/- 0.05 vs. 0.45 +/- 0.08, P < 0.05). The P-gp level increased in RMG-I-H cells compared with that in RMG-I cells both in vivo and in vitro (P < 0.05). Expressions of MDR-1, MRP-1, MRP-2, PKC-alpha, and Topo I mRNA decreased by the time in RMG-I-H cells treated with anti-Lewis y monoclonal antibody (all P < 0.05), while mRNA expressions of those genes in the control group did not statistically change (P > 0.05). In addition, MDR-1, MRP-1, MRP-2, PKC-alpha, and Topo I mRNA expressions were significantly lower in RMG-I-H cells treated with anti-Lewis y monoclonal antibody than those in the control group at 6 hours (all P < 0.05) and the inhibition ratios were 48.55%, 77.50%, 70.18%, 45.86%, and 46.13%, respectively.</p><p><b>CONCLUSION</b>The Lewis y antigen of the human ovarian cancer cell surface is closely correlated with the regulation on the gene expression of partial drug resistance associated proteins.</p>

Expression of matrix metalloproteinase-1 mRNA in peripheral blood mononuclear cells of systemic lupus erythematosus patients and its relationship with atherosclerosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Matrix metalloproteinase-1 (MMP-1) plays an important role in atherosclerosis. This study was to examine expression of MMP-1 mRNA in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), and to explore its relationship with atherosclerosis in SLE.</p><p><b>METHODS</b>Fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to examine the expression of MMP-1 mRNA in PBMCs in 80 SLE patients, including 39 prone to atherosclerosis (Group A) and 41 unprone to atherosclerosis (Group B). Meanwhile, 30 patients who were free of cardiovascular diseases and 30 healthy individuals were selected as disease and normal control group (Groups C and D). The changes of MMP-1 gene expression were analyzed by differences of cycle threshold (DeltaCt), with the following formula: DeltaCt = Ct(target) gene - Ct(reference) gene.</p><p><b>RESULTS</b>The expression level of MMP-1 mRNA in Group A was significantly higher than that of group B (DeltaCt = 8.64 +/- 2.43 vs DeltaCt = 12.09 +/- 2.26, t = 6.588, P < 0.01). The expression level of MMP-1 mRNA of SLE patients was significantly higher than that of Group C (DeltaCt = 10.41 +/- 2.90 vs DeltaCt = 12.29 +/- 2.51, t = 3.135, P < 0.01) and Group D (DeltaCt = 10.41 +/- 2.90 vs DeltaCt = 12.48 +/- 1.69, t = 3.675, P < 0.01).</p><p><b>CONCLUSIONS</b>In comparison to disease and control group, expression of MMP-1 mRNA in PBMCs of SLE patients was significantly elevated, and significant difference of MMP-1 mRNA expression was also found between SLE patients prone and unprone to atherosclerosis, indicating that expression of MMP-1 mRNA may be correlated with the pathogenesis and activity of atherosclerosis in SLE.</p>

Expressions of HLA class I antigen and CD8 and their clinical significance in cervical cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To examine the expressions of HLA class I antigen and CD8 in various cervical diseases and investigate their association with cervical cancer.</p><p><b>METHODS</b>The expressions of HLA class I antigen and CD8 in cervical tissues sampled from patients with cervical cancer, cervical intraepithelial neoplasia (CIN), and chronic cervicitis were detected using SP immunohistochemistry. The association of the expressions of HLA class I antigen and CD8 with the clinicopathologic indices of the patients was analyzed.</p><p><b>RESULTS</b>The positive expression rates of HLA class I antigen in cervical cancer, CIN, and chronic cervicitis were 22.6%, 100.0%, and 100.0%, and the positive expression rates of CD8 were 22.6%, 95.5%, and 100.0%, respectively. The positive rates of HLA class I antigen and CD8 were significantly lower in patients with cervical cancer (P<0.01). Patients with stage I cervical cancer had significantly higher positive rates of HLA class I antigen and CD8 than those with stage II cervical cancer (46.7% vs 0.0%, 46.7% vs 0.0%, both P<0.01). The expressions of HLA class I antigen and CD8 decreased with the progression of the clinicopathological stages, and may even become undetectable. The expressions of HLA class I antigen and CD8 were not related to the differentiation degree of the tumor or lymph node metastasis (P>0.05). A positive correlation was found between HLA class I antigen expression and CD8 expression.</p><p><b>CONCLUSION</b>The expressions of HLA class I antigen and CD8 are down-regulated or deleted in CIN and cervical cancer, and they may play important roles in the development and progression of CIN and cervical cancer.</p>

SiRNA inhibition of E6AP expression in cervical cancer cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the effect of RNA interference (RNAi) targeting E6AP on the proliferation and apoptosis of HeLa cells.</p><p><b>METHODS</b>HeLa cells were cultured and divided into 3 groups: blank control group, cells transfected with nonsense siRNA (small interference RNA), and cells transfected with specific E6AP siRNA. The expressions of E6AP mRNA and protein were detected by RT-PCR and Western blot before and after the transfection respectively. Cell proliferation was determined by methylthiazolyl tetrazolium (MTT). The cell apoptosis index was assessed by flow cytometry.</p><p><b>RESULTS</b>Upon treatment with E6AP siRNA for 24, 48 and 72 h, the expression level of E6AP mRNA decreased 33%, 72% and 70% than siRNA treated group. The protein expression levels in 48 h and 72 h E6AP siRNA groups decreased 38%, 59% comparing with those of the nonsense siRNA treated group (P < 0.05). The proliferative capacity of cells transfectd with E6AP siRNA was significantly lower than blank control group (F = 101.38, P < 0.05) and siRNA treated group (F = 38.64, P < 0.05). The apoptosis index of HeLa cells treated with E6AP siRNA was significantly higher than that of the nonsense siRNA (F = 41.48, P < 0.05) and the blank control group (F = 86.36, P < 0.05).</p><p><b>CONCLUSION</b>SiRNA targeting can effectively suppress the expression levels of E6AP mRNA, corresponding with a proliferation inhibition and an enhanced apoptosis of HeLa cells.</p>

Transfection of alpha1, 2-fucosyltransferase gene increases the antigenic expression of Lewis y in ovarian cancer cell line RMG-I / 中国医学科学院学报

<p><b>OBJECTIVE</b>To transfect human alpha1, 2-fucosyltransferase (alpha1, 2-FT) gene to ovarian cancer cell line RMG-I and investigate the antigenic expression change of Lewis y and the other related oligosaccharides.</p><p><b>METHODS</b>The expression vector pcDNA3.1(-)-HFUT-H was constructed by polymerase chain reaction (PCR) to clone human alpha1, 2-FT gene coding region. The alpha1, 2-FT gene stable high-expression cell line RMG-I-H was established by transfecting pcDNA3.1(-)-HFUT-H to ovarian cell line RMG-I. The change of alpha1, 2-FT activity in the cell line before and after the tranfection was confirmed by the determination of enzymatic activity. The changes of cell lipid and glucolipid, especially the change of type II oligosaccharide, in the cell line before and after the transfection was determined by Thin-Layer Chromatography (TLC) and TLC immunostaining method, respectively.</p><p><b>RESULTS</b>The H-1 antigen and Lewis y antigen were obviously increased in the cell line RMG-1-H, especially the latter one, which was 20 times higher than before, and the type I saccharide chain Lewis b was decreased significantly. The main lipid components on the cell membrane, cholesterol and phosphatides, showed no change in the cell lines before and after the transfection, and the neutral glycolipid also showed no obvious change.</p><p><b>CONCLUSIONS</b>The transfection of alpha1, 2-FT gene can increase the activity of alpha1, 2-FT in the cell line RMG-I and mainly increase the expression of Lewis y antigen simultaneously. The construction of RMG-I Lewis y high expression cell line provides a cell model for further study on the relationship between Lewis y antigen and biological behaviors in the ovarian cancer.</p>

Effect of intracoronary adenovirus vector encoding hepatocyte growth factor gene on hematopoietic stem cells mobilization in patients with extensive coronary heart disease / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effect of intracoronary adenovirus vector encoding hepatocyte growth factor gene (Ad(5)-HGF) on hematopoietic stem cells mobilization in patients with extensive coronary heart disease.</p><p><b>METHODS</b>Patients with extensive coronary heart disease were treated with intracoronary infusion of adenovirus vector encoding hepatocyte growth factor (Ad(5)-HGF 5 x 10(9) pfu) gene plus stent implantation (n = 9) or equal physiological saline plus stent implantation (n = 9). Angioplasty and stent implantation was performed according to standard clinical practice by the femoral approach and blood samples were drawn from each patient at baseline before PCI, 6 to 24 hours and 6 days post procedure. The number of CD34(+), CD38(+) and CD117(+) cells in peripheral blood was analyzed by flow cytometer.</p><p><b>RESULTS</b>The number of circulating CD34(+) cells in Ad(5)-HGF gene treatment group 6 hours after procedure and the number of circulating CD117(+) cells 6 days post procedure were significantly higher in Ad(5)-HGF gene treatment group than those in the control group (0.104 +/- 0.082 vs. 0.022 +/- 0.012, P = 0.021) and (0.058 +/- 0.058 vs. 0.012 +/- 0.009, P = 0.034), respectively.</p><p><b>CONCLUSION</b>Intracoronary administration of Ad(5)-HGF could mobilize hematopoietic stem cells into peripheral blood and the consequent role of this observation on myocardial regeneration warrants further detailed studies.</p>

Observation and clinical significance of adjuvant chemotherapy-induced amenorrhea in premenopausal breast cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>A retrospective analysis of 160 pre-menopausal breast cancer patients was carried out to elucidate the the menstrual outcome in those cases who had undergone adjuvant chemotherapy after surgery, and evaluate the relationship between chemotherapy-induced amenorrhea (CIA) and recurrence of the disease.</p><p><b>METHODS</b>160 pre-menopausal breast cancer patients were collected, 62/159 (39.0%) of them were node positive, 91/158 (57.6%) were ER positive, and 95/155 (61.3%) were PR positive. 111 cases had infiltrative ductal carcinoma, 26 cases had infiltrative lobular carcinoma, and 22 cases with others. In 152 cases data were collected by face-to-face interview and 8 cases by phone conversation. Types and cycles of chemotherapy regimen as well as menstrual abnormalities were recorded before, during, and after chemotherapy completion. Follow up duration was 12-72 months after chemotherapy completion for all patients.</p><p><b>RESULTS</b>107 (66.9%) developed CIA, 24 cases returned to normal menses (22.4%), 83 cases continued CIA during more than 12-month follow up (77.6%). The rate of CIA increased with age (P < 0.01). During the follow up, disease free survival (DFS) rate was 85.9% in CIA group and 79.2% in non-CIA group, with no statistically significant difference. But in hormonal receptor positive patients, DFS was 80.0% in non-CIA and 90.1% in CIA, respectively (P = 0.04), showing a significant difference. Because of the small number of died cases, no analysis of the overall outcome was carried out.</p><p><b>CONCLUSION</b>Adjuvant chemotherapy causes ovarian function suppression, and may further leading to amenorrhoea. Women who experienced amenorrhoea after chemotherapy had a significantly better disease-free survival (DFS) rate showed by univariate analysis than women who continued normal menstruation. Chemotherapy is insufficient therapy for very young patients who are in high risk with hormone responsive disease, particularly when chemotherapy fails to induce amenorrhea. Further research is needed to evaluate interventional chemotherapy to improve the quality of life in women with early stage breast cancer who experienced ovarian toxicity. The post-chemotherapy menstruation status is a clinically valuable, objective and salient marker for sufficient endocrine effect of chemotherapy in ER/PR-positive premenopausal patients.</p>

Detection of circulating hypermethylated tumor-specific RASSF1A DNA in ovarian cancer patients / 中华病理学杂志

<p><b>OBJECTIVE</b>To detect hypermethylated tumor-specific RASSF1A DNA in the circulation and its significance in ovarian cancers patients.</p><p><b>METHODS</b>Methylation-specific polymerase chain reaction (MSP) was used to study the hypermethylation of RASSF1A in preoperative serum samples from 51 ovarian cancer patients.</p><p><b>RESULTS</b>The RASSF1A gene was not methylated in peripheral blood samples from 51 normal patients and 51 patients with benign ovarian tumors. Hypermethylation of RASSF1A gene was found in circulating tumor-specific DNA in 43.1% of patients (22 out of 51 cases) with ovarian cancers (P < 0.05). There was no difference in hypermethylation of RASSF1A gene amongst various ovarian cancer subtypes (P < 0.05). On the other hand, hypermethylation of RASSF1A gene was more frequently encountered in stage III and IV than stage I and II tumors (P < 0.05). It was rarely seen in well and moderately differentiated groups, as compared with poorly differentiated group (P < 0.05).</p><p><b>CONCLUSIONS</b>There is a higher frequency of RASSF1A hypermethylation in circulating tumor-specific DNA of ovarian cancer patients. RASSF1A has been postulated to play an important role as tumor suppressor gene and can be silenced by promoter hypermethylation. This methylation correlates with clinical stage and histopathologic grade. Such observation may carry diagnostic and prognostic implications when assessing ovarian tumors.</p>

Expression and clinical significance of KISS-1 and GPR54 mRNA in endometrial carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of KISS-1 mRNA and GPR54 mRNA in endometrial carcinoma.</p><p><b>METHODS</b>The expression of KISS-1 mRNA and GPR54 mRNA in 32 patients with endometrial carcinoma, 10 patients with endometrial intraepithelial neoplasia (EIN) and 12 patients with normal endometrium was detected by reverse transcriptase polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The positive rate of KISS-1 mRNA in endometrial carcinoma, EIN and normal endometrium was 37.5%, 80.0% and 83.3% respectively (endometrial carcinoma vs EIN or normal endometrium, P < 0.05). The expression of KISS-1mRNA in patients with endometrial carcinoma was correlated with its clinical stage, myometrial invasion and lymph node metastasis (P < 0.05). In endometrial carcinoma, the more advanced clinical stage, the lower expression of KISS-1 mRNA was detected. The positive rate of GPR54 mRNA in endometrial carcinoma, EIN and normal endometrium was 78.1%, 70.0% and 66.7% respectively, with no significant statistical difference (P > 0.05). It was not correlated with the clinical stage, histology grade, myometrial invasion or lymph node metastasis (P > 0.05).</p><p><b>CONCLUSION</b>The interaction of KISS-1 and GPR54 may play an important role in inhibiting the invasion and metastasis of endometrial carcinoma.</p>

Transfection and expression of hRI gene on human umbilical blood stem cells and gene therapy for mouse melanoma / 生物工程学报

In order to explore the transfection and expression of hRI gene on human umbilical blood stem cells, and observe it's effect on the tumor growth. After enriching human umbilical cord blood CD34+ cells with a high-gradient magnetic cell sorting system (MACS), transfected them with supernatant of retrovirus containing human Ribonuclease inhibitor (hRI) cDNA. Hematopoietic progenitor clonogenic assay and PCR were used to evaluate transfection efficiency, and Western-blot and immune fluorescence were used to evaluate the expression quantity of hRI gene after transfection. Observe the effect of RI on the growth of melonoma in B16C57BL mice. The results showed that human umbilical blood CD34+ cells were highly purified by MACS, which made the purity of human umbilical blood CD34+ cells average 96.15%. hRI can be transfected on umbilical blood CD34+ cells, and the transfection efficiency was 35%. The positive expression of hRI gene on transfected CD34+ cells is identified by Western-blot and immune fluorescence assay. Mice injected with transfected CD34+ cells show a significant restraint of the tumor growth, a lower efficiency of tumor formation, a lower weight of the tumor and a longer incubation period of tumor formation with respect to the control groups. The results demonstrated the capacity of RI to inhibited the tumor growth by blocking the vasculature in tumor.

The expression and significance of glucose-6-phosphate isomerase mRNA in peripheral blood mononuclear cells of patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the mRNA level of glucose-6-phosphate isomerase(GPI)ex- pression in patients with rheumatoid arthritis(RA).Method Reverse transcription-polymerase chain reaction (RT-PCR)was applied to semiquantitatively analyze GPI mRNA expression in the peripheral blood mononu- clear cells(PBMCs)of 30 active RA patients,30 stable RA patients,30 patients with other rheumatic disease and 30 healthy subjects.Results There was statistically significant differences between patients with RA and other rheumatic diseases(P

Inhibition of invasive and chemotactic abilities of SKOV3 cells bv human epithelial growth receptor-2 small interfering RNA / 中华妇产科杂志

Objective To investigate the effects of RNA interference(RNAi)targeting human epithelial growth receptor-2(HER-2)gene on the invasive and chemotactic ability of SKOV3 cells. Methods Glyeeraldehyde-3-phosphate dehydrogenase(GAPDH)was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted(HER-2 siRNA Ⅰ,HER-2 siRNA Ⅱ,HER-2 siRNA Ⅲ)sequence were selected in the coding region of HER-2 mRNA.Transfection of HER-2 siRNA was conducted with lipofeetamine 2000 in ovarian carcinoma cell line SKOV3.The HER-2 gene expression was assessed by real- time PCR and western blot assays.Changes of invasive and chemotaetie capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal.Results Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose,the expression of GAPDH protein was decreased.GAPDH protein gray value in control group, different doses(0.5,1.0,1.5,2.0 ?g)GAPDH siRNA interference groups were 0.6855?0.0259,0.5698 ?0.0275,0.4542?0.0296,0.3341?0.0178 and 0.1816+0.0180,respectively.There was a significant difference in each group(F=198.126,P