Tian J.,Nanfang Hospital
Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2011
To construct the recombinant plasmid pcDNA3.0-RGC32 and evaluate the effect of the response gene to complement-32 (RGC32) on cell cytoskeleton in vitro. The full-length cDNA of RGC32 was obtained by RT-PCR and inserted into the eukaryotic expression vector pcDNA3.0 to generate the recombinant plasmid pcDNA3.0-RGC32. After transfection of the recombinant plasmid into SW480 cells, the expression of RGC32 in the cells was detected by Western blotting. The cytoskeleton of SW480 cells was visualized before and after the transfection, and the changes in the cell migration ability was assessed by wound-healing assay. The recombinant plasmid pcDNA3.0-RGC32 was successfully constructed. The expression of RGC32 was significantly increased in SW480 cells after transfection with pcDNA3.0-RGC32. Before the transfection, the microfilaments of SW480 cells were few and short without obvious polarity, but after the transfection, the microfilaments were increased and elongated with also an obvious polarity, and the invasive structures of lamellae and lamellipodia occurred. The migration ability of the cells was enhanced after transfection with pcDNA3.0-RGC32. Overexpression of RGC32 can cause the reorganization of cytoskeleton and promotes the cell migration, which can be an important mechanism of RGC32 in promoting cancer metastasis.
Zhou Q.Z.,Nanfang Hospital
Zhonghua nan ke xue = National journal of andrology | Year: 2010
OBJECTIVE: To appraise the effect of single- and two-layer Percoll density gradient centrifugation in sperm separation. METHODS: Twenty semen specimens underwent single-(50%) and two-layer (90% and 45%) density gradient centrifugation, respectively. The sperm class analyzer (SCA) was used to analyze sperm density, motility and dynamic parameters and round cell density before and after the treatment. RESULTS: After separation, the sperm recovery rate of the single-layer method was (65.5 +/- 12.8)%, significantly higher than that of the two-layer method (P < 0.01). The percentages of grade a sperm of the single- and two-layer method were significantly higher than pre-treatment (P < 0.05, P < 0.01), that of the single-layer was significantly lower than that of the two-layer method (P < 0.05), but the percentage of grade c sperm of the former was significantly higher than that of the latter (P < 0.05). Compared with pre-treatment, the percentage of grade a + b sperm of the two-layer method was significantly higher (P < 0.05), while that of the single-layer method showed no significant difference (P > 0.05), and the round cell density of both the methods was significantly lower (P < 0.05, P < 0.01), with no significant differences between the two methods (P > 0.05). CONCLUSION: The single-layer method yields a higher rate of sperm recovery and causes little change in the sperm motility, while the two-layer method effects a lower rate and significantly improves sperm motility. Both the methods can efficiently separate sperm from round cells, and each has its own advantages and its application value in in vitro treatment of sperm.
Li J.P.,Nanfang Hospital
Zhonghua nan ke xue = National journal of andrology | Year: 2010
To explore the effects of low-dose oral tadalafil on self-esteem, confidence and sexual relationship in ED patients. We treated 17 ED patients with oral tadalafil at the low dose of 5 mg once daily for 12 weeks, and used the paired t test to compare their scores on The Self-Esteem and Relationship Questionnaire (SEAR) and IIEF-5 and the results of nocturnal penile tumescence (NPT) obtained by nocturnal electrobioimpedance volumetric assessment (NEVA) before and after the medication. The scores on SEAR and IIEF-5 were significantly increased (P < 0.01) and NPT markedly improved (P < 0.05) after tadalafil treatment as compared with the baseline. Low-dose oral tadalafil once daily can significantly improve the self-esteem, confidence, sexual relationship satisfaction and NPT of ED patients.
Guo D.,Nanfang Hospital
Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials | Year: 2010
To investigate the effects of matrine on proliferation, cell cycle, apoptosis rate and beta-catenin-dependent transcriptional activity in cultured hepatoma cell line Hep3B. Cell viability was analyzed by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis rate wene determined by flow cytometry analysis, beta-catenin-dependent transcriptional activity was assayed with Dual-Luciferase Reporter System. 72 h of matrine (50 - 800 mg/L) exposure significantly suppresses Hep3B cells growth in a concentration-dependent manner (P<0.01), the 50 percent inhibitory concentration (IC50) was 312.53 mg/L. A higher proportion of apoptotic cells in matrine-treated group than in control groups (21.73 +/- 1.66% vs. 4.39 +/- 1.93%) are shown. Cell proportions in G0/G1 phase were respectively 74.48% and 57.39% in matrine-treated group and control group. Cell proportions in S phase wene respectively 12.94% and 27.67%. Beta-catenin reporter activity was also decreased by matrine treatment in a concentration-dependent manner in Hep3B cells (P<0.05 or P<0.01). Through inhibition of beta-catenin-dependent transcription, matrine induces cell cycle arrest and apoptosis, and subsequently suppresses proliferation of hepatoma cells.
Yu K.,Nanfang Hospital
Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2013
To study the relationship between sub-health status and the health-promoting lifestyle of employees. A total of 5316 employees in a company in Guangdong were surveyed using sub-health measurement scale version 1.0 (SHMS V1.0) and the health-promoting lifestyle profile (HPLP-II). The former scale included 3 subscales of somatic sub-health, psychological sub-health and social sub-health, and the latter included 6 subscales of self-actualization, healthy responsibility, physical exercise, nutrition, interpersonal sensitivity and stress treatment. The total healthy rate was 12.86% among the employees, with 76.76% and 10.5% in sub-health and disease states. The mean scores of HPLP-II was 115.95∓21.468 in the total population surveyed, 134.23∓24.72 in healthy employees and 114.69∓19.25 in the patients. There was a significant difference in the grades of health-promoting lifestyle between sub-healthy and healthy employees (P<0.05) as well as in the scores of HPLP-II and the scores of the 6 subscales (P<0.05). An appreciable correlation was found between sub-health status and the 6 subscales, and self-realization, physical exercise and stress management showed significant inverse correlation with sub-health status. Sub-health status is related to health-promoting lifestyle, and self-realization, physical exercise and management are the protective factors that influence sub-health status.
Gao G.H.,Nanfang Hospital
Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2010
To evaluate the clinical efficacy of infliximab in the treatment of moderate and severe active rheumatoid arthritis (RA). This randomized double-blind II/III clinical trial involved 30 patients with moderate and severe active RA, who were randomly allocated into 3 groups (groups A, B, and C) at the ratio of 3:1:1. At weeks 0, 2, 6, and 14, the patients in groups A and C received infliximab or placebo, and those in group B had placebo at week 14 with a stable background dose of methotrexate. The indicators for efficacy evaluation included the proportions of ACR20/50/70 of the responders and DAS28. The sharp scores of the hand joints were recorded before and after the treatment. Twenty-nine patients completed the clinical trial (18 in group A, 5 in group B, and 6 in group C). At week 14, the proportions of ACR20/50/70 in the 3 groups reached 83.33%, 60%, and 33.33%, respectively (P<0.05), as compared to 100%, 100%, and 33.33% at week 18 (P<0.05). The other indicators for clinical efficacy evaluation also suggested similar clinical improvement of the patients (P=0.000). The proportions of the patients with DAS28<3.2 and DAS28<2.6 were significantly different. Compared to the baseline, the Sharp scores in group A showed no significant changes at week 18 (P>0.930), while those in group C exhibited significant radiographic progression (P<0.044). Infliximab produces good short-term therapeutic effect against moderate and severe active RA and may help arrest the radiographic progression of the diseases, which can be more obvious in patients with moderate severity.
Mao X.M.,Nanfang Hospital
Zhonghua nan ke xue = National journal of andrology | Year: 2011
To study the differentially expressed genes in asthenospermia to gain a deeper insight into the molecular mechanisms of the disease. We analyzed the differentially expressed genes in asthenospermia using GATHER, PANTHER and ToppGene online bioinformatics tools. Our bioinformatics mining and analyses revealed that the differentially expressed genes in asthenospermia played important roles in the cellular protein and macromolecular metabolism, protein modification, cell death, cell apoptosis and apoptosis induction. Asthenospermia patients experience a decline in sperm activity and the basic life activities of sperm simultaneously, and are also prone to cell apoptosis or death. Such differentially expressed genes as KIF3B, MYO15A, KIF6, KIF26B, KIF3A, DNHD2, DMN, DYNC2H1, STARD9, MYOHD1, and TPM1, which are involved in cytoskeletal structure, microtubule movement and cell movement, may be associated with asthenospermia, and therefore deserve further studies.
Su L.,Nanfang Hospital
Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2011
To assess the diagnostic value of 8 equations using different variables for determining the estimated glomerular filtration rate (eGFR) in patients with cardiovascular diseases. GFR was estimated in 208 patients with cardiovascular diseases by (99m)Tc-DTPA dynamic renal imaging, and the eGFR was derived from 8 equations using different variables. In patients with chronic kidney disease (CKD) stages 1-3, the eGFR calculated suing serum creatinine (SCr)-based equation was better correlated to GFR estimated by (99m)Tc-DTPA renal imaging than that derived from cystatin C (Cys C)-based equations, whereas in patients with CKD stages 4 and 5, the estimates by the latter equation showed a better correlation to GFR. Compared with (99m)Tc-DTPA renal imaging, MDRD-based equation and simple MDRD equation resulted in a higher eGFR in patients with CKD stages 4 and 5, the Rule equation had a lower eGFR in CKD stages 1 and 2, the Macisaac equation yielded a higher eGFR in CKD stages 2-5, and the Tan equation showed a higher eGFR in CKD stages 2 and 3. In patients with mild renal dysfunction, the Scr-based equation had a higher AUC(ROC) than Cys C-based equation, which was reversed in patients with severe renal dysfunction; the AUC(ROC) of the two equations were comparable in patients with moderate renal dysfunction. Compared with (99m)Tc-DTPA renal imaging, the modified MDRD equation and Arnal-Dade equation showed no significant difference in the eGFR in patients with CKD stages 1-5. Modified MDRD equation (or simple MDRD equation) and Arnal-Dade equation are superior to other calculation methods for estimating the GFR in Chinese patients with cardiovascular disease.
Zhou L.,Nanfang Hospital |
Zhou L.,University of Pittsburgh |
Li Y.,University of Pittsburgh |
He W.,University of Pittsburgh |
And 6 more authors.
Journal of the American Society of Nephrology | Year: 2015
Activation of β-catenin, the intracellular mediator of canonical Wnt signaling, has a critical role inmediating podocyte injury and proteinuria. However, the underlying mechanisms remain poorly understood. Here, we show that β-catenin triggers ubiquitin-mediated protein degradation of Wilms' tumor 1 (WT1) and functionally antagonizes its action. In mice injected with adriamycin, WT1 protein was progressively lost in glomerular podocytes at 1, 3, and 5 weeks after injection. Notably, loss of WT1 apparently did not result from podocyte depletion but was closely associated with upregulation of β-catenin. This change in WT1/β-catenin ratio was accompanied by loss of podocyte-specific nephrin, podocalyxin, and synaptopodin and acquisition of mesenchymal markers Snail1, α-smooth muscle actin, and fibroblast-specific protein 1. In vitro, overexpression of β-catenin induced WT1 protein degradation through the ubiquitin proteasomal pathway, which was blocked by MG-132. WT1 and β-catenin also competed for binding to common transcriptional coactivator CREB-binding protein and mutually repressed the expression of their respective target genes. In glomerular miniorgan culture, activation of β-catenin by Wnt3a repressed WT1 and its target gene expression. In vivo, blockade of Wnt/β-catenin signaling by endogenous antagonist Klotho induced WT1 and restored podocyte integrity in adriamycin nephropathy. These results show that β-catenin specifically targets WT1 for ubiquitin-mediated degradation, leading to podocyte dedifferentiation and mesenchymal transition. Our data also suggest that WT1 and β-catenin have opposing roles in podocyte biology, and that the ratio of their expression levels dictates the state of podocyte health and disease in vivo. Copyright © 2015 by the American Society of Nephrology.
Zhang Y.,Nanfang Hospital
Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2012
To investigate the anti-apoptotic effect of low molecular weight heparin (LMWH) in the placenta of rats with preeclampsia-like symptoms. Thirty pregnant Wistar rats were randomized equally into 3 groups and received subcutaneous saline injection (control group), 200 mg/kg L-NAME injection to induce preeclampsia symptoms (PE group), or L-NAME with 40 μl/kg LMWH injections (LMWH group). The blood pressure, urine protein, the number of pups and the weight of the fetuses and placenta were measured, and the placental apoptotic index was measured by TUNEL assay. The expression of cleaved caspase-3, Bcl-2 and Bax in the placenta were examined by Western blotting. Compared with the control group, L-NAME injections caused significantly increased blood pressure and urine protein (P<0.05), which were significantly lowered by LWMH (P<0.05). The number and weight of normal pups were significantly lower in PE group than in the control group (P<0.05) and LMWH group (P<0.05); in LMWH group, the weight of pups was still lower than that of the control group (P<0.05) but the number of normal pups was comparable (P>0.05). The LMWH group showed a significantly lower placental apoptosis index than the PE group (P<0.05) with also significantly lower cleaved caspase-3 and Bax and higher Bcl-2 expressions (P<0.05). The apoptosis index and expressions of cleaved caspase-3, Bcl-2 and Bax protein were similar between LMWH group and normal pregnant group (P>0.05). LMWH can alleviate preeclampsia-like symptoms and decrease the apoptosis in the placenta of rats possibly by enhancing Bcl-2 and suppressing Bax expressions.