Résumé
Objective To investigate the effects of cinobufacini on TGF-β1-induced epithelial-to-mesenchymal transition of human colorectal carcinoma cells in vitro.Methods The cultured colorectal carcinoma cell line(SW480) was divided into the control group,TGF-β1 (10 ng/mL) individual treatment group and co-treatment groups with TGF-β1 (10 ng/mL) + cinobufacini (2.5,5,10,20,40,80 mg/mL),which were cultured in vitro for 48 h.The proliferation of the cells were measured by CCK8 assay.The morphological changes were observed by inverted phase contrast microscope.The ability of cell invasion and migration was detected by Transwell assay.The mRNA and protein expressions of E-cadherin and Vimentin were detected with QRT-PCR and Western Blot.Results (1)Compared with the TGF-β1 (10 ng/mL) individual treatment group,TGF-β1 (10 ng/mL) + cinobufacini (10,20,40,80 mg/mL) co-treatment groups significantly had significantly proliferation inhibitory effect on SW480 (P＜0.05).(2) Compared with the normal control group,TGF-β1 individual treatment group and TGF-β1 (10 ng/mL) + cinobufacini(2.5 mg/mL) group exhibited classical mesenchymal phenotype,while the TGF-β1 (10 ng/mL) + cinobufacini (5 mg/mL) co-treatment group showed classical epithelial phenotype.(3) The ability of invasion and migration in the TGF-β1(10 ng/mL)+ cinobufacini(2.5,5 mg/mL) co-treatment group were significantly weakened compared with the TGF-β1 individual treatment group (P＜0.01).(4) QRT-PCR and Western Blot results indicated that compared with the normal control group,the Vimentin expression in the in the TGF-β1 individual treatment group was significantly increased and the E-cadherin expression was significantly decreased.Furthermore,compared with the TGF-β1 individual treatment group and control group,the Vimentin expression level in the TGF-β1 (10 ng/mL) + cinobufacini (2.5,5 mg/mL) groups was significantly decreased and E-cadherin expression was significantly increased.Conclusion Cinobufacini can inhibit TGF-β1-induced cell proliferation in human colorectal carcinoma SW480 cells,and its mechanism may be related with promoting E-cadherin expression increase,meanwhile decreasing the vimentin expression,thus inhibiting the EMT process induced by TGF-β1.
Résumé
Objective To observe the clinical efficacy of comprehensive rehabilitation therapy in treating post-stroke shoulder- hand syndrome.Method Sixty patients with post-stroke shoulder-hand syndrome were randomized into a treatment group and a control group, 30 cases in each group. The treatment group was intervened by acupuncture plus correct posture, rehabilitation training, wax disc method, and intermediate-frequency medicinal penetration therapy; the control group was only by acupuncture plus correct posture. Before and after intervention, the limb function, pain, swelling degree, and activities of daily life were measured, and the clinical efficacies were compared between the two groups.Result The total effective rate was 93.3% in the treatment group versus 66.7% in the control group, and the difference was statistically significant (P<0.01). In the treatment group, the indexes (Fugle-Meyer Assessment of upper limb, movement range of shoulder, Global Perceived Effect, Visual Analogue Scale, and swelling score) were significantly changed after intervention (P<0.01). After treatment, the indexes of the treatment group were significantly different from those of the control group (P<0.01).Conclusion Comprehensive rehabilitation therapy can significantly improve pain, swelling, motor function of the affected limb, and activities of daily life in post-stroke shoulder-hand syndrome.
Résumé
Four weeks after SD diabetic rats were induced by streptozotocin,skin thickness was obviously reduced with obscure multilayer epithelium features.Moreover,the thickness of epidermic layers in diabetic rat skin was significantly thinner than that ofnornlal rat skin at the eighth week[(0.016±0.006 vs 0.041±0.007)mm,P<0.01].The percentage of G2/M phase cells in epidermic layers of diabetic group was significantly lower than that in the normal group.At the twelfth week,skin microangiopathy was easily detected in the diabetic group.The blood levels of advanced glycation end products(AGEs)and malonialdehyde were significantly increased and glutathione decreased in diabetic rats compared with control rats(aU P<0.01),along with the increased contents of local glucose and AGEs in the skin of diabetic rats.These results suggest that the local accumulation of glucose and AGEs seems to be one of the important mechanisms in the pathogenesis of diabetic skin lesions.
Résumé
Objective To screen the differentially expressed genes between papillary thyroid carcinoma (PTC) and normal thyroid tissues using cDNA microarray. Methods mRNA from both PTC and normal thyroid tissues were reversely transcribed to cDNAs with the incorporation of fluorescent dUTP (Cy-5 or Cy-3) to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray, which was scanned for the fluorescent signals and showed difference between two tissues. Two among the differentially expressed genes were verified by RT-PCR and immunohistochemistry. Results In PTC 48 genes were upregulated while 17 were downregulated. The results of RT-PCR and immunohistochemistry were consistent with that of the gene chips. Conclusion cDNA microarray technique is effective in screening the differentially expressed genes between 2 different kinds of tissue. The obtained genes are mainly related with extracellular matrix, cytokine, signal transduction and so on.
Résumé
Objective To observe nitric oxide synthase (NOS) activity and the expresssion of inducible NOS (iNOS) mRNA in cultured calf aortic endothelial cells treated with various concentrations of insulin. Methods Cultured calf aortic endothelial cells were incubated with different concentrations of insulin for 24 h. NOS activity was determined by colorimetry and iNOS mRNA was measured by semi-quantitative RT-PCR. Results NOS activity and the expression of mRNA in calf aortic endothelial cells at pharmacological concentration (10 -7 mol/L) of insulin were significantly increased than those at physiological concentration (10 -10 mol/L) of insulin (P
Résumé
With the development of modern medicine,to see a doctor is no longer a simple process of diagnosis or taking drugs.Human have deeply recognized medical science and health from a simple biomedical model to a social,psychological and biomedical model.On the one hand,people seek help from medical science.On the other hand,people fear that malpractice could occur.It has been in a love-hate mentality for people to seek medical and legal relief.In the new era,physician-patient relationship needs to be recognized again from medical staff,patients and legal angles.The combination of medicine and law is the requirement of development of the times.No matter how strong the law is,and no matter how much we hope the legislation is from strength to strength,moderation is still the virtues of the law.Medical perspective is still required in the examination of medical activity.
Résumé
AIM: To investigate the melatonin receptor (MR) gene and protein expression in the human peripheral blood lymphocytes. METHODS: Total RNA of human peripheral lymphocytes was isolated by single step method of acid guanidinium-thiocyanate-phenol-chloform, mt 1 and MT 2 mRNA were determined by the reverse transcription-polymerase chain reaction(RT-PCR). Immunohistochemistry was applied to identify and localize mt 1 and MT 2 protein. RESULTS: 1 1 kb mt 1 mRNA was detected by Northern blot, but 1 1 kb MT 2 mRNA was not detected. The mt 1 cDNA fragment of the expected size of 370 bp was determined and the MT 2 cDNA fragment of the expected size of 320 bp was not determined by RT-PCR. Sequencing result indicated that the positive product coincided with the cDNA of human mt 1. The mt 1 protein was observed by immunohistochemistry. These buffy positive granules were scattered with some areas stronger than the others, and primarily located in cytoplasm and membrane, with rare location in nucleus; meanwhile the MT 2 protein was not observed. CONCLUSION: These results demonstrated the mt 1 mRNA and protein expression in human peripheral leukocytes. It is indicated that melatonin has directly immune-regulative effects on peripheral lymphocytes.
Résumé
AIM: To explore the histochemical changes o f diabetic skin and the pathogenesis of impaired wound healing in diabetes. METHODS: 54 male Sprague-Dawley (SD) rats weighing 200-220 g wer e randomized into control and STZ-induced diabetic groups. The shaved skin speci mens from the back of rats were collected in 4, 8 and 12 weeks post STZ-inductio n, respectively. Hematoxylin-eosin dye was used for histological examination. Me a nwhile, the skin glucose contents were measured by Beckman's autoanalyzer. Skin AGEs concentrations were assessed by detecting total fluorescence in tissue coll agen and immunohistochemistry assay. RESULTS: The skin thickness in diabetic animals was decreased, w ith the features of multilayer epithelium structure disappeared in epidermis and collagen fibers atrophied, swollen and degenerated in dermis; The inflammatory responses in the dermis of diabetic animals were increased obviously. The result s also revealed that skin glucose contents in diabetic rats [(2.64?1.03)mg/ g skin] were 2-3 times higher than those in the controls [(0.74?0.33)mg/g s kin] (P
Résumé
To investigate the melatonin receptor(MR) gene and protein expression in human HUT78 cell line, total RNA of HUT78 cells was isolated by single-step method of acid guanidinium-thiocyanate-phenol-chloform, and mt 1 and MT 2 mRNA were determined by reverse transcription-polymerase chain reaction(RT-PCR). The Envision method was applied to immunohistochemistry to identify and localize mt 1 and MT 2 protein. The mt 1 cDNA fragment of the expected size of 370bp was determined, but the MT 2 cDNA fragment of the expected size of 320bp was not determinable by RT-PCR. Sequencing result indicated that the positive product coincided with the cDNA of human mt 1. The mt 1 protein was observed by immunohistochemistry. These buffy positive granules were scattered with some areas stronger than the others, and were primarily located in cytoplasm and membrane, with rare location in nucleus; meanwhile the MT 2 protein was not observed. These results demonstrated the mt 1 mRNA and protein expression in HUT78 cell line. It is indicated that melatonin has directly immune-regulative effects on T lymphocyte, The changes of MR in physiological and pathological stages need to be investigated.