4.Morphological study of co-culture of Schwann cells with small intestinal submucosa in vitro
Yan SU ; Chang-Qing ZHANG ; Kai-Gang ZHANG ; Xue-Tao XIE ; Bing-Fang ZENG ;
Chinese Journal of Microsurgery 2000;0(02):-
Objective To observe adhesion and growth of Sehwann cells(SCs) on small intestinal submueosa(SIS) and study the bioeompatibility of SIS with SCs.Methods The SCs of SD neonate rat were isolated and cultured in vitro,then were seeded on prepared SIS.At different times,the adhesion,growth and proliferation of SCs on SIS were observed by phase contrast microscope,histological examination,scanning e- lectron microscope and transmission electron microscope.Results By phase contrast microscope,SCs grew well on the edge of SIS after 3 and 5 days.SCs adhered tightly on the surface of SIS after 5 days through histo- logical examination.By scanning electron microscope,on the surface of SIS,SCs grew and adhered actively, prominence of cells body were obvious.They connected end-to-end with each other or arranged in clusters. Protein granules were secreted on cells surface.By transmission electron microscope,SCs grew in good condi- tion and adhered tightly on the surface of SIS.At the interface of SCs and SIS,prominence was seen to contact with SIS in the bottom of cell body.Conclusion SCs are able to adhere and grow well on the surface of SIS.As a scaffold,SIS has good biocompatibility with SCs.
5.Clinical Observation of Catgut-embedding Therapy Based on Respiration-induced Reinforcing and Reducing for Treatment of Simple Obesity with Spleen Deficiency and Dampness Retention
Miaomiao LI ; Jinxia NI ; Jie WANG ; Xue FANG ; Buyi SU ; Xiaona WU
Journal of Guangzhou University of Traditional Chinese Medicine 2017;34(4):534-538
Objective To observe the therapeutic effect of catgut-embedding (CE) therapy based on respiration-induced reinforcing and reducing and electro-acupuncture (EA) therapy in treating simple obesity with spleen deficiency and dampness retention. Methods Sixty simlpe obesity patients with spleen deficiency and dampness retention were randomized into CE group and EA group, 30 cases in each group. The acupoints selected for the two groups were the same, and the points were Zhongwan, Shuifen, Qihai, Guanyuan, Tianshu, Liangmen, Daheng, Fujie, Quchi, Xuehai, Yinlingquan, Fenglong, and Ashi. CE group was given CE therapy with the needling for CE therapy referred to the respiration-induced reinforcing and reducing method, and EA group was given EA therapy for 2 continuous treatment courses, 4 weeks constituting one course. Body mass and body mass index (BMI) of the two groups before and after treatment were observed, and the clinical efficacy was also evaluated after treatment. Results (1) After treatment for 2 courses, body mass and BMI of the two groups were obviously decreased(P<0.05 compared with those before treatment), but the differences between the two groups were insignificant (P > 0.05). (2) The total effective rate of CE group was 90.0% and that of EA group was 86.7%, and the difference between the two groups was insignificant (P > 0.05). Conclusion The therapeutic effect of CE therapy based on respiration-induced reinforcing and reducing in treating simple obesity with spleen deficiency and dampness retention is similar to that of EA therapy, and the patients can choose anyone of them for loosing body weight according to the preference.
6.Construction and expression in vitro of an RU486 inducible vector carrying DsRed protein.
Jian CHEN ; Xuchao XUE ; Guoen FANG ; Changqing SU ; Qijun QIAN
Chinese Journal of Biotechnology 2008;24(8):1458-1463
The regulation of a target gene expression is very important in gene therapy. However, constitutive or inappropriate expression of the genes with traditional expression system may interfere with the effect of the gene therapy, even may lead to lethal side effect. We constructed an RU486 inducible eukaryotic vector carrying DsRed protein and evaluated its regulatable effect in vitro. The single vector named PDC-RURED was constructed with molecular biological methods, which contained DsRed gene, promoter and RU486-inducible system. To minimize any potential interference, we spaced the two transcriptional elements with a 1.6 kb insulator. The vector was identified by different enzyme restrictions, sequencing analysis and PCR assay. We demonstrated the regulatable expression of this vector after transfection in HEK293 cells by fluorescence microscopy and flow cytometry. In the absence of RU486, no significant DsRed protein activation was observed, whereas in the presence of RU486 up to 40 fold activation of the DsRed protein was observed in cultured cells. The data show that the novel eukaryotic expression plasmid vector can be used to regulate the expression level of genes of interest in appropriate time under the control of RU486. This inducible expression vector provides a powerful tool for the research of gene regulation and gene therapy.
Cell Line
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Fluorescent Dyes
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metabolism
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Genetic Therapy
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methods
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Genetic Vectors
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genetics
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Humans
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Kidney
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cytology
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Luminescent Proteins
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biosynthesis
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genetics
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Mifepristone
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pharmacology
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Promoter Regions, Genetic
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genetics
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Transfection
7.The changes of inducible nitric oxide synthase activity and apoptosis-related gene expression in endotoxemia-induced rat diaphragm.
Ying-Yan FANG ; Su-Dong GUAN ; Xiao-Lei GUO ; Hong-Wei YE ; Hua-Xue WANG ; Qin GAO
Chinese Journal of Applied Physiology 2013;29(3):209-212
OBJECTIVETo study the changes of inducible nitric oxide synthase (iNOS) activity and apoptosis-related genes Bcl-2, Bax and caspase-3 mRNA expressions in endotoxemia-induced rat diaphragm injury and analyze the related apoptosis mechanism.
METHODSThirty-two male SD rats were randomly divided into 4 groups (n = 8): control group (saline 0.5 ml ip), endotoxin 24 h, 48 h and 96 h group (endotoxin 12 mg/kg ip, animals were killed either 24, 48 or 96 h after injections). Body weight were measured, the ratio between diaphragm weight and body weight, activities of constitutive nitric oxide syntheses (cNOS), iNOS and succinate dehydrogenase (SDH) were also measured. The expressions of Bcl-2, Bax and caspase-3 mRNA were detected by RT-PCR analysis.
RESULTSEndotoxin induced significant reductions in diaphragm mass in endotoxin 96 h group (P < 0.05). Endotoxin increased diaphragm cNOS or iNOS activities, and they were significantly higher in endotoxin 96 h group than those in endotoxin 24 h and 48 h groups, diaphragm SDH activity was reduced, and it was lower in endotoxin 96 h group than that in endotoxin 24 h and 48 h groups (P < 0.01). Endotoxin significantly increased Bax and caspase-3 mRNA expressions, and they were higher in endotoxin 48 h and 96 h groups than those in endotoxin 24 h group (P < 0.01). Endotoxin significantly reduced Bcl-2 mRNA expression and the ratio of Bcl-2/Bax, and they were lower in endotoxin 48 h and 96 h groups than those in endotoxin 24 h group (P < 0.01).
CONCLUSIONiNOS is activated in endotoxemia-induced rat diaphragm injury. It damages mitochondria, upregulates Bax expression and downregulates Bcl-2 expression, then induces caspase-3 related apoptotic pathway. These changes may cause diaphragm injury and atrophy.
Animals ; Apoptosis ; Caspase 3 ; metabolism ; Diaphragm ; metabolism ; physiopathology ; Endotoxemia ; metabolism ; Gene Expression ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism
8.Expression of estrogen receptors alpha and beta in human tongue squamous cancer cell and influence of beta-estradiol on the proliferation of tongue cancer cell.
Ya-Li LIU ; Su-Min CHI ; Yun-Long ZHU ; Yan-Qing ZHONG ; Cai-Fang XUE
Chinese Journal of Applied Physiology 2003;19(1):65-69
AIMTo observe the expression of estrogen receptors alpha and beta in human tongue squamous cancer line Tca8113 cell, and to study the influence of beta-estradiol (beta-E2) on the proliferation and cell cycle of cultured Tca8113 cell.
METHODSImmunocytochemistry and RT-PCR methods were used to observe the expression of estrogen receptors (ER) in human tongue squamous carcinoma line Tca8113 cell. 3H-TdR incorporation and cell cycle analysis were used to examine the change of proliferation and DNA synthesis of Tca8113 cell.
RESULTSER-alpha and ER-beta mRNA were expressed in human tongue squamous cancer cell, and the expression of ER-beta was weaker than that of ER-alpha. beta-Estradiol at 10(-8) mol/L - 10(-6) mol/L could increase the proliferation of human tongue squamous carcinoma cell in a dose dependent manner (P < 0.01). beta-E2 (10(-6) mol/L) could increase the proportion of cells in S phase and G2 phase from 23.5% up to 37.7%. The effect of estradiol on the proliferation of cultured human tongue squamous cancer line Tca8113 cell could be inhibited by Tamoxifen.
CONCLUSIONThere are ER-alpha and ER-beta expression in human tongue squamous cancer line Tca8113 cell, and beta-estradiol promotes the proliferation and cell cycle of cultured human Tca8113 cell.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; metabolism ; Estrogen Receptor beta ; metabolism ; Humans ; Tamoxifen ; pharmacology ; Tongue Neoplasms ; metabolism ; pathology
9.Relationship and clinical significance of KiSS-1, nuclear factor kappa B (NF-kappaB), p50, and matrix metalloproteinase 9 expression in breast cancer.
Xue YI ; Chang-ying LI ; Su-hua ZHANG ; Xian-hua WANG ; Zhan-qing LI ; Fang YANG
Chinese Journal of Pathology 2008;37(4):238-242
OBJECTIVETo study the expression of KiSS-1, nuclear factor kappa B (NF-kappaB) p50 and matrix metalloproteinase 9 (MMP-9) in breast cancer tissue and the relationship with clinicpathological factors.
METHODSImmunohistochemical staining for KiSS-1, NF-KappaBp50, and MMP-9 protein was performed in 152 cases of human breast tissue [92 cases of BC, 30 cases of epithelial hyperplasia, and 30 cases of peritumoral breast tissue (PMT)] and 54 cases of axillary lymph node metastases. In-situ hybridization for KiSS-1 mRNA was done in 50 cases of breast cancer, and 20 cases of PMT.
RESULTS(1) The expression of KiSS-1 gene was significantly higher in well-differentiated breast cancer than in PMT, and this expression progressively decreased with decreasing degree of tumor differentiation, increasing pathological grade, TNM stage and the presence of lymph node metastases. The expression of KiSS-1 gene in lymph node metastasis was markedly lower than the corresponding primary tumor. There was correlation between the expression of KiSS-1 mRNA and KiSS-1 protein in breast cancer group. (2) The expression of NF-kappaKBp50 and MMP-9 increased progressively with decreasing degree of tumor differentiation, increasing TNM stage, large tumor size ( >2 cm) and the presence of lymph node metastases.
CONCLUSIONSThe expression of KiSS-1 protein showed negative correlation with that of NF-kappaBp50 and MMP-9 respectively. MMP-9 protein expression was positively correlated with NF-kappap50 protein expression. These suggest that the genes of KiSS-1, NF-kappaBp50 and MMP-9 could be involved in the progression and metastasis of breast cancer.
Breast Neoplasms ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Kisspeptins ; Lymphatic Metastasis ; physiopathology ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Statistics as Topic ; Tumor Suppressor Proteins ; genetics ; metabolism
10.Epidemiological survey on pre-hypertension and hypertension prevalence among adolescents aged 11 to 17 years in Shanghai.
Bin HONG ; Xue-juan JIN ; Yan-ling SU ; Lei XIAO ; Qian-fang CAI ; Hui-fei ZOU ; Jun-bo GE
Chinese Journal of Cardiology 2012;40(5):427-431
OBJECTIVETo assess the prevalence of pre-hypertension and hypertension as well as the association with overweight/obesity in Shanghai adolescents.
METHODSSchool children aged 11 - 17 years in four schools in Shanghai were included in this survey. All students were visited in May 2010 at school by trained nurses or physicians who administered a questionnaire and carried out anthropometric measurements. Anthropometric measurements included height, weight, heart rate and blood pressure. Repeat measurements were performed in school children with elevated blood pressure within one month. The pre-hypertension and hypertension was defined on the basis of the 2004 National High Blood Pressure Education Program Working Group definitions. The overweight and obesity was defined using cutoff points recommended by Working Group of Obesity, China (WGOC).
RESULTSThere were 4175 school children aged 11 - 17 years [mean (15.0 ± 1.9) years, 52.3% (n = 2183) girls and 47.7% (n = 1992) boys]. There were 72.5% (n = 3025) participants with normal blood pressure [ girls (n = 1666) 76.3% and boys (n = 1361) 68.3%], 18.0% (n = 750)participants with pre-hypertension [14.2% (n = 310) for girls and 22.1% (n = 440) for boys], 8.3% (n = 346) participants with stage 1 hypertension [8.2% (n = 179) for girls and 8.3% (n = 165) for boys] and 1.3% (n = 54) participants with stage 2 hypertension [1.3% (n = 28) for girls and 1.3% (n = 26) for boys]. The prevalence of overweight and obesity was 6.7% (n = 147) and 3.0% (n = 66) for girls and 13.7% (n = 273) and 6.3% (n = 125) for boys. After adjusting for gender, age, parental history of hypertension and physical activities, multivariable logistic regression analysis showed that the overweight [adjusted odds ratio and 95% confidence interval: 1.42 (1.16 - 1.75)] and obesity [adjusted odds ratio and 95% confidence interval: 2.35 (1.78 - 3.11)] were independent predictors of elevated blood pressure in this cohort.
CONCLUSIONSThe prevalence of elevated blood pressure is common in adolescents in Shanghai. Overweight and obesity are closely associated with the elevated blood pressure in this cohort.
Adolescent ; Child ; China ; epidemiology ; Epidemiological Monitoring ; Female ; Humans ; Hypertension ; epidemiology ; Male ; Obesity ; epidemiology ; Overweight ; epidemiology ; Prevalence ; Risk Factors