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Effect of glucagon-like peptide-1 agonist on expression of glucose transporter 4 in the skeletal muscle of rats with impaired glucose tolerance

Guangxia XI ; Yuanxing YANG ; Qizheng SUN ; Lingxia ZHAO ;

Journal of Endocrine Surgery.2013;7(3):187-190. doi:10.3760/cma.j.issn.1674-6090.2013.03.004

Objective To investigate the effect of glucagon-like peptide-1 agonist (exendin-4) on expression of glucose transporter 4 (GLUT4) in the skeletal muscle of rats with impaired glucose tolerance (IGT).Methodis 54 Wistar rats were randomly assigned into normal glucose tolerance group (NGT group,n =18) and impaired glucose tolerance group(IGT group,n =36).The rats in NGT group were fed with routine diet and the rats in IGT group were fed with high-sugar high-fat diet.At the 12th week,IGT models were tested successful.Then,half of the rats were allocated to intervention group (Ex group) and the rest were set as IGT control group.The rats in Ex group were subject to exendin-4 subcutaneous injection (5 μg/kg,twice daily).Each rat in NGT group and IGT control group was given the same volume of saline as injection.FBG and 2 h BG were measured before intervention and after 4 weeks.The expression of GLUT4 mRNA and GLUT4 in the skeletal muscle were respectively measured by real time quantitative polymerase chain reaction and immunohistochemistry.Inter-group comparison was conducted using analysis of variance (ANOVA) and least square deviation-test (LSD-t).Results Before intervention,compared with NGT-group,the 2 h BG in IGT control group and Ex group were higher,the GLUT4 mRNA of skeletal muscle in IGT control group and Ex group were lower (respectively P ＜ 0.05).The skeletal muscle cells in IGT control group and Ex group were less colored while the skeletal muscle cells in NGT group were colored extensively,and more colored granules.After 4 weeks of exendin-4 intervention,compared with IGT control group and Ex group of non-intervention,the 2 h BG level in Ex group was lower and the expression level of GLUT4 mRNA of skeletal muscle was higher (respectively P ＜ 0.05).After intervened with exendin-4 for 4 weeks,the GLU protein mainly expressed in cytoplasma of skeletal muscle cells.Its expression was higher in Ex group than in IGT group and in Ex group before intervention.Conclusion Exendin-4 may up-regulate the expression of GLUT4,increase glucose intake of the skeletal muscle,and reduce postprandial blood sugar.

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Detection of micro RNA in cervical cancer tissue and its clinical significance

Wenbo CHEN ; Liquan WANG

Journal of Endocrine Surgery.2014;(6):495-499. doi:10.3760/cma.j.issn.1674-6090.2014.06.016

Objective To investigate the expression level of miR-21, miR-146a and miR-224 and their clinical significances in cervical cancer .Methods miR-21, miR-146a and miR-224 were extracted from the normal cervical tissues and cervical cancer tissues , and their expression levels were quantified by real-time quan-titative polymerase chain reaction analysis .t test was used to analyze the expression difference of the two tissues and examine the associations between miR expression and the clinicopathological characters .The overall survival was analyzed by log-rank test, and survival curves were plotted according to Kaplan-Meier.Results miR-21 ex-pression levels of cervical cancer tissues and normal cervical tissues were 11.02 ±4.83 and 10.36 ±4.79, and miR-146a expression levels were 24.68 ±7.92 and 22.43 ±7.25, and there was no significant difference for both miR-21 and miR-146a expression between groups (P>0.05).miR-224 expression levels were 5.78 ±1.03 and 3.35 ±0.93 in cervical cancer tissues and normal cervical tissues , and the difference had statistical significance (P<0.001).For cervical cancer tissues, the miR-224 levels were significantly higher in tumors less differentia-ted, in advanced FIGO stage and with lymph nodes metastasis (P<0.05).miR-224 levels showed obvious corre-lation with survival time and the median survival time was shorter in patients with higher expression level of miR -224(P <0.05).COX multivariate analysis showed miR-224 expression level was an independent factor in patients'survival and prognosis ( P<0.05 ) .Conclusion miR-224 is independent predictor for aggressive pro-gression and poor prognosis in cervical cancer , and it might serve as a biomarker for predicting clinical outcome of cervical cancer patients .

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Correlations of the expression of HMGA1 protein with the aggressiveness and recurrence of pituitary ade-noma

Huiting OU ; Dewen YAN ; Mingming LIU ; Jian LI ; Weihua YIN ; Min ZUO ; Wei YIN

Journal of Endocrine Surgery.2014;(6):486-490. doi:10.3760/cma.j.issn.1674-6090.2014.06.014

Objective To observe the expression of high mobility group A 1(HMGA1)protein in pituitary adenoma(PA)and explore its relationship with the aggressiveness and recurrence of PA .Methods The labeling index(LI)of HMGA1, detected by immunohistochemistry staining , was compared between aggressive and non-ag-gressive PA, recurrent and non-recurrent PA.Results The expression of HMGA1 were observed in all 52 cases of PA and homogenously distributed in nuclear of tumor cells .The LI of HMGA1 showed no significant difference between functioning and nonfunctioning PA (P=0.79), as well as among different immunophenotypical variants of PA(P=0.28).The expression of HMGA1 was significantly higher in the aggressive PA than in the nonaggres-sive PA(0.50 ±0.20 vs 0.24 ±0.17, P=0.000).Patients with a recurrent PA had a higher value of HMGA 1 than patients with a non-recurrent PA(0.60 ±0.20 vs 0.24 ±0.18, P=0.000).The expression of HMGA1 in-creased stepwise with the tumor size, as the highest was present in macro-sized PA(0.52 ±0.20)and the lowest was in micro-sized PA(0.18 ±0.17).A similar trend was found in Hardy's grade I(0.18 ±0.17),II(0.30 ± 0.20)and III-IV(0.50 ±0.20)(P=0.003).The area under the ROC curve was 0.918(>0.90).When the LI reached 49.2%, the predicting specificity was 90.5%and the sensitivity was 71.4%.Conclusions The LI of HMGA1 has a positive correlation with the tumor size , aggressiveness and recurrence of PA and also has a valua-ble predicting efficacy for PA recurrence .The characteristics of HMGA 1 in PA can help pathologists make repro-ducible evaluation on prognosis of PA .

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Expression of caudal related homeodomain transcription 2 in colon cancer and its relationship with metas-tasis

Jian ZHANG ; Rongchao YING ; Wei WEI ; Jingjing XIANG ; Huicheng JIN ; Shenglin MA

Journal of Endocrine Surgery.2014;(6):479-481. doi:10.3760/cma.j.issn.1674-6090.2014.06.012

Objective To study the expression of caudal related homeodomain transcription 2 ( CDX2 ) protein in human colon cancer , and study the relationship between protein expression and colon cancer metasta -sis.Methods With immunohistochemical technique in peroxidase notation , pathological specimens of 80 cases of colon cancer were selected , including 47 cases of distal metastasis , and 33 cases without cancer metastasis . CDX2 protein expression in cancer tissue and the related data were statistically analyzed .Results Patients'sex, age and location of colon tumors had no statistical relation with CDX 2 protein expression(P>0.05).CDX2 pro-tein was statistically different in well-differentiated(100.00%), moderately-differentiated(80.00%), and poor-ly-differentiated(33.33%) tumor tissues.The difference had statistical significance (P<0.05), The expression of CDX2 protein reduced with the reduction in the degree of colorectal adenocarcinoma differentiation , rs =0.217, indicating that CDX2 protein expression was positively correlated with the malignant degree of colon canc -er.Positive CDX2 protein expression was significantly lower in metastatic colon cancer tissues (72.3%) than in non-metastatic tissues ( 90.9%) , and the difference had statistical significance .Lymph node metastasis could down regulate CDX2 protein expression in colorectal cancer .Conclusion CDX2 protein is positively correlated with the differentiation degree of colon cancer and distal metastasis can down regulate CDX 2 protein expression .

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Expression and clinical significance of zinc finger protein 217 in human pancreatic cancer

Tingting JI ; Qingying TAN ; Shenghua PAN ; Rongzhou LI ; Na LIN

Journal of Endocrine Surgery.2014;(6):475-478. doi:10.3760/cma.j.issn.1674-6090.2014.06.011

Objective To investigate the expression and clinical significance of zinc finger protein 217 (ZNF217)in human pancreatic cancer.Methods 43 cases with pancreatic cancer undergoing surgery in the PLA 117 Hospital and People's Hospital of Ruian City from Apr .2011 to May.2014 were enrolled in the study . The pancreatic cancer and the corresponding tumor-adjacent tissues were collected .Real-time quantitative PCR (qRT-PCR)was applied to detect ZNF217 mRNA expression in pancreatic cancer (n=43)and the corresponding tumor-adjacent normal tissues.The protein expression of ZNF217 was measured by immunohistochemistry(IHC). The relationship between the expression of ZNF 217 and clinical features was analyzed by pearson chi-square test . Results The expression level of ZNF217 mRNA and protein was significantly higher in pancreatic cancer tissues than in adjacent normal tissues(P<0.05).The high expression of ZNF217 protein was positively correlated with perineural invasion, tumor size, lymphatic metastasis and advanced TNM stage (P<0.05).Conclusions The expression of ZNF217 is significantly higher in pancreatic cancer tissues than in tumor-adjacent normal tissues , and the upregulation of ZNF 217 is associated with clinicopathological features of tumor malignance .ZNF217 may become a new marker and effective therapeutic target in early diagnosis of pancreatic cancer .

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Effect of high-flux hemodialysis on mineral and bone metabolism of patients with diabetic kidney disease undergoing maintenance hemodialysis

Xiangdong LIN ; Zhenfen HU ; Xiao JIN

Journal of Endocrine Surgery.2014;(6):456-458. doi:10.3760/cma.j.issn.1674-6090.2014.06.005

Objective To investigate the effect of high-flux hemodialysis on mineral and bone metabo-lism( MBD) in diabetic kidney disease patients with maintenance hemodialysis .Methods 60 cases with diabetic kidney disease undergoing maintenance hemodialysis were studied and they were randomly divided into two groups according to the number table method .Patients in each group were treated using either high-flux dialyzer or low-flux dialyzer for 6 months.Blood calcium(Ca), blood phosphorus(P), calcium*phosphorus(Ca*P), alkaline phosphatase(AKP), parathyroid hormone(PTH), and 25 hydroxy vitamin D3(25-OH-D3),etc were deter-mined at the start and end of the study .Results In high flux dialysis group , serum P, serum Ca*P, and PTH levels was(1.52 ±0.50)mmol/L,(3.05 ±1.19)pg/L and(368.61 ±235.32)pg/L respectively, which was sig-nificantly decreased than before treatment ((1.78 ±0.55)mmol/L,(3.94 ±1.31),(427.45 ±288.93)pg/L) (P<0.05), while serum Ca, and 25-OH-D3 levels were(2.3 ±0.16)mmol/L and(21.64 ±8.51)nmol/L respectively, which were increased significantly than before treatment ((2.12 ±0.18)mmol/L,(16.77 ±7.69) nmol/L)(P<0.05), yet in low flux dialysis group serum Ca, P, AKP, PTH, and 25-OH-D3 didn't change sig-nificantly before and after treatment ( P>0.05) .Conclusion High-flux dialyser provides a better effect on MBD for patients with diabetic kidney disease undergoing maintenance hemodialysis .

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Comparison of different multivessel revascularization methods in type 2 diabetes mellitus patients accom-panying with coronary artery disease

Hong ZHANG ; Kai HOU ; Hongliang CONG ; Ximing LI ; Zuocheng LI

Journal of Endocrine Surgery.2014;(6):452-455. doi:10.3760/cma.j.issn.1674-6090.2014.06.004

Objective To compare two different revascularization methods in type 2 diabetes mellitus (T2DM) patients with multivessel coronary artery disease treated by percutaneous coronary intervention (PCI) and coronary artery bypass graft ( CABG) .Methods T2DM patients with multivessel disease undergoing success-ful PCI or CABG were enrolled in the study .They were diagnosed by coronary angiography ( CAG) in Tianjin Chest Hospital from May 2009 to May 2010 whose.The patients were followed up for 3 years.The information of patients including physical performance , clinical features , and laboratory examination results were collected .The major ad-verse cardio cerebral events(MACCE)including death, myocardial infarction(MI), revascularization, angina pecto-ris, heart failure, and stroke were collected.Results During the 3 years of follow-up, MACCE(31.58% vs 17.68%, P<0.01), death(4.82%vs 1.10%, P<0.05), MI(4.39%vs 1.10%, P<0.05), angina pectoris (17.27%vs 10.50%, P<0.05)occurred more frequently in PCI group than in CABG group .Conclusion Evi-dences now tend to support CABG for revascularization in T 2DM patients with multivessel disease .

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Effects of embryonic fibroblasts on the aging-related phenotypes and dysfunction of diabetic ulcer fibro-blasts

Liu LIU ; Xue ZHANG ; Wentao ZHANG ; Shuhua NING

Journal of Endocrine Surgery.2014;(6):447-451. doi:10.3760/cma.j.issn.1674-6090.2014.06.003

Objective To observe the effects of human embryonic fibroblasts ( FBs ) on the biological characteristics of diabetic FBs in vitro , and to explore the possible therapeutic mechanism on diabetic ulcers . Methods Diabetic FBs were isolated and cultured with embryonic FBs (experimental group), adult FBs(control group 1)and alone(control group 2)in transwell chambers.Diabetic FBs growth curves in the coculture systems were described.The cell shape was observed by microscopy with HE staining .On the 4th and 7th day of cocul-ture, diabetic FBs were isolated to measure the expression levels of hydroxyproline and transforming growth factor -beta 1(TGF-β1)in the supernatant of monoculture .The cell senescense was displayed by β-galactosidase histo-chemical staining on the 3th, 5th and 7th day of coculture.Eight patients with diabetic foot ulcers who were can-didates for embryonic FBs constructs after failed traditional treatment were included in the study .The pre and post self-control study was conducted by observing the wound healing process before and after embryonic FBs treat -ment.Results Diabetic FBs in the experimental coculture system displayed more typical pattern and faster grow -ing rate.The level of hydroxyproline and TGF-β1 in the supernatant of diabetic FBs monoculture was significantly higher in the experimental group than in the control group after treated 4 or 7 days ( hydroxyproline:4 d, 7 d :compared with control group 1:P=0.023,P=0.007;compared with control group 2: P=0.007, P=0.003;TGF-β1 4 d, 7 d:compared with control group 1:P=0.000, P=0.000;compared with control group 2: P=0.000, P=0.000).The expression of SA-β-gal tended to be more apparent in control group 1 and 2 than in the experimental group on the 3rd,5th,and 7th day.All of the 8 cases with diabetic ulcers were cured by using em-bryonic FBs constructs .Conclusion Embryonic FBs can promote the proliferation of diabetic FBs , correct cell senescence and increase the production of hydroxyproline and TGF-β1,which maybe enhance the process of dia-betic wound healing .

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Expression of hepatic glucose transporter-2 and glucokinase after gastric bypass in rats with spontaneous type 2 diabetes mellitus

Jian XU ; Shan LIN ; Jiajun YIN ; Min YIN ; Wei WANG ; Zhequn NIE ; Yadong WANG ; Yulong HAN ; Huigeng ZHAO

Journal of Endocrine Surgery.2014;(6):443-446. doi:10.3760/cma.j.issn.1674-6090.2014.06.002

Objective To investigate the expression of hepatic glucose transporter-2 ( GLUT-2 ) and glu-cokinase(GCK)after gastric bypass(GBP)in type 2 diabetes mellitus(T2DM)GK rats and to discuss the mecha-nism of GBP improving insulin resistance .Methods 30 male GK rats aged 8 weeks were randomly divided into 3 groups:the operation group(10 rats), the sham operation group(10 rats)and the diet control group(10 rats), and 10 male SD rats aged 8 weeks were made as blank control group .The levels of fasting plasma glucose ( FPG) and fasting insulin(FINS)were measured and compared among the 4 groups before and 1,2 and 4 weeks after the operation and then the rats were sacrificed to retrieve the liver .The expressions of GLUT-2 and GCK mRNA and protein were detected by PT-PCR and Western blot respectively .Results As for GK operation group ,FPG levels at the 1st, 2nd,and 4th week after surgery ((11.06 ±0.52) mmol/L,(7.49 ±0.34) mmol/L,(5.20 ±0.08) mmol/L)respectively)were all lower than that before surgery (all P<0.05),and FINS level at the 4th week after surgery increased from(11.90 ±0.75)mmol/L to(14.20 ±1.23)mmol/L(P<0.05).The expressions of GLUT-2 and GCK mRNA and protein significantly increased in GK operation group 4 weeks after surgery ( P<0.01 ) . Conclusion The expressions of hepatic GLUT-2 and GCK in insulin signal transmission in T2DM rats are up-regu-lated after GBP , which enhance the signal transmission of insulin receptor , and improves the insulin sensibility .

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Construction of recombinant expression plasmid pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and production of the lentivirus in the immunotherapy for prostate cancer

Kuo YANG ; Ting ZHANG ; Yong XU ; Jiwu CHANG ; Yan LIU ; Zhihong ZHANG

Journal of Endocrine Surgery.2009;3(5):296-303. doi:10.3760/cma.j.issn.1674-6090.2009.05.003

Objective Our hypothesis was if we rendered host' s cytotoxic tumor lymphocytes insensitive to TGF-β,these immune cells could be able to overcome the TGF-β mediated immunosuppression and reject the tumor.We aimed to develop a lentivirus mediated gene transfer program incorporating a herpes simplex virus thymidine kinase(HSV-tk)in our dominant negative TGF-β type Ⅱ receptor(TβRⅡDNglytk)expression vector.So we first need to construct the lentiviral pLenti6/V5-D-TOPO vector containing TβRⅡDNglytk and produce the recombinant lentivirus as the transfection vector.Methods PCR were used to amplify the genes TβRⅡDN and HSV-tk from the respective plasmids.Then the genes were linked by recombinant PCR technology to construct the fusion gene TβRⅡDNglytk and control vector TRANSglytk.According to the operation manual from the Invitrogen company,TOPO cloning technology was used to construct the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk.Both of the constructed plasmids were verified by sequencing.ViraPowerTM Lentiviral System and 293 FT cells provided by Invitrogen were used to produce the recombinant lentivirus vector,the tillers of the lentivirus were determined by 293 cells.Results The construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk were completed succefully.The DNA sequencing results showed that both the plasmids were constructed correctly.Using them we successfully produced infectious lentivirus vectors with appropriate tilters.Conclusions Using TOPO cloning technology in the construction of the plasmids of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and pLenti6/V5-D-TOPO(R)-TRANSglytk and recombinant PCR in the fusion genes is feasible.Recombinant PCR combine with TOPO cloning technology can be a simple,highly efficient and rapid way to construct lentiviral vector.The production of infectious lentivirus with appropriate tilters using 293FT is suitable and feasible.The construction of pLenti6/V5-D-TOPO(R)-TβRⅡDNglytk and production of the infectious lentivirus will lay a foundation for immunotherapy of prostate cancer.

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