1.Influence of jiaweibugan decoction on c-jun expression of sciatic nerve in experimental diabetic rats
Juan HUANG ; Zeqi CHEN ; Lili XIONG ; Jieyu LI
Journal of Chinese Physician 2008;10(11):1443-1446
Objective To observe the effect of Jiaweibugan decoction on c-jun mRNA expression of sciatic nerve in experimental di-abetic rat and explore the preventive and therapeutic effects of Jiaweibugan decoction on peripheral neuropathy in experimental diabetic rats. Methods The diabetic rat model, was established by streptozotocin (STZ). The rots were killed on the 4th or 8th week from the beginning of treatment, and c-jun mRNA of sciatic nerve was detected by reverse-transcriptase polymerase chain reaction (RT-PCR). Serum SOD was determined by xanthine oxidase method. Results The results of RT-PCR showed that c-jun mRNA expression of the model group on the 4th or 8th week was higher than that of the normal control group (P <0.05) , while those in model rats treated by Jiaweibugan decoction were markedly lower than those in non-treated model rats(P <0.05). The results showed that SOD of the model group on the 4th or 8th week was lower than that of the normal control group (P < 0.05) , while those in model rats treated by Jiaweibugan decoction were markedly higher than those in non-treated model rats(P < 0.05). Conclusion Jiaweibugan decoction has a preventive and therapeutic effect on peripheral neuropathy in experimental diabetic rats.
2.Influence of jiaweibugan decoction on VEGF expression of sciatic nerve in experimental diabetic rats
Jianyu FU ; Zeqi CHEN ; Juan HUANG ; Lili XIONG ; Xingping DAI
Journal of Chinese Physician 2008;10(8):1028-1031
Objective To observe the effect of jiaweibugan decoction on VEGF expression of sciatic nerve in experimental diabetic rat and explore the preventive and therapeutic effects of iiaweibugan decoction on peripheral neumpathy in~xperimental diabetic rats.Methotis The diabetic rat model was established by streptozotocin(STZ).The rats were killed on the 4th or 8th week from the beginning of treatment respectively,and VEGF mRNA of sciatic nerve was detected by reverse-transcriptase polymerase chain reaction(RT-PCR).Resuits The results of RT-PCR showed that VEGF expression ofthe model group and treated group on the4th or8th week WaS higherthan that of the normal control group(P<0.05).Compared with the model group,VEGF expression in the diabetic rats treated byjiaweibugan decoction increased markedly at the end of4th week(P<0.05),while decreaSed significantly at the end of8th week(P<0.05).Conclusion Jiaweibugan decoction has a proventive and therapeutic effect on peripheral neumpathy in experimental diabetic rats.
3.Influence of jiaweibugan decoction on the expression of JNK mitogen-activated protein kinase in sciatic nerve of experimental diabetic rats
Juan HUANG ; Zeqi CHEN ; Lili XIONG ; Jieyu LI
Journal of Chinese Physician 2009;11(2):156-159,169
Objective To observe the effect of jiaweibugan decoction on the expression of p-JNK mitogen-activated protein kinase in sciatic nerve of experimental diabetic rats and explore the preventive and therapeutic effects of Jiaweibugan decoction on peripheral neuropa-thy in experimental diabetic rats. Methods The diabetic rat model was established by streptozotocin (STZ). The rats were killed on the 4th or 8th week from the beginning of treatment, and the expression of p-JNK mitogen-activated protein kinase in sciatic nerve was detected by immunohistochemistry. Serum SOD was determined by xanthine oxidasemethod. Results The immunohistochemistry results showed that the expression of p-iNK in the model group on the 4th or 8th week was higher than that in normal control group (P<0.05) , while p-JNK in jiaweibugan decoction group was markedly lower than that in model rats(P<0.05). The results showed that SOD in the model group on the 4th or 8th week was lower than that in the normal control group (P<0.05) , while SOD in jiaweibugan decoction group was markedly high-er than that in model rats(P<0.05). Conclusion Jiaweibugan decoction has a preventive and therapeutic effect on peripheral neuropathy in experimental diabetic rats.
4.Construction and expression of the fusional genes Hsp65-Esat6 expressing plasmid of mycobacterium tuberculosis
Junwu LI ; Zeqi HUANG ; Cuichan YAO ; Shuguang ZHOU ; Xiaodong LI ; Dong SONG ; Qinghua HUANG
Chinese Journal of Tissue Engineering Research 2007;11(29):5836-5839
BACKGROUND:There is a linker between Hsp65 and Esat6 coding a segment of water repellent polypeptide.It is very gentle and easy to be folded,which is profitable to translate the keno-folding correctly between the two proteins and to make the space structure of fusional proteins consistent with those two native ones.Then,a correct structure is formed and the immunogenicity of the fusional proteins is improved.OBJECTIVE:This study was to clone the fusional genes Hsp65-Esat6 from mycobacterium tuberculosis(MTB)H37Rv, then to construct into a eukaryotic expressing vector which contains an enhanced green fluorescent protein(EGFP) reporter gene,and finally to identify the expression of Hsp65 protein and Esat6 protein by IHC methods.DESIGN:Single sample experiment.SETTING:Department of Microbiology & Immunology,Medical College of Jinan University.MATERIALS:Plasmid pVAE was donated by Professor Cao from South China University of Technology.MTB H37Rv,E.coil DH5 α,expressing plasmid pEGFP-C1 and Hela cells were donated by Doctor Hu Ping.METHODS:The experiment was conducted at the Department of Microbiology & Immunology and the Medical Experimental Center of Jinan University between September 2005 and June 2006.The whole-genome was extracted from MTB H37Rv by molecular cloning technique,and used it as template to amplify Hsp65 (no terminator) gene by polyrnerase chain reaction (PCR),to recombine it with pEGFP-C1 vector after purification to construct pEGHsp65(no terminator)recombination vector.pVAE vector was used as template to amplify Linker-Esat6 gene(with terminator)by PCR, and then to recombine it with pEGHSP65(no terminator)to construct an eukaryotic expressing vector pEGHsp65-Esat6 with the fusional genes Hsp65-Esat6 inside.Finally,genes Hsp65-Esat was checked by molecule biology methods such as PCR, restriction endonuclease and DNA sequencing.Hela cells were transfected with pEGHsp65-Esat6 and the expression of EGFP and the efficiency of transfection were observed.The expressions of Hsp65 protein and Esat6 protein were detected by IHC methods.MAIN OUTCOME MEASURES:①The size of pEGHsp65-Esat6.②The DNA sequencing result of the pEGFP-C1.③The expression of EGFP in the transfected Hela cells.④The IHC results of Hsp65 protein and Esat6 protein in Hela cells.RESULTS:①The size of pEGFP-C1 was 4.7 kb,that of pEGHsp65 was 6.4 kb,and that of pEGHsp65-Esat6 was 6.7 kb.There were differences between their speeds In agarose electrophoresis.②The results showed that it was the same as reported Hsp65 sequence and Esat6 sequence of MTB H37Rv.③After transfecting pEGHsp65-Esat6 for 24 hours,EGFP was found in 30% of Hela through Laser scanning confocal microscope. But there was no EGFP in non-transfected Hela.④Hsp65 protein and Esat6 protein with biological activities were detected in transfected Hela cells by IHC methods.CONCLUSION: Using Hsp65 and Esat6 as immunogens,we have successfully cloned and constructed a eukaryotic expressing vector which contain fusionaI genes Hsp65-Esat6 of MTB H37Rv and EGFP.
5.Clinical analysis of islet autoantibodies,hypersensitivity-CRP and renal function in elderly type 2 diabetic patients
Zhilan JIANG ; Aihong DENG ; Zeqi HUANG ; Caiyun HE ; Yongfen JIAN ; Yongjian SHEN
Chongqing Medicine 2013;(24):2861-2863
Objective To investigate the clinical value in changes of serum glutamic acid decarboxylase antibody (GAD-Ab) ,islet cell antibodies(ICA) ,insulin autoantibodies (IAA) and high-sensitivity C-reactive protein (hs-CRP) and renal function in elderly type 2 diabetic patients .Methods 122 cases of endocrine inpatient in our hospital had been diagnosed with type 2 diabetes were chosen from January 2012 to December 2012 .They were divided into islet autoimmunity antibody positive group (n=21) and islet autoimmunity antibody negative group (n=101) according to the antibody test results ,Fasting C-peptide(FCP) ,2-hour postprandial C-peptide(2 h CP) ,glycosylated hemoglobin(HbA1c) ,high-sensitivity-CRP(hs-CRP) and renal function[urea (UREA) ,creatinine (Cr) ,microalbuminuria(urinary mALB) ,urinary β2-microglobulin (urinary β2-MG)]were detected .Test results were statistically analyzed and compared .Results At least one Islet autoimmune antibodies were found in 21 cases of 122 elderly patients with type 2 diabetes .The positive rate was 17 .21% .GAD-Ab was detected positive in 14 cases(11 .47% ) ,ICA was detected positive in 10 ca-ses(8 .19% ) ,IAA was detected positive in 1 case(0 .82% ) ,Two antibodies were detected positive together in 4 patients(3 .27% ) , Three antibodies were not detected positive together .The levels of hs-CRP ,UREA and Cr in Islet autoantibodies positive group were higher then in islet autoimmunity antibody negative group ,the difference was statistically significant (P<0 .05) .The levels of FCP ,2 h CP ,HbA1c ,urinary mALB and urinary β2-MG in both group ,the difference was not statistically significant (P>0 .05) . Conclusion Chronic inflammation and the appearance of islet autoantibodies are closely related to the damage of islet cell function . It has a higher value in monitoring complications and efficacy through understanding islet autoantibodies ,inflammation and changes in renal function in elderly type 2 diabetes .
6.Effects of apolipoprotein (a) on vascular smooth muscle cells proliferation and cell signal transduction pathway
Chunying WEI ; Menghong WANG ; Yunfeng WEI ; Zeqi ZHENG ; Jingtian PENG ; Jun HUANG ; Yuan WEN ; Zhiyong WU
Chinese Journal of Pathophysiology 2009;25(11):2122-2125
AIM: To investigate the effect and the mechanism of apolipoprotein (a) [apo (a) ] on proliferation of vascular smooth muscle cells ( VSMCs). METHODS: All VSMCs used in experiments were serial subcultured from primary cells and were identified by immunohistochemistry staining of a - actin. Cell growth assay was observed as cell counting and MTT assay. Western blotting was also employed to detect the related mechanism. RESULTS: All cells used in experiments were confirmed as VSMCs. Although apo (a) enhanced VSMCs proliferation, this effect was attenuated by anti -integrin α_vβ_3, LM609.Use these reagents alone had no effect on VSMCs growth. The results of Western blotting demonstrated that focal adhesion kinase (FAK) was activated by apo (a) and the expression of total or phosphorylated transforming growth factor β_1 (TGF -β_1) was also decreased. However, these effects described above were all blocked by LM609.CONCLUSION: Apolipoprotein (a) enhances VSMCs proliferation and this effect is mediated by integrin α_vβ_3, which activates FAK and attenuates TGF - β_1 and phospho -TGF - β_1 expression.
7.Effect of devated plasma lipoprotein (a) on restenosis after stent implantation
Chunying WEI ; Yunfeng WEI ; Menghong WANG ; Zeqi ZHENG ; Jingtian PENG ; Jun HUANG ; Bingong LI ; Yuan WEN ; Zhiyong WU
Clinical Medicine of China 2009;25(10):1027-1029
Objective To investigate the association between plasma lipoprotein (a) [LP (a)] concentra-tion and in-stent restenosis after coronary stent implantation. Methods 152 patients with successful elective coro-nary stont implantation and percutancous transluminal coronary angioplasty (PICA) undergoing foUow-up angiogra-phy were retrospectively analyzed. These patients were divided into restenosis group( n = 29) and no-restenosis group (n = 123 ). The serum LP (a) levels of all patients were also investigated. The general clinical data were analyzed. Multivariate logistic regression was used for statistical analysis. Results We compared the serum Lap (a) levels, smoking and diabet in the two groups, and there was a statisticaLly significant difference between the restenosis group and no-restenosis group(P<0.05). Multivariate logistic regression showed that the elevation of Lap (a) level re-mained as an independent predictor of restenosis (RR =2. 648,95% CI 1. 066-6. 575,P <0. 05). Other risk fac-tors,such as smoking(P =0.023) ,diabet(P =0. 036) and the type of stent(P = 0.011 ) were also correlated with restenosis. Conclusions High plasma LP (a) concentration is an independent predictor of stent restenosis after stent implantation.
8.Genotypes and phenotypes of rare thalassemia in Foshan, Guangdong Province
Lin ZOU ; Shanghua CHEN ; Weixian XIE ; Zeqi HUANG ; Caiyun HE ; Yongfen JIAN
Chinese Journal of Endemiology 2020;39(11):791-795
Objective:To analyze the gene mutation types and haematological characteristics of αβ compound thalassemia, non-delectable α-thalassemia and Hemoglobin H Disease (HbH disease) in Foshan.Methods:Using the method of retrospective analysis, we selected the population who had been tested for thalassemia gene in Foshan Second People's Hospital Affiliated to Southern Medical University from January 2011 to November 2019. Sysmex XT-5000 automatic hematology analyzer was used for routine blood analysis. α-, β- thalassemia genes were detected by PCR + diversion hybridization method.Results:A total of 4 563 people were tested, of which 1 829 were diagnosed as thalassaemia through genetic diagnosis. αβ compound thalassaemia was detected in 81 cases with a positive rate of 1.8%; non-delectable α-thalassemia was detected in 18 cases with a positive rate of 0.4%; HbH disease was detected in 23 cases with a positive rate of 0.5%. The most common genotypes of αβ compound thalassemia were -- SEA/αα\β41-42/βN (17.3%, 14/81), -α 3.7/αα\β41-42/βN (14.8%, 12/81), -- SEA/αα\β654/βN (11.1%, 9/81). The main manifestations of hematology were normal to mild anemia (93.8%, 76/81). Only β-thalassemia with double heterozygotes and α-thalassemia showed severe anemia. αα CS/αα\βN/βN genotypes were common in the local non delectable α-thalassemia (50.0%, 9/18), and the non delectable α-thalassemia was characterized by non-positive phenotype or typical small-cell hypochromatosis in hematology. The genotypes of local HbH patients were -α 3.7/-- SEA\βN/βN (65.2%, 15/23), and simple HbH manifested as moderate anemia (87.0%, 20/23). Patients with HbH disease and β-thalassemia had normal or mild anemia (13.0%, 3/23). Conclusions:The genotypes of αβ compound thalassemia in Foshan area are diverse and complex, and hematology mainly manifests as mild anemia or normal. Non-delectable α-thalassaemia is common in the genotype of αα CS/αα\βN/βN, and clinical manifestations are asymptomatic gene carriers. The genotype of local HbH patients is mainly -α 3.7/-- SEA\βN/βN, and the hematology mainly shows moderate anemia.
9.Isotoosendanin exerts inhibition on triple-negative breast cancer through abrogating TGF-β-induced epithelial-mesenchymal transition via directly targeting TGFβR1.
Jingnan ZHANG ; Ze ZHANG ; Zhenlin HUANG ; Manlin LI ; Fan YANG ; Zeqi WU ; Qian GUO ; Xiyu MEI ; Bin LU ; Changhong WANG ; Zhengtao WANG ; Lili JI
Acta Pharmaceutica Sinica B 2023;13(7):2990-3007
As the most aggressive breast cancer, triple-negative breast cancer (TNBC) is still incurable and very prone to metastasis. The transform growth factor β (TGF-β)-induced epithelial-mesenchymal transition (EMT) is crucially involved in the growth and metastasis of TNBC. This study reported that a natural compound isotoosendanin (ITSN) reduced TNBC metastasis by inhibiting TGF-β-induced EMT and the formation of invadopodia. ITSN can directly interact with TGF-β receptor type-1 (TGFβR1) and abrogated the kinase activity of TGFβR1, thereby blocking the TGF-β-initiated downstream signaling pathway. Moreover, the ITSN-provided inhibition on metastasis obviously disappeared in TGFβR1-overexpressed TNBC cells in vitro as well as in mice bearing TNBC cells overexpressed TGFβR1. Furthermore, Lys232 and Asp351 residues in the kinase domain of TGFβR1 were found to be crucial for the interaction of ITSN with TGFβR1. Additionally, ITSN also improved the inhibitory efficacy of programmed cell death 1 ligand 1 (PD-L1) antibody for TNBC in vivo via inhibiting the TGF-β-mediated EMT in the tumor microenvironment. Our findings not only highlight the key role of TGFβR1 in TNBC metastasis, but also provide a leading compound targeting TGFβR1 for the treatment of TNBC metastasis. Moreover, this study also points out a potential strategy for TNBC treatment by using the combined application of anti-PD-L1 with a TGFβR1 inhibitor.