1.Expression of transient receptor potentials of vanilloid subtype 1 and pain in endometriosis
Nan SONG ; Jinhua LENG ; Jinghe LANG
Chinese Journal of Obstetrics and Gynecology 2012;47(5):333-336
Objective To investigate the expression of transient receptor potential vanilloid subtype 1 (TRPV1) in uterosacral ligament and its correlation with pain in endometriosis.Methods Total of 54 patients undergoing endometriotic lesions excision in uteroscaral ligament by laparoscopy due to pelvic pain were enrolled in this study.According to visual analogue scale(VAS) scores,27 patients with VAS 5 - 10 were in group A and 27 patients with VAS 0 - 4 were in group B.In the mean time,20 patients with dysmenorrhea without endometriosis (VAS:0 -4 ) were matched as group C.Specimens (including the sacro-ligaments of 20 women without endometriosis) were immunostained with specific antibodies of TRPV1.Western blot and real time PCR were performed to detect TRPV1 expression in endometriosis lesions and control group.Results( 1 ) Immunohistochemnistry:the positive area of TRPV1 was found in endometriotic lesions in uterosacral ligament in group A,B and tissue of uterosacral ligament group C.The semi-qualitification of TRPV1 expression were 3 in group A, 1 in group B and 1 in group C by immunohistochemistry staining.There was significantly different expression between group B and group A ( P =0.005 ) or group C ( P =0.027 ).(2) mRNA expression:the expression of TRPV 1 was 1.84 in group A,0.80 in group B,0.24 in group C,respectively.With higher VAS scores,the expression of TRPV1 exhibited increasing trends.The expression of TRPV1 mRNA was higher in group A than thai in group B ( P =0.022).There was statistically different expression between group B and group C ( P =0.031 ).( 3 ) Western blot:the expression of TRPV1 protein was 0.63 in group A,0.19 in group B,0.02 in group C.There was significant differences between group A and group B ( P =0.022 ),and between group B and group C (P < 0.01 ).Conclusion The expression of TRPV1 was correlated with the degree of pain in patient with endometriosis.
2.The relationship between soluble CD147 and the risk factors of atherosclerosis in rheumatoid arthritis
Honghong YUE ; Ping ZHU ; Nan LENG ; Zhenbiao WU ; Hongming LI
Chinese Journal of Rheumatology 2008;12(8):515-518
Objective To study the relationship between soluble CD147 (sCD147) level in peripheral blood and serum lipid level and explore the effect of sCD147 on atherosclerosis in rheumatoid arthritis (RA). Methods The level of sCD147 in 36 patients with RA,36 patients with coronary artery disease (CAHD) and 30 healthy volunteers was detected by enzyme linked immunosorbent assay (ELISA) .The disease activity score (DAS28) in RA patients was evaluated and the correlation between sCD147 level and DAS28 score was analyzed.The serum lipid level of RA patients was detected by an automatic biochemical analyzer and the cor relation between sCD147 level and serum lipid level was analyzed.Results The level of sCD147 in serum of RA patients was significantly higher than that in patients with CAHD and healthy volunteers,sCDI47 level in the RA group with high DAS28 score was significantly higher than that with low or medium DAS28 score.In RA patients,elevated total cholesterol (TC) and triglyceride (TG) level was positively correlated with serum sCDI47 level (r=0.84,P<0.05;r=0.87,P<0.05;while slightly elevated,normal TC and normal TG had no correlation with serum sCDI47 level (r=0.41,P=0.21;r=0.14,P=0.57;r=0.49,P=0.87).Elevated or slight ly elevated LDL-C was positively correlated with serum sCD147 level (r=0.86,P<0.05;r=0.81,P<0.05), while no correlation could be found in the group with normal LDL-C level (r=0.78,P=0.22).The high density lipoprotein-cholesterol (HDL-C) level decrease in RA patients had no correlation with serum sCD147 level (r--0.04,P=0.96;r=0.13,P--0.87).Conclusion sCD147 may be involved in the pathogenesis of RA and associate with disease activity.Elevated sCD147 level may be associated with abnormal serum lipid in RA.
3.Clinical analysis of 6 patients with drug-induced lupus
Nan ZHANG ; Xiaomei LENG ; Xinping TIAN ; Yan ZHAO ; Xiaofeng ZENG
Chinese Journal of Internal Medicine 2016;55(3):211-215
Objective To improve the understanding of drug-induced lupus (DIL) and the differences from systemic lupus erythematosus (SLE).Methods Clinical manifestation and treatment of patients with definite DIL were retrospectively analyzed.Results Six patients with DIL were enrolled in this study,including 4 females and 2 males.Two patients were diagnosed after receiving interferon,one after soluble tumor necrosis factor receptor fusion protein,one after propylthiouracil,one after penicillamine,and one after levofloxacin.High titer of antinuclear antibody was identified in all six patients,including 3 with positive anti-dsDNA antibody.One patient had positive anti-Sm antibody.One patient had positive anti-RNP antibody.One patient had anti-nucleosome antibody.One patient had anti-histone antibody.One patient had antimitochondrial antibodies-M2,and one patient had anticardiolipin antibodies.Conclusion Patients with DIL are not as severe as those with SLE.After cessation of suspected drugs and administration of standard treatment,the clinical outcome of DIL is satisfying.
4.Practice and insights of a healthcare alliance development: experiences of a hospital in Xinjiang
Ling TANG ; Zhiwei LENG ; Yuehong TANG ; Nan ZHANG ; Xiaoming JIANG
Chinese Journal of Hospital Administration 2017;33(8):569-573
The paper covered dominant models and organization of healthcare alliances in Xinjiang, illustrating the hospital group model, synergy development model, focused partnership support model, three-level integration model, and other business models.As described by the authors, healthcare alliances in Xinjiang, thanks to telemedicine, have achieved initial success by means of disciplines support, primary care human resources, new technologies and new service spreading, and promotion of appropriate medical techniques, in such aspects as regional medical cooperation, population benefits and medical resources sharing.
5.Expression of mitogen-activated protein kinase and its upstream regulated signal in human hepatocellular carcinoma.
Jiye ZHU ; Xisheng LENG ; Nan DONG ; Yannan LIU ; Guangming LI ; Ruyu DU
Chinese Journal of Surgery 2002;40(1):1-16
OBJECTIVETo detect protein expression of ERK(1), ERK(2), JNK(1), p38 and MEK(1), MEK(2) in human hepatocellular carcinoma and adjacent non-neoplastic liver.
METHODSIn 16 surgically resected hepatocellular carcinoma and para-carcinoma tissues, Western blotting was used to detect expression of ERK(1), ERK(2), JNK(1), p38 and MEK(1), MEK(2).
RESULTSIn all cases, ERK(1), ERK(2), p38 expression in hepatocellular carcinoma was significantly higher than that in para-carcinoma: integral optic density (IOD) of ERK(1) was 300 +/- 98 in carcinoma and 98 +/- 48 in para-carcinoma tissues (t = 2.519, P < 0.01); IOD of ERK(2) was 587 +/- 83 in carcinoma and 232 +/- 96 in para-carcinoma tissues (t = 2.745, P < 0.01); IOD of p38 was 270 +/- 85 in carcinoma and 107 +/- 88 in para-carcinoma tissues (t = 2.491, P < 0.01). JNK(1) expression in hepatocellular carcinoma was significantly lower than that in para-carcinoma; IOD of JNK(1) was 111 +/- 93 in carcinoma and 292 +/- 109 in para-carcinoma tissues (t = 2.473, P < 0.01). Protein levels of MEK(1) and MEK(2) in carcinoma were significantly higher than in para-carcinoma. IOD of MEK(1) was 1 418 +/- 244 in carcinoma and 806 +/- 90 in para-carcinoma tissues (t = 2.546, P < 0.01). IOD of MEK(2) was 1 041 +/- 122 in carcinoma and 468 +/- 40 in para-carcinoma tissues (t = 2.861, P < 0.01).
CONCLUSIONSERK(1), ERK(2), MEK(1) and MEK(2) in the signal transduction pathway for cell proliferation are significantly overexpressed and the expression of JNK(1) is lower in hepatocellular carcinoma. Their unbalance is one of the important reasons for the over growth and infinite proliferation of the hepatocellular carcinoma cell. The p38 and JNK(1) may be activated by different pathway.
Adult ; Aged ; Carcinoma, Hepatocellular ; enzymology ; Enzyme Activation ; Female ; Humans ; JNK Mitogen-Activated Protein Kinases ; Liver Neoplasms ; enzymology ; MAP Kinase Kinase 1 ; Male ; Middle Aged ; Mitogen-Activated Protein Kinase Kinases ; analysis ; Mitogen-Activated Protein Kinases ; metabolism ; Protein-Serine-Threonine Kinases ; analysis
6.LBP regulates PI3 K/Akt/eNOS signaling pathways in ovariectomized rat myocardium to exert antioxidative effect
Ning YU ; Fang YANG ; Xue LENG ; Ni ZHANG ; Junyan WANG ; Nan SONG
Chinese Journal of Pathophysiology 2016;32(8):1370-1375
AIM:To observe the influence of Lycium barbarum polysaccharide (LBP) on the PI3K/Akt/eNOS signaling pathways in ovariectomized rat myocardium .METHODS:Female SD rats (n=30) were divided into sham oper-ation group , ovariectomized group , progynova group , high-dose LBP group and low-dose LBP group .The serum levels of estradiol, lactate dehydrogenase (LDH) and creatine kinase (CK) were measured by ELISA.The myocardial contents of H2 S and oxidative stress injury-related indicators were also detected .The morphological changes of the myocardium were observed with HE staining.The expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium was deter-mined by Western blot .RESULTS: Compared with sham operation group , the serum level of estradiol , the content of H2 S, the activity of GSH-Px, and the expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium in ovariectomized group were all decreased , and the levels of ROS and MDA in the myocardium were increased (P<0.05). The serum levels of LDH and CK were also increased .The arrangement of the myocardial cells was disordered , and the in-tercellular space was also increased in the ovariectomized group .Compared with ovariectomized group , the serum level of estradiol, the myocardial levels of H2S and GSH-Px, and the protein levels of eNOS and phosphorylated Akt were all in-creased in high dose group, while the levels of ROS and MDA in the myocardium were decreased (P<0.05).The serum levels of LDH and CK were also decreased .The morphological changes of the rat myocardium were improved in high dose group.CONCLUSION: LBP prevents and treats postmenopausal cardiovascular lesions through regulating PI 3K/Akt/eNOS signaling pathways in ovariectomized rats .
7.Inhibition of HepG2 cell growth by LHRH-MPG△NLS/CDK2-siRNA nanoparticles
Xinxiao QIAO ; Nan SHAO ; Xia DONG ; Lanxia LIU ; Dunwan ZHU ; Xigang LENG
International Journal of Biomedical Engineering 2014;37(2):85-88
Objective To detect the size distribution and Zeta potential of LHRH-MPG△NLS/CDK-siRNA nanoparticles,to observe the effect of different solvents on the nanoparticle size,and to investigate the inhibitory effect of nanoparticles on HepG2 cell growth.Methods LHRH-MPG △NLS and CDK2-siRNA were mixed by continuous stirring to form nanoparticles at different N/P ratios (10/1,20/1 and 40/1).The size distribution and Zeta potential of LHRH-MPG△NLS/CDK2-siRNA nanoparticles were detected by dynamic light scattering,and the stability of the nanoparticles in normal saline,10% glucose and pure water was discussed.Finally,the inhibitory effect of the nanoparticles on HepG2 cells was determined by CCK8 kit.Results The mean size of the nanoparticles was within 200 nm,and the Zeta potentials were (70±5) mV (N/P=10/1),(120±5) mV (N/P=20/1) and (130±5) mV (N/P=40/1),respectively.The size of the nanoparticles in normal saline was significantly increased,which demonstrated that strong electrolytes had a great impact on the nanoparticles size.When nanoparticle concentration was 200 nmol/L,LHRH-MPG△NLS/CDK2-siRNA nanoparticles (N/P=10/1) showed significantly inhibitory effect on HepG2 cell growth.Conclusions The mean size of the LHRH-MPG△NLS/CDK2-siRNA nanoparticles was within 200 nm,which was ideal for cellular uptake.The Zeta potential of nanoparticles revealed that nanoparticles could be stable in aqueous solution,while strong electrolytes would affect nanoparticle size.When nanoparticle concentration was 200 nmol/L,LHRH-MPG△NLS/CDK2-siRNA nanoparticles (N/P=10/1) showed significantly inhibitory effect on HepG2 cell growth.
8.Serum level of soluble interleukin-2 receptors in hepatitis B patients
Jie HAN ; Long-Bang CHEN ; Hui XIE ; Nan LENG ; Shu-Kuan WANG ;
Chinese Journal of Immunology 1985;0(06):-
The level of serum soluble interleukin-2 receptor(sIL-2R)was measured in 103 patientswith hepatitis B and 26 hepatitis B virus(HBV)carriers by enzyme-linked assay.The sIL-2Rconcentration were elevated significantly in each type of hepatitis B patients and HBV carriers,compared with control group(P
9.Directed evolution of D-lactonohydrolase by error prone PCR and DNA shuffling.
Zhi-Qiang LIU ; Zhi-Hao SUN ; Pu ZHENG ; Yong LENG ; Jia-Nan QIAN
Chinese Journal of Biotechnology 2005;21(5):773-781
D-lactonohydrolase is useful in the procedure of resolution of racemic pantolactone to produce D-pantolactone, but the activity and stability under low pH of the wild type enzyme is not satisfactory enough to be applied to industrial production. The expected properties of wild type enzyme were enhanced by directed evolution. According to the formation of products and pH indicators, a screening system was designed. After three sequential error prone PCR and one round DNA shuffling followed by screening, Mut E-861, the best mutant with improved activity and stability under low pH situation was obtained. Gene analysis of the Mut E-861 mutant indicated that the mutant enzyme had A352C, G721A mutations and a silent mutation of position 1038. Moreover, the activity and stability of Mut E-861 were determined. The results showed that the activity of this mutant was 5.5-fold higher than that of wild type, and the stability under low pH was improved at no expense of D-lactonohydrolase activity. After incubated at pH 6.0 and pH 5.0 the activity of D-lactonohydrolase could be retained 75% to 50%, however, compared with 40% to 20% for wild type.
Carboxylic Ester Hydrolases
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biosynthesis
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genetics
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DNA Shuffling
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Directed Molecular Evolution
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Enzyme Stability
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Escherichia coli
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enzymology
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genetics
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Mutagenesis, Site-Directed
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Mutant Proteins
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genetics
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metabolism
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Polymerase Chain Reaction
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methods
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Protein Engineering
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Saccharomyces cerevisiae
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enzymology
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genetics
10.Screening of a novel CRM1 targeting inhibitor and its effects on the proliferation and growth of extranodal NK/T cell lymphoma
Meishuo LIU ; Haina WANG ; Feida LONG ; Nan LENG ; Yongliang YANG
Journal of China Pharmaceutical University 2020;51(1):44-51
The purpose of this study was to screen out the novel chromosome maintenance protein 1(CRM1)covalent targeting inhibitors by computer-assisted drug design(CADD), and to study their effects on the proliferation of extranodal nature killer/T cell lymphoma(ENKTL). A novel CRM1 inhibitor LFS-829 was designed based on the molecular structure of LFS-01 by means of ADME/T and covalent docking. The target binding of LFS-829 with CRM1 was analyzed by MALDI-TOF mass spectrometry. The effects of LFS-829 on the proliferation of extranodal NK/T cell lymphoma SNK6 and HANK-1 cells were detected by CCK-8. The cell morphology was observed by live cell workstation. Immunofluorescence experiments were used to analyze the effect of LFS-829 on nuclear export function of CRM1. The changes of NF-κB signaling pathway under different concentrations of LFS-829 were analyzed by Western blot, dual luciferase reporter gene assay and enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry, and the expression of proteins related to apoptosis pathway was detected by Western blot. Tests of peripheral blood mononuclear lymphocyte(PBMC)toxicity, platelet toxicity and mouse acute toxicity were done to make sure that it is not harmful to human. LFS-829 could bind covalently to the cysteine residue of the hydrophobic active pocket of CRM1. LFS-829 could selectively kill SNK6 and HANK-1 cells, with IC50 of 366 nmol/L and 158 nmol/L in 72 h, respectively, and cell morphology was significantly changed. LFS-829 at 800 nmol/L significantly inhibited the nuclear export function of CRM1, promoted nuclear assembly of IκB-α, down-regulated the transcriptional activity of NF-κB signaling pathway, significantly up-regulated the expression of apoptotic pathway protein p53, cleaved Caspase 3 and cleaved Caspase 9 and induced apoptosis, with no obvious killing effect on PBMC or platelets. It did not cause substantial tissue damage to mice at the high dose of 300 mg/kg, which shows its great prospect of future application.