1.An investigation on the relationship between sulfonylurea receptor 1 gene intron 24-3t/c polymorphism and blood lipids in Chinese population
Yan FENG ; Qiongfang LI ; Changzhong CHEN
Chinese Journal of Diabetes 2000;0(05):-
Obeject To study the association of sulfonylurea receptor (SUR) 1 gene and blood lipids level.Method We investigated the SUR1 gene intron 24 3t/c polymorphism by polymerase chain reaction (PCR) and appropriate restriction enzyme (PCR RFLP) in 132 couples of type 2 diabetic case control and 282 type 2 diabetic pedigrees. The statistical methods were t test, multiple line regression and family based association test (FBAT).Result In the control group, the level of total cholesterol (TC), low density lipoprotein (LDL) and Apo B were higher in the cc genotype than that in the tt and the tc genotype. FBAT showed that sulfonylurea receptor 1 gene intron 24 3t/c polymorphism was significantly associated with TC, ApoB and BMI.Conclusion Sulfonylurea receptor 1 gene intron 24 3t/c polymorphism is associated with blood lipid level in north Chinese population.
2.Effect of fetal liver AFT024 cells on multidrug resistant gene 1 transfection efficiency and in vitro expansion of CD34~+ cells derived from umbilical cord blood
Hualing ZHANG ; Zeqing WEN ; Shoumin LAN ; Changzhong LI ; Jianfeng LI
Chinese Journal of Cancer Biotherapy 1994;0(01):-
Objective:To investigate the influence of fetal liver AFT024 cells on the transfection efficiency of multidrug resistant gene 1(MDR1)and the in vitro expansion of CD34+ cells derived from umbilical cord blood.Methods:CD34+ cells were isolated from human umbilical cord blood by MACS CD34 Progenitor Cell Isolation Kit and co-cultured with AFT024 cells(AFT024 group)or cultured alone(control group)for 7 days.During the subsequent 14 days,retrovirus carrying MDR1 gene was supplemented twice a week to transfect CD34+ cells.On the 7th,14th and 21st day after culture,the number of total nucleated cells(TNC)was counted,the ratio of CD34+ cells was assayed by flow cytometry(FCM)and the number of CD34+ cells was calculated,and colony-forming cells(CFC)were counted by methylcellulose cultures.RT-PCR method was used to detect the level of MDR1 mRNA in the transfected cells.The expression and function of P-glycoprotein(P-gp)were evaluated by FCM assay and Rhodamine-123 efflux assay,respectively.The gene transfection efficiency was calculated by drug-resistant colony-forming cells assay.Results:(1)The MDR1 mRNA level in AFT024 group than that in control group.The gene transfection efficiency in AFT024 group was significantly higher than that in control group(46.0% vs 15.2%,P0.05).On the 14th day,the expansion fold of TNCs in control group was significantly higher than that in AFT024 group(P0.05).The expansion folds of CD34+ cells and CFCs in the AFT024 group were significantly higher than that of the control group(P
4.Study on the treatment of high dose mifepristone and progesterone in endometrial carcinoma
Changzhong LI ; Zeqing WEN ; Shoumin LAN ; Jiayao WANG ; Ying LIU ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To investigate the effect of high dose mifepristone and high dose progesterone in the treatment of patients with endometrial carcinoma and to explore the possible mechanisms associating with them Methods Thirty untreated patients diagnosed as endometrial carcinoma through dilation and curettage of the uteri were divided into 3 groups at random Each group was given medroxyprogesterone acetate(MPA),(500 mg/day) or mifepristone(MIF),(100 mg/day)or MIF(100 mg/day)+ MPA(500 mg/day)for 5 days respectively On the sixth day, hysterectomy was performed on these patients The endometrial cancer specimen of post hysterectomy was compared with the one of pre administrating The morphologic changes of the endometrial cancer cells were observed through light microscope Immunohistochemistry assay (SP method) was applied to determine the localization and immunoreactive intensity of proliferating cell nuclear antigen(PCNA), estrogen receptor (ER), progesterone receptor (PR), B cell leukemia lymphoma 2 (bcl 2), bcl 2 associated X protein(bax) and CD 44 v6 Results Better differentiation degree and active excretion were observed in all of the post hysterectomy endometrial specimen In the same time, apoptosis of carcinoma cells was observed The most significant changes were seen in the MIF+MPA group In the MPA group,the pre treatment and post treatment expression of PR(2 9?1 1,1 6?0 8),ER(2 8?0 9,1 4?0 9),PCNA(0 84?0 10,0 60?0 12),bcl 2(0 236?0 089,0 157?0 981) and CD 44 v6 (4 6?1 8,2 5?1 9) were all decreased(all P 0 05) In the MIF+MPA group, the expression of PR(3 2?1 0,0 8?0 8),ER(2 7?0 9,0 7 ?0 9 ),PCNA(0 81?0 09,0 25?0 09),bcl 2(0 225?0 091,0 066?0 009)and CD 44 v6(4 5?1 9,2 7?1 6) were all decreased(all P
5.Role of DC-SIGN induced signaling pathway in the activation of HIV-1 virus
Jie LI ; Changzhong JIN ; Linfang CHENG ; Fumin LIU ; Nanping WU
Chinese Journal of Biochemical Pharmaceutics 2016;36(12):41-45
Objective To explore the mechanism of latent human immunodeficiency ciency virus type 1 (HIV-1) infection is unclear, especially in dendritic cells (DC).We hypothesized that DC-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) binds with HIV-1 may activate HIV-1 provirus.Methods We generated a model by transfecting 293T cells with a DC-SIGN expression plasmid and a HIV-1 5'long terminal repeat (LTR) reporter plasmid, and then stimulated the 293T cells with HIV-1 gp120 protein, wild-type HIV-1 and VSV-G-pNL4.3 pseudotype virus ( without gp120 protein).CEM-Bru cells were transfected with the DC-SIGN expression plasmid and stimulated by HIV-1 gp120 protein.Then HIV-1 replication was detected.The involvement of the ERK, p38 and NF-κB pathways signaling in this response were determined by inhibiting the pathways specifically and detecting the phosphorylation of the signaling kinase.Results The HIV-1 5'LTR was reactivated by HIV-1 gp120 in DC-SIGN-expressing 293T cells.After HIV-1 gp120 protein stimulation of the mold of CEM-Bru cells, the increasing expression of HIV-1 Tat mRNA and HIV-1 p24,which implies early and late HIV-1 provirus replication was reactivated by the HIV-1 gp120/DC-SIGN stimulation.HIV-1 gp120/DC-SIGN stimulation reactivates latent HIV-1 provirus via the NF-κB signal pathway.Conclusion HIV-1 gp120/DC-SIGN stimulation reactivates latent HIV-1 provirus via the NF-κB signal pathway.
6.Impact on ovarian reserve function by different homostasis methods during laparoscopic cystectomy in treatment of ovarian endometrioma
Changzhong LI ; Deying WEI ; Fei WANG ; Hongqing WANG ; Chunrun YANG
Chinese Journal of Obstetrics and Gynecology 2013;(1):11-15
Objective To investigate the impact on ovarian reserve function by different hemostasis methods during laparoscopic surgery in treatment of ovarian endometrioma.Methods From September 2008 to February 2010,162 cases with ovarian endometrioma undergoing laparoscopic surgery in Shandong Provincial Hospital were enrolled in this study.At the 3rd day of the menstrual cycle before surgery and the 1 st,3rd,6th and 12th cycle after surgery,serum FSH and anti-mullerian hormone(AMH) and ultrasound basal antral follicle count (AFC) and peak systolic velocity (PSV) were examined and compared.Based on hemostasis method,those patients were divided into 3 groups,including 54 cases in bipolar hemostasis,54 cases in ultrasonic scalpel hemostasis and suture after excision of endometrioma.Results (1) Before surgery:no significant different factors among three groups before surgery were observed,including age,size of endometrioma,the level of FSH,AMH,AFC,PSV (P > 0.05).(2) Ovarian reserve function after surgery:①FSH:at the 1st,3rd,6th and 12th month follow-up,the FSH in the bipolar group was (11.7 ±4.0),(9.9 ± 4.0),(9.5 ± 4.3),(9.5 ± 3.9) U/L,and the FSH in ultrasonic scalpel group was (11.4 ±4.3),(9.7 ± 4.0),(9.2 ± 3.7),(9.9 ± 4.6) U/L,were significantly higher than (9.3 ± 3.8),(6.7 ±3.0),(6.5 ± 3.2),(6.4 ± 2.2) U/L in suture group respectively (all P < 0.05).()AMH:at the 1 st,3rd,6th and 12th month follow-up,the AMH in the bipolar group was (1.8 ±0.9),(1.8 ± 1.0),(1.9 ±1.0),(2.0 ± 1.0) μg/L,and the AMH in the ultrasonic scalpel group was (1.6 ±± 0.8),(1.8 ± 1.0),(2.0 ± 1.1),(2.1 ± 1.0) μg/L,which were significantly lower than (2.8 ± 1.7),(2.9 ± 1.6),(3.0 ±1.3),(3.2 ± 1.5) μg/L in suture group,respectively (all P < 0.05).③AFC:there was no significant difference of APC among the three groups in the 1st month after surgery.However,at the 3rd,6th and 12th month follow-up,the AFC of 4.8 ± 1.4,5.9 ± 1.5,6.1 ± 1.5 in the suture group was significant higher than 3.7 ± 1.4,4.1 ± 1.4,4.0 ± 1.5 in bipolar group and 3.6 ± 1.3,4.0 ± 1.1,3.9 ± 1.5 in ultrasonic group,respectively (all P < 0.05).④PSV:at the 1 st,3rd,6th and 12th month follow-up,the PSV of the bipolar group(7.9 ±3.5),(8.1 ±3.3),(8.4 ±3.1),(8.6±3.0) cm/s in bipolar group and (8.1 ±3.5),(8.0 ± 3.0),(7.9 ± 3.2),(8.0 ± 2.9) cm/s in ultrasonic group were significant lower than (10.9 ± 3.3),(12.0 ± 3.2),(11.8 ± 3.0),(12.1 ± 4.1) cm/s in suture group,respectively.(allP<0.05).Conclusions Bipolar or ultrasonic scalpel hemostasis during laparoscopic excision of ovarian endometrioma is associated with a significant reduction in ovarian reserve.Electrocoagulation of the ovarian tissue should be avoided.
7.Construction of Iuciferase reporter vectors harboring DC-SIGN promoters without AP-1 or ETS-1 transcription factor binding site and detection of their activity
Changzhong JIN ; Jie LI ; Hangping YAO ; Nanping WU
Chinese Journal of Microbiology and Immunology 2009;29(12):1075-1079
Objective To detect the role of AP-1 or ETS-1 transcription factor binding sites (TFBS) in activity of DC-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) pro-moter. Methods Genomic DNA was extracted from peripheral blood. Complete DC-SIGN promoter and those without AP-1 or ETS-1 TFBS were amplified by PCR using designed primers. The PCR products were digested with MLu Ⅰ and Bgl Ⅱ and then ligated into MLu Ⅰ and Bgl Ⅱ digested pGL-3/Basic and pGL-3/En-hancer vectors. Transfection in Hacat and 293 cells was accomplished with Trans Fast liposome. Activity of luciferase was detected after 48 h. Results The DC-SIGN promoters without AP-1 or ETS-1 TFBS and the recombined pGL-3 vectors were correctly constructed. The activity of DC-SIGN promoters without AP-1 was reduced 20% (293) and 10% (Hacat), which was 40%-50% with enhancer. The activity of DC-SIGN pro-moters without ETS-1 was nearly vanished, no matter with or without enhancer. Conclusion ETS-1 TFBS, not AP-1 TFBS, plays an important role in activity of DC-SIGN promoter.
8.Preliminary study on inhibitory effects of sub-MIC matrine alone or in combination with erythromycin On Staphylococcus epidermidis biofilm
Yan GUAN ; Huana ZHOU ; Jingjin SHI ; Li LIU ; Chun LI ; Yun YUN ; Changzhong WANG
Chinese Journal of Microbiology and Immunology 2013;(2):86-90
Objective To observe the inhibitory effects of sub-MIC matrine alone and in combination with erythromycin on Staphylococcus epidermidis biofilms and their influences on morphological changes of the biofilms.Methods Minimum inhibitory concentrations (MIC) of matrine and erythromycin against Staphylococcus epidermidis were determined by the serial dilution method,antibacterial activity of matrine combined with erythromycin against planktonic S.epidermidis was evaluated by the checkerboard method.S.epidermidis biofilms were constructed in vitro,XTT reduction assay was used to evaluate influences of sub-MIC matrine alone and in combination with erythromycin on metabolism and adhesion of S.epidermidis biofilms,and scanning electronic microscope(SEM) was applied to observe the morphological and the structural changes of the biofilms.Results The MIC of erythromycin to S.epidermidis was 7.8125 μg/ml,while the MIC of matrine was greater than 1000 μg/ml,besides,a synergistic effect between erythronmycin and matrine on planktonic S.epidermidis was shown (FIC<0.5).The sub-MIC matrine had no significant inhibitory effect on adhesion of S.epidermidis,and also the combination of the two agents was better than was used alone.However,the sub-MIC matrine had inhibitory effects on metabolism and morphology of S.epidermidis biofilms,and the combination of the two agents was weaker than was used alone.Conclusion Both the sub-MIC matrine and erythromycin had a significant inhibitory effect on S.epidermidis biofilm formation.Combination of the two agents showed synergistic effects on plankton and adhesion of S.epidermidis,but showed no synergistic effect on metabolism and morphology of the biofilms.
9.Progress in study of self-assembling peptides.
Yuanwei CHEN ; Changzhong ZHANG ; Tianquan LI ; Changxiu WAN
Journal of Biomedical Engineering 2006;23(1):209-211
Self-assembly of peptides is ubiquitous in the body of creatures. The molecules of peptides combine with each other to form proteins with different functions through self-assembly. The formation of a specific conformation of one type of protein is owing to the self-assembly of its compositive amino acids. So, researchers can design self-assembly of peptides at the molecular level and can control its formation and configuration. It has the potential for application in the preparation of new medicines and biomaterials. In recent years, self-assembling peptides have been increasingly high-lighted and used to simulate the function of natural biomolecules, to synthesize peptide-medicine, and to serve as the carriers of medicine.
Biocompatible Materials
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chemical synthesis
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10.Association of GYS1 and beta(3)-AR gene with postprandial hyperglycemia and serum uric acid in type 2 diabetes mellitus.
Guoying WANG ; Qiongfang LI ; Tianhua NIU ; Changzhong CHEN ; Xiping XU
Chinese Medical Journal 2002;115(9):1308-1311
OBJECTIVETo determine the relationships of Met416Val and XbaI polymorphism of muscle glycogen synthase (GYS1) gene and Trg64Arg variant of the beta(3)-adrenergic-receptor (beta(3)-AR) gene with type 2 diabetes mellitus (DM) and its intermediate phenotypes in the Chinese population.
METHODSPolymerase chain reaction-oligonucleotide ligation assay and restriction fragment length polymorphism assay were used to evaluate the GYS1 and beta(3)-AR gene polymorphisms in 102 pairs of case-control Chinese spouses.
RESULTSSubjects with Met416Val variant had a significantly higher 2-hour post-glucose level than subjects without this variant had in diabetic group (P = 0.032). The Met416Val polymorphism of GYS1 gene was not significantly associated with the risk of type 2 DM (adjusted OR = 1.67; 95% CI: 0.73 - 3.81, P = 0.223). Subjects with Trp64Arg variant had a significantly higher serum uric acid level than subjects without this variant had in diabetic group (P = 0.034). The combination of BMI and Arg64 allele carrier of the beta(3)-AR gene increased the diabetic risk over four-fold (adjusted OR = 4.00; 95% CI: 1.53 - 10.45, P = 0.005).
CONCLUSIONSIn the Chinese population, Met416Val polymorphism is identified in a subgroup of diabetic subjects with high 2-hour post-glucose. It will explain why some diabetic patients appear to be genetically predisposed to developing high postpradial glucose level. The presence of the Arg64 allele in the beta(3)-AR gene may predispose patients to higher serum uric acid level.
Adult ; Aged ; Alleles ; Body Mass Index ; Diabetes Mellitus, Type 2 ; blood ; genetics ; Glycogen Synthase ; genetics ; Humans ; Hyperglycemia ; genetics ; Middle Aged ; Polymorphism, Genetic ; Postprandial Period ; physiology ; Receptors, Adrenergic, beta-3 ; genetics ; Uric Acid ; blood