2.Effects of recombinant gene lentivirus containing LIM mineralization protein-1 on proliferation effect and expression of bone marrow mesenchymal stem cells in rats.
Chang-sheng LIANG ; Chuan XIANG ; Zeng-yong WEI ; Hui-ming HOU ; Ying-ze QIN ; Xiao-chun WEI
China Journal of Orthopaedics and Traumatology 2013;26(12):1023-1027
OBJECTIVETo explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC.
METHODSSix clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot.
RESULTS1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa).
CONCLUSIONThere is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Cytoskeletal Proteins ; genetics ; metabolism ; Female ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; LIM Domain Proteins ; genetics ; metabolism ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; virology ; Osteoporosis ; genetics ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley
5.Combined assay of serum prostate specific antigen and chromogranin A helps diagnosis of prostate cancer.
Chang-Cheng SUN ; Wei ZHENG ; Chui-Ze KONG ; Xia WANG ; Chun-Ming YANG
National Journal of Andrology 2006;12(10):904-909
OBJECTIVETo evaluate the significance of the combined assay of chromogranin A (CgA) and prostate specific antigen (PSA) in the diagnosis of prostate cancer.
METHODSSerum CgA and PSA were detected by ELISA technique in 55 cases of prostate cancer (PCa), 25 cases of benign prostate hyperplasia (BPH), and 50 cases of normal subjects (control).
RESULTSThe serum CgA level in the PCa group was significantly higher than those in the control and BPH groups (P < 0.05), and increased with clinical stages. The parallel and serial tests associated with serum PSA and CgA raised the rate of detection of prostate cancer.
CONCLUSIONThe combined assay of serum PSA and CgA is of significant clinical value in raising the rate of diagnosis of prostate cancer, as well as in staging and prognosing the disease.
Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Case-Control Studies ; Chromogranin A ; blood ; Enzyme-Linked Immunosorbent Assay ; Humans ; Male ; Middle Aged ; Prognosis ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; diagnosis ; Sensitivity and Specificity
6.Comparison of clinical efficacy of orthotopic ileal neobladder versus orthotopic sigmoid neobladder
Jian-Song WANG ; Hong-Yi XU ; Yong-Fu SHI ; Hui ZHAN ; Jong-Ming LI ; Ze-Hui LI ; Yi-Gang ZUO ; Delin YANG ; Chao WANG ; Chang-xing KE ; Ming-xia DING ; Ru-ping YAN
Chinese Journal of Urology 2000;0(12):-
Objective To compare the clinical efficacy of orthotopic ileal neobladder versus ortho- topic sigmoid neobladder.Methods The data of 96 patients who had undergone orthotopic ileal neoblad- der and 68 patients who had undergone orthotopic sigmoid neobladder were retrospectively analyzed.The perioperative condition,urinary continence,urodynamics,and pouch-related complications were compared between the 2 groups.Results Of all the 164 patients,12(7.3%)were lost to follow-up.The mean fol- low-up was 46(2-86)months in orthotopic ileal neobladder group,and 42(4-78)months in orthotopic sigmoid neobladder group.There was no significant difference in intraoperative blood loss and postoperative urinary continence between the 2 approaches(P>0.05).However,compared with sigmoid neobladder group,ileal neobladder group had longer operative time and postoperative recovery time,and got a bigger pouch(P<0.05).The early and late pouch-related complication rates of ileal neohladder group were 16. 7% and 29.2%,which were higher than those of sigmoid neobladder group.During the follow-up,tumor recurred in 3 cases of ileal neobladder group,but none in sigmoid neobladder group.Conclusions Ortho- topic ileal neobladder and sigmoid neobladder are similar in operative difficulties,and both can achieve satis- factory clinical results.Compared with ileal neobladder,sigmoid neobladder has shorter operative time, quicker recovery and lower rate of pouch-related complications,thus is a preferred procedure.
7.Two single nucleotide polymorphisms on chromosome 3 and the risk of prostate cancer in Chinese men.
Lin ZHOU ; Ming LIU ; Jin HUANG ; Yi-Ge YANG ; Dong WEI ; Xiao-Hong SHI ; Yao-Guang ZHANG ; Xin WANG ; Chang-Hu ZHOU ; Xin CHEN ; Ze YANG ; Jian-Ye WANG
National Journal of Andrology 2011;17(8):682-687
OBJECTIVETo investigate the correlation of the common variant single nucleotide polymorphisms (SNP) on chromosome 3 with the incidence and related risk factors of prostate cancer (PCa) in Chinese men.
METHODSUsing the case-control meth- od, we included 124 PCa patients in the PCa group and 111 age- and gender-matched cancer-free healthy subjects as normal controls. We detected the distribution of allele and genotype frequencies of the SNP rs10934853 and rs2660753 with the polymerase chain reaction-high resolution melting curve (PCR-HRM) combined with gene sequencing, analyzed the cumulative effect of the risk genotypes of these two independent variants, and determined the correlation between different genotypes of these two SNPs and clinically related risk factors in the PCa patients.
RESULTSAs for the genotypes of rs10934853, there were 28 cases of AA (22.8%), 46 cases of CC (37.4%), and 49 cases of AC (39.8%) in the PCa patients, as compared with 24 (22.0%), 34 (31.2%) and 51 (46.8%) in the healthy controls. As regards the genotypes of rs2660753, there were 13 cases of AA (11.0%), 59 cases of GG (50.0%) and 46 cases of AG (39.0%) in the PCa patients, in comparison with 9 (8.8%), 47 (45.6%) and 47 (45.6%) in the controls. No significant differences were found in the distribution of the genotype and allele frequencies of rs10934853 and rs2660753 between the two groups (P = 0.520 & 0.582). Analysis on the cumulative effect of the risk genotypes of rs10934853 and rs2660753 showed a slightly higher risk of PCa (OR = 1.831 & 1.968) in the two groups with risk genotypes than in the one with wild types (P > 0.05). Different genotypes of rs10934853 and rs2660753 were not correlated with clinically related risk factors of the PCa patients (P > 0.05).
CONCLUSIONSNP rs10934853 and rs2660753 on chromosome 3 are not obviously correlated with PCa in Chinese patients, and may not be a genetic risk factor of PCa.
Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Chromosomes, Human, Pair 3 ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Prostatic Neoplasms ; epidemiology ; genetics ; Risk Factors
8.Severity of sepsis is correlated with the elevation of serum high-mobility group box 1 in rats.
Li-chao HOU ; Ming-zhe QIN ; Li-na ZHENG ; Yan LU ; Qiang WANG ; Dao-rong PENG ; Xin-ping YU ; Yu-chang XIN ; Gen-lin JI ; Li-ze XIONG
Chinese Medical Journal 2009;122(4):449-454
BACKGROUNDSepsis is a leading cause of death in the intensive care units. The late inflammatory cytokine, high-mobility group box 1 (HMGB1), plays a critical role in sepsis. In the present study, we investigated the association between the serum HMGB1 levels and the severity of organ injury in the lipopolysaccharide-induced sepsis in rats.
METHODSTo produce an animal model of sepsis with different degree of organ injury, animals were treated with three different doses of lipopolysaccharide (4, 8 and 16 mg/kg), and the animals in control group were treated with the same volume of the vehicle (saline). The levels of serum HMGB1 were measured at 0, 2, 4, 8, 16, 24, 32 and 48 hours after lipopolysaccharide (LPS) or vehicle injection, meanwhile the biochemical and histopathological indicators for the severity of organ injury were assessed.
RESULTSThe level of HMGB1 had a positive, high correlation with the abnormal changes of serum cardiac troponin I, alanine aminotransferase, aspartate aminotransferase, creatinine and blood urea nitrogen, as well as the pathologic scores of heart, lung, liver and kidney.
CONCLUSIONSThe level of serum HMGB1 is highly correlated with the severity of sepsis in rats, suggesting that HMGB1 could serve as a valuable adjunct in the diagnosis and management of sepsis.
Animals ; HMGB1 Protein ; blood ; Lipopolysaccharides ; therapeutic use ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sepsis ; blood ; drug therapy ; pathology
9.Differentially expressed genes between a fertile patient and an infertile patient in a large Chinese androgen insensitivity syndrome pedigree.
Lu-yi YE ; Zhi-min ZHAO ; Ming-hua WANG ; Shi-jing LUO ; Cheng-chang SU ; Rong-mei ZHANG ; Ze-wei LUO ; Jiu-cun WANG
Acta Academiae Medicinae Sinicae 2006;28(2):219-224
OBJECTIVETo screen and identify differentially expressed genes between a fertile patient and another infertile patient who belonged to a large Chinese pedigree affected with androgen insensitivity syndrome (AIS).
METHODSWe constructed the forward and reversed subtracted libraries using genital skin fibroblasts (GSF), which were obtained from the fertile patient MJ and infertile patient ZGJ, as tester respectively. Candidate clones were screened with colony in situ hybridization, dot blot, and Southern blot analysis step by step and conformed with Northern blot analysis. The potential positive clones were sequenced and the homology of the sequences was analyzed.
RESULTSThe forward and reversed subtracted libraries containing differentially expressed pattern of two GSF cell lines were constructed. Two positive clones identified by Northern blot were obtained in the reversed subtracted library. Eleven candidate clones from the two libraries that failed to hybridize with both RNA populations were obtained simultaneously, which might represent differentially expressed low abundance transcripts. Sequencing results and homology analysis demonstrated that the two positive clones were significantly homologous with the genes of autotaxin-t and calcium binding protein calcyclin (S100A6), respectively.
CONCLUSIONSTwo positive clones and eleven clones showing no hybridization signals may represent differentially expressed genes between the two GSFs. This finding may be useful to elucidate the molecular mechanisms leading to phenotypic variation and preserved fertility of the AIS pedigree.
Androgen-Insensitivity Syndrome ; complications ; genetics ; Blotting, Northern ; Fertility ; genetics ; Fibroblasts ; cytology ; Gene Expression Profiling ; Gene Library ; Genitalia, Male ; cytology ; Humans ; In Vitro Techniques ; Infertility, Male ; etiology ; genetics ; Male ; Nucleic Acid Hybridization ; methods ; Pedigree ; Polymerase Chain Reaction ; Skin ; cytology
10.Relationship between heat stress protein 70 gene polymorphisms and the risk of acute mountain sickness.
Fang-Ze LI ; Fang ZHOU ; Chang-Zheng JIANG ; Su-Yong SUN ; Mei-An HE ; Shu-Yu ZHANG ; Rong LIAO ; Hua-Song ZENG ; Yi-Ming WU ; Tang-Chun WU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(6):413-415
OBJECTIVETo investigate the relationship between heat stress proteins 70 (HSPs70) gene polymorphism and the risk of acute mountain sickness.
METHODSFifty-six soldiers with acute mountain sickness and 173 soldiers without that were chosen as cases and controls. HSP70-1, HSP70-2 genotypes were analyzed by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique.
RESULTSThe HSP70-1 polymorphism was similar in two groups. The genotype frequency of HSP70-2 B/B in acute mountain sickness group (23.2%) was significantly higher than that in the control (6.9%, P < 0.05, OR = 4.02).
CONCLUSIONThere is a significantly increased association of HSP70-2 B/B genotype with the risk of acute mountain sickness. Individuals with HSP70-2 B/B genotype may have weaker adaptive ability than those without this genotype under altitude stress. The results contribute to provide scientific bases for finding these individuals in specified occupational people, ensuring their health and enhancing work efficiency.
Acute Disease ; Adolescent ; Adult ; Altitude ; Altitude Sickness ; epidemiology ; genetics ; Genotype ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Male ; Polymorphism, Genetic ; Young Adult