1.Prokaryotic expression,identification,primary purification of human prostate specific membrane antigen
Hai HUANG ; Jian HUANG ; Tianxin LIN ; Al ET
Chinese Journal of Urology 2001;0(08):-
Objective To describe the cloning of cDNA of prostate specific membrane antigen (PSMA),construction of the recon of PSMA,induction of the expression of 6 ? His PSMA fusion protein from PSMS recon,and purification of the expression products. Methods The total RNA was extracted from prostate cancer tissues.The site of EcoR I lying in the middle of PSMA’s cDNA was used to perform the reverse transcriptase and polymerase chain reaction of PSMA in two different parts,which were then linked into the carrier of pEt 30(a),thus the full cDNA of PSMA and the pronucleus expression carrier pET 30(a) PSMA were obtained.The pET30a (+) PSMA recon was transformed into the Escherichia coli BL 21 and the engineering bacteria expressing protein PSMA was gotten. With the induction of IPTG, the recon was expressed. The expression products were purified with Ni NTA Agarose resin, then the purer protein PSMA was obtained. The antigenicity and specificity of the expressed PSMA were evaluated with Western blotting and SDS PAGE. Results The wholly right base sequence of PSMA’s cDNA was successfully cloned.PSMA protein was expressed and purified.This protein had better antigenicity and specificity. Conclusions This study provides experimental basis for the function study of PSMA and scanning of the phage antibody library.
3.Evaluation system and model for evaluating development strategies of military preventive medicine based on SWOT analysis
Hai LIN ; Guangwei CHEN ; Chunji HUANG ; Changkun LUO
Military Medical Sciences 2017;41(5):390-393,397
Objective To provide an index system and model for evaluating the development strategies of military preventive medicine (MPM).Methods SWOT was used to analyze the internal and external conditions of the development strategies of MPM,and the evaluation index system was constructed.The weight of each evaluation index was determined based on the analytic hierarchy process (AHP) and expert consultation.The evaluation model of MPM development strategies and confrontation matrix were constructed based on SWOT analysis.The effectiveness of the index system and model was evaluated through empirical research.Results The evaluation index system was constructed,which included four grade-one indexes (R&D quality,difficulty,requirement and support) and sixteen grade-two indexes (research level,academic status,etc.).The advantage and disadvantage models,opportunity threat model and SWOT evaluation model were established.The confrontation matrix of development strategies for MPM was developed.Conclusion This evaluation index system and model can accurately evaluate MPM development strategies and provide reference for formulating development strategies of MPM.
4.Posterior indirect reduction and internal fixation and laminectomy in the treatment of thoracolumbar vertebrae burst fracture complicated with spinal cord injury
Chen HUANG ; Qionghua FENG ; Bo CAO ; Wei LIN ; Hai XU
Journal of Regional Anatomy and Operative Surgery 2017;26(3):204-208
Objective To explore the clinical effect of posterior indirect reduction and internal fixation and laminectomy in the treatment of thoracolumbar vertebrae burst fracture complicated with spinal cord injury.Methods Eighty patients with thoracolumbar vertebrae burst fracture and spinal cord injury treated in our hospital from March 2014 to March 2015 were selected as the objects,and they were divided into reset group and laminectomy group with forty cases in each group according to surgical method.All the patients were followed up for 1 year,the lumbar function of two groups at 1 week and 1 year after operation were observed respectively,and the pain degree was observed in 1month,3 months and 6 months after operation.The amount of bleeding,operation time,hospitalization time and fracture healing time were observed.Neurological function was assessed by classification criteria of the American Spinal Cord Injury Association(ASIA),and incidence of complications was figured in the two groups.Results The anterior heights of the injured vertebra were higher than those before the operation,and the Cobb's angles were lower than those before the operation,the differences were significant(P < 0.05);while there was no significant differences in the anterior heights of the injured vertebra between the two groups at 1 week and 1 year after operation(P > 0.05).VAS scores of the two groups after 1 month,3 months and 6 months decreased significantly when compared with the preoperative scores(P < 0.05),and VAS scores of each time in the reset group were significantly lower than those in the laminectomy group(P < 0.05).The amount of bleeding,operation time,hospitalization time and fracture healing time in the reset group were less than those in the laminectomy group (P < 0.05).The neurological function recovery of the two groups were significantly improved when compared with that before the operation(P <0.05).There was no significant difference in recovery of neurological function between the two groups(P > 0.05).The complication rate was 7.50% in the reset group,lower than 12.50% of the laminectomy group,the difference was significant (P < 0.05).Conclusion Posterior indirect reduction and internal fixation of lamina both have a certain effect in the treatment of thoracolumbar vertebrae burst fracture complicated with spinal cord injury.But posterior indirect reduction has less complications and less amount of bleeding,which is beneficial to postoperative recovery.
5.Study on Association between Interleukin-1 Beta Gene Polymorphism and Childhood Asthma
xiao-fen, ZHAO ; hai-lin, LI ; yong-kun, HUANG
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To evaluae possible association between interleukin-1 beta (IL-1?) gene exon 5 polymorphism and childhood asthma.Methods The study was conducted in two different groups: asthmatic children(n=55) and healthy children(n=35). The IL-1? gene exon 5 polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP). Results Frequencies of CC,CT and TT genotypes were 92.7%,7.3%,0, and frequencies of C,T allele were 96.4%,3.6% in asthmatic group. However, frequencies of CC,CA and AA genotypes were 85.7%,14.3%,0, and frequencies of C,T allele were 92.9% ,7.1% in healthy group. There were no significant difference in distribution of genotypes and allele frequencies between two groups.Conclusion IL-1? gene exon 5 polymorphism may not be associated with childhood asthma.
6.Correlation of Platelet-Activating Factor Acetylhydrolas Gene Polymorphism and Bronchial Asthma in Children
hai-lin, LI ; fei, CHENG ; yong-kun, HUANG
Journal of Applied Clinical Pediatrics 2006;0(20):-
0.05).Conclusion PAF-AH-Ala379Val gene mutation is unrelated to bronchial asthma in children.
7.Association between Polymorphism of Glutathione S-Transferease P1 Gene and Susceptibility in Childhood Asthma
hai-lin, LI ; jing-jing, XIONG ; yong-kun, HUANG
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To investigate the polymorphism of glutathione S-transferease P1 gene(GSTP1) and the association between the mutation and susceptibility in childhood asthma.Methods The distribute frequency of Ile105/Ile105,Ile105/Val105 and Val105/Val105 ge-notype in GSTP1 of 51 children with asthmatic and 40 normal children were studied with polymerase chain reaction-restriction tragment length polymorphism(PCR-RFLP).Results The frequencies of Ile/Ile Ile/Val,Val/Val genotype were 66.7%,27.4% and 5.9%,the frequencies of Ile,Val allele were 80.4% and 19.6% in the asthmatic group.But the frequencies of Ile/Ile,Ile/Val,Val/Val genotype were 90.0%,7.5% and 2.5%,the frequencies of Ile,Val allele were 93.8%,6.2% in control group.The frequencies Ile/Val,Val/Val genotype and Val allele in asthmatic group were more than that in control group.A significant difference was found in the frequency distribution of GSTP1 genotypes between two groups(?2=6.947 P
9.Effects of tetramethylpyrazine on fibrosis of atrial tissue and atrial fibrillation in a canine model of congestive heart failure induced by ventricular tachypacing.
Yazhou LIN ; Chunxuan XU ; Yulian DENG ; Lin CHEN ; Hai HUANG ; Jian DU
Journal of Integrative Medicine 2006;4(1):35-8
To explore the effects of tetramethylpyrazine (TMP) on fibrosis of atrial tissue and atrial fibrillation in a canine model of congestive heart failure (CHF) induced by ventricular tachypacing.
10.Culture and differentiation of bone marrow mesenchymal stem cells on bladder acellular matrix
Zuoqiang LIU ; Hai HUANG ; Jian HUANG ; Tianxin LIN ; Kewei XU ; Zhenghui GUO ; Chun JIANG ; Jinli HAN
Chinese Journal of Tissue Engineering Research 2008;12(14):2780-2784
BACKGROUND:Smooth muscle cells and transitional epithelial cells were traditionally used to construct tissue-engineered bladder and to perform double-sided implantation of scaffold.However,double-sided implantation is difficult to perform,because smooth muscle cells are difficult to isolate or culture in vitro and passage is limited.OBJECTIVE:To verify the feasibility of tissue-engineered bladder reconstruction with bone marrow mesenchymal stem cells(BMSCs)and bladder acellular matrix(BAM).DESIGN:A basic empirical study.SETTING:Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University.MATERIALS:Experiments were performed at the Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University from March 2006 to Mav 2007.The laboratory was the Opening Laboratory of Hospital Affiliated to Health Department of China.One-month old SD rats of either sex,weighting 80-100 g were provided by Animal Experimental Center of Sun Yat-sen University.Fresh porcine bladders were offered by Animal Experimental Center of Southern Medical University.METHODS:Whole bone marrow culture and successive adherence method was used to culture rat BMSCs in vitro.Flow cytometry was employed to detect surface antigen.Eradicator washing method was applied to prepare porcine BAM and measure its purity and characteristies.Third passage of BMSCs were inoculated in BAM and cultured in a medium containing vascular endothelial growth factor(VEGF165)(25 ng/L)in vive and in vitro to test compatibility.Cells cultured alone were considered to be controls for the in vivo trial,and materials non-implanted with cells were considered to be controls for in vitro trial.Suitable microenvironment was simulated to induce the differentiation of BMSCs.Four weeks and eight weeks later,compound materials were respectively removed to perform tissue section test.Simultaneously,immunohistochemistry keratin staining was conducted to examine regeneration of epithelial cells.MAIN OUTCOME MEASURE:Biocompatibility of BMSCs and BAM.RESULTS:①BMSCs were cultured by whole bone marrow method.Flow cytometry demonstrated that third passage of cells were positive for CD29(99.43%).②BAM had good biological characteristics.Homogen matrix and byssoid collagen appeared under a microscope.Compatibility trials showed good compatibility of BMSCs and BAM and well-growth cells.③Four weeks later,histological section test confirmed inflammatory cell infiltration,closely-arranged collagen and elastic fiber.Immunohistochemistry keratin staining showed lamellar and discontinuous simple epithelium.Eight weeks later,no inflammatory cell infiltration was found,and closely-arranged collagen and elastic fiber were detected.Immunohistochemistry keratin staining showed lamellar and continuous multiple epitheliums.CONCLUSIoN:With good compatibility,BMSCs and BAM appear to be an ideal material for bladder tissue engineering.