2.Expression, purification and preliminary activity study of recombinant hepatocyte growth factor protein in E.coli
Xiaojiao ZHENG ; Zhou GAO ; Rongrong SHEN ; Hang ZHAO ; Dong CEN ; Jianping LUO ; Jianxin Lü ; Renzhi PEI ; Shixuan HUA
Chinese Journal of Microbiology and Immunology 2012;(11):967-971
Objective To prepare hepatocyte growth factor(HGF) recombinant protein and confirm its activity preliminarily according to building HGF gene prokaryotic expression vector and transforming into E.coli.Methods Clone HGF inserted into the vector pET-26b(+) to construct prokaryotic expression vector pET-26b(+)-HGF and transform into E.coli Rosseta(DE3).The transformed bacteria induced by IPTG was purified through Ni-NTA resin affinity chromatography frozen-drying after renaturation.Results HGF gene recombinant prokaryotic expression vector pET-26b(+)-HGF was constructed successfully.E.coli Rosseta(DE3) which was transformed into pET-26b(+)-HGF expresses the target protein as the form of inclusion bodies,accounting for 38% of the total bacterial proteins,and confirmed by Western blot.HGF protein which was purified by Ni-NTA resin affinity chromatography,has a purity of about 95%,and can promote proliferation,migration,and inhibition of apoptosis for human non-small cell lung cancer cell line A549 cells after interaction.Conclusion HGF gene recombinant prokaryotic expression vector pET-26b (+)-HGF was constructed and expressed in transformed E.coli Rosseta(DE3) successfully.They resumed their recombinant HGF protein structure after purification and renaturation,and had biological activity confirmed by in vitro studies.
3.Stress area of the mandibular alveolar mucosa under complete denture with linear occlusion at lateral excursion.
Ya-Lin LÜ ; Hang-di LOU ; Qi-Guo RONG ; Jian DONG ; Jun XU
Chinese Medical Journal 2010;123(7):917-921
BACKGROUNDThe rocking and instability of a loaded complete denture (CD) during lateral excursion reduce the bearing area under the denture base, causing localized high stress concentrations. This can lead to mucosal tenderness, ulceration, and alveolar bone resorption, and the linear occlusion design was to decrease the lateral force exerted on the denture and to ensure denture stability. But it is not known how the bearing areas of linear occlusal CDs (LOCDs) and anatomic occlusal CDs (AOCDs) differ. The purpose of this study was to analyze and compare the distributions of the high and low vertical stress-bearing areas in the mandibular alveolar mucosa under LOCDs and AOCDs at lateral excursion.
METHODSComputerized tomography (CT) and finite element analysis were used to establish three-dimensional models of an edentulous maxilla and mandible with severe residual ridge resorption. These models were composed of maxillary and mandibular bone structure, mucosa, and the LOCD or AOCD. Lateral excursion movements of the mandible were simulated and the vertical stress-bearing areas in the mucosa under both mandibular CDs were analyzed using ANSYS 7.0.
RESULTSOn the working side, the high stress-bearing (-0.07 to -0.1 MPa) area under the LOCD during lateral excursion was smaller than that under the AOCD, while the medium stress-bearing (-0.03 to -0.07 MPa) area under the LOCD was 1.33-fold that under the AOCD. The medium stress-bearing area on the non-working side under the LOCD was 2.4-fold that under the AOCD. Therefore, the overall medium vertical stress-bearing area under the LOCD was 20% larger than that under the AOCD.
CONCLUSIONSDuring lateral excursion, the medium vertical stress-bearing area under a mandibular LOCD was larger and the high vertical stress-bearing area was smaller than that under an AOCD. Thus, the vertical stress under the LOCD was distributed more evenly and over a wider area than that under the AOCD, thereby improving denture stability.
Aged ; Computer Simulation ; Dental Occlusion ; Dental Stress Analysis ; Denture, Complete ; Female ; Finite Element Analysis ; Humans ; Mandible ; physiology ; Stress, Mechanical
4.Detection of hematopoietic chimera by real-time fluorescent quantitative PCR with erythrocyte Kidd blood group gene.
Shu CHEN ; Xian-Guo XU ; Ying LIU ; Xiao-Zhen HONG ; Fa-Ming ZHU ; Hang-Jun LÜ ; Li-Xing YAN
Journal of Experimental Hematology 2012;20(3):676-678
This study was aimed to establish the real-time fluorescent quantitative PCR (RT-qPCR) with erythrocyte Kidd blood group gene for detecting the hematopoietic chimera and to investigate the feasibility of this method. The TaqMan MGB probes and special primers were designed on basis of difference of erythrocyte Kidd blood group alleles, the hematopoietic chimerism was detected by RT-qPCR, the DNA chimerism was simulated by means of dilution of multiple proportions, and the sensitivity analysis was performed. The results showed that the RT-qPCR with erythrocyte Kidd blood group gene could effectively distinguish JK*A and JK*B alleles. There was no significant difference between the theoretic value and the practical measured value by this method (P > 0.05). As 156 donor's cells could be discriminated from 10(4) chimeric cells, this method may effectively detect donor's cells with correlation coefficient 0.998. It is concluded that the established RT-qPCR with erythrocyte Kidd blood group gene shows the feasibility for quantitative detection of hematopoietic chimera, and may be used to quantitatively detect chimera in a certain range.
Chimera
;
Erythrocytes
;
Humans
;
Kidd Blood-Group System
;
genetics
;
Real-Time Polymerase Chain Reaction
5.Survey of studies on drug abstinence with acupuncture in recent 10 years.
Xiao-Ge SONG ; Hang LÜ ; Xing-Hui CAI ; Rong-Jun ZHANG
Chinese Acupuncture & Moxibustion 2012;32(7):669-672
The effect of acupuncture on substance withdrawl syndromes and craving relapse prevention of the recent 10 years were reviewed as well as its mechanism. The therapeutic effect and the possible mechanism were analyzed on the basis. From the three aspects of anti protracted abstinence symptoms, craving relapse prevention and mechanism of acupuncture, the development tendency and the prospect of application on drug withdrawl with acupuncture were expected. And it is proposed that clinical observation of acupuncture intervention on craving should be developed, the mechanism of acupuncture impact on cognitive behavior, blocking study and memory processing related to drug addiction should be explored, so as to further give play to the advantages of acupuncture on anti-drug addiction.
Acupuncture Therapy
;
Humans
;
Substance-Related Disorders
;
therapy
;
Time Factors
6.Circumcision with no-flip Shang Ring technique for adult males: analysis of 168 cases.
Jun-Hao LEI ; Liang-Ren LIU ; Xiao LÜ ; Si-Hang CHENG ; Ying-Chun CAI ; Yong-Ji CHEN ; Qiang WEI ; Yu-Chun ZHU
National Journal of Andrology 2014;20(4):320-324
OBJECTIVETo observe the clinical effects of the no-flip procedure with the Chinese Shang Ring when circumcising adult males with redundant prepuce or phimosis, and to discuss its advantages and disadvantages.
METHODSUsing the no-flip Shang Ring technique, we performed circumcision for 167 adult males aged 18 -72 (mean 27.8) years with redundant prepuce or phimosis, and analyzed the clinical data, including the operation time, postoperative complications, ring-removal time, and postoperative appearance of the penis.
RESULTSComplete follow-up data of 94 cases (56.29%) were obtained. The mean operation time was (5.03 +/- 0.71) minutes and the average ring-removal time was (18.83 +/- 6.70) days. The primary postoperative complications were edema (35 cases [37.23%] at 2 weeks and 9 cases [9.57%] at 4 weeks), including 2 severe cases (2.13%), and infection (3 cases [3.19%]). The pain scores were 2.01 +/- 2.46 during the procedure and 4.52 +/- 2.53 at 24 hours postoperatively. Slipping of the outer ring occurred in 1 case, and delayed removal of the ring in 30 cases (31.91%).
CONCLUSIONAdult male circumcision with the no-flip Shang Ring technique is recommended for its short operation time, simple procedure, fewer postoperative complications, less pain, and better incision appearance.
Adult ; Aged ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Edema ; etiology ; Humans ; Male ; Middle Aged ; Operative Time ; Pain, Postoperative ; etiology ; Penile Diseases ; etiology ; Penis ; abnormalities ; surgery ; Phimosis ; surgery ; Postoperative Complications ; Postoperative Period ; Prostheses and Implants ; Young Adult
8.Discrimination of alleles in HLA-C*07:01:01G and HLA-C*07:02:01G groups through detection sequences in exons 1 to 7 of HLA-C locus by using polymerase chain reaction sequence-based typing.
Hang-Jun LÜ ; Wei ZHANG ; Jun-Jun HE ; Yan-Min HE ; Wei WANG ; Zhe-Dong HANG ; Nan-Yin CHEN ; Fa-Ming ZHU ; Li-Xing YAN
Journal of Experimental Hematology 2012;20(1):178-181
This study was aimed to discriminate the alleles in the HLA-C*07:01:01G and HLA-C*07:02:01G groups and analyze their associations with HLA-B locus. Samples previously typed as HLA-C*07:01:01G and HLA-C*07:02:01G were collected. The nucleotide sequences in exons 1 to 7 of the HLA-C locus were sequenced by polymerase chain reaction sequence-based typing (PCR-SBT) and HLA-B genotyping was also preformed by PCR-SBT in these samples. The results showed that 4 samples (30.8%) were confirmed as HLA-C*07:01:01 and 9 samples (69.2%) were HLA-C*07:06 among 13 samples previously typed as HLA-C*07:01:01G. Linkage disequilibrium (LD) analysis showed that HLA-C*07:06 allele was strongly related with HLA-B*44:03. All samples were typed as C*07:02:01 among 102 individuals previously typed as C*07:02:01G. LD analysis showed that C*07:02:01 was strongly related with HLA-B*51:01, B*46:01, B*39:01, B*40:01, B*38:02, B*15:02 alleles. It is concluded that HLA-C*07:01:01 and HLA-C*07:06 alleles are confirmed in the HLA-C*07:01:01G group and HLA-C*07:02:01 is a preferred allele in the HLA-C*07:02:01G.
Alleles
;
Base Sequence
;
Exons
;
HLA-B Antigens
;
genetics
;
HLA-C Antigens
;
genetics
;
Humans
;
Molecular Sequence Data
;
Polymerase Chain Reaction
;
methods
;
Sequence Analysis, DNA
9.Anti-rejection effect of ethanol extract of Poria cocos wolf in rats after cardiac allograft implantation.
Guo-wei ZHANG ; Hong-yu LIU ; Qiu-ming XIA ; Jun-quan LI ; Hang LÜ ; Qing-hua ZHANG ; Zhi-fa YAO
Chinese Medical Journal 2004;117(6):932-935
BACKGROUNDA living fetus within the maternal uterus provides an example of allogene tolerance in mammals. Poria cocos Wolf is the main component of many Chinese medicinal combination drugs that have therapeutic effects on recurrent spontaneous abortion and that can maintain pregnancy until delivery. It was hypothesized that this herbal medicine can also prolong allograft survival after organ transplantation. Here, in an in vivo study, we report the anti-rejection effect of the ethanol extract of Poria cocos Wolf (EEPCW) in rats after cardiac allograft implantation.
METHODSTen normal rats were healthy controls. Eighty rats receiving homologous heart transplants were divided into 4 groups of 20 rats each based on type of treatment: olive oil 8 ml.kg(-1).d(-1), EEPCW 25 mg.kg(-1).d(-1), EEPCW 50 mg.kg(-1).d(-1) or cyclosporin A 5 mg.kg(-1).d(-1). Allograft survival was observed in 10 rats from each group. On the seventh day post transplantation, pathological lesions and percentages of CD3+, CD4+, and CD8+ lymphocytes and the CD4+/CD8+ ratio in peripheral blood were assessed in another 10 rats from each group and in 10 normal rats.
RESULTSThe survival time of donor hearts in the two EEPCW groups was significantly prolonged, to (15.9 +/- 2.4) days and (30.0 +/- 0.0) days, respectively, compared with (6.7 +/- 0.8) days in the control group. Pathological lesions in the two EEPCW groups were also less severe, and the percentages of CD3+, CD4+, and CD8+ lymphocytes and CD4+/CD8+ ratio were significantly lower in the EEPCW groups.
CONCLUSIONSAcute rejection of heart transplants and cellular immune reaction can be effectively suppressed using the EEPCW. Taking advantage of novel immunosuppressants derived from Chinese medicinal herbs used to treat abnormal pregnancy provides a hopeful road for future research and treatment in organ transplantation.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Graft Rejection ; prevention & control ; Graft Survival ; drug effects ; Heart Transplantation ; Immunosuppressive Agents ; pharmacology ; Male ; Polyporales ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar
10.Sequence analysis of a novel HLA-A * 2459 allele.
Wei ZHANG ; Jun-Jun HE ; Qin-Feng LÜ ; Wei WANG ; Zhe-Dong HANG ; Fa-Ming ZHU ; Li-Xing YAN
Journal of Experimental Hematology 2007;15(5):1090-1092
This study was aimed to investigate the molecular genetics basis of a novel allele HLA-A * 2459 in Chinese population. DNA was extracted from whole blood by PEL-FREEZ DNA extraction kit. The amplification of HLA-A exons 2 - 4 of the proband was preformed by allele specific primer PCR and the amplified product was sequenced bidirectionally with primers. The sequencing results showed HLA-A alleles of the proband as A * 1101 and the novel allele. The sequences of the novel allele have been submitted to GenBank (DQ313255, DQ313256, DQ313257). After Blast HLA analysis, the novel allele showed only one nucleotide differences with HLA-A * 24020101 at nucleotide position 527 T to C in exon 3. This results in an amino acid changes from Val to Ala at codon 152. In conclusion, this allele is a novel one and has been officially named HLA-A * 2459 by the WHO Nomenclature Committee.
Alleles
;
Asian Continental Ancestry Group
;
genetics
;
Base Sequence
;
Blood Donors
;
China
;
HLA-A Antigens
;
genetics
;
Humans
;
Molecular Sequence Data
;
Sequence Analysis, DNA