1.An experimental study on the reconstruction of hepatic blood flow with flow-restricted arterio-portal shunt
Yongliang CHEN ; Xiaoqiang HUANG ; Zhiqiang HUANG
Medical Journal of Chinese People's Liberation Army 1982;0(03):-
Objective To study the effect of flow-restricted arterio-portal shunt(APS)on the liver.Methods Experimental model of APS and flow-restricted APS were reproduced in rats,and its effects on hepatic blood flow(HBF)and portal venous pressure(PVP)were observed,and the changes in liver structure were investigated six months after the operation.Results Compared with that of pre-operation,both HBF and PVP in APS group at the sixth month after operation changed significantly(P0.05).The PVP changed significantly(P0.05).No obvious lesions were found in the liver by histological examination.Significant differences of both PVP and HBF were found at the sixth month between the two groups(P
2.CONSTRUCTION AND IDENTIFICATION OF THERAPEUTIC DOUBLE-PLASMID HBV DNA VACCINE
Xiaoqiang HE ; Guangming CHEN ; Yin HUANG
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
To develop an effective therapeutic HBV DNA vaccine, two eukaryotic expressing plasmids, namely pcDNAS 2 ?S and pcDNAIIF , which respectively encoding HBV middle envelope protein preS 2 ?S and human IL 2/IFN ? fusion protein, were constructed by DNA recombinant technique. After identification by restriction analysis and DNA sequencing, the constructed recombinant plasmids were transfected into COS 7 cells in vitro with transfection reagent Lipofectamine. The expressed product in supernatant was quantified by ELISA , and the biologic activities of IL 2 and IFN ? were assayed by CTLL 2 cell line proliferation and cytopathogenic inhibition, respectively. The results showed that secretive expression of preS 2 ?S and hIL 2/hIFN ? fusion protein peaked at 48h after transfection, and the expression levels were HBsAg(P/N)=7 63, IL 2=10 35ng/ml, and IFN ?=7 90ng/ml. The IL 2 and IFN ? activities in the culture supernatant of COS 7 cells transfected with pcDNAIIF were 998U/ml and 249U/ml, respectively. These results suggested that the recombinant plasmids pcDNAS 2 ?S and pcDNAIIF were correctly constructed and the transfected cells could express secretive active protein.
3.Adult reconstruction plate of humeral shaft for 32 children with femoral shaft fracture:Follow-up analysis
Xiaoqiang LIU ; Zongxiong CHEN ; Wanzong WANG
Chinese Journal of Tissue Engineering Research 2010;14(17):3193-3196
BACKGROUND:Femoral shaft fracture is common in children Various methods of treatment can be used successfully,depending On the age of the child and the type of fracture.OBJECTIVE:To evaluate the feasibility of adult reconstruction plate of humeral shaft for pediatric femoral shaft fracture.METHODS:A total of 32 children with femoral shaft fracture were treated with adult reconstruction plate of humeral shaft in First Department of Orthopedics,Fuzhou General Hospital of Fujian Medical University,including 20 males and 12 females aged 6.7years(5-8 years).All patients were treated with incision reduction and adult reconstruction plate of humeral shaft.The incision length should be made as short as possible according to the fracture type The result of surgery was determined using clinical and radiographic examinations The pain condition was evaluated using visual analog scale method before and 3 days following surgery.RESULTS AND CONCLUSION:The patients were followed up for 1-2 years(average 1.5 years).All fractures were healed 1-2months postoperatively,and the internal fixator was removed 4-8 months postoperatively.The affected limb was shortened0.6-1.5 cm(average 1.1 cm)in 3 cases.Overgrowth was obsewed in the other patients by 0.3-1.2 cm The average overgrowth length was(0.64±0.312)cm in 29 patients.At 3 days postoperatiVely,the mean subjective pain was significantly reduced,and range of motion was improved compared with the day before surgery There was no infection or implant displacement or re-fracture.It is feasible to use adult reconstruction plate of humeral shaft for paediatric femoral shaft fracture.
4.Three advantages of using traditional Chinese medicine to prevent and treat tumor.
Changquan LING ; Xiaoqiang YUE ; Chen LING
Journal of Integrative Medicine 2014;12(4):331-5
Traditional Chinese medicine (TCM), an important component of complementary and alternative medicine, has evolved over thousands of years with its own unique system of theories, diagnostics and therapies. TCM has been increasingly used in the last decades and become well known for its significant role in preventing and treating cancer. We believe that TCM possesses advantages over Western medicine in specific aspects at a certain stage of cancer treatment. Here we summarize the advantages of TCM from three aspects: preventing tumorigenesis; attenuating toxicity and enhancing the treatment effect; and reducing tumor recurrence and metastasis.
5.Research into the management model of hospital information systems
Min CHEN ; Jianguo HU ; Xiaoqiang ZHANG
Chinese Journal of Hospital Administration 2001;0(08):-
As the development of hospital information systems in the third-tier hospitals of the first grade in China gradually deepens, there has appeared an urgent need to formulate standards for the management of such systems. The paper offers proposals for the structure, composition and main responsibilities of the centers for hospital information management, posts setting and post responsibilities in the centers, the number of allocated personnel in the centers and the number of personnel required for various posts, and management standards and operating procedures in the centers.
6.Construct the recombinant plasmid of recombinant human basic fibroblast growth factor
Lihong FAN ; Xueqing CHEN ; Kequan CHEN ; Xiaoqiang YANG ; Zhenshu ZHANG
Journal of Chinese Physician 2012;14(5):593-595,599
Objective To obtain the sequence of recombinant human basic fibroblast growth factor (rhbFGF) with endonuclease sites of BamHI and Pst Ⅰ by PCR based gene assembly and construct the recombining vector of rhbFGF gene by TA cloning technique.Methods The rhbFGF gene sequence which was designed for lactococcus with endonuclease sites of BamHI and Pst Ⅰ was divided into 22 oligonucleotides by DNASTAR 6.0 (bFGFl-bFGF22).The 22 oligonucleotides were spliced by PCR based gene assembly to get the rhbFGF eDNA with endonuclease sites of BamHI and Pst Ⅰ.The PCR product was inserted into the PMD18-T VECTOR.The recombining vector were converted to the competent E.coli TOP10.The clones generated from LAB were analyzed by miniprep isolation from LAB host.They were identified by the restriction enzyme cuuing and sequencing.Results The rhbFGFcDNA synthesized by PCR based gene assembly with endonuclease sites of BamHI and Pst Ⅰ was verified by 2% agarose electrophoresis.The recombining vector of rhbFGF gene by TA cloning technique was identified by enzyme digestion and gene sequencing.Conclusions The TA cloning vector of recombining hbFGF with endonuclease sites of BamHI and Pst Ⅰ was constructed successfully.
7.Expression, purification Orai2 protein and preparation, application it's polyclonal antibody
Xiaoqiang XIA ; Yiyuan CUI ; Fanghua LI ; Mina CHEN
Chinese Journal of Immunology 2009;25(11):965-968
Objective:To prepare GST-Orai2 fusion protein and to prepare polyclonal antibody against Orai2 by immunizing rabbits.To further investigate the function of Orai2,a transmembrane protein,this antibody was used to identify the Orai2 conditional gene knockout mice.Methods:ORF of Orai2 was amplified by PCR and subcloned into pGEX-6p-1 vector.After transforming BL21 (DE3) competent cells,we succeeded in inducing the expression of GST-Orai2 fusion protein using IPTG.Then the GST-Orai2 protein was purified by immobilized Glutathione affinity chromatography and identified by SDS-PAGE.A New England rabbit was immunized with the prepared fusion protein in Freund's adjuvant to prepare specific antibody.Finally,the prepared antibody was identified by Western blot by checking its titer and specificity.Furthermore,we took use of the prepared Orai2 antibody to identify Orai2 conditional gene knockout mice,with comparing to the wildtype ones in the same cage.Results:The purity of purified GST-Orai2 reached to 90% and the concentration was 0.35 mg/ml by BCA kit.We could detect Orai2 protein even in dilution of 1:10,000.Also,the prepared polyclonal antibody agianst Orai2 could detect both overexpressed and endogenous Orai2 protein in mouse-brain,without crossing reaction with Orai1.As well,we found that the Orai2 protein expression was of obvious reduction in Orai2 conditional gene knockout mice,compared with the wildtype ones in the same cage.Conclusion:We successfully obtain the purified GST-Orai2 fusion protein and prepare specific and highly sensitive polyclonal antibody against Orai2.The antibody can be used to detect overexpressed and endogenous Orai2 protein inmouse-brain specifically,and to identify Orai2 conditional gene knockout mice,without any crossing reaction with Orai1.Our work contributes a lot to the future investigation of functions of Orai2.
8.DNA vaccine induced cellular immunity in healthy mice and HBsAg serum conversion in HBV transgenic mice
Fuqiang YANG ; Guangming CHEN ; Xiaoqiang HE ; Al ET
Chinese Journal of Immunology 1985;0(06):-
Objective:To investigate the feasibility and its action of the therapeutic DNA vaccine for treatment of HBV infection.Methods:By genetic recombinant technique,have constructed 2 eukaryotic expressing plasmids namely pS2.S and pFP,encoding HBV envelope middle protein(preS2+HBsAg) and human leukocyte cytokines fusion protein(IL 2/IFN ?) genes respectively and evaluated its efficacy for inducing cellular immunity in healthy BALB/c mice and HBsAg serum conversion in HBV transgenic(Tg) mice after immunization of the plasmids by intramuscular administration.Results:1.The T cell proliferation by in vitro HBsAg stimulation closely correlated with HBsAg concentration,with its stimulation index(SI=5.6?0.9) in pS2.S immunized group,being significantly higher than that (2.0?0.5) of the control.The IL 2 and IFN ? secretion level in supernatant of the DNA vaccine immunized spleen cell culture were significantly higher than that of the control,while the IL 4 secretion level was not affected.2.The dendritic cells(DCs) extracted from the local draining lymph node(LN) after DNA vaccination induced a HBsAg specific T cell proliferation with its SI(4.20) being higher than that(2.55) of the control.3.In each group of high dose pS2.S and the middle dose combined with pFP inoculation,the HBsAg serum conversion occurred in one HBV Tg mouse,with the serum anti HBs level increasing as the time prolonged,while the serum HBsAg level of the rest mice were significantly lower than that of the control.Conclusion:The results suggest that HBV DNA vaccine can effectively induced cellular immunity in healthy mice;and the preliminary results of the immunized HBV Tg mice may provide an experimental evidence for further investigation of DNA vaccine for its use in treatment of HBV infection.
9.Construction of cloning vector of c-myc-TFF2 fusion gene by PCR based gene assembly
Rui XING ; Xiaoqiang YANG ; Shaorong ZHANG ; Xueqing CHEN ; Zhenshu ZHANG
Journal of Xi'an Jiaotong University(Medical Sciences) 1982;0(04):-
Objective To assemble fusion gene of c-myc tagged human trefoil factor family 2(hTFF2)(in vitro) and construct a cloning vector of this fusion gene.Methods Based on amino acid sequence of hTFF2 and codon of lactococcus lactis,cDNA of htff2 sequence was designed and extended at their 5' ends with a sequence encoding the c-myc tag;and the sequence of fusion gene c-myc-htff2 was designed.According to restriction enzyme sites of pBluescript Ⅱ sk(+),the SalⅠ and BamHⅠ were arranged at 5′ and 3′ ends of the fusion gene respectively.Instructed by DNAWORKS program,the fusion gene sequence of c-myc-htff2 was designed as 14 oligonucleotides that overlapped each other.The target gene fragment of cmyc-htff2 fusion gene was obtained by the means of polymerase chain reaction(PCR) based gene assembly.Then,the c-myc-htff2 was subcloned into vector of pBluescript II sk(+) for identification of the fusion gene by restricting enzyme excision and DNA sequencing.Results Assembly of c-myc-TFF2 fusion gene(in vitro) and construction of pBS-TFF2 had been completed successfully;and DNA sequencing showed that the sequence of synthetic gene accorded with the expectation.Conclusion With the help of DNAWORKS program,the design and assembly of oligonucleotides (in vitro) is effective to synthesize a target gene,and the cloning vector of c-myc-htff2 fusion gene has been successfully constructed.
10.Effects of irbesartan on the aldosterone escape in essential hypertension
Yufeng CHEN ; Xiaoqiang TAN ; Tao YE ; Liuping HE
Chinese Journal of Primary Medicine and Pharmacy 2008;15(10):1587-1588
Objective To investigate the influence provided by irbesartan on aldosterone escape during an-giotensin-converting enzyme inhibitor(ACEI) therapy in patients with essential hypertension (EH). Methods 166 EH patients were treated with ACEI. Plasma angiotensin Ⅱ ( Ang Ⅱ ) and aldosterone(Ald) concentrations were meas-ured by radioimmunoassay following the therapy of 3 months treatment. Aldosterone escape was estimated by the plas-ma Ald level after 3 months treatment. Patients with aldosterone escape were given irbesartan for 6 months. The aldo-sterone escape were observed following the therapy with irebesartan at 1,3,6 months. Results Following the treat-ment of irbesartan for 1 month, plasma Ang Ⅱ and Ald concentrations significantly decreased (P < 0.01 ). However, it began to increase after 3 and 6 months treatment, still lower than that of the therapy not given irbesartan ( P < 0.01 ).The levels of plasma Ang Ⅱ and Aid were higher in 6 months than in 1,3 months ( P < 0.01 ), but there are no differ-ences between 1 and 3 months(P>0.05). Conclusion Irbesartan can reduce aldosterone escape during 6 months.But aldosterone escape may be occur again after 6 months treatment of irebesartan.