1.Synthesis of DNA Encoding Botulinum Neurotoxin Receptor syt II and Its Fusion Expression in E.coli
Jing SHI ; Hui WANG ; Shi-Zhong BAO ; Xiao-Jun HOU ; Jun YIN ;
China Biotechnology 2006;0(07):-
In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.
2.Effects of intensive insulin therapy on the intestinal permeability in sepsis patients
Bin SHI ; Hong GUO ; Bin ZHANG ; Chao YIN ; Qiliang HOU ; Yongqiang CAO ; Xinzhong WEN
Parenteral & Enteral Nutrition 2010;17(1):11-12,16
Objective: To discuss the effect of intensive insulin therapy on D-lactic acid and diamine oxidase level in patients with sepsis.Methods: 48 patients was divided into control group and conventional group randomly.Content of D-lactic acid and diamine oxidase in serum was detected with absorption spectrometry before and after therapy.Result: D-lactic acid and diamine oxidase in patients decreased significantly in control group compared to that of conventional group.Conclusion: Insulin can depress interstinal permeability and ameliorate sepsis symptom resulted from bacterial translocation from intestine.
3.HO-1 overexpression induced by CoPP in donors can prolong the survival of transplanted allogeneic islets of rats
Ling HOU ; Zhuzeng YIN ; Ying XIANG ; Junhua LI ; Shi CHEN ; Gang CHEN ; Xiaoping LUO
Chinese Journal of Organ Transplantation 2009;30(4):211-214
Objective To investigate the effects of the overexpression of HO-1 induced by CoPP in the donor on the survival of transplanted allogeneic islets of rats and the mechanism.Methods (1) Brown Norway rats were randomly divided into control group, and CoPP-induced group receiving intraperitoneal injection of CoPP (2.5 mg/kg) at 3rd and 1st day prior to islet isolation.By using the cytoimmunofluorescenee and Western blot, the expression of HO-1 in isolated islets was detected.The insulin level in the supernatant of the cultured islets stimulated with glucose was determined by ELISA.(2) Lewis male rat diabetic models were established by a single intravenous injection of alloxan, and then randomly divided into CoPP group and control group.Islets were transplanted under the left kidney capsule of each diabetic recipient.The survival time after transplantation, and pathological changes following rejection of the islet grafts were analyzed.Results The HO-1 was highly expressed in the islets isolated from CoPP-treated rats by cytoimmunofluorescence and Western blot.After stimulation with 16.7 mmol/L glucose, the insulin concentration in Copp-treated and Copp-untreated groups was (46.60± 1.13) and (19.01 ± 1.49) mIU/L respectively (P<0.05).The insulin concentration in Copp-treated and Copp-untreated groups in islets stimulated with 5.6 mmol/L glucose was (15.65 ± 0.89) and (12.28 ± 0.89) mU/L respectively (P>0.05).The stimulated index in Copp-treated and Copp-untreated groups was (2.98 ± 0.10) and 1.55 + 0.01 respectively (P< 0.05).The survival time of islets allograft in Copp-treated and Copp-untreated groups was separately (12.20±5.67) and (5.60± 1.14) days respectively (P<0.05).Histological analysis revealed the presence of more islands of insulin-positive cells and considerably fewer lymphocytes or inflammatory infiltration than the controls.Conclusions CoPP could induce the HO-1 expression of islets, and improve their function.Over-expression of HO-1 in islets could prolong survival time of islets allograft.
4.A study of the treatment method of cerebral artery dissection
Rong YIN ; Xiangqun SHI ; Zhiqiang ZHANG ; Xuemei HOU ; Shaoju SHAO ; Yanping LIU ; Weimin WANG ; Hongbo LUO
Chinese Journal of Radiology 2012;46(7):640-644
ObjectiveTo explore the best treatment method of cerebral artery dissection.Methods This study included eight patients who were definitely diagnosed as cerebral artery dissection by the cerebral angiography in our department of neurology during Oct.2009 and Nov.2011.They were all treated by the anticoagulation or anti-platelet methods.Some patients received the stent therapy.All patients' were followed for at least three months. The treatment effect was assessed by NIHSS,mRS and by the cerebral angiography.ResultsSix patients had carotid artery dissection,2 had vertebral artery dissection.Four patients were given anticoagulant therapy and the other 4 were given anti-platelet therapy. The reexamination by angiography 10-14 days after admission showed that in 3 patients,the stenosis was aggravated or the infarction occurred.They were diagnosed as having repeated transient ischemic attack (TIA) during pharmacotherapy and received stents for treatment.There was no TIA and cerebral infarction in the follow-up period after individualized therapy.Mean NIHSS scores of 8 patients between pre and post treatment were 5.9,1.6 respectively. Mean mRS scores pre and post treatment were 2.5,0.9respectively.Conclusion The treatment for patients with cerebral artery dissection should be individuated.The patients in acute stage should get anticoagulation,anti-platelet therapy and angiography re-examination.According to the clinical manifestation and cerebral angiography,the next step for the treatment should be done.
5.Construction and Screening of a Phage Display Library of Repertoire Single Chain Fv Antibody from Mouse Immunized with BoNTB/Hc
Xiu-Qing YANG ; Hui WANG ; Jing SHI ; Kun CAI ; Xiao-Jun HOU ; Shi-Zhong BAO ; Jun YIN ;
Microbiology 1992;0(06):-
To produce antibodies capable of neutralizing botulinum neurotoxin type B(BoNT/B),We cloned the carboxy-terminal end of Hc containing the major determinants responsible for specific toxin,induced and purifed.The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were amplified individually from the spleen cell mRNA by RT-PCR and joined as a single-chain Fv(scFv)DNA fragment.These fragment were cloned into the phagemid pCANTAB5E and the phage display library was constructed.Results showed that the high affinity scFv was obtained after 4 rounds of panning,with its DNA sequence conforming to that of mouse antibody.
6.Granulocytic sarcoma: a clinical and pathologic analysis of ten cases.
Zhen-bo HOU ; Huai-yin SHI ; Xiao LIANG ; Xin-mei WANG
Chinese Journal of Pathology 2012;41(5):331-334
OBJECTIVETo investigate the clinical and pathological features, differential diagnosis of granulocytic sarcoma.
METHODSThe clinical manifestations, histopathological features, immunohistochemistry, treatment and prognosis were analyzed retrospectively in 10 cases of granulocytic sarcoma.
RESULTSThe age of patients ranged from 10 to 56 years (means = 35.8 years). The male-to-female ratio was 1.5:1. Histologically, the malignant cells of granulocytic sarcoma grew in a diffuse pattern. The cytoplasm was scanty, with eosinophilic fine granularity in some cells. The nuclei were round or focally irregular, and had finely dispersed chromatin. The mitotic figures were visible. Immunohistochemical stains for MPO, CD43, CD117, CD34 and CD99 were positive.
CONCLUSIONSGranulocytic sarcoma can occur in patients of all ages with a male predominance. The diagnosis of granulocytic sarcoma is assisted by the cytochemical stain for naphthol-ASD-chloroacetate esterase and/or immunophenotypic analyses for MPO, CD43, CD117, CD34, CD99. These stains aid in the distinction of granulocytic sarcoma from: lymphoblastic lymphoma, Burkitt lymphoma, diffuse large B-cell lymphoma, small round cell tumours, particularly in children, and blastic plasmacytoid dendritic cell neoplasm.
12E7 Antigen ; Adolescent ; Adult ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Burkitt Lymphoma ; metabolism ; pathology ; Cell Adhesion Molecules ; metabolism ; Child ; Dendritic Cells ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Leukosialin ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Middle Aged ; Peroxidase ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-kit ; metabolism ; Retrospective Studies ; Sarcoma, Myeloid ; metabolism ; pathology ; Skin Neoplasms ; metabolism ; pathology ; Young Adult
7.Prognostic analysis of acute motor axonal neuropathy
Shi-Fang HOU ; Xian-Hao XU ; Yin-Hong LIU ; Huo ZHANG ; Xiang WANG ; Jian YIN
Chinese Journal of Neuromedicine 2009;8(5):496-499
Objective To investigate the clinical features of Guillain-Barre syndrome (GBS) and clarify the long-term prognosis of acute inflammatory demyelinating polyneuropathy (AIDP) and acute motor axonal neuropathy (AMAN). Methods We conducted an analysis of the clinical data of 50 patients with GBS treated in our hospital between 2003 and 2007. According to the eleetrophysiological criteria, the eases were classified into AIDP (n=24) and AMAN eases (n=22), with 4 eases of unknown classification. The patients unable to walk upon discharge were followed up for more than 6 months, and the clinical features and prognosis of the two groups were compared. Results The age, gender, cranial nerve involvement, Hughes grade at the initial neurological examination and at the time of symptom peak did not differ significantly between the patients with AIDP and AMAN (P>0.05), and most of the AMAN patients had a good recovery. The number of patients capable of walking at one month after the onset was comparable between the two groups (P>0.05). In AMAN group, the percentages of patients with slow recovery and those having rapid recovery were significantly higher than those in AIDP group (P<0.05). Conclusion The clinical recovery of AMAN patients can be either rapid or prolonged, and rigorous immunotherapy should be administered to achieve early recovery and ensure more favorable outcomes of the patients.
8.Dynamic Changes of Hydrogen Sulfide in Cortical Tissues of Neonatal Rats with Hypoxia-Ischemia Brain Damage
cai-li, REN ; hong-gang, ZHAO ; lei, LIU ; wan-li, ZHEN ; shi-qing, WANG ; xiao-feng, YIN ; zhi-hui, HOU ; dong-liang, LI
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To explore the dynamic changes of hydrogen sulfide(H2S)in the pathological course in cortical tissues at diffe-rent times of hypoxia-ischemia brain damage(HIBD).Methods Fifty-six healthy 7-day-old Sprague-Dawley newborn rats were randomly assigned into 7 groups(n=8):normal group,sham-operated group,HIBD 12 h group,HIBD 24 h group,HIBD 48 h group,HIBD 72 h group,and HIBD 7 d group.HIBD rat models were established by ligating the left common carotid artery,after 2-4 h,followed by exposuring to hypoxia(80 mL/L oxygen and 920 mL/L nitrogen)for 2 h.The achievement of HIBD model was determined by the change on behaviour of neonatal rats.There were no treatment on the normal group,and the left common carotid artery was only separated in the sham group.The left cortical tissues in the experimental group were removed at 12,24,48,72 h,and 7 d after HIBD.H2S amounts in cortical tissues at different times after HIBD were measured by biochemical methods.Results H2S level in cortical tissues in HIBD 12 h group increased significantly compared with sham-operated group(P
10.Intrauterine HBV infection: risk factors and impact of HBV DNA.
Yu-zhu YIN ; Xiao-wei CHEN ; Xiao-mao LI ; Hong-ying HOU ; Zhong-jie SHI
Journal of Southern Medical University 2006;26(10):1452-1454
OBJECTIVETo investigate the risk factors of intrauterine hepatitis B virus (HBV) infection and the impact of HBV DNA on the infection.
METHODSThe serum levels of HBsAg, HbsAb, HBeAg, HBeAb, HBcAb and HBV DNA were determined in blood samples from 230 HBsAg-positive pregnant women and their newborns by enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative PCR (FQ-PCR), respectively. The newborns acquiring HBV infection via intrauterine transmission were selected as the case group and others as the control group. The risk factors for intrauterine HBV infection were analyzed by non-conditional logistic regression model.
RESULTSSix infants were found to be HBsAg-positive, and 18 HBV DNA-positive, and 3 of them were positive for both HBsAg and HBV DNA. The rate of intrauterine HBV infection was 9.6% (22/230). The grade of HBV DNA level was identified as the only risk factor of intrauterine HBV infection by non-conditional logistic regression model, with odds ratio (OR) of 1.57 (95% confidence interval 1.12-2.21). Of the 119 pregnant women positive for HBV DNA, 18 were diagnosed as having intrauterine HBV infection, and the likeliness of the infection significantly increased for a maternal serum HBV DNA level > or =10(7) copies/ml (chi(2)=7.92, P<0.05).
CONCLUSIONThe grade of serum HBV DNA level is the predominant risk factor for intrauterine HBV infection in pregnant women, and for those with serum HBV DNA lever > or =10(7) copies/ml, the chance for intrauterine HBV infection can be significantly increased.
DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B ; transmission ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Logistic Models ; Odds Ratio ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy Complications, Infectious ; blood ; virology ; Risk Factors