2.Expression and Purification of Enterovirus Type 71 Polyprotein P1 using Pichia pastoris system
Xue HAN ; Xiaoling YING ; Hao HUANG ; Shili ZHOU ; Qi HUANG
Virologica Sinica 2012;27(4):254-258
Enterovirus type 71(EV71) causes severe hand-foot-and-mouth disease(HFMD) resulting in hundreds of deaths of children every year; However,currently,there is no effective treatment for EV71.In this study,the EV71 poly-protein(EV71-P1 protein) gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GS115.The EV71 P1 pretein with a molecular weight of 100 kD was produced and secreted into the medium.The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70%.The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits.We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.
3.Purification of L-amino acid oxidase from Naja atra venom and its effects on endothelial cells
Zhou CHEN ; Jianjun HUANG ; Ling XUE ; Yunlu XU ; Lishan LIN
Chinese Pharmacological Bulletin 2003;0(08):-
Aim To purify L-amino acid oxidase(LAAO) from the venom of Naja atra and study its effect on endothelial cells.Methods The NAV-LAAO was purified by ion-exchange chromatography and affinity chromatography.The MTT assay and Western blot were used to detect the viability and apoptosis of HUVEC.The tubule-forming was used to study the angiogenesis of cells.Results The NAV-LAAO was purified successfully from the venom of Naja atra.The molecular weight of NAV-LAAO was determined to be 58 ku by SDS-PAGE.NAV-LAAO effectively inhibited the growth and tubule-forming of HUVEC,and the 50% inhibitory concentration(IC50) was 21.42 mg?L-1.Compared with control,the levels of caspase-3 and caspase-8 increased in HUVEC treated with NAV-LAAO.Conclusions The NAV-LAAO is purified successfully from Naja atra venom by ion-exchange chromatography and affinity chromatography.The NAV-LAAO inhibits the growth and tubule-forming capacity of HUVEC in a dose-dependent manner.
4.Inhibitive effects of trichostatin A on telomerase activity of HL-60 cells and expression of subunit hTERT during apoptosis
Yongming ZHOU ; Keying XUE ; Yanhong CHEN ; Jun LIU ; Shiang HUANG
Chinese Pharmacological Bulletin 1987;0(02):-
Aim To investigate the inhibitive effects of trichostatin A(TSA) on telomerase activity of HL-60 cells and expression of subunit hTERT during apoptosis in vitro and its mechanism.Methods The proliferative activity of HL-60 cells was assessed using morphology and MTT assay.Cell apoptosis was confirmed using Flow Cytometer.Telomerase activity was examined using TRAP-ELISA.The expression status of telomerase subunits was analyzed using RT-PCR.Results A time-and dose-dependent inhibition was detected in HL-60 cells treated with TSA.After 48 h TSA(300 nmol?L~(-1)) treatment,the apoptotic rate detected using cytometric assay(Annexin V/PI double staining)of HL-60 cells was 42.6%.Telomerase activity and expression level of hTERT and the key subunit of telomerase decreased at 24-hour after TSA treatment.No significant changes were observed in the expression of hTR,hTP and the other two subunits of telomerase.Conclusion TSA inhibits telomerase activity and induces apoptosis in HL-60 cells.The underlying mechanism might be related to the down regulation of hTERT transcription.
5.An analysis of visual quality after Epi-LASIK
Lu ZHOU ; Zhenping HUANG ; Chunyan XUE ; Yuan XIA
Journal of Medical Postgraduates 2003;0(12):-
Objective: Epi-LASIK appeared to be effective and safe in the treatment of high myopia,but some patients complain about visual symptoms postoperation.The purpose of this study was to evaluate the high order aberrations(HOAs) of the vision-decreased eyes treated by Epi-LASIK,and to analyze the causes of postoperative glare,scieropia and monocular diplopia.Methods: We evaluated 76 eyes of 49 patients treated by Epi-LASIK with a questionnaire three months after surgery,and from them selected 34 eyes of 27 patients who scored less than 80 points and complained of postoperative visual symptoms,including glare(25 eyes in 20 patients),scieropia(7 eyes in 5 patients)and monocular diplopia(2 eyes in 2 patients).Another 42 symptom-free eyes of 22 patients treated by Epi-LASIK were included in the control group.The root mean square(RMS) of total HOAs,coma and SA were determined with the iTrace Visual Function Analyzer from the central 6.0 mm zone,and the modulation transfer function(MTF) was simulated with the iTrace 3.1 software.Results: Significant differences were found in the RMS of total HOAs,coma and SA between the eyes with visual symptoms and those without,and the values of ablation zone decentration were significantly higher in the former than in the latter(P
6.Correlation between overexpression of TLR4/MyD88 signaling and intestinal graft-versus-host disease after allogeneic bone marrow transplantation in mice
Shuai XING ; Xue ZHANG ; Xia HUANG ; Ping ZHOU
Chinese Journal of Organ Transplantation 2014;35(7):431-435
Objective To explore the expression of TLR4/MyD88/NF-κB signaling and its correlation with the progression of acute intestinal graft-versus-host disease (iGVHD) after allogeneic bone marrow transplantation in mice.Method Recipient BALB/c female mice were lethally irradiated and were reconstituted within 4-6 h with a transplant of bone marrow cells (1 × 107) and different amounts of splenocytes (1 × 107,n =12 or 2 × 107,n =12) from MHC-mismatched C57BL/6 donors to induce iGVHD,and 6 healthy BALB/c mice served as controls.A globe survey observation of GVHD by survival,clinical manifestation,and histological detection was performed.RT-PCR,immunohistochemistry,and Western blotting technology were used to detect the mRNA and protein levels of TLR4,MyD88 and NF-κB p65 in the small intestine tissue.Result The exacerbation of iGVHD was associated with the increasing dose of allogeneic spleen lymphocytes.The mRNA expression of TLR4,MyD88 and NF-κBp65 was increased as the iGVHD progressed.All of them in severe iGVHD model were significantly increased as compared with the healthy controls (P<0.05).The expression of corresponding proteins had the same tendency as mRNA.All of the three genes expression was not only positively correlated with each other,but also with the clinical GVHD score:TLR4 (R =0.814,P<0.001),MyD88 (R=0.828,P<0.001),and NF-κB p65 (R=0.568,P =0.034).Conclusion Excessive activation of TLR4/MyD88/NF-κB signaling pathway does exist in iGVHD,and the enhanced levels of gene transcription and translation are positively correlated with the deterioration of iGVHD.
7.Primary gastric inflammatory myofibroblastic tumor:a clinicopathologic study
Yiqiang LIU ; Dan XU ; Xiaozheng HUANG ; Lixin ZHOU ; Weicheng XUE
Chinese Journal of Clinical and Experimental Pathology 2015;(2):164-168
Purpose To investigate the clinicopathologic features, diagnosis and differential diagnosis of primary gastric inflammatory myofibroblastic tumor ( IMT) . Methods Four cases of gastric IMTs were studied by clinicopathologic analysis, immunohistochemistry and in situ hybridization, and the related literature was reviewed. Results In four cases there are two males and two females, age range from 21 to 51 years old, and tumor size ranged from 1. 5 to 6. 5 cm in the greatest dimension. Histologically, these tumors were composed of varied spindle cells and chronic inflammatory cells, in a myxoid or hyalinized stroma. Occasionally, there were calcifica-tion and ossification areas. Most of the spindle cells had bland appearance and a minority of the tumor cells showed mild atypia. One to two mitotic figures were recognized in 10 high power fields ( HPFs) in 1 to 2 patients. Smooth muscle actin staining was observed in all tumors and ALK staining observed in two tumors. One tumor focally expressed CD34. S-100, desmin, CD68, CD117 and DOG1 was negative in all IMTs. The patients were followed up from 24 to 66 months, and none of them had tumor relapsed or metastasis. Conclu-sions Primary gastric IMTs have an intermediate behavior, and a few cases have malignant potential. It should be distinguished from other spindle cell lesions similar to IMT.
8.Effects of Yisui Jiedu Recipe on JAK2-STAT5 signal transduction pathway in bone marrow hematopoietic cells from patients with myelodysplastic syndrome-refractory anemia.
Shengli TIAN ; Yongming ZHOU ; Tao HUANG ; Zhizhong XUE ; Wei HE
Journal of Integrative Medicine 2008;6(2):185-9
To investigate the effect of Yisui Jiedu Recipe (YSJDR), a compound traditional Chinese herbal medicine, on cytokines and their corresponding just another kinase 2-signal transducers and activators of transcription 5 (JAK2-STAT5) signal transduction pathway in bone marrow hematopoietic cells from patients with myelodysplastic syndrome-refractory anemia (MDS-RA).
9.The changes of the intercellular spaces of squamous epithelium of lower esophagus in gastroesophageal reflux disease
Yan XUE ; Li-Ya ZHOU ; San-Ren LIN ; Yonghui HUANG ;
Chinese Journal of Digestion 2001;0(12):-
Objective To observe the changes of the intercellular spaces of squamous epithelium of lower esophagus in gastroesophageal reflux disease(GERD).Methods Eleven outpatients with GERD symptoms more than 3 months [6 with nonerosive reflex disease(NERD)and 5 with erosive esophagitis(EE)]and 5 healthy volunteers were recruited.All of them underwent endoscopy and 24-hr ambulatory pH monitoring.Biopsies were taken in lower esophagus(2 cm above Z-line)for electron microscope examination.Results Intercellular spaces of esophageal epithelial cell in volun teers,NERD patients and EE patients were (0.374?0.073)?m,(1.308?0.079)?m and (1.332?0.144)?m respectively,with significant differences between the control group and the NERD or EE group.There was no difference between NERD group and EE group.Conclusions Dilated intercellular spaces were seen in both NERD and EE cases,which was significantly different from the control cases.
10.Reconstruction of soft tissue defects at finger tip with relay flaps pedicled by perforator from digital artery.
Zhou XIAO ; Xue MINGYU ; Xu YAJUN ; Qiang LI ; Huang JUN
Chinese Journal of Plastic Surgery 2015;31(6):422-425
OBJECTIVETo investigate the application of relay flaps pedicled by perforator from digital artery for reconstruction of soft tissue defects at finger tip.
METHODSFrom Mar. 2012 to Jun. 2014, 9 cases with soft tissue defects at finger tip were reconstructed with relay flaps at one side of finger pedicled by perforator from digital artery. The flap size ranged from 1.3 cm x 1.6 cm to 1.6 cm x 2.2 cm. The defects at donor sites were covered by adjacent web perforator V-Y advanced flaps.
RESULTSAll the 18 flaps in 9 cases survived completely with primary healing both in recipient and donor sites. The patients were followed up for 5 months to 2 years ( average, 12 months) with good elasticity and cosmetic results. No pain happened in the treated finger. The 2-point discrimination distance was 7-8 mm in fingertip flaps, and 10-12 mm in web perforator flaps. Hand function was graded as excellent in 7 cases, good in 2 cases, based on ATM assessment. The affected fingers had normal temperature and cold-resistance during winter. The width and depth of web in the donor site were not affected.
CONCLUSIONSThe relay flaps pedicled by perforator from digital artery can be applied for reconstruction of soft tissue defects at finger tip. The procedure is easy with satisfactory results and reservation of main artery. No skin graft is necessary for closure of defects on donor sites.
Arteries ; Elasticity ; Finger Injuries ; surgery ; Fingers ; blood supply ; Follow-Up Studies ; Humans ; Perforator Flap ; transplantation ; Time Factors ; Transplant Donor Site ; Wound Healing