1.Stable surface expression of a gene for Helicobacter pylori toxic porin protein with pBAD expression system.
Zhixiang, PENG ; Xi, WEI ; Zhengmei, LIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):435-8
Helicobacter pylori (H. pylori) infection causes peptic and duodenal diseases in humans. Among a 32-protein family of outer membrane proteins, a porin-like protein, HopE, has been a subject of note, mainly for its conservative nature among H. pylori, and for its potential as a vaccine candidate. To achieve stable surface expression of this host cell-toxic protein, hopE gene was introduced into pBAD expression system. After induction with arabinose, all 15 randomly-chosen E. coli LMG 194 colonies from 3 successive passages could express HopE protein, while only 1 from 5 E. coli colonies that contained lac operon-regulated plasmid encoding hopE gene could express HopE. Indirect immunofluorescence confirmed the expression of HopE on E. coli cell surface.
2.Visualization Study of Virtual Human Tongue in Speech Production
Zhixiang CHEN ; QInqin XIN ; Yuexiu ZHU ; Ziqiong LIN ; Lin WANG
Chinese Journal of Rehabilitation Theory and Practice 2013;19(10):993-997
Pronunciation obstacle is one of the characteristics of hearing and speech disabled persons. As the main organ of pronunciation,tongue plays an important role in pronunciation training. If the pronunciation visualization is applied to rehabilitation training, this can make the hearing and speech disabled persons intuitively watch the change of the tongue in the process of pronunciation, which may promote the rehabilitation training. On the basis of tongue anatomical structure and movement characteristics, the common movements of tongue in pronunciation are realized after the establishment of three-dimensional tongue muscle model and the relevant data of the tongue Xray images and electropalatography. Using this kind of visualization technology, we can help correction and rehabilitation for the hearing and speech disabled persons.
3.Significance of peripheral CD_(34)~+ cell count on the harvest of mobilized peripheral hematopoietic stem cells
Wei TANG ; Lin WANG ; Weili ZHAO ; Zhixiang SHEN ; Jiong HU
Journal of Leukemia & Lymphoma 2010;19(5):265-268
Objective Autologous hematopoietic stem cell transplantation (Auto-HSCT) has been widely used in hematological malignancies.To mobilize and harvest sufficient number of peripheral CD_(34)~+ cells is one of key issues for auto-HSCT. Peripheral CD_(34)~+ cell numeration has been used as an indicator for apheresis while we mostly rely on the peripheral WBC or MNC count. In this study, we try to evaluate the association of peripheral CD_(34)~+ count to the CD_(34)~+ cells number in the apheresis product and to find out a potential threshold. Methods From Jan 2007 to Dec 2009, a total of 57 apherosis for auto-HSCT were analysed. All patients were mobilized by cyclophophamide (CTX) plus G-CSF(5-10μg/kg) regimen. The apheresis were performed with COBE SPECTRA VERSION 6 and CD_(34)~+ count of both peripheral and apheresis products were analysed by flow cytometry. Results The median number of MNC in apheresis products was 4.6(0.3-10.5)×10~8/kg with median CD_(34)~+ cells at 2.4(0.16-34.9)×10~6/kg. The peripheral CD_(34)~+ count was the only parameter associated with the MNC and CD_(34)~+ cell numbers in the apheresis products while the WBC number was irrelevant to the results of apheresis. Our data showed that when the peripheral CD_(34)~+ count reach 15/μl, the efficacy of a single apheresis significantly improved with 81 % and 60 % reached 1 and 2×10~6 CD_(34)~+ cells/kg respectively and the total number of MNC and CD_(34)~+ cells were significantly superior to apheresis with peripheral CD_(34)~+ cells <15/μl, thus indicated that CD_(34)~+ ≥15 /μl can be used as the threshold for apheresis. Furthermore, the ROC analysis demonstrated that CD_(34)~+ cells ≥25(26.5-28.6) /μl is the best indicator level for a successful single apheresis. Conclusion Our study clearly showed that peripheral CD_(34)~+ cell count is a key indicator of apherosis. CD_(34)~+ cells at 15/μl can be used as the threshold to start apheresis in the clinical setting.
4.The carcinogenic effect of UVB sensitive miR-365 in cutaneous squamous cell carcinoma
Meijuan ZHOU ; Haibo HUANG ; Zhixiang LIN ; Zhenhua DING
Chinese Journal of Radiological Medicine and Protection 2014;34(11):813-816,866
Objective To investigate the carcinogeic role of miR-365 in cuntanerous squamous cell carcinoma (cSCC).Methods Normal HaCaT cells were divided into control and irradiation groups,the later was exposed by UVB irradiation (50 J/m2).MicroRNA expression profiles of the two groups were analyzed by microRNA array.The expression variations of miR-365 in HaCaT,A431,Tca8113 and HSC-1 cells were validated by qRT-PCR analysis.The colony-forming and invasion capacities were dectected by colony forming assay and Transwell migration assay in vitro,respectively.HaCaTpre-miR365-2 highly expressing miR-365 was constructed by retroviral vector infection.Tumorigenicity evaluation was carried out by subcutaneously inject of the cells at the right back flank of nude mice.Results There were 30 microRNAs differentially expressed in HaCaT cells after UVB irradiation and miR-365 was one of the most sensitive miRNAs(as high 6.7 times as control).Expression of miR-365 in all the cSCC cell lines A431,Tca8113 and HSC-1 were significantly higher than that in HaCaT cell,in which the maximum was A431 (15.67 ±1.12 times,P < 0.01),and the minimum was TcaS113 (4.72 ± 0.85 times,P < 0.05).Knockdown of miR-365 in cSCC cell lines significantly inhibited the colony forming ability (t =13.68,P < 0.05) and cell migration (t =19.98,P < 0.05) in vitro.HaCaT cells overexpressing miR-365 by transient transfection significantly increased the ability of colony formation (t =7.11,P < 0.05) and cell migration (t =22.03,P <0.05) in vitro.In addition,HaCaTpre-miR-365-2 cell line stably expressing miR-365 could successfully establish tumors in nude mice.Conclusions MiR-365 is an oncogene for cutaneous squamous cell carcinoma.
5.Risk factors of disease progression and adverse pregnancy outcome in patients with systemic lupus erythematosus during pregnancy
Weinian LI ; Xiaoyan CAI ; Xiaojun LIN ; Chun TANG ; Jinghua YE ; Zhixiang HE ; Fangfei LI
Chinese Journal of General Practitioners 2017;16(3):214-218
Objective To investigate the risk factors of disease progression and adverse pregnancy outcome in patients with systemic lupus erythematosus (SLE) during pregnancy.Methods Clinical data of 118 pregnant women with SLE admitted from June 2004 to October 2015 were retrospectively analyzed.The patients were divided into selective pregnant group (group A,n =72 cases) and non-selective pregnant group (group B,n =46) according to the disease activity of SLE.The disease progression and pregnancy outcomes were compared between two groups.Results The various system damages occurred in group B,including hematological damage in 16 cases,kidney damage in 19 cases,erythra in 10 cases,arthritis in 10 and serositis in 12 cases;while the corresponding cases in group A were 10,14,6,4 and 4(x2 =7.133,P=0.008;x2 =6.658,P =0.010;x2 =4.304,P =0.038;x2 =7.030,P =0.008;x2 =10.095,P =0.001).SLE exacerbation occurred in 28 cases (74%) of group B and 12 cases (17%) of group A (x2 =34.944,P =0.000).The logistic regression analysis showed that hypocomplementemia,proteinuria,SLEDAI score before pregnancy and positive anti-dsDNA antibody were the risk factors of SLE disease exacerbation during pregnancy.The maternal complications occurred in group B,including pregnancyinduced hypertension in 7 cases,preeclampsia in 10 cases and infections in 11 cases/times;while the corresponding cases (case/time) in group A were 2,6 and 4 (x2 =4.526,P =0.033;x2 =4.304,P =0.038;x2 =8.525,P =0.004).There were 14 cases of therapeutic induced labor,7 case of stillbirth and 27 cases of total fetal loss in group B,while the corresponding cases in group A were 2,0 and 4 (x2=18.317,P =0.000;x2 =9.080,P =0.003;x2 =40.920,P =0.0300).The logistic regression analysis showed that positive anticardiolipin antibody,proteinuria,SLEDAI score before pregnancy and renal dysfunction during pregnancy were risk factors of fetal loss.There were 87 cases of successful delivery (73.7%),the successful delivery rates were 41.3% (19/46) in group B and 94.4% (68/72) from group A,respectively.The infant complications occurred in group B,including premature birth in 15 cases,low birth weight in 13 cases,neonatal jaundice in 5 cases and mild asphyxia in 5 cases;while the corresponding cases in group A were 24,18,4 and 2 (x2 =11.442,P =0.001;x2 =11.395,P =0.001;x2 =4.664,P =0.031;x2 =8.035,P =0.005).Conclusion SLE patients whose disease conditions are not well controlled would lead to higher percentage of disease deterioration during pregnancy and worse pregnancy outcomes.
6.A non-controlled, multicenter open-label study to evaluate the safety, tolerability and efficacy of caspofungin in the treatment of invasive candidiasis and esophageal candidiasis
Dongfang LIN ; Jianmin WANG ; Yunsong YU ; Mingzhe HAN ; Zhixiang SHEN ; Shiduo SONG ; Yingyuan ZHANG
Chinese Journal of Infection and Chemotherapy 2014;(5):375-381
Objective This study was designed to evaluate the safety ,tolerability and efficacy of intravenous caspofungin for treatment of invasive candidiasis and esophageal candidiasis in Chinese adults .Methods This was a non-controlled ,multicenter ,candidiasis .All the 63 patients were included in the safety set (SS) and the full analysis set (FAS) .In the SS ,19 SAEs occurred in 14 patients .All these SAEs were unrelated to caspofungin .There were 73 caspofungin-related non-serious AEs in 31 patients (49 .2% ) .Five patients (7 .9% ) had both clinical AEs and laboratory abnormalities .Eight patients (12 .7% ) had clinical AEs (mainly rashes) ,and 27 patients (42 .9% ) had laboratory abnormalities ,mainly increases in liver enzymes alanine transaminase and aspartate transaminase and reduction in blood potassium .About 91 .7% of the clinical AEs were mild to moderate .Treatment was discontinued in 1 patient (1 .6% ,1/63) due to AEs .The overall efficacy was 58 .1% (36/62) in the FAS and 70 .0% (35/70) in the per-protocol set (PPS) .In the FAS ,the therapeutic efficacy was 57 .6% (34/59) for invasive candidiasis and 66 .7% (2/3) for esophageal candidiasis .In the PPS , the therapeutic efficacy was 68 .8% (33/48 ) for invasive candidiasis and 100% (3/3 ) for esophageal candidiasis .Conclusions The AEs of caspofungin were mostly mild to moderate in the treatment of invasive candidiasis and esophageal candidiasis in Chinese adults .Only one patient terminated therapy due to drug-related AE .Caspofungin is safe and effective for the treatment of invasive candidiasis and esophageal candidiasis in Chinese adults .
7.Expression and significance of B lymphocyte stimulator and BAFF-receptor in peripheral blood mononuclear cells of patients with systemic lupus erythematosus.
Zhixiang HE ; Xiaoyan CAI ; Xiaojun LIN ; Yanli XU ; Chun TANG ; Jinghua YE ; Shuguang QIN ; Junzhou FU
Clinical Medicine of China 2008;24(11):1075-1077
Objective To investigate the expression and significance of B lymphocyte stimulator (Blys) and its receptor BAFF (BAFF-R) in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE). Methods The expression of Blys and BAFF-R was measured by flow cytometry in 90 pa-tients with SLE,which was compared with that of 45 healthy controls. The relationships between the expression of Blys, BAFF-R and other laboratory parameters as well as disease activity were analyzed. Results The expression of Blys and BAFF-R in PBMCs from patients with SLE was significantly elevated compared to healthy controls (P <0.001), so did the active group (P < 0.001) and inactive group (P < 0.001 and P < 0.01). The expression of Blys in PBMCs from active SLE patients was higher than that of inactive patients (P <0.05). However,there was no statisti-cal difference of BAFF-R between the two groups. The expression of Blys in PBMCs was positively related to SLEDAI (r =0.728,P <0.001) ,IgG and IgM(r=0.691,P<0.001 and r =0.453,P<0.01) ,but negatively related to C3 and CA (r = -0.510, P < 0.001 and r = -0.312, P < 0.05). The expression of Blys in dsDNA positive group was higher than those of dsDNA negative group (P < 0.01). The expression of Blys and BAFF-R in Cl qAb positive group was higher than those of ClqAb negative group as well (P <0.01). Conclusion The expression of Blys and its receptor BAFF-R in PBMCs from SLE is elevated ,which may reflect the disease activity and is related to the pro-duction of autoantibody. They might be involved in the pathogenesis of SLE.
8.Diagnosis and management of rectal gastrointestinal stromal tumor in 19 cases
Hongwei LIN ; Yongfu SHAO ; Dongbing ZHAO ; Haizeng ZHANG ; Tiecheng WU ; Zhixiang ZHOU ; Yuliang WAN ; Jianxiong WU ; Ping ZHAO
Chinese Journal of General Surgery 1993;0(03):-
Objective To investigate the clinical and pathologic characteristics of rectal gastrointestinal stromal tumor (GIST),and to evaluate the management of rectal GIST. Methods The clinical and pathological data of 19 cases of rectal GIST in recent 19 years were studied retrospectively. Results The diagnosis of 19 cases of rectal GIST were identified by surgery and pathology. Most rectal GISTs were spindle cell type. Immunohistochemical analysis displayed positive reactivity for CD117(100%) and CD34(73. 7% ). Graded by aggressive behavior there were 4 cases of high risk, 3 cases of intermediate risk, 5 cases of low risk and 7 cases of very low risk. Conclusions Rectal GISTs have a low prevalence and have no specific symptom in the early stage. Most tumors are low risk in aggressive behavior. It is difficult to get an accurate pathological diagnosis before operation and difficult to decide whether a sphincter preserving procedure is justified however trans-anal local resection is the therapy of choice for low risk submucosal rectal GIST(
9.Stable Surface Expression of a Gene for Helicobacter pylori Toxic Porin Protein with pBAD Expression System
PENG ZHIXIANG ; WEI XI ; LIN ZHENGMEI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):435-438
successive passages could express Hope protein, while only 1 from 5 E. coli colonies that contained lac operon-regulated plasmid encoding hopE gene could express HopE. Indi-rect immunofluorescence confirmed the expression of HopE on E. coli cell surface.
10.Serum vitamin D levels in patients with systemic lupus erythematosus and their relationship with disease activity
Qinyan LU ; Xiaoyan CAI ; Xiaojun LIN ; Chun TANG ; Jinhua YE ; Weinian LI ; Zhixiang HE ; Xinying LI ; Rongwei ZHANG
The Journal of Practical Medicine 2015;(7):1123-1125
Objective To detect the serum 25- OH-D levels in patients with systemic lupus erythematosus (SLE) and investigate the function of vitamin D in the pathogenesis of SLE. Methods Eighty SLE patients including 40 healthy donors and 40 RA (rheumatoid arthritis) patients were enrolled in this study. Serum 25-OH-D levels were detected with ECL method. Results (1)Serum 25-OH-D levels in patients with active SLE were lower than those with inactive SLE patients and healthy controls. (2)Serum 25-OH-D levels were negatively correlated with SLEDAI (Systemic Lupus Erythematosus Disease Activity Index) scores (r = -0.45,P < 0.01) and 24-hours urinary protein excretion (r = -0.32,P < 0.05). Conclusion Serum 25-OH-D levels in SLE patients decrease and have close relationship with disease activity and renal lesions.