1.Characterization and clinical significance of the Smith antigen
Journal of Medical Postgraduates 2004;0(01):-
Systemic lupus erythematosus(SLE) is an autoimmune disease characterized by the production of autoantibodies against a wide spectrum of nuclear, cytoplasmic,and cell membrane autoantigens.Among them,the anti-Smith antibodies are specific for SLE and thus have been included as a criteria for diagnosis.They are not only a diagnostic marker but also a reliable measure of disease activity in SLE,we intend to describe the biological functions and clinical significance of the Smith antigen and the detection of anti-Smith antibodies.
2.Secreted expression of recombinant human autoantigen Sm B' in methylotrophic yeast Pichia Pastoris
Xiangyue YANG ; Xiaopeng LAN ; Fuying FENG ; Wenbing WU ; Zhongyong ZHU
Chinese Journal of Laboratory Medicine 2008;31(10):1178-1181
Objective To clone and express human autoantigen Sm B'in methylotrophie yeast Pichia Pagtoris.Methods The gene Sm B' was cloned bv PCR The PCR product wag inserted into the vector pPIC9k.The recombinant plasmid pPIC9k.Sm B' was transformed into yeast Sm D1168 by electroporation.The positive clones were screened in MD plates.The high copy number transformants were rapidly selected by using G418 and were induced by methan01.Supematants after induction were analyzed by SDS-PAGE and western blot.Sera collected from thirty patients with SLE.thirty patients with mixed connective tissue disease(MCTD)and thirty healthy volunteers were detected by immunodot and immunoblot.Results The PCR product wag about 700 bD in size which Wag in accordance with predicted 657 bp.The pPIC9k-Sm B'showed the same seqencing result with GenBank's report and restriction enzyme analysis confirmed our prediction.The pPIC9k-Sm B' positive clone produced a 32 000 protein which had natural immunogenicitv of human autoantigen Sm B'by SDS-PAGE and western blot.The positive rate of immunodot and IBT were 46.7%(42/90)and 51.1%(46/90),respectively.The agreement between immunodot and IBT was very close(Kappa value=0.911 2,P<0.01).Conclusion Successfully cloning and expression of human autoantigen Sm B' in methylotmphic yeast Pichia Pagtoris hid a foundation for further research work.
3.Risk factor analysis for 30 - day mortality in patients with malignant hilar obstruction after percutaneous transhepatic biliary stent deployment
Mingwu LI ; Wenbing WU ; Zhanxin YIN ; Guohong HAN
Journal of Interventional Radiology 2014;(9):788-791
Objective To analyze the risk factors of 30-day mortality in patients with malignant hilar obstruction (MHO) after percutaneous transhepatic biliary metal stent deployment. Methods One hundred and fifty-nine consecutive patients with MHO caused by cholangiocarcinoma or gallbladder carcinoma were enrolled in this study. Percutaneous transhepatic biliary stent (PTBS) implantation was carried out in all the patients. Independent predictors for 30-day mortality were evaluated by logistic regression analysis. Covariates that were incorporated into the multivariate analysis were the variables that reached statistical significance (P < 0.1) in univariate analysis. Two-tailed, P value of less than 0.05 was considered to be statistically significant. Results The 30-day mortality of patient with MHO after metal stent deployment was 9.4%. Univariate analysis indicated that the differences in WBC (OR = 1.224.95%CI [1.07 - 1.44], P < 0.01), INR (OR=78.75, 95%CI [5.02-1 235.70], P<0.01), PT(OR=1.55, 95%CI [1.18-2.04], P<0.01), BUN (OR=1.19, 95%CI [1.02- 1.38], P < 0.05), CRE(OR = 1.02, 95%CI [1.000 - 1.041], P < 0.1) and lymph nodes metastasis(OR = 0.334. 95%CI[0.105 - 1.131], P < 0.1) were statistically significantly between 30-day mortality group and non-30-day mortality group. Multivariate analysis showed that statistically significant differences in WBC (OR = 1.19, 95%CI[1.026 - 1.380], P < 0.05), INR(OR = 151.5, 95%CI [3.13 - 5 440.7], P < 0.05) and CRE (OR = 1.025, 95%CI [1.002 - 1.048], P < 0.05) also existed palliative treatment for patients with malignant hilar obstruction. Active preoperative measures to improve hepatic and renal functions as well as to control infection are necessary in order to reduce 30-day mortality.
4.Detection of human coronaviruses NL63, 229E, HKU1 and OC43 in children with acute respiratory tract infection in Fnzhou, China
Yanan WU ; Xiaoqing WU ; Xiuya ZENG ; Junping CHEN ; Jianlin ZHOU ; Yunhuan CHEN ; Wenbing WU
Chinese Journal of Infectious Diseases 2012;30(1):53-57
Objective To establish real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) for human coronaviruses(HCoV)-NL63,HCoV-HKU1,HCoV-OC43 and HCoV-229E,and to investigate the prevalence of the four coronaviruses in children with acute respiratory tract infection(ARTI) in Fuzhou area.Methods Totally of 538 nasopharyngeal swabs were collected from pediatric patients with ARTI,including 289 specimens from children with acute upper respiratory tract infection (AURTI) and 249 from acute lower respiratory tract infection (AURTI) during three consecutive winter-spring seasons from December to April of 2006 - 2009 in Fuzhou area.All the specimens were subjected to FQ-PCR specific for HCoV-NL63,HCoV-HKU1,HCoV-OC43 and HCoV-229E,respectively.The enumeration data were analyzed by chi square test.Results The FQ-PCR methods were established for detecting HCoV-NL63,HCoV-HKU1,HCoV-OCA3 and HCoV-229E.The intraassay coefficient of variation (CV) and interassay CV were both ≤ 1.6%.The coronaviruses were detected in 41 (7.6%) children with ARTI,including HCoV-NL63 in 8 (1.5%)children (1 with AURTI,7 with ALRTI),HCoV-229E in 5 (0.9%; 1 with AURTI,4 with ALRTI),HCoV-HKU1 in 6 (1.1%; 1 with AURTI,5 with ALRTI),and HCoV-OC43 in 22(4.1%; 13 with AURTI,9 with ALRTI).The four coronaviruses were detected during each of the three winter-spring seasons and the positive rates of different periods were not significantly different (P>0.05).The HCoV-OC43 positive rate was significantly higher than HCoV-NL63,HCoV-229E and HCoV-HKU1 (x2 =6.721,10.979,9.387; respectively; all P<0.01).ConclusionsIt is suggested that the four coronaviruses might be important virus pathogens in children with ARTI in Fuzhou,China.And detection of them is needed for etiology and epidemiology evaluations for children with ARTI.
5.Differential expression of ADD3 splicing isoforms between colorectal cancer and normal mucosa tissues
Min TAO ; Liangxiang HUANG ; Pengwei CAI ; Long JIN ; Wenbing WU ; Changqing ZENG ; Yi HUANG ; Yanan WU
Chinese Journal of Pathophysiology 2016;32(3):451-457
AIM:To investigate the relationship between the expression of adducin 3 (ADD3) and its splicing isoforms and colorectal cancer (CRC).METHODS:The expression of ADD3, ADD3-Ia and ADD3-Ib in 50 pair of CRC tissues , 20 pairs of colorectal polyp tissues , and 2 CRC cell lines SW480 and SW620 before and after oxaliplatin or fluoroura-cil intervention were detected by real-time PCR.The cell activity was determined by MTT assay , the cell migration ability was evaluated by wound-healing assay , and the cell invasion ability was measured by Transwell assay .RESULTS:The expres-sion levels of ADD3 and ADD3-Ib were decreased in the CRC tissues as compared with the normal mucous (P<0.01), and ADD3-Ia/Ib ratio was increased in the CRC tissues (P<0.01).The expression level of ADD3-Ia was higher in T3-4 group than that in T1-2 group (P<0.05).Reduced expression of ADD3, ADD3-Ia and ADD3-Ib in colorectal polyps was observed compared with the normal tissues (P<0.01).Compared with the SW480 cells, the expression levels of ADD3-Ia and ADD3-Ib were lower (P<0.05) and the ADD3-Ia/Ib ratio was higher (P <0.01) in the SW620 cells.After treated with oxalipla-tin or fluorouracil, the cell activity, migration and invasion in the SW620 and SW480 cells were weakened accompanied by the increases in the expression levels of ADD 3, ADD3-Ia and ADD3-Ib to various certain extents .CONCLUSION:In CRC there is a tendency that ADD3-Ib reduction leads to ADD3 decrease, accompanied by an increased ADD3-Ia/Ib ratio.The expression changes of ADD 3 and its splicing isoforms in the CRC may be relevant to its invasion ability .
6.Expression of eEF1A2 in hepatocellular carcinoma
Yi HUANG ; Funan QIU ; Dunyan CHEN ; Yanan WU ; Feng LI ; Xiaoli HUANG ; Wenbing WU
Chinese Journal of Pathophysiology 2015;(12):2144-2150
eEF1A2 might be a putative oncoprotein in HCC .eEF1A2 over-expression has noticeable effects on the HCC cell prolifera-tion enhancement , differentiation inhibition , and cell cycle acceleration through the G 0/G1 phase to S phase and G 2/M phases.
7.Simultaneous Content Determination of Multiple Indicator Ingredients in Diefferent Grades of Phellodendron amurense Decoction Piece by HPLC
Shanshan WU ; Lin HU ; Xiaomeng GONG ; Mengqi LI ; Zhimin CHEN ; Changjiang HU ; Wenbing LI
China Pharmacy 2016;27(15):2135-2137
OBJECTIVE:To establish a method for the simultaneous content determination of berberine hydrochloride,phello-dendrine hydrochloride and magnoflorine in Phellodendron amurense decoction piece,and to campare the contents of the 3 ingredi-ents in different grades of P. amurense decoction piece. METHODS:HPLC was performed on the column of Phenomenex Luna C18 with mobile phase of acetonitrile-0.05 mol/L KH2PO4 (gradient elution) at a flow rate of 1 ml/min,the detection wavelength was 280 nm,the column temperature was 30 ℃,and injection volume was 5 μl. RESULTS:The linear ranges were 0.387 0-7.740 μg for berberine hydrochloride(r=0.999 9),0.044 4-0.888 0 μg for phellodendrine hydrochloride(r=0.999 8)and 0.048 0-0.960 0 μg for magnoflorine(r=0.999 9);RSDs of precision, stability and reproducibility tests were lower than 3%, recoveries were 95.61%-103.22%(RSD=2.80%,n=6),96.18%-102.80%(RSD=1.84%,n=6) and 97.93%-102.78%(RSD=1.84%,n=6). CONCLUSIONS:The method is simple and accurate,and can be used for the contents determination of berberine hydrochloride, phellodendrine hydrochloride and magnoflonine in P. amurense. The contents of berbenine hydrochloride and phellodendrine hydro-chloride in the first-grade decation piece are higher than those in the second-grade decoction piece,and the content of magnoflorine in both decoction pieces shows no discernible differences.
8.Determination of plasma neutrophil gelatinase associated lipocalin and its clinical significance in sepsis
Meimiao LIU ; Wenbing QIU ; Lanchun CHEN ; Zhaoxi WU ; Weixin TANG ; Xuetao YU
Chinese Journal of Primary Medicine and Pharmacy 2016;23(6):826-829,830
Objective To investigate concentration of plasma neutrophil gelatinase -associated lipocalin ( NGAL) and its clinical significance in ICU populations with sepsis.Methods Fourteen ICU patients diagnosed with sepsis and twenty-four patients with non-sepsis were enrolled in the study,and seventeen healthy adults were selected as healthy control.Blood samples were drawn from patients to measure NGAL, and Acute Physiology and Chronic Health Evaluation ( APACHE ) II and Sequential Organ Failure Assessment ( SOFA ) were performed, the outcome was recorded.Plasma NGAL concentration was measured by enzyme -linked immunosorbent assay ( ELISA) .Results On admission to ICU, the pNGAL concentrations of septic and non -septic patients were (131.4 ±116.4)ng/mL and (48.7 ±30.8)ng/mL respectively,which of healthy control was (39.07 ±12.74)ng/mL.The pNGAL was significantly higher in sepsis than non-sepsis and healthy control ( t=-3.280,3.313,P=0.003,0.021 respectively).Otherwise,no differences of white blood cell (WBC) count [(12.6 ±5.7) ×109/L vs (15.6 ±5.8) ×109/L] and SOFA scores [(7.7 ±3.3)points vs (8.6 ±3.4)points] were found in septic and non-septic patients (t=-1.554,-0.802,P=0.129,0.428).On the admission to ICU,areas under the receiver oper-ating characteristic curves ( ROC curves) of NGAL and WBC for predicting sepsis were 0.740 [95%confident inter-val(CI) 0.566,0.913,P =0.015] and 0.345 ( 95%CI 0.158,0.533,P =0.116) respectively.If pNGAL of 99.4ng/mL as the threshold for predicting sepsis, the sensitivity and the specificity was 50.0% and 85.2%. Conclusion The concentrations of NGAL in septic patients significantly increased compared with that in non-sepsis and in healthy adults.And the NGAL levels maybe another sensitive biochemical marker for predicting infection.
9.The effects of exercises on the expression of matrix metalloproteinase-9 in perihematomal brain tissue after intra cerebral hemorrhage in rats
Duozi WANG ; Fuqiang GUO ; Hongyan ZHANG ; Hao SUN ; Xiangrong SUN ; Xianrong ZENG ; Jinyu WANG ; Wenbing WU ; Fuqiong PAN
Chinese Journal of Physical Medicine and Rehabilitation 2009;31(8):509-513
Objective To study the effects of exercises training on the recovery of neurological function and the expression of matrix metalloproteinase-9 (MMP-9) in perihematomal brain tissue after intra cerebral hemorrhage (ICH) in rats. Methods Sixty-four male adult Sprague-Dawley rats were randomly divided into exercises group and control group. ICH model was induced by autobiood injection. The rats in exercises group were given balance, grasping and walking exercises every day. The rats in control group exercised freely in standard cages. Neurological function in both groups was measured at the 3rd, 7th, 14th and 21st d. All the rats were sacrificed and the concentration of MMP-9 was measured with immunohistochemical method and in situhybridization. Results In both groups neurological function scores was lowest at the 3rd d, were highest at the 21st d. There was no significant difference between two groups at the 3rd d, but at the 7th, 14th and 21st d the differences between two groups were significant ( P < 0.05 ). In exercises group, MMP-9 expressed weakly at the 3rd d, and peaked at the 7th d, then declined gradually. In control group, MMP-9 expression peaked at the 3rd d, then decreased slowly. MMP-9 protein expression was in accord with mRNA. MMP-9 expression in exer-cises group was significantly different from control group on the 3rd and 7th d (P < 0.05 ) , but not at the 14th and 21st d ( P > 0.05 ). Conclusions Early exercises can inhibit the expression of MMP-9 that could play a role in protecting neurons. Daily exercises can stimulates the expression of MMP-9 so as to have a positive role in midstage of disease. MMP-9 may be involved in tissue remodelling and vascular repairing, which prompt neu-rofunction recovery.
10.Research and development on microwave sterilization.
Xiaoli XI ; Daocheng WU ; Gang WANG ; Wenbing WANG
Journal of Biomedical Engineering 2002;19(2):334-343
This paper reviews recent research and development on microwave sterilization, including the experiments of sterilization effects, the influence of microwave radiating on the media parameters, the development of microwave sterilizers and the study of sterilization mechanism. Many results show that the method of microwave sterilization is more easy-to-perform, economical and reproducible than the conventional autoclaving method. It is necessary to study the sterilization conditions through the experiments for different media and bacteria. The research on sterilization mechanism, especially non-thermal sterilization mechanism is still a hotpoint.
Bacteria
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radiation effects
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Microwaves
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Research
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trends
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Sterilization
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methods