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1.
China Journal of Chinese Materia Medica ; (24): 3066-3073, 2023.
Article in Chinese | WPRIM | ID: wpr-981437

ABSTRACT

This study aimed to investigate the effect of Wenyang Zhenshuai Granules(WYZSG) on autophagy and apoptosis of myocardial cells in rats with sepsis via regulating the expression of microRNA-132-3p(miR-132-3p)/uncoupling protein 2(UCP2). Sixty SD rats were randomly divided into modeling group(n=50) and sham operation group(n=10). The sepsis rat model was constructed by cecal ligation and perforation in the modeling group. The successfully modeled rats were randomly divided into WYZSG low-, medium-and high-dose groups, model group and positive control group. Rats in the sham operation group underwent opening and cecum division but without perforation and ligation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of rat myocardial tissue. Myocardial cell apoptosis was detected by TdT-mediated dUTP nick end labeling(TUNEL) assay. Real-time quantitative polymerase chain reaction(RT-qPCR) was performed to detect the expression of miR-132-3p and the mRNA expressions of UCP2, microtubule-associated protein light chain 3(LC3-Ⅱ/LC3-Ⅰ), Beclin-1 and caspase-3 in rat myocardial tissue. The protein expressions of UCP2, LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 in myocardial tissue were detected by Western blot. Dual luciferase reporter assay was used to verify the regulatory relationship between miR-132-3p and UCP2. The myocardial fibers of sepsis model rats were disordered, and there were obvious inflammatory cell infiltration as well as myocardial cell edema and necrosis. With the increase of the WYZSG dose, the histopathological changes of myocardium were improved to varying degrees. Compared with the conditions in the sham operation group, the survival rate and left ventricular ejection fraction(LVEF) of rats in the model group, positive control group and WYZSG low-, medium-and high-dose groups were decreased, and the myocardial injury score and apoptosis rate were increased. Compared with the model group, the positive control group and WYZSG low-, medium-and high-dose groups had elevated survival rate and LVEF, and lowered myocardial injury score and apoptosis rate. The expression of miR-132-3p and the mRNA and protein expressions of UCP2 in myocardial tissue in the model group, positive control group and WYZSG low-, medium-and high-dose groups were lower, while the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 were higher than those in the sham operation group. Compared with model group, the positive control group and the WYZSG low-, medium-and high-dose groups had an up-regulation in the expression of miR-132-3p and the mRNA and protein expressions of UCP2, while a down-regulation in the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3. WYZSG inhibited excessive autophagy and apoptosis of myocardial cells in septic rats and improved myocardial injury, possibly by regulating the expression of miR-132-3p/UCP2.


Subject(s)
Animals , Rats , Rats, Sprague-Dawley , Caspase 3 , Beclin-1/genetics , Stroke Volume , Ventricular Function, Left , Apoptosis/genetics , Autophagy/genetics , Heart Injuries , MicroRNAs/genetics
2.
Chinese Journal of Industrial Hygiene and Occupational Diseases ; (12): 292-295, 2022.
Article in Chinese | WPRIM | ID: wpr-935795

ABSTRACT

Objective: To analyze the incidence characteristics of occupational diseases in Guangzhou from 2010 to 2020, provide scientific basis for formulating occupational disease prevention and control policies. Methods: In January 2021, based on the data of occupational diseases in Guangzhou reported in the Information Monitoring System of Occupational Diseases and Occupational Health, descriptive epidemiological method was used to analyze the types and characteristics of occupational diseases in Guangzhou from 2010 to 2020. Results: A total of 1341 cases of 38 kinds of occupational diseases in 9 categories were reported in the past 11 years. The incidence of occupational pneumoconiosis, occupational otolaryngology and oral diseases and occupational chemical poisoning ranked the top three, accounting for 38.1% (511/1341) , 30.5% (409/1341) and 16.2% (217/1341) of the total cases respectively. The cases of pneumoconiosis in welders and silicosis accounted for 47.7% (244/511) and 34.4% (176/511) of the cases of occupational pneumoconiosis respectively. The cases of noise deafness accounted for 99.8% (408/409) of occupational otorhinolaryngology oral diseases. Acute occupational chemical poisoning cases accounted for 26.7% (58/217) of the occupational chemical poisoning cases, in which dichloroethane poisoning cases ranked the first, accounting for 79.3% (46/58) . Chronic occupational chemical poisoning cases accounted for 73.3% (159/217) of the occupational poisoning cases, in which benzene and lead poisoning cases ranked the top two, accounting for 79.2% (126/159) and 17.6% (28/159) respectively. Conclusion: Pneumoconiosis, silicosis, noise deafness, benzene poisoning, lead poisoning, dichloroethane poisoning should be supervised and managed as key occupational diseases in Guangzhou.


Subject(s)
Humans , Benzene , China/epidemiology , Deafness , Ethylene Dichlorides , Incidence , Lead Poisoning , Occupational Diseases/epidemiology , Pneumoconiosis/epidemiology , Silicosis
3.
Frontiers of Medicine ; (4): 829-841, 2021.
Article in English | WPRIM | ID: wpr-922513

ABSTRACT

Astrocytes are an abundant subgroup of cells in the central nervous system (CNS) that play a critical role in controlling neuronal circuits involved in emotion, learning, and memory. In clinical cases, multiple chronic brain diseases may cause psychosocial and cognitive impairment, such as depression and Alzheimer's disease (AD). For years, complex pathological conditions driven by depression and AD have been widely perceived to contribute to a high risk of disability, resulting in gradual loss of self-care ability, lower life qualities, and vast burden on human society. Interestingly, correlational research on depression and AD has shown that depression might be a prodrome of progressive degenerative neurological disease. As a kind of multifunctional glial cell in the CNS, astrocytes maintain physiological function via supporting neuronal cells, modulating pathologic niche, and regulating energy metabolism. Mounting evidence has shown that astrocytic dysfunction is involved in the progression of depression and AD. We herein review the current findings on the roles and mechanisms of astrocytes in the development of depression and AD, with an implication of potential therapeutic avenue for these diseases by targeting astrocytes.


Subject(s)
Humans , Alzheimer Disease , Astrocytes , Depression , Neurons
4.
Chinese Journal of Biotechnology ; (12): 1443-1449, 2021.
Article in Chinese | WPRIM | ID: wpr-878646

ABSTRACT

We introduce the portfolio assessment into the classroom teaching reform in the curriculum of Cell Engineering, a specialty course in bioengineering & biotechnology. We established a complete classroom evaluation system that was divided the classroom assessment system of portfolio into four stages including the preparation stage, training stage, implementation stage and exhibition stage. We also discuss the feasibility and necessity of implementing the portfolio evaluation method in the course of cell engineering, the construction of evaluation system, and the key points and matters needing attention in the implementation process. The classroom reform is very productive, not only the classroom atmosphere has been activated, students' learning initiative and autonomy has been enhanced, but also the students' ability to analyze and solve professional problems related to cell engineering technology has been improved. The implementation of classroom teaching reform of this course can provide reference for other similar professional courses in colleges and universities.


Subject(s)
Humans , Cell Engineering , Curriculum , Learning , Students , Universities
5.
Practical Oncology Journal ; (6): 206-210, 2019.
Article in Chinese | WPRIM | ID: wpr-752840

ABSTRACT

Objective The aims of this study were to determine the expression of CXCR family proteins in various subtypes and adjacent tissues of breast cancer and its relationship with prognosis,and to provide a reference for clinical diagnosis,treatment and prognosis of breast cancer. Methods The mRNA expressive profiles of CXCR family proteins in paracancerous tissues and different subtypes of breast cancer tissues were obtained from the TCGA(The Cancer Genome Atlas)database. The prognostic survival analysis of each differentially expressed protein was obtained using the PRECOG website. Results Except for CXCR1,the expression of CX-CR family proteins in breast cancer tissues and adjacent tissues was statistically different( P<0. 05). CXCR2P1,CXCR3,CXCR4, CXCR5 and CXCR6 were highly expressed in breast cancer tissues,CXCR2 and CXCR7 were lowly expressed in breast cancer tis-sues. The expressive levels of CXCR3,CXCR4 and CXCR7 were associated with the prognosis of patients. Conclusion The expres-sions of CXCR3,CXCR4 and CXCR7 in breast cancer tissues and adjacent tissues is significantly different,and its expression is related to the prognosis of breast cancer patients,which may be a potential target for molecular diagnosis or targeted therapy of breast cancer.

6.
Journal of Clinical Hepatology ; (12): 2805-2808, 2019.
Article in Chinese | WPRIM | ID: wpr-778692

ABSTRACT

At present, radical resection remains the best treatment strategy for liver metastasis of colorectal cancer, and patients with large or multiple tumors often face the problem of insufficient residual liver volume after hepatectomy, which may lead to serious complications including liver failure after surgery. Associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) is a novel two-step hepatectomy that allows rapid proliferation of the residual liver after primary surgery and can improve the resectability of liver tumors and reduce the risk of postoperative liver failure. Although ALPPS can achieve R0 resection or tumors and prolong patients’ survival time, there are still controversies over its application due to high incidence rates of complications and mortality rate. With reference to related studies in China and foreign countries, this article reviews the current application status and clinical value of ALPPS in liver metastasis of colorectal cancer.

7.
Journal of Experimental Hematology ; (6): 439-444, 2019.
Article in Chinese | WPRIM | ID: wpr-774296

ABSTRACT

OBJECTIVE@#To explore whether tumor suppressor gene Foxo1 and PTEN play a critical role in the tumorigenesis of mouse natural killer-cell lymphoma.@*METHODS@#NKp46-iCre mice were crossed with mice carrying floxed Foxo1 alleles (Foxo1) as well as floxed PTEN alleles (PTEN) to generate mice in which Foxo1 and PTEN in NK cells were knock-out, referred as Foxo1PTEN. The growth and development of the mice and tumor formation were observed. The flow cytometry was used to detect the percentages of NK cells in main lymphatic organs. B16F10 metanoma model of tumor metastasis was utilized to investigate NK cell-mediated tumor surveillance in vivo after NK cells special deletion of Foxol and PTEN.@*RESULTS@#The mouse model with NK cell-special Foxo1 and PTEN double knockout was established. Compared with control group (Foxo1PTEN mice), Foxo1PTEN mice were born alive and appeared to be healthy over a period of 46 weeks. No spontaneous tumor formation was observed at this stage. There were no significant differences in NK cell percentages of gated lymphocytes from various organs including blood, bone marrow, peripheral lymph node and spleen between Foxo1PTEN mice and Foxo1PTEN mice [PB: 4.76%±0.46% vs 4.17%±0.64% (P>0.05, n=8); BM: 1.13%±0.23% vs 1.31%±0.10% (P>0.05, n=8) ; LN: 0.50%±0.10% vs 0.85%±0.20% (P>0.05, n=8); SP: 4.41%±0.65% vs 3.50%±0.24% (P>0.05, n=8)]. B16F10 melanoma metastasis model of tumor was established, No differences in median survival time were observed in the 2 types of mice (P>0.05, n=13).@*CONCLUSION@#The simultaneous deletion of the Foxo1 and PTEN genes may not plays significant role in the tumorigenesis of mouse natural killer-cell lymphoma and NK cell-mediated tumor surveillance in vivo.


Subject(s)
Animals , Mice , Cell Transformation, Neoplastic , Forkhead Box Protein O1 , Genes, Tumor Suppressor , Killer Cells, Natural , Lymphoma , Mice, Knockout
8.
The Journal of Practical Medicine ; (24): 1947-1950, 2018.
Article in Chinese | WPRIM | ID: wpr-697862

ABSTRACT

Objective To express and purify GRIK3 intracellular region protein in prokaryotic cell system. Methods The histamine(His)tagged GRIK3 intracellular region recombinant plasmid pCzn1-GRIK3-in-tra was constructed and transformed into Escherichia coli(E.coli). The His-GRIK3-intra recombinant protein was expressed after the induction with IPTG. The target protein was purified by Ni-NTA affinity chromatography column. Results The prokaryotic expression plasmid pCzn1-GRIK3- intra was successfully constructed and the GRIK3 intracellular region protein in E.coli were efficiently expressed after the induction with IPTG. The purified target pro-tein was successfully obtained by Ni-NTA affinity chromatography column. Conclusion The successful construc-tion of prokaryotic expression plasmid expressing GRIK3 intracellular region gene and preparation of high purity GRIK3 intracellular region protein have paved the way for further exploration of the intracellular signal transduction mechanism of membrane protein GRIK3.

9.
Chongqing Medicine ; (36): 1904-1907, 2018.
Article in Chinese | WPRIM | ID: wpr-692038

ABSTRACT

Objective To explore the risk factors of nosocomial infections to provide a reference for prevention and control of nosocomial infection.Methods The data of 680 patients with nosocomial infection in this hospital from January 2015 to December 2016 were retrospectively collected.The characteristics of nosocomial infection were analyzed from the aspects of age,hospitalization department,infection site,etc.Results A total of 680 cases of nosocomial infection accounted for 1.67%(680/40 657) of all hospitalization patients,including 316 cases(nosocomial infection rate was 2.47%) of 61-80 years old.The infection rate was the highest in ICU(24.90%),followed by the nephrology department(4.99%).For infection sites,the maximal cases-times in infection site was lower respiratory tract(413 case-times),followed by urinary tract and superficial incision(each 68 case-times).The pathogenic detection rate was 45.03%,and 190 strains of pathogens were detected.The top five places were Escherichia coli(47 strains),Klebsiella pneumoniae(28 strains),Pseudomonas aeruginosa (23 strains),Staphylococcus aureus (22 strains) and Bauman Acinetobacter (16 strains).Logistic regression analysis showed that age larger than 60 years old,hospitalization time longer than 60 d,indwelling urinary catheter,peripherally inserted central catheter(PICC),tracheotomy and enteral nutrition were the risk factors for nosocomial infection occurrence.Conclusion Hospital should put the emphasis on preventing nosocomial infection of lower respiratory tract and strengthen the nosocomial infection management of key departments like ICU.

10.
Chinese Journal of Pathophysiology ; (12): 206-211, 2018.
Article in Chinese | WPRIM | ID: wpr-701103

ABSTRACT

AIM:To investigate the enhancing effect of quercetin on the 5-fluorouracil-induced apoptosis in gastric cancer.METHODS: MTT assay was conducted to evaluate the effect of quercetin on the 5-fluorouracil-induced death of gastric cancer cell line BGC-823.Co-immunoprecipitation and Western blot analysis were performed to detect the expression of c-Jun and Bcl-xL,phosphorylation of c-Jun,activation of caspase-9 and caspase-3,and release of cytochrome C in BGC-823 cells treated with quercetin and 5-fluorouracil.The apoptosis of BGC-823 cells treated with quercetin and 5-fluorouracil was analyzed by flow cytometry.RESULTS: Adjuvant therapy of quercetin significantly enhanced the 5-fluorouracil-induced death of BGC-823 cells.Meanwhile, quercetin decreased the half maximal inhibitory concentration (IC50)of 5-fluorouracil to BGC-823 cells.Quercetin treatment significantly inhibited the expression of c-Jun,and inhibited the 5-fluorouracil-induced phosphorylation of c-Jun and the interaction between c-Jun and activating transcription factor 2 (ATF2).Subsequently,quercetin inhibited the up-regulation of Bcl-xL induced by 5-fluorouracil in the BGC-823 cells. However,transfection with c-Jun plasmid abolished the promoting effect of quercetin on 5-fluorouracil-induced cell death. In addition, quercetin promoted 5-fluorouracil-induced release of cytochrome C from mitochondria and caspase-dependent apoptosis in BGC-823 cells.CONCLUSION:Quercetin treatment enhances 5-fluorouracil-induced mitochondrial apoptosis in BGC-823 cells through c-Jun/ATF2/Bcl-xL pathway.

11.
The Journal of Practical Medicine ; (24): 4038-4042, 2017.
Article in Chinese | WPRIM | ID: wpr-665461

ABSTRACT

Objective CRISPR/Cas9 genome-editing technique provides an novel method for whole genome editing in eukaryotic cells.Recently,we found that gene subtype library with smaller size and focused pur-pose is more economical and practical. In this study,we aimed to target kinases,a group of pivotal cell signal transducers,to construct a kinase knock-out library using CRISPR/Cas9 technique.The construction strategy wll al-so be discussed. Methods 10 sgRNA was designed for each kinase target.After oligo pool synthesis by semicon-ductor chip,the oligos were eluted from the chip. The oligo templates were amplified and cloned into Cas9 vector and transformed into Stble3 competent cells.Monoclonal colonies were selected for DNA sequencing. Results(1) GO analysis of 507 cell kinases showed that the cell kinases took part in a wide range of cell signaling.(2)The sgRNA pool with about 140 bp in length was successfully amplified by using oligo pool as the template and univer-sal PCR primers.(3)In 40 identified library clones,34 clones were sequenced successfully. Among them,the DNA sequencing results of 25 samples were completely consistent with the designed target sequences.But there are some mutations in the primers of 9 samples.Failure in bacteria shaking,DNA sequencing and other factors were ex-isted in the other clones. Conclusion The CRISPR/Cas9 kinase knock-out library can be widely used for screen-ing the important kinases which may mediate cell proliferation,metastasis,drug resistance and autophagy.This li-brary will play an important role in clarifying the development of disease associated with kinases.

12.
The Journal of Practical Medicine ; (24): 3814-3818, 2017.
Article in Chinese | WPRIM | ID: wpr-697537

ABSTRACT

Objective To compare different diagnosis values of age-specific procalcitionin (PCT) and C-reactive protein (CRP) on early-onset neonatal sepsis (EONS) during the first 72h of life.Methods From September 2008 to December 2015,96 neonates (including 2 confirmed sepsis and 94 clinical sepsis) without severe complications were chosen as the EONS group and 170 non-infectious newborns as the control group.A total of 605 blood samples were collected from all 266 newborns.Serum concentration of PCT and CRP were measured in both the EONS group and the control group at each age over the first 72 h of life.The diagnostic value of PCT and CRP within 1 ~ 12 h and 13 ~ 24 h and 25 ~ 48 h and 49 ~ 72 h of life was evaluated by calculating the cut-off values,sensitivity,specificity,and the area under the receiver operating characteristic curve (ROC).Results PCT and CRP levels of neonates within each age in the EONS group were significantly higher than those in the control group during the first 72h of life.(all P < 0.05).Within 1 ~ 12 h,13 ~ 24 h,25 ~ 48 h and 49 ~ 72 h of life,the cutoff value of PCT was 0.45 μg/L (sensitivity 84.2%,specificity 74.4%),1.885 μg/L (sensitivity 73.5%,specificity 75%),0.995 μg/L (sensitivity 82.4%,specificity 74.1%) and 0.51 μg/L (sensitivity 83.3%,specificity 79.2%) respectively;that of CRP3 185 mg/L (sensitivity 68.4%,specificity 82.1%),6.29 mg/L (sensitivity 58.8%,specificity 89.7%),8.615 mg/L (sensitivity 54.3%,specificity 93.9%) and 10.27 mg/L (sensitivity 59.1%,specificity 100%) respectively,and the area under ROC of PCT for the diagnosis of EONS was 0.767,0.754 and 0.755,and 0.8 respectively;that of CRP 0.773,0.8,0.815 and 0.789 respectively.Conclusions There are age-specific cut-off values of PCT and CRP in the diagnosis of EONS without severe complications during the first 72 h of life.Both PCT and CRP are moderately accurate for the diagnosis of EONS.PCT may be more helpful for the early diagnosis of EONS for its higher sensitivity but CRP presents higher specificity.

13.
Chinese Journal of Behavioral Medicine and Brain Science ; (12): 252-256, 2016.
Article in Chinese | WPRIM | ID: wpr-670231

ABSTRACT

Objective To explore the time course of response inhibition function in juvenile delin-quents with antisocial personality characteristics.Methods The healthy control group ( n=21),juvenile delinquents with antisocial personality characteristics ( CD +AP ) ( n=18) and juvenile delinquents ( CD) ( n=18) were selected in current study by recording the event-related potentials in a Go/Nogo task.N2 and P3 components of event-related potentials were analyzed.Results Behavioral results showed that Nogo cor-rection rate of control group ((93.13±2.71)%) were significantly higher than CD group ((87.51±2.82)%, P<0.01) and CD +AP group((85.63±2.45)%, P<0.01).In CD+AP group,the amplitude of the N2nogo ( (-1.82±1.64)μV) was significantly lower than control group ( (-6.36±2.93)μV, P<0.01) ,and the am-plitude of the P3nogo ((5.52±2.79)μV) was significantly decreased than healthy control ((11.26±4.92)μV, P<0.01).In CD group,the amplitude of P3nogo ((5.20±3.17)μV) was significantly reduced than healthy control ((11.26±4.92)μV, P<0.01).Conclusion N2nogo and N2d are associated with the early phases of response inhibition and reflected response conflict.P3nogo and P3d are associated with the late phases of response inhibition and monitored inhibitory control.These data suggest that CD+AP participants exhibited im-paired response conflict and inhibitory control.This may be associated with persistent antisocial behavior.

14.
Singapore medical journal ; : 58-64, 2015.
Article in English | WPRIM | ID: wpr-244712

ABSTRACT

<p><b>INTRODUCTION</b>Patients with rheumatoid arthritis (RA) and osteoarthritis (OA) may require total hip replacement (THR) or total knee replacement (TKR). The present study aimed to compare the demographic characteristics and medical costs of RA and OA patients from Taiwan who underwent either THR or TKR.</p><p><b>METHODS</b>The medical records of patients who had undergone THR or TKR from 1 January 1996 to 31 December 2010 were obtained from the Taiwan National Health Insurance Research Database (NHIRD). In all, we found 49 and 146 RA patients who received THR and TKR, respectively, and 1,191 and 6,574 OA patients who received THR and TKR, respectively. The gender, age, Charlson comorbidity index (CCI), hospital grade, age at registration in the catastrophic illness dataset, and medical utilisation costs of the different groups were compared.</p><p><b>RESULTS</b>There were statistically significant differences in age, CCI score, drug costs and surgery costs between RA and OA patients. Joint replacement incidence was lower in RA patients than in OA patients, and among patients who underwent THR, total medical costs incurred were higher for RA patients than OA patients. RA patients who underwent THR incurred a significantly greater total medical utilisation cost in the outpatient department (3 months before surgery and 12 months after surgery) than OA patients who underwent THR.</p><p><b>CONCLUSION</b>Analysis of Taiwan NHIRD with regard to patients who had undergone either THR or TKR indicated that RA patients were younger than OA patients, and that significantly more medical resources were used for RA patients before, during and after hospitalisation for these procedures.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Age Factors , Arthritis, Rheumatoid , General Surgery , Arthroplasty, Replacement, Hip , Economics , Methods , Arthroplasty, Replacement, Knee , Economics , Methods , Databases, Factual , Health Care Costs , Hospitalization , Length of Stay , Osteoarthritis , General Surgery , Sex Factors , Taiwan , Treatment Outcome
15.
Chinese Journal of Experimental and Clinical Virology ; (6): 89-91, 2013.
Article in Chinese | WPRIM | ID: wpr-318096

ABSTRACT

<p><b>OBJECTIVE</b>To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.</p><p><b>METHODS</b>The hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.</p><p><b>RESULTS</b>The expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.</p><p><b>CONCLUSION</b>The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.</p>


Subject(s)
Bacterial Proteins , Genetics , Blotting, Western , Carrier Proteins , Genetics , Escherichia coli , Genetics , Haemophilus influenzae type b , Genetics , Immunoglobulin D , Genetics , Lipoproteins , Genetics , Plasmids , Recombinant Proteins , Solubility
16.
Journal of Southern Medical University ; (12): 742-745, 2012.
Article in Chinese | WPRIM | ID: wpr-269007

ABSTRACT

<p><b>OBJECTIVE</b>To prepare a monoclonal antibody (mAb) against the fusion protein preM/EIII of West Nile virus (WNV) for clinical detection of WNV.</p><p><b>METHODS</b>Sp2/0 cells were fused with the spleen cells of BALB/c mice immunized with the recombinant fusion protein preM/EIII expressed in E. coil to obtain the hybridoma cell line that secreted preM/EIII mAb. The hybridoma cells were injected into the peritoneal cavity of BALB/c mice and the ascites was collected and purified. The specificity and titer of the obtained mAb were determined using ELISA and Western blotting.</p><p><b>RESULTS</b>One hybridoma cell line secreting preM/EIII mAb, named ME1, was obtained. The titer of the purified mAb was 10(-6). Identified as a mAb of the Ig subclass IgG1, ME1 was capable of specific reactions with preM/EIII protein and WNV without cross-reactions with other viruses such as JEV, SLEV, YFV and DENV. The accuracy of clinical testing of MNV with ME1 was 97.78%.</p><p><b>CONCLUSION</b>The mAb against preM/EIII obtained have a high specificity and accuracy in clinical testing of MNV and can be used in clinical diagnosis of MNV infection.</p>


Subject(s)
Animals , Mice , Antibodies, Monoclonal , Allergy and Immunology , Blotting, Western , Cross Reactions , Hybridomas , Allergy and Immunology , Mice, Inbred BALB C , Viral Fusion Proteins , Allergy and Immunology , West Nile virus , Allergy and Immunology
17.
Chinese Journal of Behavioral Medicine and Brain Science ; (12): 1022-1024, 2012.
Article in Chinese | WPRIM | ID: wpr-429820

ABSTRACT

Objective To explore the effect of weightlessness simulation on processing speed of mental rotation of males in 72 h head-down tilt,in order to further reveal human performance changes caused by weightlessness.Methods During 72 h weightlessness simulation (9 am day 1 to 9 pm day 4,head-down tilt started from 9 pm day 1),16 male subjects were processed speed of rotation data byhand picture mental rotation (test 8 times),using self cross-references design,and the variation of processing speed of mental rotation by duration was analyzed.Results The linear regression analysis showed,the reaction time of each time point for each subject as the independent variable,the angle of rotation as the dependent variable,with the extension of simulated weightlessness time.The intercept(constant term of the regression equation),indicating the processing speed of non-rotation,were gradually increased with HDT time,but didn(t) reached significantly difference (F(3,14) =0.551,P =0.650) ;the slope(Regression coefficient of the regression equation),indicating the processing speed of rotation.were gradually increased with HDT time,and reached the significantly difference (F(3,14) =4.338,P =0.009).The value of intercept and slope was like upside-down U in three days,and reached the highest value at day 2.Conclusion Weightlessness simulation have affection on the speed of mental rotation,especially on the rotation speed.

18.
Chinese Journal of Laboratory Medicine ; (12): 507-512, 2010.
Article in Chinese | WPRIM | ID: wpr-379791

ABSTRACT

Objective To explore the diagnostic value of GP-/3 protein in gene detection in the patient of primary hepatic carcinoma, to discuss the joint roles of serum GP73 and AFP, and provide a novel method for the diagnosis for PHC and screening for high-risk population. Methods ELISA was used to detect the serum level of GP73 and AFP in 73 cases of PHC, 13 cases of hepatic cirrhosis, 32 cases of hepatitis and 62 cases of health people. SYBR Green real time fluorescence quantitative PCR was used to detect the relative value of GP73 mRNA in the peripheral blood cells of each group. Comparative Ct method was used to evaluate the relative expression levels. Eight cases of normal liver tissues and 8 cases of PHC tissues were detected at the same time to compare the relative expression levels. Results Kruskal-Wallis test showed that the serum levels of GP73 and AFP had significant differences between four groups(H value were 89. 6 and 52.0, P < 0. 01) and the whole blood GP73 mRNA had no significant differences(H =4. 33, P > 0. 05). Mann-Whitney test showed that the serum levels of GP73 had significant differences among PHC groups[166. 7 (162. 7-231.8) μg/L] and liver cirrhosis[57. 3 (46. 6-113. 6) μg/L], hepatitis[29. 6(26. 2-54. 5) μg/L], health group[25.1 (20. 8-29. 4) μg/L] (U value were 246, 297, 349, P < 0. 01).The A FP levels of the four groups were 380. 9 (258.5-503.2) μg/L, 3.8 (1.3-14. 5) μg/L, 5. 1 (2. 4-7. 8)μg/L and 2. 8(2. 2-5.7) μg/L. It also showed significant differences (U value were 246,419 and 790,P <0. 01). The GP73 mRNA expression of PHC liver tissues(12. 64) was significant higher than normal liver tissues (1.00). The critical values for GP73 and AFP was determined to be 123. 2 μg/L and 10. 6 μg/L through the 8OC curves. Under the critical value the sensitivity of GP73 and AFP were 65.8% and 53.4% ,and the specificity of CP73 and AFP were 95.3% and 92. 5% respectively. Joint detection could increase the sensitivity up to 79. 5%, and achieve the high specificity of 90. 7%. Conclusions As a new diagnositic marker of primary hepatic carcinoma, GP73 protein has the very good sensitivity and specificity. The GP73 mRNA in the whole blood sample could not be used for the diagnosis of PHC. But it woule be a good molecular marker for diagnosis of PHC in the liver tissue sample. The joint detection of GP73 and AFP could improve PHC diagnostic performance, and provide an effective approcach to the PHC high-risk screening.

19.
Chinese Journal of Epidemiology ; (12): 795-799, 2010.
Article in Chinese | WPRIM | ID: wpr-341033

ABSTRACT

Objective To study the change of special antibodies titer IgG, IgM and nucleocaspid to SARS coronavirus (CoV) and observing the expression of stomach and enteric involvement on SARS-CoV infection by monoclonal antibody against N protein of SARS-CoV in the 7- year recovery period among family clustering cases of severe acute respiratory syndrome. Methods Special antibody titer to SARS-CoV of 14 patients from 5 different families and their 10 kinfolks continuously tested by IFA and antigen-capturing ELISA methods. Samples were taken in the 1st-7th year periods after SARS patients infected by SARS-CoV, being diluted and measured on it titers of three kinds of antibodies. Immunochemical staining with monoclonal antibody (mAb) against N protein of SARS-CoV was used to determine the stomach and enteric tissues among 5 SARS patients with their nucleocaspid antibody titer ascended obviously after 1st-7th year. Results When testing the IgG antibody titer of the 14 SARS patients by IFA method, the average titer was 1/71 (95%CI:1/58-1/85) in the 1st year, but began to descend in the following years, and the IgG antibody of the most SARS patients disappeared in the 7th year. Regarding the IgM titer, it disappeared in most of the SARS patients 1 year later. The average value of nucleocaspid antibody titer was 1/146 (95% CI:1/122-1/171) in the 1st year, and it descended as the IgG antibody titer did. In 5 cases, differences appeared.The nucleocaspid antibody titer was between 1/156 and 1/210 in 3 cases, and 2 cases were normal.Immunochemical staining with mAb against N protein of SARS-CoV was identified in the stomach and enteric tissues of 5 SARS patients with the nucleocaspid antibody titer increased significantly, 1st-7th year later. The five patients were detected by gastroscopy detection and cell immunohistochemistry test. 3 cases showed N protein antibody positive in the serum, and positive immunohistochemical expression in most of the cytoplasm in the gastric tissue mucous gland epithelial cells. 1 case also expressed in the intestinal tissue slurry columnar epithelium and interstitial cells. The other two cases showed negative on both serum N protein antibody and immunohistochemical expression. The biopsy results of the 5 patients were as follows: 1 case diagnosed as "signet-ring cell carcinoma of the stomach and rectum multiple transfer", 1 case of gastric polyp, 1 case of superficial antral gastritis and 2 cases were normal. Conclusion By testing the special IgG, IgM, nucleocaspid antibody to SARS-CoV of the 14 family clustering cases , we found that they all decreased in the 7th year, and most of them disappeared. The nucleocaspid antibody titer was related to pathogenetic condition. SARS-CoV was proved to be still present in stomach and enteric tissues of SARS patients with the nucleocaspid antibody titer increased significantly after the 7th year.

20.
Chinese Journal of Experimental and Clinical Virology ; (6): 119-121, 2010.
Article in Chinese | WPRIM | ID: wpr-316947

ABSTRACT

<p><b>OBJECTIVE</b>To prepare Vero cell-adapted influenza H5N1 virus strain by Genetic Reassortment and produce influenza H5N1 vaccine using Vero cell as a substrate.</p><p><b>METHODS</b>Embryonated specific pathogen-free (SPF) hen's eggs and Vero cells were co-infected with Vero cell-adapted influenza virus A/Yunnan/1/2005 Va(H3N2) and A/Anhui/1/2005 (H5N1) via reverse genetics. The reassortant was screened with goat antibody against strain A/Yunnan/1/2005 Va(H3N2) and identified for subtype by hemagglutination-inhibition (HI) assays and gene analysis of HA and NA.</p><p><b>RESULTS</b>A Vero cell-adapted influenza H5N1 virus strain was obtained, and there was no significant difference in serum antibody titers of monovalent inactivated vaccine reassorted before and after (F = 0.857, P > 0.05).</p><p><b>CONCLUSION</b>The Vero cell-adapted influenza virus of epidemic strain may be reassortment between Vero cell-adapted and epidemic strains.</p>


Subject(s)
Animals , Chick Embryo , Antibodies, Viral , Allergy and Immunology , Chlorocebus aethiops , Influenza A Virus, H5N1 Subtype , Genetics , Allergy and Immunology , Influenza Vaccines , Genetics , Allergy and Immunology , Recombination, Genetic , Vero Cells
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