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1.
Chinese Journal of Cellular and Molecular Immunology ; (12): 742-747, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1009425

RESUMO

Objective To prepare and identify rabbit anti-cyclin dependent kinase 6 (CDK6) antibody. Methods The recombinant pET21a (+)/CDK6 was successfully constructed, then the recombinant plasmid was transformed into E.coli BL21 (DE3) competent cells and was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) for protein expression, which was detected by SDS-PAGE and Western blot analysis. The expressed protein was purified by nickel-chelating nitrilotriacetic acid (Ni-NTA) agarose and then analyzed by SDS-PAGE. Japanese white rabbits were immunized with purified CDK6 protein for many times every two weeks. The blood was collected at 0, 2, 4 and 6 weeks after immunization, and serum was separated from blood. The titer was detected by indirect ELISA. Western blot analysis, immunofluorescence assay and immunohistochemistry were employed to determine the specificity. Results High purity CDK6 protein and high specificity of rabbit anti-CDK6 antibody were successfully prepared. The titer of CDK6 rabbit serum antibody reached 1:30 000 after immunization, which could specifically recognize the CDK6 protein expressed in cervical cancer cell line and cervical cancer tissues. Conclusion The high titer and specificity of rabbit anti-CDK6 antibody is successfully prepared.


Assuntos
Animais , Feminino , Humanos , Coelhos , Anticorpos , Especificidade de Anticorpos , Western Blotting , Quinase 6 Dependente de Ciclina , Ensaio de Imunoadsorção Enzimática , Neoplasias do Colo do Útero
2.
Chinese Journal of Medical Genetics ; (6): 1181-1184, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1009273

RESUMO

OBJECTIVE@#To present on a prenatally diagnosed case with complex structural rearrangements of chromosome 8.@*METHODS@#Chromosome karyotyping, chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH) were carried out for a fetus with increased nuchal thickness.@*RESULTS@#The karyotype of the amniotic fluid sample showed extra materials on 8p. FISH revealed a centromeric signal at the terminal of 8p with absence of telomeric signal. CMA revealed partial deletion of 8p23.3 [(208049_2256732)×1], partial duplication of 8p23.3p23.2 [(2259519_3016818)×3], and partial duplication of 8q [8q11.1q12.2(45951900_60989083)×3].@*CONCLUSION@#The complex structural rearrangements of chromosome 8 in this case has differed from the commonly seen inv dup del(8p).


Assuntos
Feminino , Gravidez , Humanos , Cromossomos Humanos Par 8/genética , Hibridização in Situ Fluorescente , Rearranjo Gênico , Diagnóstico Pré-Natal , Centrômero
3.
International Journal of Cerebrovascular Diseases ; (12): 700-705, 2022.
Artigo em Chinês | WPRIM | ID: wpr-989143

RESUMO

Studies have shown that plasma apolipoprotein B (ApoB) has a good predictive value for ischemic stroke and plays an important role in the prevention and treatment of ischemic stroke. More and more guidelines and consensus opinions began to recommend ApoB as a routine intervention target. This article reviews the biological characteristics, clinical detection advantages, and role and treatment prospect of ApoB in the prevention and treatment of ischemic stroke.

4.
Chinese Journal of Medical Education Research ; (12): 929-933, 2022.
Artigo em Chinês | WPRIM | ID: wpr-955567

RESUMO

Objective:To investigate the post competency of "Academic-Practical" of clinical nursing teachers, and analyze related influencing factors.Methods:A total of 312 "Academic-Practical" and "Non-Academic-Practial" clinical nursing teachers from The Affiliated Hospital of Southwest Medical University were surveyed by the Clinical Nursing Teacher Post Competency Evaluation Questionnaire. SPSS 20.0 software was used for t test, chi-square test and rank sum test. Results:The average self-evaluation scores of post competency of "Academic-Practical" clinical nursing teachers were (4.26±0.41) points, which were higher than those of the "Non-Academic-Practical" teachers [(3.19 ±0.50) points], showing good post competency. There were significant differences in the scores of professional quality (17.39±1.54), professional attitude (21.75±2.21), professional ability (21.14±2.31), teaching ability (50.39±5.93), interpersonal coordination ability (25.57±3.04), and personality characteristics (17.27±2.04) between the "Academic-Practical" and "Non-Academic-Practical" teachers (all P<0.01). And there were significant differences in self-evaluation post competency scores of "Academic-Practical" teachers in "with or without teacher qualification certificate" ( P=0.001), "whether she/he is the backbone of the department" ( P=0.002), degree ( P=0.001), age ( P<0.001), positional title ( P<0.001) and working year ( P<0.001) (all P<0.01). But there was no significant difference in gender ( P=0.735) and "whether she/he is a specialized nurse" ( P=0.335). Conclusion:"Academic-Practical" and "Non-Academic-Practical" medical teachers should take the post competency as the core orientation, adopt the "Ladder" mode of training and management, and constantly improve the training plan of post competencey.

5.
Chinese Journal of Emergency Medicine ; (12): 1491-1497, 2022.
Artigo em Chinês | WPRIM | ID: wpr-954570

RESUMO

Objective:To investigate the predictive value of serum D-dimer combined with myocardial injury markers on admission for early identification of high-risk patients with acute myocarditis.Methods:Patients hospitalized for acute myocarditis in China-Japan Friendship Hospital were retrospectively enrolled from 2010 to 2021. Patients were divided into the high D-dimer level group and low D-dimer level group according to the median value of D-dimer measured by immunoturbidimetry within 24 h of admission. In-hospital adverse events were defined as death, cardiogenic shock, malignant ventricular arrhythmia and new-onset heart failure. Multivariate logistic analysis was used to explore the independent predictors of in-hospital adverse events, and receiver operating characteristic curve was used to evaluate the predictive value.Results:A total of 106 patients were analyzed, including 52 high level D-dimer patients and 54 low level D-dimer patients, with an average age of (36±16) years, and 62.3% were male. Compared with the low D-dimer level group, patients in the high D-dimer level group had lower mean systolic blood pressure [(114±21) mmHg vs. (121±14) mmHg] and diastolic blood pressure [(71±13) mmHg vs. (76±10) mmHg], higher heart rate [(97±26) beats/min vs. (79±15) beats/min], higher C-reactive protein levels [6.82 (1.61, 20.05) mg/dL vs. 1.30 (0.13, 8.93) mg/dL] and creatinine levels [86.95 (67.63, 117.83) μmol/L vs. 68.80 (60.18, 81.93) μmol/L] on admission. The proportion of patients having QRS interval >120 ms on electrocardiogram was higher in high D-dimer level group (25.0% vs. 7.4%). There was no significant difference in patients with positive myocardial injury biomarkers between the two groups. The incidence of in-hospital adverse events was higher in the high D-dimer level group (67.3% vs. 22.2%, P<0.001). Multivariate logistic analysis showed that serum D-dimer levels and elevated myocardial injury markers on admission were independently associated with in-hospital adverse events. The area under the curve (AUC) of elevated serum D-dimer level on admission for predicting in-hospital adverse events was 0.781 (95% CI: 0.690-0.873), the sensitivity was 74.5%, and the specificity was 71.2%. When combined with positive cardiac biomarkers, the AUC was 0.831 (95% CI: 0.752-0.910) with a sensitivity of 80.9% and a specificity of 78.0%. Conclusions:Elevated D-dimer level on admission can predict the risk of in-hospital adverse events in patients with acute myocarditis. The combination of cardiac injury biomarkers can improve the predictive value.

6.
Chinese Journal of Laboratory Medicine ; (12): 444-448, 2022.
Artigo em Chinês | WPRIM | ID: wpr-934394

RESUMO

Objective:To develop a self-made plasma quality control material for non-invasive prenatal testing (NIPT) and evaluate its performance.Methods:139 NIPT-negative maternal plasmas stored in the genetic department of Shaoxing maternal and child health hospital from January 1, 2019 to June 30, 2021 were divided into male groups (19 cases) and female groups (120 cases) according to the neonatal gender. 9360 cases from September 2020 to September 2021 were enrolled as clinical validation cases.First step, 200 μl plasma from a 47 years-old non-pregnant healthy women was used as a matrix. Different amounts (0.1, 0.2, 0.5, 2.5, and 5 μl) of positive DNA from fetal chromosome aneuploidy (T21, T18, T13) detection kit were added. The appropriate volume of positive DNA was 0.5 μl according to the test results. Second step,Plasma in male and female group was treated as matrix. 0.5 μl positive DNA was added per 205 μl. Plasma matrix from female group showed good repeatability and the sensitivity was 100%.Third step, evaluate the self-made plasma quality control material, including storage stability, matrix uniformity and repeatability, and the effect of different batch numbers of positive DNA, by calculating Z score and the CV of fetal DNA concentration (FF).Results:Plasma matrix from female group showed good repeatability and the sensitivity was 100%, while the sensitivity of male group was only 84%. The CV of FF in female matrix was 3.9% in the repetitive experiments. After adding 0.5 μl positive DNA, the mean FF of self-made positive plasma quality control was 5.63%±0.42%, Z values>6, and the CV was 7% after storage of three months. Considering the concentration variation of positive DNA in different lots, 1 μl of positive DNA should be added when the FF of positive DNA is lower than 10%.Used in 9360 clinical cases from September 2020 to September 2021, all positive plasma quality control materials showed positive results, and the positive predictive value of trisomy 21 was 100%.Conclusions:The NIPT self-made positive plasma quality control material has been successfully developed in this study. The preliminary experimental results show that it has good repeatability and stability, which is suitable for clinical application.

7.
Chinese Journal of General Practitioners ; (6): 66-69, 2022.
Artigo em Chinês | WPRIM | ID: wpr-933699

RESUMO

Trousseau′s syndrome(TS) is a complication of cancer-associated thrombosis caused by hypercoagulability. A 58-years female patient admitted to the Affiliated Hospital of Qingdao University on October 2020 and diagnosed with Trousseau′s syndrome was reported. This was a patient with pancreatic malignant tumor. On the second day of admission, the mouth angle was distorted and the speech was vague. Craniocerebral MR showed multiple DWI high signals in the brain parenchyma, and brain MR enhancement showed no abnormal enhancement in the brain parenchyma. The patient was considered to be Trousseau′s syndrome. Using "Trousseau′s syndrome" and "cerebral infarction" as key words, the relevant literature was searched in CNKI, Wanfang and PubMed databases from January 2011 to June 2021, total of 76 cases of Trousseau′s syndrome complicated with cerebral infarction were reported in the literature. Among 77 cases (including one in this study) 36 were males and 41 were females, with a median age of 63 years old. The most common tumor type was lung adenocarcinoma (24 cases, 31.2 %). The mean D-dimer level was (17.3±12.8) mg/L, Craniocerebral CT or MRI showed that 57 cases (74.0 %) had bilateral multiple lesions; and 56 cases received anticoagulant therapy. A total of 68 patients were followed up, with a median survival time of 90 days, and one year overall survival rate was 32.6 %. The study indicates that for cerebral infarction with significantly elevated D-dimer level and multiple vascular involvement, malignant tumors should be considered.

8.
Chinese Journal of Medical Genetics ; (6): 76-80, 2022.
Artigo em Chinês | WPRIM | ID: wpr-928366

RESUMO

OBJECTIVE@#To report on a case of mosaicism 13q inversion duplication, analyze its mechanism, and discuss the correlation between its genotype and phenotype.@*METHODS@#Amniotic fluid and umbilical cord blood were collected at 23 and 32 weeks of gestation, respectively. Combined with G-banding chromosome karyotyping analysis, single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) were used to confirm the result.@*RESULTS@#The karyotype of the fetus was determined as 47,XY,+inv dup(13)(q14.3q34)/46,XY. After careful counseling, the couple decided to continue with the pregnancy, and had given birth to a boy at 40 weeks' gestation. Except for a red plaque (hemangioma) on the nose bridge, no obvious abnormality (intelligence to be evaluated) was discovered.@*CONCLUSION@#To provide reference for clinical genetic counseling and risk assessment, the location and proportion of new centromere formation should be fully considered in the case of mosaicism 13q inversion duplication.


Assuntos
Feminino , Humanos , Masculino , Gravidez , Amniocentese , Inversão Cromossômica/genética , Hibridização Genômica Comparativa , Feto , Hibridização in Situ Fluorescente , Mosaicismo , Diagnóstico Pré-Natal
9.
Journal of Central South University(Medical Sciences) ; (12): 384-389, 2022.
Artigo em Inglês | WPRIM | ID: wpr-928981

RESUMO

Nevus-like basal cell carcinoma syndrome (NBCCS) is a rare autosomal dominant disease characterized by the occurrence of multiple maxillofacial keratocysts, basal cell carcinoma, child medulloblastoma, and various skeletal and soft tissue dysplasia. In 2020, a patient with NBCCS dominated by facial basal cell carcinoma was admitted to Xiangya Hospital of Central South University. The patient was an elderly woman. Ten years ago, the systemic mass appeared, especially on the face, but it was not treated. Later, these masses gradually increased in volume and number, and showed invasive properties. The nasal mass was broken and suppurated, seriously affecting the patient's life quality. The patient came to the hospital to improve the symptoms. Staphylococcus aureus and Providencia rettgeri were cultured in the patient's nasal secretions. Nasal sinus enhanced MRI showed that the subcutaneous soft tissue of the right cheek and the anterolateral mucosa of the left nasal cavity were invaded, indicating multiple malignant skin lesions. After admission, local anesthesia was performed and some masses were removed. Pathological examination of the mass showed basal cell carcinoma. After general anesthesia, multiple masses were resected. The postoperative pathological examination showed that multiple basal cell carcinoma invaded the deep dermis near subcutaneous fat layer. Combined with the results of clinical and immunohistochemical examination, the patient was diagnosed as NBCCS. There were no clear tumor thrombus in the vessel and no nerve invasion. No recurrence or new tumor was found after 1 year follow-up. The incidence rate of NBCCS is low and clinical symptoms are different. The patient's life quality is poor and the patient needs long-term individualized treatment.


Assuntos
Idoso , Criança , Feminino , Humanos , Síndrome do Nevo Basocelular/cirurgia , Carcinoma Basocelular/cirurgia , Síndrome do Hamartoma Múltiplo , Imageamento por Ressonância Magnética
10.
Chinese Journal of Endemiology ; (12): 535-539, 2021.
Artigo em Chinês | WPRIM | ID: wpr-909047

RESUMO

Objective:To investigate the expressions of transforming growth factor-β1 (TGF-β1), aquaporin (AQP) and osteopontin (OPN) in renal injury induced by fluorosis in rats.Methods:According to body weight (80 - 100 g), forty-eight SD rats were divided into control group (normal saline), low fluoride group (10 mg/kg) and high fluoride group (20 mg/kg) by random number table, 16 rats in each group (half males and half females). The rats were exposed to fluoride by intraperitoneal injection of sodium fluoride. After the rats were treated with fluoride for 12 weeks, the rats were killed by femoral artery bloodletting and the renal tissue was taken. The contents of TGF-β1, AQP and OPN in serum of rats with fluorosis were analyzed by enzyme-linked immunosorbent assay (ELISA). The expressions of TGF-β1, AQP and OPN in renal tissue of rats with fluorosis were analyzed by quantitative real-time PCR (qRT-PCR) and immunocytochemistry.Results:The serum levels of TGF-β1, AQP and OPN in high fluoride group [(26.42 ± 6.09), (378.60 ± 36.84) μg/L, (603.45 ± 64.32) pg/ml] were higher than those in control group [(2.41 ± 0.42), (157.41 ± 15.26) μg/L, (182.45 ± 30.63) pg/ml] and low fluoride group [(13.15 ± 3.26), (245.65 ± 23.21) μg/L, (359.47 ± 55.26) pg/ml, P < 0.05]. The levels of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). The expressions of TGF-β1, AQP and OPN mRNA in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the expressions of TGF-β1, AQP and OPN mRNA in low fluoride group were higher than those in control group ( P < 0.05). The positive expression scores of TGF-β1, AQP and OPN in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the positive expression scores of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). Conclusion:TGF-β1, AQP and OPN are highly expressed in the renal tissue of rats with fluorosis, which can be used as indicators to judge the damage of renal aggregation system caused by fluorosis.

11.
Chinese Journal of Endemiology ; (12): 259-262, 2021.
Artigo em Chinês | WPRIM | ID: wpr-883705

RESUMO

Objective:To investigate the association between selenoprotein P (Sepp1) and Brucella infection at gene and protein levels. Methods:In this case-control study, 32 patients with chronic brucellosis (referred to as brucellosis) in Jiuyuan District and Bayan Obo Mining District of Baotou City from June to September 2019 were selected as brucellosis group, and 30 healthy people with the same epidemiological environment as the brucellosis group were selected as control group. The two groups were Han nationality. The single nucleotide polymorphism (SNP) of Sepp1 gene(rs7579) in 32 brucellosis patients and 30 healthy controls were tested by amplification refractory mutation system-PCR (ARMS-PCR). And the Sepp1 protein content was measured by enzyme linked immunosorbent assay (ELISA).Results:Thirty-two cases of brucellosis group, including 6 females and 26 males, aged (50.312 ± 5.035) years; 30 cases of control group, including 4 females and 26 males, aged (49.994 ± 5.098) years. There were no significant differences in sex ratio and age between the two groups (χ 2=0.336, t = 1.744, P > 0.05). There were no significant differences in the frequency distribution of G, A allele and GG, GA, and AA genotypes of rs7579 locus of Sepp1 gene between brucellosis group and control group (χ 2=0.263, 0.942, P > 0.05). In stratified analysis, there was a statistically significant difference in the frequency distribution of genotypes between female brucellosis group and control group (GG: 0/6, GA: 2/6, AA: 4/6; GG: 4/4, GA: 0/4, AA: 0/4, P < 0.05). And the level of Sepp1 protein in brucellosis group was significantly lower than that in control group (13.71±0.32, 19.26±0.69, t = 20.316, P < 0.05), low level of Sepp1 was a risk factor for brucellosis ( OR = 1.512, 95% CI: 1.290 - 1.687). Conclusions:The SNP of Sepp1 gene (rs7579) plays a role in Brucella infection, G allele may be the protective factor for women. The low level of Sepp1 protein is also associated with brucellosis.

12.
Chinese Journal of Medical Genetics ; (6): 891-894, 2020.
Artigo em Chinês | WPRIM | ID: wpr-826462

RESUMO

OBJECTIVE@#To explore the value of BACs-on-Beads (BoBs) for the practice of prenatal diagnosis.@*METHODS@#The results of chromosomal karyotyping and BoBs of 1773 prenatal samples were compared. Microdeletions and microduplications detected by BoBs were subjected to chromosome microarray analysis (CMA) with informed consent from patients.@*RESULTS@#BoBs has detected 46 cases of common aneuploidies involving chromosomes 13, 18, and 21, and 16 cases involving X and Y chromosomes. For 4 fetuses with normal results by BoBs, karyotyping analysis of amniotic fluid sample suggested low percentage mosaicisms (< 20%). BoBs has detected none of the 9 common microdeletions, but 14 male fetuses with Xp22 microdeletions and 5 with other microdeletions/microduplications. In 10 cases, the couples had chosen CMA verification, and the results were all consistent.@*CONCLUSION@#As a rapid diagnostic technique, BoBs has a high accuracy for common aneuploidies, and is capable of discovering certain chromosome microdeletions and microduplications. The difficulty lies in the inability to detect low proportion mosaicisms and the consultation following detection for male fetuses carrying Xp22 microdeletions.

13.
Chinese Journal of Physical Medicine and Rehabilitation ; (12): 685-689, 2020.
Artigo em Chinês | WPRIM | ID: wpr-871205

RESUMO

Objective:To explore the suitability of using Rasch analysis to transform scores on the Activities and Participation component of the ICF Rehabilitation Set.Methods:Between January and May 2017, five hundred and fifteen post-acute rehabilitation patients were recruited in 13 research centers through quota sampling. Specially-trained health professionals used the Activities and Participation component of the ICF Rehabilitation Set to collect data. Rasch analysis was performed using RUMM 2030 software to test for goodness of fit, threshold estimates, and local dependency.Results:Of the five hundred and fifteen participants, 279 (54.2%) had a neurological condition, 119 (23.1%) a cardiopulmonary condition, 49 (10.0%) a musculoskeletal condition, and 68 (13.2%) some other condition. After Rasch analysis, the Activities and Participant component of the ICF Rehabilitation Set was divided into two domains: Activities, and Participation with 9 categories under activities and five under participation.Conclusions:After rescoring the response options, deleting four poorly-fitting categories and generating one testlet, it was found that the activities and participation component the ICF Rehabilitation Set can be divided into two outcome measures to be applied in clinical practice.

14.
Chinese Journal of Hepatobiliary Surgery ; (12): 173-176, 2020.
Artigo em Chinês | WPRIM | ID: wpr-868795

RESUMO

Objective:To evaluate the efficacy of laparoscopic splenectomy plus pericardial devascularization combined with gastroscopic (double endoscopy) treatment of patients with cirrhosis and portal hypertension presenting with bleeding esophagogastric varices.Methods:To retrospectively analyze 108 patients who presented with bleeding esophageal and gastric varices at the First Hospital of Jiaxing from March 2013 to March 2018. Of 108 patients, there were 61 males and 47 females, with an average age of 61 years. According to the disease and desires of patients and family members, 28 patients underwent laparoscopic splenectomy plus devascularization (the laparoscopic group), 43 endoscopic treatment (the endoscopic group) and 37 double endoscopic treatment (the double endoscopic group). The liver function, renal function, hemoagglutination and degrees of recurrence of the three groups were compared after operation.Results:The renal function, coagulation function, HbA1c in the double endoscopic group was significantly better than that in the other two groups ( P<0.05). In the laparoscopic group, there were 4 patients who presented with rebleeding within 36 months, compared with 3 in the endoscopic group, and no patients in the combined group. At 36 months after operation, gastroscopy performed in the laparoscopic group showed mild varices in 8(28.6%) patients, moderate in 9(32.1%), and severe in 11(39.3%). In the endoscopic group, there were 7(16.3%) patients with mild, 26(60.5%) with moderate, and 10(23.2%) with severe. In the double endoscopic group, there were 32(86.5%) patients with mild and 5(13.5%) with moderate. The degrees of recurrence and postoperative esophageal and gastric varices rebleeding in the double endoscopic group were significantly better than those in the laparoscopic group and the endoscopic group ( P<0.05). Conclusion:Laparoscopic combined with endoscopic treatment was more effective in patients with cirrhosis and portal hypertension who presented with bleeding esophageal varices.

15.
Chinese Journal of Behavioral Medicine and Brain Science ; (12): 655-659, 2019.
Artigo em Chinês | WPRIM | ID: wpr-754179

RESUMO

Objective To examine the reliability and validity of the Connor-Davidson resilience scale (CD-RISC) among coal miners,and explore factors structure and essence. Methods A questionnaire survey was conducted among 731 coal miners by using CD-RISC,Maslach burnout inventory( MBI-GS) and general self-efficacy scale( GSES). Results Twenty-five items of CD-RISC were retained based on the re-sults of exploratory factor analysis,including three factors of stress resistance,sense of competence and optim-ism. CD-RISC factors were significantly correlated with each other,and the correlation coefficient ranged from 0. 623 to 0. 777(P<0. 01). There was also a significant correlation between CD-RISC factors and total score, and the correlation coefficient ranged from 0. 837 to 0. 939(P<0. 01). The fitting index of confirmatory factor analysis were χ2/df=3. 76<5,GFI=0. 884,CFI=0. 909,AGFI=0. 862>0. 8,RMSEA=0. 065,proved that the measured data fitted well with the hypothesized three-factor model. Internal consistency reliability coeffi-cient of the total scale,stress resistance,sense of competence and optimism factors were 0. 942,0. 920,0. 868 and 0. 765,repectively. The CD-RISC scale was positively correlated with GSES and negatively correlated with MBI-GS and its factors named emotional exhaustion and depersonalization. Conclusion The CD-RISC of 25 items for coal miners possesses good reliability and validity.

16.
Chinese Journal of Comparative Medicine ; (6): 101-105, 2018.
Artigo em Chinês | WPRIM | ID: wpr-703371

RESUMO

Objective To purify marmoset serum IgG, prepare and identify the antiserum and the rabbit anti-marmoset antibody IgG-HRP (horseradish peroxidase). Methods Using SDS-PAGE analysis to identify the serum IgG from HiTrapTM Protein G. The antiserum titer was determined by double immunodiffusion assay. The rabbit anti-marmoset IgG was labeled with HRP by improved sodium periodate method. ELISA and western blotting were used to evaluate the concentration and specificity of rabbit anti-marmoset IgG-HRP. Results The purity of purified marmoset serum IgG determined by SDS-PAGE was higher than 95% , and the anti-serum titer of the anti-marmoset IgG polyclonal antibody was 1∶64. The concentration of rabbit anti-marmoset IgG-HRP identified by direct ELISA was 1∶256 000, and that by western-blotting was 1∶15 000, with a strong specificity. Conclusions The IgG-HRP marker antibody is prepared and the specificity and concentration are identified by ELISA and western blotting. It reserves the resources for the detection system of marmoset pathogens and the molecular immunological testing system.

17.
Chinese Journal of Comparative Medicine ; (6): 98-102, 2018.
Artigo em Chinês | WPRIM | ID: wpr-703325

RESUMO

Objective To analyze the effect of transport and storage conditions on the detection of pathogenic nucleic acid MHV, Reo-3, MNV in laboratory mouse cecal contents samples. Methods MHV, Reo-3 and MNV were mixed with mouse cecal contents and used as reference samples,respectively. They were placed in the lysis buffer of RNA extraction reagent(buffer AVL)or normal saline, and stored at 4℃ and room temperature(22℃-25℃). RNA of these samples was extracted at 1,2,3,7,and 14 days. Then the amount of nucleic acid in samples was detected by real-time fluorescence quantitative PCR. Results A greater decrease of the amount of nucleic acid was observed when the samples were placed in normal saline than that kept in buffer AVL. The amount of nucleic acid in samples stored at 4℃ was found to be higher than that stored at 25℃ room temperature. The amount of nucleic acid in the samples which were kept in buffer AVL at 4℃ for 3 days was higher than 50%,still detectable in the samples kept for 7 days,and undetectable at 14 days. Conclusions Mouse cecal content samples are preferably stored in the lysis buffer of RNA extraction reagent and transported at 4℃ for the detection of MHV, Reo-3, and MNV nucleic acid. It is better to complete the detection test within 3 days.

18.
Shanghai Journal of Acupuncture and Moxibustion ; (12): 525-528, 2017.
Artigo em Chinês | WPRIM | ID: wpr-512971

RESUMO

Objective To observe the clinical efficacy of electroacupuncture plus moxibustion in treating infantile cerebral palsy. Method Eighty eligible patients with infantile cerebral palsy were randomized into a treatment group of 42 cases and a control group of 38 cases. The control group was intervened by physiotherapy (PT), while the treatment group was given electroacupuncture at scalp acupoints, body acupuncture, and moxibustion along the Governor Vessel in addition to the treatment given to the control group. The two groups were treated once a day, 6 months in total. The comprehensive ability and the Gross Motor Function Measure (GMFM) were evaluated before and after the interventions in the two groups, and the clinical efficacies were compared.Result After 6-month treatment, the total effective rate was 92.9% in the treatment group versus 78.9% in the control group, and the total effective rate of the treatment group was superior to that of the control group (P<0.05). The motor function and activities of daily living were significantly improved in both groups after the interventions (P<0.05,P<0.01); after the treatment, there were significant differences in comparing the motor function and activities of daily living between the two groups (P<0.05). The GMFM scores of each domain were significantly changed after the treatment in both groups (P<0.05); after the treatment, there were significant differences in comparing the GMFM scores of each domain between the two groups (P<0.05).Conclusion Compared with being used alone, conventional PT can more significantly benefit the improvement of motor function of cerebral palsy patients when electroacupuncture and moxibustion were added.

19.
Chinese Journal of Immunology ; (12): 85-89, 2017.
Artigo em Chinês | WPRIM | ID: wpr-508447

RESUMO

Objective:To investigate the activation of peripheral basophils from patients with rheumatoid arthritis ( RA) and its mechanisms. Methods:The activation markers including CD203c and the proportion of IL-4 positive rate of peripheral basophils from RA patients and healthy controls were detected by flow cytometry. Serum levels of IgE in RA patients and healthy controls were detected by electrochemilu minescence immunoassay. Basophils were negatively isolated and co-cultured with or without purified IgE,anti-IgE as positive control,then,the expression of CD203c and propotion of IL-4 positive basophils were detected by flow cytometry. Results:The expression of CD203c and IL-4 positive rate of basophils from RA patients were higher than that of healthy controls (P<0. 05). Serum levels of IgE in RA were higher than that of healthy controls (P<0. 05). After co-cultured with isolated IgE from RA patients,basophils negatively isolated from healthy controls were activated,and higher expression of CD203c and proportion of IL-4 (P<0.05). Conclusion:Basophil activation is related with development of RA,and its activation is mainly mediated by IgE. Targeting basophil and its activation pathway would be expected to provide new strategies for the treatment of RA.

20.
Chinese Journal of Endemiology ; (12): 713-716, 2016.
Artigo em Chinês | WPRIM | ID: wpr-502211

RESUMO

Objective This study was designed to compare the mRNA expression of Toll-like receptors 2 (TLR2) and Toll-like receptors 4 (TLR4) in monocytes between brucellosis cases and healthy controls,to explore the role and value of TLR2 and TLR4 in the pathogenesis of brucellosis.Methods From March to June in 2015,a total of 155 brucellosis cases at different clinical stages were diagnosed in Ulanqab Center for Endemic Disease Prevention and Control and 50 healthy controls were selected as research subjects.Subjective clinical symptoms of all patients were observed.Serological detection of all cases was done using serum tube agglutination test (SAT).The mRNA expression of TLR2 and TLR4 in peripheral blood cell was detected by real-time PCR technique.Results Fever,hyperhidrosis,weak,bone ache and hepatosplenomegaly were found in patients of acute (69 people),subacute (34 people) and chronic (52 people) periods.The incidences of fever and hyperhidrosis in acute and subacute periods were higher than that in chronic period,and the differences were statistically significant (x2 =58.427,26.190,all P < 0.01).The incidence of bone ache in chronic period was higher than that in acute period,and the difference was statistically significant (x2 =9.264,P < 0.01).In the agglutination titer of 1:800 and higher,the positive rate of chronic period was lower than those in acute and subacute periods,and the differences were statistically significant (x2 =14.302,8.682,all P < 0.01).The mRNA expression levels of TLR2 and TLR4 were different among different clinical periods of brucellosis patients,and the differences were statistically significant (F =17.502,24.931,all P < 0.01).The expression levels of TLR2 mRNA (8.67 ± 2.39) and TLR4 mRNA (12.38 ± 3.87) in acute period of brucellosis patients were higher than those of subacute period (5.21 ± 1.76,7.62 ± 2.21),chronic period (1.25 ± 0.47,1.72 ± 0.55) and healthy control (1.17 ± 0.23,1.43 ± 0.62),and the differences were statistically significant (all P < 0.01).The expression levels of TLR2 and TLR4 mRNA in subacute period of brucellosis patients were higher than those of chronic period and healthy controls,and the differences were statistically significant (all P < 0.01).The expression levels of TLR2 and TLR4 mRNA were not significantly different (all P > 0.05) between chronic period and healthy controls.Conclusion TLR2 and TLR4 may be the specific recognizing receptors participated in the immune response in brucellosis and associated with the occurrence and development of brucellosis.

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