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Objective To investigate the prevalence of thyroid nodules in women participating in physical examination in Beijing and analyze the influencing factors. Methods The data of physical examination (height, weight, blood pressure, blood glucose, etc.) and questionnaire survey (activity intensity, eating habits, etc.) of women in Beijing in 2016 were collected, and the influencing factors of thyroid nodules were analyzed by multivariate logistic regression. Results A total of 4 732 women were included in this study. The prevalence of thyroid nodules was 49.6%. Multivariate logistic regression analysis showed that compared with women aged 18-29 years, OR value was 1.769 (95% CI =1.489 ~ 2.102) for women aged 30 ~ 59 years, and OR value was 4.716 (95% CI = 3.577- 6.216) for women aged 60 years and over. Compared with the balanced diet, the OR value was1.237（95%CI=1.056-1.450）for vegetarian diet. Compared with the normal weight, the OR value was 1.331（95%CI=1.153-1.537）for the overweight. Compared with the healthy women, the OR value was 1.405 （95%CI=1.146-1.723）for hypertension, the OR value was 1.184（95%CI=1.040-1.347）for hyperlipidemia, and the OR value was 1.779（95%CI=1.178-2.687）for diabetes, while the OR value was 1.183（95%CI=1.018-1.376）for women with mammary gland nodules, and the OR value was 1.376（95%CI=1.201-1.575）for women with uterine leiomyoma. Compared with the education degree of high school, technical secondary school, technical school and below, the OR value was 0.648（95%CI=0.522-0.806）for college or undergraduate, and the OR value was 0.564（95%CI=0.440-0.723）for graduate students and above. Conclusion The prevalence of thyroid nodules in women in Beijing is at a high level. Age, vegetarian diet, overweight, hypertension, hyperlipidemia, diabetes, mammary gland nodules and uterine leiomyoma are risk factors for thyroid nodules. Education level is a protective factor for the prevalence of thyroid nodules.
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Objective To explore food allergies in children with asthma in urban areas. Methods A total of 1 462 children with asthma who were treated in Mianyang Central Hospital of Sichuan Province from January 2018 to January 2019 were enrolled as the observation group, and 1,828 children who underwent physical examination in the same hospital at the same time were selected as the control group. The types and proportions of common food allergies were summarized and calculated, and the types and proportion of clinical symptoms in the two groups were statistically analyzed. Results A total of 219 children with food allergy were found in the observation group, and the prevalence rate was 14.98%. In the control group, 72 children with food allergy were found, and the prevalence rate was 3.94%. The difference between the two groups was statistically significant (χ2=5.036, P=0.024). The peanut allergy rate in the observation group was significantly higher than that in the control group, while the fruit allergy rate was significantly lower than that in the control group. The differences were statistically significant (P<0.05). The incidence of itch, lip and mucous membrane swelling, sneezing and shock were significantly higher in the observation group than in the control group, while the incidence of skin itching and rash was significantly lower than that in the control group. The differences were statistically significant (P<0.05). Conclusion The prevalence of food allergy in children with bronchial asthma is higher than that in non-bronchial asthma children, and the common food allergies and clinical symptoms of bronchial asthma are significantly different from those of non-bronchial asthma children. The clinicians can make a preliminary diagnosis based on the common food allergies and clinical symptoms of children.
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Objective To establish a one-step recombinase polymerase amplification (RPA) method for pathogen screening and rapid detection in the field targeting for five hemorrhagic fever related viruses (Zaire ebola virus, Sudan ebola virus, Marburg virus, Lassa virus and Yellow fever virus). Methods The specific nucleic acid (NA) fragments of each virus were selected as target genes by genome sequence analysis, and the primers and probes for RPA assays were designed according to the sequence. A series of diluted template genes were used for RPA detection to determine the sensitivity. The hemorrhagic fever-related viral nucleic acids were used for RPA detection to determine the specificity. The amplification experiments were carried out at different temperature ranging from 37℃ to 42℃ to validate the reaction temperature range. Results The RPA reaction systems of the five hemorrhagic fever viruses could effectively amplify the target genes, the sensitivities were between 1.5×102 and 1.5×103 copies. No cross reactions existed with the other hemorrhagic fever-related viral genes. Meanwhile, RPA assay could effectively amplify the target genes at 37-42℃. Conclusion The isothermal RPA assays of five hemorrhagic fever viruses are established, which may amply target genes fast and react at a wide temperature range, and be potentially useful for in field pathogens detection.
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Objective To investigate the value of brain natriuretic peptide and heart failure ultrasound in the diagnosis of early chronic heart failure.Methods One hundred and forty-six patients with chronic heart failure and seventy-five healthy people with normal heart function were involved.According to NYHA classification,patients with chronic heart failure were divided into grade Ⅰ,Ⅱ,Ⅲ and Ⅳ,including grade Ⅰ twenty six,grade Ⅱ forty three,grade Ⅲ forty eight and grade Ⅳ twenty nine patients.Results There was no significant difference in the general information between patients with chronic heart failure and healthy people with normal heart function (P ＞ 0.05),including age,sex,ALT,AST,fasting blood glucose,total cholesterol,triglyceride,serum creatinine and CRP.The BNP of the two groups were compared,and we found the BNP of grade Ⅱ (181.19 ±48.06),Ⅲ (279.26 ±49.45) and Ⅳ (882.34 ± 388.71) patients was significantly higher than healthy people (54.12 ± 25.87),and with the severity of heart failure,BNP increased gradually.There was statistically significant the difference (P ＜ 0.05).but there was no significant difference between the patients of grade Ⅰ (77.14 ± 24.32) and healthy people (P ＞ 0.05).Compared with healthy people(0.78 ± 0.42),HFEI was significantly higher in patients with chronic heart failure.The HFEI of grade Ⅰ (1.58 ± 0.89),Ⅱ (2.64 ± 1.07),Ⅲ (4.47 ± 1.39) and Ⅳ (6.33 ± 1.61) were compared,we found HFEI increased gradually with the severity of heart failure,and the difference was statistically significant (P ＜ 0.05).From the analysis between BNP and HFEI,we found that BNP was significantly positively correlated with HFEI in patients with chronic heart failure (r =0.935,P =0.000).From ROC curve analysis,the area under the ROC curve of early chronic heart failure was 0.970,showing that plasma BNP combined with HFEI could be accurately diagnosed early chronic heart failure.Conclusion We find plasma brain natriuretic peptide and heart failure echocardiography index can be used in early diagnosis of chronic heart failure.
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Objective To investigate the relationship between pulmonary function,serum inflammatory factors and coronary artery disease in elderly patients with hypertension.Metbods One hundred and fifty-two patients with hypertensive disease were involved.According to whether the patients had coronary artery disease,the patients were divided into hypertension with coronary artery disease group included 84 patients and non-combined coronary artery disease group included 68 patients.Results Age (75.23-± 7.49years vs 71.42 ± 8.21years),triglyceride (1.59 ± 0.41mmol/L vs 1.41 ± 0.63mmol/L) serum creatinine (82.42 ± 39.27μmol/L vs 70.39 ±25.76μmol/L),blood urea nitrogen (7.85 ± 4.31mmol/L vs 6.12 ± 2.49mmol/L),IL-2 (567.32 ± 191.53pg/d1 vs 501.26 ±214.74pg/dl),IL-6 (3.95 ± 2.92pg/dl vs 3.04 ± 2.61pg/dl) and CRP (1.75 ± 1.84mg/dl vs 1.04 ± 2.01 mg/dl) of hypertension with coronary artery disease group were higher than hypertension non-combined coronary artery disease group,however FEV1％ (83.42％ ± 24.57％ vs 92.15％ ± 19.38％) and FEV1/FVC (62.91 ± 13.65 vs 70.24 ± 9.42) were more lower.There was significant difference (P ＜ 0.05).From the multiple linear regression analysis of relevant factors,we found that FEV1 ％ (P =0.005),FEV1/FVC (P =0.003),IL-2 (P =0.012) and CRP (P =0.009) was independently associated with hypertension complicated with coronary artery disease.From the analysis between IL-2,CRP and FEV1％,FEV1/FVC of hypertensive patients with coronary artery disease,we found that IL-2 was significantly negatively correlated with FEV1 ％ (r =-0.391,P =0.000) and FEV1/FVC (r =-0.571,P =0.000),and CRP was also was significantly negatively correlated with FEV1％ (r =-0.437,P =0.000) and FEV1/FVC (r =-0.559,P =0.000).Conclusion We find that pulmonary function and serum inflammatory factors are the risk factors of the coronary artery disease in elderly patients with hypertension.
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Objective To investigate the relationship between pulmonary function,serum inflammatory factors and coronary artery disease in elderly patients with hypertension.Metbods One hundred and fifty-two patients with hypertensive disease were involved.According to whether the patients had coronary artery disease,the patients were divided into hypertension with coronary artery disease group included 84 patients and non-combined coronary artery disease group included 68 patients.Results Age (75.23-± 7.49years vs 71.42 ± 8.21years),triglyceride (1.59 ± 0.41mmol/L vs 1.41 ± 0.63mmol/L) serum creatinine (82.42 ± 39.27μmol/L vs 70.39 ±25.76μmol/L),blood urea nitrogen (7.85 ± 4.31mmol/L vs 6.12 ± 2.49mmol/L),IL-2 (567.32 ± 191.53pg/d1 vs 501.26 ±214.74pg/dl),IL-6 (3.95 ± 2.92pg/dl vs 3.04 ± 2.61pg/dl) and CRP (1.75 ± 1.84mg/dl vs 1.04 ± 2.01 mg/dl) of hypertension with coronary artery disease group were higher than hypertension non-combined coronary artery disease group,however FEV1％ (83.42％ ± 24.57％ vs 92.15％ ± 19.38％) and FEV1/FVC (62.91 ± 13.65 vs 70.24 ± 9.42) were more lower.There was significant difference (P ＜ 0.05).From the multiple linear regression analysis of relevant factors,we found that FEV1 ％ (P =0.005),FEV1/FVC (P =0.003),IL-2 (P =0.012) and CRP (P =0.009) was independently associated with hypertension complicated with coronary artery disease.From the analysis between IL-2,CRP and FEV1％,FEV1/FVC of hypertensive patients with coronary artery disease,we found that IL-2 was significantly negatively correlated with FEV1 ％ (r =-0.391,P =0.000) and FEV1/FVC (r =-0.571,P =0.000),and CRP was also was significantly negatively correlated with FEV1％ (r =-0.437,P =0.000) and FEV1/FVC (r =-0.559,P =0.000).Conclusion We find that pulmonary function and serum inflammatory factors are the risk factors of the coronary artery disease in elderly patients with hypertension.
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Objective To prepare quality control samples for St.Louis encephalitis virus(SLEV)molecular detection by constructing pseudovirus containing target sequences of SLEV.Methods According to the principles of armored RNA technique, the prM gene sequence of SLEV was cloned into the prokaryotic expression vector to generate recombinant plasmid pSE380-MS2-SLEV.Then, recombinant E.coli transformed with the corresponding plasmid was induced with IPTG to produce recombinant pseudovirus particles.The particles were purified by chloroform and further characterized by double enzyme digestion and transmission electron microscopy.The temperature sensitivity experiments and quantitative RT-PCR were performed to validate the potential of these pseudovirus particles as quality control samples.Results PCR amplification and sequencing analysis confirmed that the prM gene sequence of SLEV was cloned into vector pSE380-MS2.Transmission electron microscopy showed that homogenous spherical particles with a diameter of about 25 nm were produced upon IPTG induction.The SLEV genomic RNA within the pseudovirus particles was resistant to DNaseⅠand RNase A digestion, and remained stable for 20 days at 37℃.These samples were validated with quantitative RT-PCR for SLEV.Conclusion The RNase-resistant and stable pseudovirus particles containing prM fragment of SLEV are constructed successfully, which can be used as positive quality control samples for RNA extraction and molecular detection.
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Objective To construct the pseudovirus containing nucleic acid(NA)fragments of Marburg virus,Zaire Ebola virus,Sudan Ebola virus,Lassa fever virus and Yellow fever virus by using a lentiviral vector system in order to provide a reference standard for the detection of the five viruses.Methods The gene fragments of the above five viruses were synthesized in vitro,connected into a single gene by fusion PCR technique,and cloned into lentiviral vectors with its auxiliary vector.After co-transfecting into 293T cells,the supernatants were collected on 48 h and 72 h post transfection. The naked NA was cleaned from the supernatants using DNase and RNase digestion before pseudotype virus was purified and concentrated.After the NA of the pseudotype virus,were extracted normal PCR and real-time PCR were conducted. Results Sequence analysis showed that the five target genes in vitro synthesis were properly connected and inserted into lentivirus vectors.Using the NA of the pseudotype virus as the template,both normal PCR and real-time PCR could sensitively amplify the target gene with the primers and probes of the above five,viruses respectively.The result indicated that the pseudovirus particles containing the five kinds of hemorrhagic fever virus target genes were successfully packaged. Conclusion The pseudovirus particles containing gene fragments of five viruses are constructed,which can be used as a common reference standard for NA detection.
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Objective To explore the changes of cytokines in myasthenia gravis children and the effect of glucocorticoid on the serum levels of them.Methods Forty cases of myasthenia gravis children were the observation group which was divided to group one (pre-glucocorticoid therapy)and group two (post-glucocorticoid therapy),and 20 cases of healthy children in the same period as the normal control group.The serum levels of cytokines INF-γ,TGF-β1,IL-10 and IL-18 of the observation group one and two and the normal control group were detected by ELISA and were compared between the observation group one and the normal control group and the observation group one and two.Results The serum levels of cytokine INF-γand IL-18 were higher and IL-10 and TGF-β1 were lower in the observation group than in the normal control group,the differences were statistically significant (t =4.45 ~16.72,P 0.05).Conclusions Cytokines INF-γ,TGF-β1,IL-10 and IL-18 are involved and probably play different roles in the pathogene-sis of myasthenia gravis in children;Glucocorticoid could affect the secretion of cytokines IFN-γ,TGF-β1 and IL-18 of myasthenia gravis children,which would ultimately to achieve the aim of interfering and con-trolling the clinical symptom of myasthenia gravis in children.
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Objective To express and purify the recombinant nucleoprotein fragments of hemor-rhagic fever with renal syndrome ( HFRS) virus and to evaluate their diagnostic efficacy by using array-ELISA technology .Methods The target genes encoding nucleoprotein fragments of HFRS virus were amplified by PCR, and then inserted into prokaryotic expression vectors to construct the recombinant plasmids of pET -32a (+)/Pn and pET-32a(+)/Pc.The plasmids were transformed into E.coli BL21 ( DE3) to induce the ex-pression of nucleoprotein fragments by IPTG and the expressed products were purified by affinity chromatog -raphy using Ni-NTA agarose.The specificity and sensitivity of the recombinant antigens were evaluated by the assay of array-ELISA using commercial colloidal gold assay kit as a comparison .Results The recombi-nant nucleoprotein fragments of HFRS virus were correctly expressed in E.coli and highly purified by affinity chromatography .Array-ELISA showed that 13 of 16 suspected serum samples were positive by using the His-Pn protein as diagnostic antigen , consistency with the commercial colloidal gold assay kit reaching 94%. Conclusion The recombinant His-Pn protein expressed in E.coli cells could be used for specific serodiag-nosis of HFRS virus as its high antigenicity and sensitivity .The array-ELISA is an effective assay for the de-tection of virus at protein level .
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Objective To express and purify Japanese encephalitis virus ( JEV) EDⅢprotein and evaluate the possibility of using it as a candidate antigen in JEV diagnostic kit .Methods PCR primers spe-cific for the gene encoding JEV EDⅢprotein were designed and used to amplify the gene fragment by RT-PCR.The cloned gene fragment was then inserted into pET-30a (+) to construct the recombinant expression plasmid.The transformed E.coli BL21 carrying expression plasmid were induced by IPTG to express JEV EDⅢ protein.The expressed JEV EDⅢprotein and a control antigen of tick-borne encephalitis virus protein were deposited in small spots to set up ELISA microarray .The serum samples from patients with Japanese encephalitis and healthy people were detected by Array-ELISA.The results obtained by Array-ELISA were compared with those by using indirect immunofluorescence assay .Results The gene fragment encoding JEV EDⅢprotein was successfully cloned and expressed in E.coli BL21.The recombinant protein could be used in Array-ELISA assay for the detection of serum samples from patients with Japanese encephalitis and healthy subjects .The results were consistent with those by using indirect immunofluorescence assay.Conclusion The recombinant JEV EDⅢprotein can be used as a candidate antigen for the diagnosis of JEV infection .
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Objective To identify the infection and the replication of Tick-borne encephalitis virus(TBEV) in human neuroblastoma cells.Methods After being inffected with TBEV,the cell culture supernatant of human neuroblastoma cell line SK-N-SH was collected and assayed at different time points.Byusing real-time RT-PCR and plaque assay to measure the titer of virus in the supernatant,the replication andproliferation of TBEV in human neuroblastoma cell was identified.Meanwhile,the morphological change of SK-N-SH after TBEV infection was also visualized by observation under microscope and immunmquorescenceassay.Results Real-time RT-PCR and plaque assay both demonstrated that TBEV could replicate effectively in SK-N-SH cells,the peak titer could reach 2.92× 107 PFU/ml on 3 days post-inoculation.And significant morphological change occured on infected SK-N-SH cells after 2 days post inoculation.By immunofluorescence assay,the virus particles could be detected and visualized.Conclusion TBEV can replicate andproliferate effcctively and cause significant cell morphological changes in human neuroblastoma cell SK-N-SH,which demonstrated that SK-N-SH could be a suitable cell model for TBEV culture.
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ObjectiveTo develop an antibody-array system for multiple detection of antibodies against Japanese B encephalitis virus (JEV),Tick-borne encephalitis virus (TBV),Dengue virus ( DENV ),West Nile virus (WNV),Western equine encephalitis virus (WEEV) and East Equine encephalitis virus (EEEV).MethodsRecombined antigens were spotted on array as capture antigens.Specific antibodies were detected by using a sandwich ELISA format.Rabbit antiserum was employed to select and confirm the specificity of antigens and to optimize the conditions of the assay.The detection efficiency of the system was validated by 40 clinical suspected serum samples and compared with the relative ELISA assays.ResultsEleven recombined antigens were selected as diagnostic antigens with high specificity.Better detection could be achieved when scale of antigen concentrations were within 0.125-0.900 mg/ml and the serum dilutions were 1:100-1:1000.When detecting the 26 clinical suspected TBE serum samples,20 were IgG positive (76.9％),and 17 were IgM positive (65.3％) which was 96.1％ and 84.6％ consistent with the relevant ELLSA tests,the 8 clinical suspected JEV serum samples,4 were IgG positive (50.0％),and 5 were IgM positive (62.0％),which was 86.3％ and 90.1％ consistent with the relevant ELLSA tests.As for the 22 DEN serum samples,13 were IgG positive (60％) and 15 were IgM positive (68％) which was 85％ and 93％ consistent with ELISA.The specificity of the assay was 100％ and the sensitivity was higher than the relative ELISAs.ConclusionThe developed antibody-array is highly specific and reliable,which could be used for the detection of antibodies against the 6 arboviruses.
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Objective To evaluate the prevalence, associated risk factors and the impact on health related quality of life of overactive bladder (OAB) syndrome in Beijing adult women. Methods In the Community-based, cross-sectional study, 2973 women aged over 18 years who lived in urban and suburban communities respectively in Beijing were interviewed through Voiding Function Questionnaire, using a stratified system sampling approach. The women meeting the diagnostic criteria of OAB using the International Continence Society (ICS) definition were further interviewed through King's health questionnaire(KHQ) to estimate the impact of OAB on health related quality of life in Beijing adult women. Results A total of 2379(80.0%) women with complete data were included in this study. The overall prevalence of OAB was 4.7% (112/2379)and demonstrated an significant increasing with advancing age (P＜0.01). The prevalence of OAB was 2.0% in urban and 8. 1 % in suburban. In multiple logistic models, age, BMI, region of residence and anxiety level were associated risk factors for OAB. The result of King's health questionnaire showed the symptoms of OAB affect general health perception and sleep and energy severely. Conclusions The prevalence of OAB in Bei-jing adult women is 4.7%, lower than that of most reports in Occidental women, increases with advancing age, higher BMI and higher anxiety level, and is higher in suburban than in urban. The symptoms of OAB have a detrimental effect on quality of life.
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Objective To evaluate the reliability and validity of the Chinese version of the King's Health Questionnaire (KHQ) in patients with overactive bladder(OAB).Methods The original English KHQ was translated into Chinese and linguistically validated following the Cross-cultural adaptation of health-related quality of life measures. Patients recruited randomly from urology clinics were scheduled for two visits with 2 weeks apart, and they were surveyed through the Chinese version of the KHQ. Internal consistency reliability was assessed by Cronbach's α test;Test-retest reliability was examined among stable patients using Intraclass correlation coefficient (ICC) and Spearman's rank correlation statistical analyses. Content validity was estimated by Spearman's rank correlation statistical analyses. A factor analysis was conducted to validate the underlying factor structure of the Chinese version of the KHQ. Results A total of 48 OAB patients who met the criteria participated the study, and 40 patients (7 men, 33 women) completed the questionnaires twice. All the subscales and domains of the KHQ showed high levels of internal consistency (Cronbach's a: 0.718-0. 924) , moderate to excellent test-retest reliability (ICC:0.551-0.923,P<0.01) and acceptable construct validity. The content validity was moderate to excellent except for the Social Limitations domain. Conclusion Psychometric testing supports the reliability and validity of the Chinese version of the KHQ as an OAB-specific measure of HR QOL.
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Objective To evaluate the Flavivirus specific monoclonal antibody(McAb) 2A10 as detective antibody for simultaneously identify tick borne encephalitis virus( TBEV), Japanese encephalitis virus( JEV), dengue ( DEN )-2, DEN-4 and yellow fever virus ( YFV ) by antibody microarray technique.Methods The antibody microarray was developed by spotting TBEV, JEV, DEN-2, DEN-4 and YFV specific McAb on chip as capture antibodies. After incubating with cultured viral supernatants of the above viruses, CY3 labeled detective antibody 2A10 was added to the chips. After reaction, the antibody microarray was scanned and the results were analyzed. By comparing the signal intensities of different spots on chips,the detecting titre and sensitivity of 2A10 for Flavivirus were determined, and the value of 2A10 in detection of Flavivirus was evaluated. Results The hybridization results demonstrated that the titre of 2A10 for Flavi2A10 was specific for Flavivirus and could be used as universal detective antibody for Flavivirus on antibody microarray.
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The goal of this work is to understand the difference between vertical structure and horizontal structure on data management. We take adverse event report as an example to find out the relationship between the storage space and the factors related to the trail. From the study we find out the difference of storage space between horizontal structure and vertical structure goes up with the increase of the storage space of each adverse event record and the number of adverse event allowed to record in the horizontal structure and goes down with the increase of the happening rate and the average times of the adverse event. When using the most widely used study design, vertical structure always takes less storage space than the horizontal structure. This vertical structure database is more suitable for the data management than the horizontal structure database.