Objective To establish a method for the isolation of Zika virus and to gather experi-ences for viral isolation. Methods Suckling mice at age 1-3 days were inoculated with serum samples posi-tive for Zika virus through intracranial injection. All mice were sacrificed 6 days after the injection. Viral nu-cleic acids were extracted from brain, heart, liver, spleen, lung, kidney, muscle, skin and intestine tissue samples and analyzed by real-time RT-PCR. The supernatants of brain tissues positive for Zika virus were used for subculturing. Nested PCR was performed to amplify the NS5 gene of the isolated virus. The se-quences of NS5 gene were analyzed by using MEGA6. 0 software. Results All of the tissue samples were positive for Zika virus. Higher viral loads were detected in heart and brain tissue samples with cycle thresh-old (Ct) values of 24. 4 and 25. 3, respectively. The second generation of Zika virus was identified in suck-ling mice brain tissues 2 days after infection by using real-time RT-PCR. The amplified product of nested PCR was 972 bp in length. Sequencing analysis showed that the isolated Zika virus ( GDZ16002 strain) be-longed to the Asian lineage. Conclusion A strain of Zika virus was successfully isolated in China by using intracranial injection via a suckling mouse model. The isolated Zika virus belonged to the Asian lineage.

Objective To provide scientific evidences for Zika virus detection by clarifying the means by which Zika virus was discharged and the duration of corresponding processes. Methods Various samples of Zika cases were collected at different times and detected by using real-time RT-PCR. The positive samples were inoculated into cells and suckling mice through intracranial injection. The whole genome se-quences of those isolated Zika virus strain were sequenced and the results were further analyzed by comparing with the sequences of Zika virus from GenBank. Results The positive rates of Zika virus in urine, saliva and serum samples were 82. 4% (14/17), 82. 4% (14/17) and 52. 9% (9/17) respectively. The longest period of detected presence of Zika virus was found in urine samples amongst the three types of samples, fol-lowed by saliva and serum samples. Six Zika virus strains were isolated from 9 positive serum samples. Phy-logenetic analysis showed that the six genomes of Zika virus all belonged to Asia lineage, but located in two branches by Samoa and Venezuela strains. Conclusion This study indicated that urine, saliva and serum all could be used as the samples for routine detection of Zika virus. Urine and saliva samples showed higher detection rates of Zika virus RNA in comparison to serum samples, while Zika virus could be easily isolated from positive serum samples. Suckling mice were better for Zika virus isolation than cell lines.

Objective To assess the clinical therapeutic effects and safety of Jiangtang Shuxin decoction (JTSXD) on diabetic patients complicated with chronic heart failure (CHF),and to search for its possible function mechanisms.Methods A prospective randomized controlled study was conducted,80 diabetic patients complicated with CHF [New York Heart Association (NYHA) functional class Ⅱ-Ⅲ] admitted into the Department of Traditional Chinese Medicine (TCM) or of Cardiology in Affiliated Hospital of Guangxi Youjiang National Medical College from October 2015 to September 2016 were enrolled,they were assigned to an observation group and a control group by randomized method with a computer,and finally 77 patients (39 cases in observation group and 38 cases in control group) completed this trial.The patients in control group received standardized routine western medical treatment,while the observation group was additionally administered JTSXD (including ingredients:astragalus 15 g,ginseng 10 g,radix ophiopogonis 15 g,radix rehmanniae 15 g,comus 10 g,rhizome coptidis 8 g,peach kernel 10 g,salvia mitiorrhiza 10 g,magnoliaceae 10 g,yam 15 g) on the basis of conventional therapy.The therapeutic course for all the patients in both groups was 2 months.Before and after treatment,the 6-minute walking distance (6MWD) was assessed;the TCM syndrome accumulated scores of the two groups were calculated;the left ventricular end-diastolic volume (LVEDV),the left ventricle ejection fraction (LVEF),the stroke volume (SV),the cardiac output (CO),and the maximum blood flow velocity of early diastolic/atrium late diastolic (E/A) were detected by echocardiography.The serum levels of glycosylated hemoglobin (HbA1c),angiotensin Ⅱ (Ang Ⅱ) and plasma B type brain natriuretic peptide (BNP) were tested with enzyme linked immunosorbent assay (ELISA);the level changes of total cholesterol (TC),triglyeride (TG),high density lipoprotein cholesteral (HDL-C) and low density lipoprotein cholesteral (LDL-C) were observed.Results Compared with the control group,after treatment in the observed group,the TCM syndrome score of palpitation,fatigue and thetotal accumulated score were all obviously decreased (palpitation score:0.9 ± 0.4 vs.1.2 ± 0.8,fatigue score:1.1 ± 0.7 vs.1.7 ± 0.8,total accumulated score:4.8 ± 1.2 vs.8.1 ± 1.8,all P ＜ 0.05);the LVEDV,the serum levels of HbA1c,Ang Ⅱ and BNP were also obviously decreased in the observed group [LVEDV (mL):136.28 ± 17.52 vs.158.82 ± 19.03,HbA1c (％):6.11±0.36 vs.6.89 ±0.32,Ang Ⅱ (ng/L):66.48 ± 17.64 vs.84.55 ± 20.39,BNP (μg/L):138.45 ± 87.55 vs.219.14±88.83,all P ＜ 0.05];The 6MWD,LVEF,SV,CO and E/A were all increased plainly in the observed group [6MWD (m):470.47 ± 79.66 vs.428.46 ± 88.56,LVEF:0.51 ±0.05 vs.0.46 ± 0.04,SV (mL):55.36 ± 2.88 vs.50.32±2.76,CO (L/min):5.74±0.91 vs.4.92±0.74,E/A:1.18±0.27 vs.0.83±0.28,all P ＜ 0.05].The degrees of decreased levels in TC,TG,LDL-C and the degrees of increased levels of HDL-C in observed group were superior to those of the control group,but there were no statistical significant differences (all P ＞ 0.05).Conclusion JTSXD shows good therapeutic effect and safety for treatment of diabetic patients accompanied by CHF (NYHA functional class Ⅱ-Ⅲ),and its mechanisms may be related to its regulation of glucose (reduction of HbA1c level),correction of lipid metabolism disorders,improvement of myocardial energy supply,inhibition of the activation of renin-angiotensin-aldosterone system (RAAS) and the secretion of BNP.

Vaccination is the most effective way to prevent influenza and severe outcoming caused by influenza viruses.Health care workers(HCW) are exposed to patients with influenza and they are at high risk of occupationally acquired influenza and of causing nosocomial infection among patients,increasing the incidence rate,the risk of severe and death of patients.Improving the influenza immunization in HCW can not only reduce the prevalence of themselves and keep a weel-oiled of health care facilities during the influenza seasons,but also reduce the risk of severe and death among patients and increase the influenza vaccine uptake in whole population.At present,the influenza immunization coverage of HCW is low.The obstacles and myths of influenza vaccine are barriers for vaccine uptake among HCW.The various strategies are critical in order to improve the influenza coverage rates of HCW.

OBJECTIVE: To determine the incidence of traffic law violations among middle school students of Hunan province and to identify the influencing factors. METHODS: Stratified sampling and cluster sampling were used to randomly select students from 96 classes of 16 middle schools. Road traffic law violations were measured through recalling the occurrence of 5 common violations in the prior year. Most of influencing factors were collected by self-designed questionnaire except for family support function and parenting that were measured by the family assessment device (FAD) and the parenting locus of control scale (PLOC), respectively. Kruskal Wallis H test and multinomial logistic regression were used to analyze the data. RESULTS: The overall incidence rate of five common traffic law violations among middle school students in Hunan province fell between 16.6%-43.3%. Except for running against traffic light or not using pedestrian crossings (8.2%), students with the other 4 traffic law violations merely accounted for less 4%. The rates of students with 1, 2, 3, 4 and 5 kinds of traffic law violations were 25.3%, 17.8%, 10.2%, 6.2% and 3.4%, respectively. Logistic regression showed that there were 3 factors (area, junior or senior high school, and single child or not), 5 factors (area, junior or senior high school, sex, single child or not, and class leader or not) and 6 factors (type of school, junior or senior high school, sex, class leader or not, family support function, and type of family education) significantly associating with the occurrence of 1, 2, and 3 or more traffic law violations, respectively. CONCLUSION Only a small proportion of students often or almost always break traffic law or break several traffic laws at the same time. Students with serious traffic law violations should be targeted by school safety education and intervention.
Accidents, Traffic ; prevention & control ; Adolescent ; Automobile Driving ; legislation & jurisprudence ; Dangerous Behavior ; Female ; Humans ; Law Enforcement ; Logistic Models ; Male ; Risk Factors ; Safety ; legislation & jurisprudence ; Sampling Studies ; Students ; psychology ; Surveys and Questionnaires

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

OBJECTIVE: To determine the prevalence and incidence of illness of two-week duration, and the factors influencing these, among residents 15 years and older in four counties of Hunan Province. METHODS: Data were sampled from four counties of Hunan Province for the Fourth National Health Service Survey. Incidence and two-week prevalence of disease were used to assess the health service needs of residents. A non-conditional, stepwise logistic regression was employed to explore the influencing factors. RESULTS: The two-week prevalence and incidence were 11.5% and 3.9%, respectively, in four counties of Hunan. The three leading diseases of two-week prevalence were: respiratory diseases, digestive diseases, and musculoskeletal diseases. Non-conditional stepwise logistic regression showed that urban residents had 0.64 times the risk of two-week illness compared with the rural residents (P< 0.05); residents in the 45-59 year age group and the 60+ year age group had 1.69 and 2.62 times the risk of two-week illness compared with residents in the 15-44 year age group, respectively (P<0.05). The widowed had 1.91 times the risk of prevalence of two-week illness contrasted to singles (P<0.05); the students had 0.29 times the risk of two-week illness contrasted to the workers (P<0.05); urban residents had 0.63 times the risk of two-week illness compared with the rural (P<0.05); the widowed had 2.37 times the risk of incidence of two-week illness compared with singles (P<0.05). CONCLUSION The majority of health service needs of residents of four counties is generated by three diseases: respiratory diseases, digestive diseases, and musculoskeletal diseases. Relatively, rural residents, the elderly, employed persons and the widowed have higher health service needs than others and deserve specific attention.
Adolescent ; Adult ; Aged ; China ; epidemiology ; Community Health Services ; statistics & numerical data ; Digestive System Diseases ; epidemiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Musculoskeletal Diseases ; epidemiology ; Respiratory Tract Diseases ; epidemiology ; Sampling Studies ; Surveys and Questionnaires ; Young Adult

Objectiv:To evaluate ultrasound, radionuclide imaging and CT in preoperative localization diagnosis of primary hyperparathyroidism (PHPT).Method:A total of 170 PHPT patients admitted to the hospital between Jan 1992 and Dec 2020 were analyzed retrospectively. The preoperative localization diagnostic efficacy of ultrasonography, radionuclide and CT alone and in combination was compared in groups.Results:The overall sensitivity of ultrasound, radionuclide and CT were 82.13%,80.43% and 75.74%. For normal positioned parathyroid adenoma: as for sensitivity of location diagnosis, ultrasound (86.67%) was higher than radionuclide (81.82%, P<0.05) and CT (80.59%, P<0.05), ultrasound/CT parallel test (94.70%, P<0.05) was higher than ultrasound alone. For specificity of location diagnosis, radionuclide (97.78%) was higher than ultrasound (91.62%) and CT (93.39%), both ultrasound/radionuclide series tests (99.00%, P<0.001)and ultrasound/CT series tests (96.94%, P<0.001) were higher than ultrasound alone. In case of ectopic parathyroid adenoma and parathyroid hyperplasia: the sensitivity and specificity of radionuclide seemed higher than ultrasound and CT. Conclusions:Ultrasound is the first choice for preoperative location diagnosis of PHPT. Ultrasound combined with radionuclide or CT can significantly improve the diagnostic efficiency of parathyroid lesions.

Objective:To compare the changes of serum calcium level before and after surgical resection in patients with primary hyperparathyroidism.Methods:Two hundred and seventy-one patients with primary hyperparathyroidism were enrolled from Dec 1992 to Dec 2020 in Beijing Jishuitan Hospital. Serum calcium concentrations were measured before operation, 20 min during surgery, then 2 weeks 1-6 months , 7-12 months and 1 year respectively after operation. The baseline data of postoperative serum calcium such as sex, age, other genetic endocrine diseases, osteopathia and urolithiasis were calculated. The generalized estimation equation was used to analyze the changes of serum calcium in different types of patients before and after operation.Results:The most common postoperative hypocalcemia occurred within 2 weeks, and it occurred frequently half a year after surgery. There was no significant difference in blood calcium between male patients ( t=0.875, P=1.000) and patients with bone lesions ( t=0.034, P=3.049) from 1 to 6 months after surgery and 2 weeks after surgery. Blood calcium level in patients aged 15-35 years old from 1 to 6 months ( t=0.239, P=1.000) , from 7 to 12 months ( t=1.380, P=0.935) and 2 weeks after surgery was not statistically different. The change of bone mineral density was correlated with the change of blood calcium after operation ( F=6.895, P=0.004). Conclusions:The incidence of hypocalcemia was the highest in patients with hyperparathyroidism 2 weeks after surgery, and the blood calcium level was stable within the normal range 1 year later. The blood calcium value of male patients was still at a lower level than that of female patients within six months after surgery. In patients with bone disease, the blood calcium value was lower and recovered slowly 2 weeks after surgery. The blood calcium value of patients aged 15-35 was at a low level within 1 year after surgery.

Objective: To investigate the sensitivity and specificity of commercial nonstructural protein 1 (NS1) testing kits for Dengue fever diagnose, and provide the evidence for diagnostic criteria revision. Methods: 300 PCR or virus isolation positive blood samples for dengue virus were collected from sentinel hospitals for dengue surveillance in Guangzhou, Dongguang and Zhongshang from May 2015 to Nov. 2016. At the same time, 308 PCR negative samples for Dengue virus were collected as control group. The information of the sample was collected using questionnaires. These samples were tested using imported and domestic ELISA and the colloidal gold-labeled kits that were widely used for detecting dengue NS1. Sensitivity, specificity and coincidence were calculated and analyzed, and Z hongshan's result was regarded as the reslut of the third part. Results: The positive group includes 133 males and 167 females, average ages are 47.2±13.3, 179, 110 and 11 of them is Dengue Ⅰ, Ⅱ and Ⅲ respectively. The negative group includes 154 males and 154 females, average ages are (40.1±11.6) years old. The sensitivity of domestic ELISA Kits (94.5%) is less than imported (99.5%), and the result has statistical significance (χ²=8.59, P=0.030), the specificity is 99.7% and 97.7% respectively; The sensitivity of imported and domestic the colloidal gold-labeled Kits is 97.5% and 96.5% respectively, both of specificities are 100%. The sensitivity and specificity of Dengue Ⅰ for NS1 test are more than 97.0%. The sensitivity of domestic ELISA and gold-labeled Kits is 90.0% and 95.0%, and the specificity is 96.8% and 100% respectively for Dengue Ⅱ test. The sensitivity of imported ELISA and gold-labeled Kits is 100% and 98.0%, and the specificity is 99.4% and 100% respectively for Dengue Ⅱ test. The result of the third party show the sensitivity and specificity of domestic ELISA and gold-labeled Kits are 90.0% and 98.0%, the differences has statistical significance (χ²=5.67, P=0.020). Conclusion NS1 testing can be used as early dengue fever diagnose for higher sensitivity and specificity.