Establishment of qRT-PCR for absolute quantitative detection of Chikungunya virus / 中国热带医学

@#Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

Comparison of aspirin treatment strategies for primary prevention of cardiovascular diseases: A decision-analytic Markov modelling study / 北京大学学报(医学版)

OBJECTIVE@#To compare the expected population impact of benefit and risk of aspirin treatment strategies for the primary prevention of cardiovascular diseases recommended by different guidelines in the Chinese Electronic Health Records Research in Yinzhou (CHERRY) study.@*METHODS@#A decision-analytic Markov model was used to simulate and compare different strategies of aspirin treatment, including: Strategy ①: Aspirin treatment for Chinese adults aged 40-69 years with a high 10-year cardiovascular risk, recommended by the 2020 Chinese Guideline on the Primary Prevention of Cardiovascular Diseases; Strategy ②: Aspirin treatment for Chinese adults aged 40-59 years with a high 10-year cardiovascular risk, recommended by the 2022 United States Preventive Services Task Force Recommendation Statement on Aspirin Use to Prevent Cardiovascular Disease; Strategy ③: Aspirin treatment for Chinese adults aged 40-69 years with a high 10-year cardiovascular risk and blood pressure well-controlled (< 150/90 mmHg), recommended by the 2019 Guideline on the Assessment and Management of Cardio-vascular Risk in China. The high 10-year cardiovascular risk was defined as the 10-year predicted risk over 10% based on the 2019 World Health Organization non-laboratory model. The Markov model simulated different strategies for ten years (cycles) with parameters mainly from the CHERRY study or published literature. Quality-adjusted life year (QALY) and the number needed to treat (NNT) for each ischemic event (including myocardial infarction and ischemic stroke) were calculated to assess the effectiveness of the different strategies. The number needed to harm (NNH) for each bleeding event (including hemorrhagic stroke and gastrointestinal bleeding) was calculated to assess the safety. The NNT for each net benefit (i.e., the difference of the number of ischemic events could be prevented and the number of bleeding events would be added) was also calculated. One-way sensitivity analysis on the uncertainty of the incidence rate of cardiovascular diseases and probabilistic sensitivity analysis on the uncertainty of hazard ratios of interventions were conducted.@*RESULTS@#A total of 212 153 Chinese adults, were included in this study. The number of people who were recommended for aspirin treatment Strategies ①-③ was 34 235, 2 813, and 25 111, respectively. The Strategy ③ could gain the most QALY of 403 [95% uncertainty interval (UI): 222-511] years. Compared with Strategy ①, Strategy ③ had similar efficiency but better safety, with the extra NNT of 4 (95%UI: 3-4) and NNH of 39 (95%UI: 19-132). The NNT per net benefit was 131 (95%UI: 102-239) for Strategy ①, 256 (95%UI: 181-737) for Strategy ②, and 132 (95%UI: 104-232) for Strategy ③, making Strategy ③ the most favorable option with a better QALY and safety, along with similar efficiency in terms of net benefit. The results were consistent in the sensitivity analyses.@*CONCLUSION@#The aspirin treatment strategies recommended by the updated guidelines on the primary prevention of cardiovascular diseases showed a net benefit for high-risk Chinese adults from developed areas. However, to balance effectiveness and safety, aspirin is suggested to be used for primary prevention of cardiovascular diseases with consideration for blood pressure control, resulting in better intervention efficiency.

A consensus on the management of allergy in kindergartens and primary schools / 中国学校卫生

Abstract@#Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.

Knowledge， attitude and practice（KAP） of adolescents visiting STD clinic and influencing factors / 上海预防医学

Objective:To understand the status and knowedge， attitude， practice（KAP） of adolescent sexually transmitted disease（STD） patients in Shanghai， and provide evidence for the development of health interventions. Methods:A cross-sectional study was conducted by anonymous interview survey using self-made questionnaires in patients during 2017.10.1-2017.12.31. Consecutive variables were analyzed using Mann-Whitney U test. Classification variables were analysed using Chi-square analysis and logistic analysis. Results:Regarding to sex and reproductive health， the knowledge awareness rate of STD patients was very low. The rate of correctly answering most questions was less than 5%. The patients were more open to risky sex， while the proportion of positive attitude towards safe sex was lower. Men were more open than women to risky sex. For example， 34.7% and 35.7% of men accepted homosexuality and one-night stand， respectively， but only 1.5% and 12.4% of women showed acceptance. There was statistically significant difference in knowledge scores of STD between different genders （P=0.020）. The knowledge score of female patients was higher than that of male patients. There were statistically significant differences in unsafe sex and preventive health care attitudes among patients of different gender （P<0.001） and sex orientation （P<0.001）. The attitudes of female patients were better than those of male patients， and the attitudes of heterosexual patients were better than those of sexual minorities. There was statistically significant difference in sexual behavior among patients with different sexual orientation （P=0.014）， and the behavior score of sexual minority patients was better than that of heterosexual patients. The influencing factors of sex and reproductive knowledge were gender （P=0.001） and native place （P=0.004）. STD knowledge level in females and Shanghai residents was higher than males and the non-residents. Conclusion:There is a large gender difference in adolescent STD groups in Shanghai. Women， floating population and sexual minorities are vulnerable groups and need more social support. More health education should be targeted on the factors affecting reproductive health knowledge， and adolescent STD prevention strategies should be formulated.

Influence of road types on road traffic accidents in northern Guizhou Province, China / 中华创伤杂志(英文版)

PURPOSE@#The increasing number of deaths due to road traffic accidents (RTAs) has attracted global attention. However, the influence of road types is rarely considered in the study of RTAs. This study evaluates the influence of different road types in RTAs in northern Guizhou to provide a basis for the formulation of evidence-based policies and measures.@*METHODS@#We obtained the data from the Zunyi Traffic Management Data Platform for the years 2009-2018. The mortality rates of RTAs were calculated. Descriptive methods and Chi-square tests were used to analyze the characteristics of road traffic collisions on different road types. We also examined the associations between the mortality rate per 10,000 vehicles and the growth of per capital gross domestic product (GDP) with Spearman's rank correlation analysis. According to the passing volume and the infrastructure, we defined different types of roads, like administrative road, functional road, general urban road and urban expressway.@*RESULTS@#In 2012, the traffic mortality rate of administrative roads was 8.9 per 100,000 people, and the mortality rate of functional roads was 7.4 per 100,000 people, which decreased in 2018 to 6.1 deaths per 100,000 people and 5.2 deaths per 100,000 people, respectively. The mortality rate per 10,000 vehicles reached the highest level in 2011 (28.8 per 10,000 vehicles and 22.5 per 10,000 vehicles on administrative and functional roads, respectively). The death rate of county roads was the highest among administrative roads (χ@*CONCLUSION@#Although our research shows that RTAs in northern Guizhou have steadily declined in recent years, the range of decline is relatively small. Many measures and sustainable efforts are needed to control road traffic death and accelerate the progress in road traffic safety in northern Guizhou.

Research progress of circular RNAs in ocular proliferative diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#Circular RNAs(circRNAs)comprise a novel class of non-coding RNAs that are found to be highly abundant in eukaryotic cells and have implicated in various cellular functions such as cell proliferation, differentiation, and apoptosis. Recent advances have suggested that dysregulated circRNAs play a critical role in the pathogenesis of several proliferative retinal diseases including proliferative vitreous retinopathy, diabetic retinopathy, age-related macular degeneration, retinopathy of prematurity, and corneal neovascularization. Here, we review current knowledge about circRNAs and summarize new insights into potential functions of some aberrantly expressed circRNAs and possible future directions in ocular proliferative diseases.

Impact of different clear corneal incision sizes on anterior corneal aberration for cataract surgery / Impacto de diferentes tamanhos de incisões em córnea clara nas aberrações da córnea anterior em cirurgia de catarata

ABSTRACT Purpose: To investigate the impact of different sizes of steep meridian clear corneal incisions for phacoemul sification on anterior corneal higher-order aberrations. Methods: Medical records of patients who underwent 2.2-mm coaxial micro-incision cataract surgery or 2.75-mm coaxial small-incision cataract surgery were retrospectively reviewed. Only patients with preexisting anterior corneal astigmatism <2.00 diopters (D) and ≥0.50 D who underwent a steep meridian clear corneal incision were included. Primary outcomes were 3rd- to 6th-order anterior corneal higher-order aberrations with an 8-mm pupil. Anterior corneal astigmatism and effective phaco time were evaluated as secondary outcomes. Preoperative and 3-month postoperative outcomes were evaluated. Results: Anterior corneal astigmatism significantly decreased after both procedures; however, there was no significant difference found in surgically induced anterior corneal astigmatism between the two procedures (p=0.146). Although the total higher-order aberrations did not significantly change after both procedures, the group comparison showed a significant difference in surgically induced total higher-order aberrations (a decrease of 0.337 ± 1.156 mm in 2.2-mm coaxial micro-incision cataract surgery and an increase of 0.106 ± 0.521 mm in 2.75-mm coaxial small-incision cataract surgery, p=0.046). Spherical aberrations significantly decreased after 2.2-mm coaxial micro-incision cataract surgery (p=0.001), whereas they did not change significantly after 2.75-mm coaxial small-incision cataract surgery (p=0.564). Coma did not significantly change after either of the procedures. Trefoil did not significantly change after 2.2-mm coaxial micro-incision cataract surgery (p=0.361), whereas it significantly increased after 2.75-mm coaxial small-incision cataract surgery (p<0.001). There was no significant difference shown in effective phaco time between the procedures. A significantly positive correlation was shown between surgically induced anterior corneal astigmatism and coma in 2.75-mm coaxial small-incision cataract surgery (r=0.387, p=0.006). There was no significant correlation found between any surgically induced higher-order aberration changes and effective phaco time. Conclusions: The results showed that 2.2-mm coaxial micro-incision cataract surgery and 2.75-mm coaxial small-incision cataract surgery did not significantly degrade the total higher-order aberrations of the anterior cornea. However, the surgically induced changes in total higher-order aberration showed a significant difference between the two procedures, with a slight reduction after 2.2-mm coaxial micro-incision cataract surgery and a slight increase after 2.75-mm coaxial small-incision cataract surgery. Phaco time and power used during surgery had no impact on corneal aberrations.

RESUMO Objetivo: Investigar o impacto de diferentes ta manhos de incisões em córnea clara com meridiano íngreme para facoemulsificação com aberrações de mais alta ordem da córnea anterior. Métodos: Foram retrospectivamente revisados os prontuários médicos de pacientes que se submeteram a cirurgias de catarata com microincisões coaxiais de 2,2 mm ou com incisões coaxiais pequenas de 2,75 mm. Foram apenas incluídos pacientes com astigmatismo preexistente da córnea anterior <2,00 dioptrias (D) e ³0,50 D, e submetidos a incisões em córnea clara com meridiano íngreme. Os desfechos primários foram aberrações da córnea anterior da 3ª à 6ª ordem com uma pupila de 8 mm. O astigmatismo da córnea anterior e o tempo efetivo de facoemulsificação foram avaliados como desfechos secundários. Os desfechos pré-operatório e pós-operatório aos 3 meses também foram avaliados. Resultados: O astigmatismo da córnea anterior diminuiu significativamente após ambos os procedimentos, mas não se encontrou nenhuma diferença significativa entre os dois procedimentos quanto ao astigmatismo da córnea anterior, induzido pela cirurgia (p=0,146). Embora as aberrações totais de mais alta ordem não se tenham alterado significativamente após ambos procedimentos, a comparação entre os grupos revelou uma diferença significativa nas aberrações totais de mais alta ordem, induzidas pela cirurgia (uma diminuição de 0,337 ± 1,156 mm na cirurgia de catarata por microincisão coaxial de 2,2 mm e um aumento de 0,106 ± 0,521 mm na cirurgia de catarata por incisão coaxial pequena de 2,75 mm; p=0,046). A aberração esférica diminuiu significativamente após cirurgia de catarata por microincisão coaxial de 2,2 mm (p=0,001), mas não se alterou significativamente após cirurgia de catarata por incisão coaxial pequena de 2,75 mm (p=0,564). A aberração de coma não mudou significativamente após qualquer dos procedimentos. O trifólio não se alterou significativamente após cirurgia de catarata por microincisão coaxial de 2,2 mm (p=0,361), mas aumentou significativamente após cirurgia de catarata por incisão coaxial pequena de 2,75 mm (p<0,001). Nenhuma diferença significativa se evidenciou quanto ao tempo efetivo de faco-emulsificação entre os dois procedimentos. Houve uma correlação positiva significativa entre o astigmatismo da córnea anterior, induzido pela cirurgia e a aberração de coma na cirurgia de catarata por incisão coaxial pequena de 2,75 mm (r=0,387, p=0,006). Não foi encontrada correlação significativa entre as alterações nas aberrações totais de mais alta ordem, induzidas pela cirurgia e o tempo efetivo de faco-emulsificação. Conclusões: Nem a cirurgia de catarata por microincisão coaxial de 2,2 mm, nem aquela por incisão coaxial pequena de 2,75 mm degradaram significativamente as aberrações totais de mais alta ordem da córnea anterior. Porém, as alterações nas aberrações totais de mais alta ordem, induzidas pela cirurgia mostraram uma diferença significativa entre os dois procedimentos, com uma ligeira redução na cirurgia de catarata por microincisão coaxial de 2,2 mm e um pequeno aumento na cirurgia de catarata por incisão coaxial pequena de 2,75 mm. O tempo de facoemulsificação e a potência utilizada durante a cirurgia não tiveram impacto nas aberrações corneanas.

Three-dimensional finite element analysis of Kirschner wire fixation configuration for supracondylar fracture of humerus fracture in children / 中国骨伤

OBJECTIVE@#To establish a new mechanical model of distal humerus in children with epiphysial cartilage, stimulate supracondylar humerus fracture and perform three dimensional finite elements, and study effect of pins numbers, pin tract, outlet height and pin configurations on stability of fixation.@*METHODS@#Three dimensional computed tomography (CT) data of 6-year-old boy with distal humerus was downloaded from picture archiving and communications systems software (PACS), the data of picture was imported into Simpleware and SolidWorks 2016 software to establish distal humerus fracture in children contained ossific nucleus of the capitellum (ONC) and distal cartilage. Normal extense supracondylar humerus fracture model was established to stimulate configurations of crossed and lateral pinning fixation, 30 N was added on the direction of flexion extension and varus valgus, while 50 N was added on the direction of internal and external turning. Stability was analyzed by displacement degree of distal fracture.@*RESULTS@#Among 2-pin configurations, 2-crossed pins were more stable against rotation forces which could resist rotation stress over 2 585 Nmm/ °, while low position through ONC of 2-divergent lateral pins were more stable, which could resist stress of 45 N /mm and 190 N /mm during the test of resistant strains and varus-valgus stress. The third pins was added into the more stable lateral 2-pins, the stability in all directions were increased obviously, and 3 crossed pins is the most stable, stress of flexion-extension, varus-valgus and internal-external turning were 198 N /mm, 395 N /mm and 6 251 Nmm/ °.@*CONCLUSION@#Two-divergent lateral pins could provide enough stability for supracondylar humerus fracture in children. In two-crossed pins, the upper border of MDJ could provide the best stability. Three-crossed pins could offer the best stability against both translation and rotation forces.

Expression of Plasma miRNA-10b in Patients with Cervical Cancer and Its Clinical Significance / 现代检验医学杂志

Objective To investigate the expression and clinical significance of plasma microRNA 10b (miRNA-10b) in patients with cervical cancer.Methods The levels of plasma miRNA-10b,SCCA and CEA were detected by RT-PCR in 168 patients with cervical cancer,68 patients with CIN (CIN group) and 45 healthy women (control group),analyzed the relation between miRNA 10b expression and clinicopathological features of cervical cancer.The sensitivity and specificity of miRNA-10b,SCCA and CEA in the diagnosis of cervical cancer were evaluated by ROC curve,and the relationship between three indexes and cervical cancer were analyzed by multivariate Logistic regression model.Correlation analysis of plasma miRNA-10b and SCCA,CEA in patients with cervical cancer was analysed by Pearson.Results The levels of miRNA-10b,SCCA and CEA in the cervical cancer group were significantly higher than those in the CIN group and the control group[miRNA-10b(2-△△Ct):5.83± 1.84 vs 2.64±0.92 and 2.38±0.75;SCCA(ng/ml):8.74±2.26 vs 1.97±0.62 and 0.61±0.15;CEA (ng/ml):5.71±2.15 vs 1.56±0.58 and 1.34±0.16,F=17.842,13.614,8.273,all P＜0.01].The level of plasma miRNA-10b expression was correlated with clinical stage,lymph node metastasis,depth of invasion and SCCA level in patients with cervical cancer(t/F=19.287,21.528,5.672,5.284,P＜0.05).Plasma miRNA-10b,SCCA and CEA and three combined diagnosis of cervical cancer of AUC (95％CI) were 0.836(0.752～0.924),0.795(0.722～0.875),0.664(0.596～0.738) and 0.882(0.794～0.958),respectively.The optimal cut-off values were 4.26,6.58 ng/ml and 4.05 ng/ml,respectively.Logistic regression analysis showed that elevated plasma miRNA-10b and SCCA levels were independent risk factors for cervical cancer[OR(95％CI)=1.816(1.629～2.483),OR(95％CI) =1.427(1.206～ 1.975)].Plasma miRNA-10b was positively correlated with SCCA in patients with cervical cancer(r=0.637,P＜0.01).Conclusion Plasma miRNA-10b will be expected to be a molecular marker for the early diagnosis of cervical cancer.The combination of SCCA and CEA can improve the diagnostic accuracy of cervical cancer.

Evaluation of mid-term follow-up after Salter innominate osteotomy in developmental dysplasia of the hip / 中华外科杂志

<p><b>OBJECTIVES</b>To evaluate the mid-term outcome after Salter innominate osteotomy in developmental dysplasia of the hip (DDH), and to observe the developmental characteristics of the hip after operation and the relationships between the mid-term outcome and radiographic parameters as well as age at operation.</p><p><b>METHODS</b>: Forty-four patients with 61 treated hips were selected. The patients were treated with Salter innominate osteotomy and followed-up for at least three years with intact serial radiographs. Radiographs taken before operation, 6 weeks, 1 year and 2 - 3 years after operation and in the latest follow-up were selected. Acetabular index (AI), Sharp acetabular angle (SAA) and center-edge angle of Wiberg (CEA) were measured and Severin classification was done according to radiographs taken in the latest follow-up.</p><p><b>RESULTS</b>The average correction of AI was 14° postoperatively. The acetabulum remodels best at 2-3 years after operation when the average AI became very close to normal. In the latest follow-up the SAA was 41° which could be regarded as normal. Postoperative CEA was on average 23° which increased to 25° 2-3 years later. In the latest follow-up, the average CEA was 26°. The ratio of excellent and good outcomes (Severin I, II) was 84%, while the ratio of moderate and poor outcomes (Severin III, IV, V, VI) was 16%. Age at operation had a negative effect on outcomes. Although 70% patients operated after age 6 had satisfactory outcomes. The Severin I, II group showed no difference in AI from III, IV, V, VI group 6 weeks after operation, but the AI of the former obviously improved 2-3 years after operation while that of the latter deteriorated. Significant difference in SAA and the CEA could be observed in the latest follow-up.</p><p><b>CONCLUSIONS</b>Salter innominate osteotomy focuses on normalizing the abnormal acetabular direction in DDH children as well as stimulating the remodeling of the acetabulum, which provides a satisfactory middle-term outcome. The acetabulum remodels rapidly during the first three years after operation when AI and CEA develops into normal. Interference should be adopted if these changes have not appeared in the first three years.</p>

Laminin and its alpha 6 integrin receptor in the regulation of human hepatocellular carcinoma cell phenotypes / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the role of integrin alpha 6 in cell attachment, spread, survival/proliferation and differentiation of human hepatocellular carcinoma (HCC) BEL-7402 cells on various substrates by monoclonal antibody against the extracellular domain of alpha 6 subunit (IA6ED McAb).</p><p><b>METHODS</b>The effect of McAb on attachment and spread of BEL-7402 cells on LN or FN substrate was examined. MTT analysis was used to examine the cell survival/proliferation, gelatin zymography to the matrix metalloproteinases (MMPs) secreted by BEL-7402 cells, and microparticle immunoabsorbent kit to AFP secretion while the cells were cultured on LN-, FN- or Matrigel-coated substrates.</p><p><b>RESULTS</b>The cell attachment, spread and survival/proliferation were inhibited. Moreover importantly, the malignant cell dedifferentiation and abnormal differentiation on LN-coated substrate were also strongly inhibited by IA6ED McAb.</p><p><b>CONCLUSION</b>LN and integrin alpha 6 regulate human HCC cell phenotypes of survival/proliferation and differentiation. The phenotypes of dedifferentiation and abnormal differentiation can be reversed by blocking the interaction between integrin alpha 6 and LN using IA6ED McAb, which may lower metastatic potency of tumor cells.</p>

Pretreatment and Determination of Dissolvable Zinc Salt in Deodorizing Cosmetics / 环境与健康杂志

Objective To explore a method for determination of dissolvable zinc salt in deodorizing cosmetics. Methods The contents of dissolvable zinc salt in deodorants extracted by pure water were detected by flame atomic absorption spectrophotometry. Results The correlation coefficient of the standard curve was more than 0.999. The detection limit, precision and recovery rates of the method were 8.2?10-3 ?g, 1.26% and 97.0%～98.5% respectively. The lowest detection limit of zinc salt was 2.7?10-2 ?g. No other interfering factors were found during detection process. Conclusion This method could meet the requirements of hygienic physico_chemical detection for cosmetics.