Biomedical and Environmental Sciences

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Bone Injury and Fracture Healing Biology (1095 day view times: 1400)

Ahmad Oryan, Somayeh Monazzah, Amin Bigham-Sadegh

2015, 28(1): 57-71. doi: 10.3967/bes2015.006

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Report on Cardiovascular Health and Diseases in China 2021: An Updated Summary (1095 day view times: 1109)

The Writing Committee of the Report on Cardiovascular Health and Diseases in China

2022, 35(7): 573-603. doi: 10.3967/bes2022.079

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In 2019, cardiovascular disease (CVD) accounted for 46.74% and 44.26% of all deaths in rural and urban areas, respectively. Two out of every five deaths were due to CVD. It is estimated that about 330 million patients suffer from CVD in China. The number of patients suffering from stroke, coronary heart disease, heart failure, pulmonary heart disease, atrial fibrillation, rheumatic heart disease, congenital heart disease, lower extremity artery disease and hypertension are 13.00 million, 11.39 million, 8.90 million, 5.00 million, 4.87 million, 2.50 million, 2.00 million, 45.30 million, and 245.00 million, respectively. Given that China is challenged by the dual pressures of population aging and steady rise in the prevalence of metabolic risk factors, the burden caused by CVD will continue to increase, which has set new requirements for CVD prevention and treatment and the allocation of medical resources in China. It is important to reduce the prevalence through primary prevention, increase the allocation of medical resources for CVD emergency and critical care, and provide rehabilitation services and secondary prevention to reduce the risk of recurrence, re-hospitalization and disability in CVD survivors. The number of people suffering from hypertension, dyslipidemia and diabetes in China has reached hundreds of millions. Since blood pressure, blood lipids, and blood glucose levels rise mostly insidiously, vascular disease or even serious events such as myocardial infarction and stroke often already occured at the time of detection in this population. Hence, more strategies and tasks should be taken to prevent risk factors such as hypertension, dyslipidemia, diabetes, obesity, and smoking, and more efforts should be made in the assessment of cardiovascular health status and the prevention, treatment, and research of early pathological changes.

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Burden of Cirrhosis and Other Chronic Liver Diseases Caused by Specific Etiologies in China, 1990−2016: Findings from the Global Burden of Disease Study 2016 (1095 day view times: 611)

LI Man, WANG Zhuo Qun, ZHANG Lu, ZHENG Hao, LIU Dian Wu, ZHOU Mai Geng

2020, 33(1): 1-10. doi: 10.3967/bes2020.001

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Objective To estimate the burden of cirrhosis and other chronic liver diseases caused by specific etiologies in China. Methods Data from the Global Burden of Disease Study 2016 (GBD 2016) were used. We evaluated the burden by analyzing age-sex-province-specific prevalence, mortality, and disability-adjusted life-years (DALYs) of 33 provinces in China. Results From 1990 to 2016, prevalence cases in thousands increased by 73.7% from 6833.3 (95% UI: 6498.0–7180.6) to 11869.6 (95% UI: 11274.6–12504.7). Age-standardized mortality and DALY rates per 100,000 decreased by 51.2% and 53.3%, respectively. Male and elderly people (aged ≥ 60 years) preponderance were found for prevalence, mortality, and DALYs. The number of prevalence cases, deaths, and DALYs due to hepatitis C virus (HCV) increased by 86.6%, 8.7%, and 0.9%, respectively. Also, age-standardized prevalence rates decreased in 31 provinces, but increased in Yunnan and Shandong. The Socio-demographic Index (SDI) values were negatively correlated with age-standardized mortality and DALY rates by provinces in 2016; the correlation coefficients were −0.817 and −0.828, respectively. Conclusion Cirrhosis and other chronic liver diseases remain a huge health burden in China, with the increase of population and the aging of population. Hepatitis B virus (HBV) remains the leading cause of the health burden in China.

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The Serum Exosome Derived MicroRNA-135a, -193b, and-384 Were Potential Alzheimer's Disease Biomarkers (1095 day view times: 462)

YANG Ting Ting, LIU Chen Geng, GAO Shi Chao, ZHANG Yi, WANG Pei Chang

2018, 31(2): 87-96. doi: 10.3967/bes2018.011

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Objective MicroRNAs (miRs) are attractive molecules to be considered as one of the blood-based biomarkers for neurodegenerative disorders such as Alzheimer's disease (AD). The goal of this study was to explore their potential value as biomarkers for the diagnosis of AD. Methods The expression levels of exosomal miR-135a, -193b, and-384 in the serum from mild cognitive impairment (MCI), dementia of Alzheimer type (DAT), Parkinson's disease with dementia (PDD), and vascular dementia (VaD) patients were measured with a real-time quantitative reverse transcriptase PCR (qRT-PCR) method. Results Both serum exosome miR-135a and miR-384 were up-regulated while miR-193b was down-regulated in serum of AD patients compared with that of normal controls. Exosome miR-384 was the best among the three miRs to discriminate AD, VaD, and PDD. Using the cut-off value could better interpret these laboratory test results than reference intervals in the AD diagnosis. ROC curve showed that the combination of miR-135a, -193b, and-384 was proved to be better than a particular one for early AD diagnosis. Conclusion Our results indicated that the exosomal miRs in the serum were not only potential biomarker of AD early diagnosis, but might also provide novel insights into the screen and prevention of the disease.

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Effects of Short-Term Forest Bathing on Human Health in a Broad-Leaved Evergreen Forest in Zhejiang Province, China (1095 day view times: 439)

MAO Gen Xiang, LAN Xiao Guang, CAO Yong Bao, CHEN Zhuo Mei, HE Zhi Hua, LV Yuan Dong, WANG Ya Zhen, HU Xi Lian, WANG Guo Fu, YAN Jing

2012, 25(3): 317-324. doi: 10.3967/0895-3988.2012.03.010

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Objective To investigate the effects of short-term forest bathing on human health.Methods Twenty healthy male university students participated as subjects and were randomly divided into two groups of 10.One group was sent on a two-night trip to a broad-leaved evergreen forest,and the other was sent to a city area.Serum cytokine levels reflecting inflammatory and stress response,indicators reflecting oxidative stress,the distribution of leukocyte subsets,and plasma endothelin-1 (ET-1) concentrations were measured before and after the experiment to evaluate the positive health effects of forest environments.A profile of mood states (POMS) evaluation was used to assess changes in mood states.Results No significant differences in the baseline values of the indicators were observed between the two groups before the experiment.Subjects exposed to the forest environment showed reduced oxidative stress and pro-inflammatory level,as evidenced by decreased malondialdehyde,interleukin-6,and tumor necrosis factor α levels compared with the urban group.Serum cortisol levels were also lower than in the urban group.Notably,the concentration of plasma ET-1 was much lower in subjects exposed to the forest environment.The POMS evaluation showed that after exposure to the forest environment,subjects had lower scores in the negative subscales,and the score for vigor was increased.Conclusion Forest bathing is beneficial to human health,perhaps through preventive effects related to several pathological factors.

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Hypertension Prevalence, Awareness, Treatment, and Control and Their Associated Socioeconomic Factors in China: A Spatial Analysis of A National Representative Survey (1095 day view times: 368)

WANG Wei, ZHANG Mei, XU Cheng Dong, YE Peng Peng, LIU Yun Ning, HUANG Zheng Jing, HU Cai Hong, ZHANG Xiao, ZHAO Zhen Ping, LI Chun, CHEN Xiao Rong, WANG Li Min, ZHOU Mai Geng

2021, 34(12): 937-951. doi: 10.3967/bes2021.130

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Objective We aimed to investigate and interpret the associations between socioeconomic factors and the prevalence, awareness, treatment, and control of hypertension at the provincial level in China. Methods A nationally and provincially representative sample of 179,059 adults from the China Chronic Disease and Nutrition Surveillance study in 2015–2016 was used to estimate hypertension burden. The spatial Durbin error model was fitted to investigate socioeconomic factors associated with hypertension indicators. Results Overall, it was estimated that 29.20% of the participants were hypertensive nationwide, among whom, 34.32% were aware of their condition, 27.69% had received antihypertensive treatment, and 7.81% had controlled their condition. Per capita gross domestic product (GDP) was associated with hypertension prevalence (coefficient: −2.95, 95% CI: −5.46, −0.45) and control (coefficient: 6.35, 95% CI: 1.36, 11.34) among adjacent provinces and was also associated with awareness (coefficient: 2.93, 95% CI: 1.12, 4.74) and treatment (coefficient: 2.67, 95% CI: 1.21, 4.14) in local province. Beds of internal medicine (coefficient: 2.66, 95% CI: 1.08, 4.23) was associated with control in local province. Old dependency ratio (coefficient: −3.58, 95% CI: −5.35, −1.81) was associated with treatment among adjacent provinces and with control (coefficient: −1.69, 95% CI: −2.42, −0.96) in local province. Conclusion Hypertension indicators were not only directly influenced by socioeconomic factors of local area but also indirectly affected by characteristics of geographical neighbors. Population-level strategies should involve optimizing supportive socioeconomic environment by integrating clinical care and public health services to decrease hypertension burden.

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Application of Nanopore Sequencing Technology in the Clinical Diagnosis of Infectious Diseases (1095 day view times: 341)

ZHANG Lu Lu, ZHANG Chi, PENG Jun Ping

2022, 35(5): 381-392. doi: 10.3967/bes2022.054

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Abstract: Infectious diseases are an enormous public health burden and a growing threat to human health worldwide. Emerging or classic recurrent pathogens, or pathogens with resistant traits, challenge our ability to diagnose and control infectious diseases. Nanopore sequencing technology has the potential to enhance our ability to diagnose, interrogate, and track infectious diseases due to the unrestricted read length and system portability. This review focuses on the application of nanopore sequencing technology in the clinical diagnosis of infectious diseases and includes the following: (i) a brief introduction to nanopore sequencing technology and Oxford Nanopore Technologies (ONT) sequencing platforms; (ii) strategies for nanopore-based sequencing technologies; and (iii) applications of nanopore sequencing technology in monitoring emerging pathogenic microorganisms, molecular detection of clinically relevant drug-resistance genes, and characterization of disease-related microbial communities. Finally, we discuss the current challenges, potential opportunities, and future outlook for applying nanopore sequencing technology in the diagnosis of infectious diseases.

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Trends in Lipids Level and Dyslipidemia among Chinese Adults, 2002-2015 (1095 day view times: 311)

SONG Peng Kun, MAN Qing Qing, LI Hong, PANG Shao Jie, JIA Shan Shan, LI Yu Qian, HE Li, ZHAO Wen Hua, ZHANG Jian

2019, 32(8): 559-570. doi: 10.3967/bes2019.074

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Objective To investigate the trends of lipid profiles and dyslipidemia among Chinese adults from 2002 to 2015. Methods Data were collected from three nationally representative cross-sectional surveys. Fasting venous blood samples were collected and serum lipids were tested by biochemical analysis and enzymatic determination. Lipid levels and the prevalence of dyslipidemia among adults were analyzed with complex sampling weighting adjustment for age and gender. Results The weighted means of TC, TG, and LDL-c significantly increased linearly from 3.93, 1.12, and 2.12 mmol/L in 2002 to 4.59, 1.41, and 2.78 mmol/L in 2010 and then to 4.63, 1.47, and 2.87 mmol/L in 2015, respectively; by contrast, HDL-c levels decreased significantly from 1.30 mmol/L to 1.26 mmol/L over the same period. Similar trends in mean non-HDL-c and lipid-related ratios were observed. The weighted dyslipidemia prevalence linearly increased; in particular, hypercholesterolemia increased from 1.6% to 5.6% and then to 5.8%, hypertriglyceridemia increased from 5.7% to 13.6% and then to 15.0%, low HDL-c increased from 18.8% to 35.5% and then to 24.9%, and high LDL-c increased from 1.3% to 5.6% and then to 7.2% (P for trend < 0.001). Conclusion Dyslipidemia increased among Chinese adults from 2002 to 2015. Development of a comprehensive strategy to decrease lipid levels in this population is urgently required.

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Cover (1095 day view times: 297)

2023, 36(7).

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The Influence of Diabetes, Hypertension, and Hyperlipidemia on the Onset of Age-Related Macular Degeneration in North China: The Kailuan Eye Study (1095 day view times: 295)

ZHANG Yong Peng, WANG Ya Xing, ZHOU Jin Qiong, WANG Qian, YAN Yan Ni, YANG Xuan, YANG Jing Yan, ZHOU Wen Jia, WANG Ping, SHEN Chang, YANG Ming, LUAN Ya Nan, WANG Jin Yuan, WU Shou Ling, CHEN Shuo Hua, WANG Hai Wei, FANG Li Jian, WAN Qian Qian, ZHU Jing Yuan, NIE Zi Han, CHEN Yu Ning, XIE Ying, JONAS JB, WEI Wen Bin

2022, 35(7): 613-621. doi: 10.3967/bes2022.081

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Objective To analyze the prevalence of dry and wet age-related macular degeneration (AMD) in patients with diabetes, hypertension and hyperlipidemia, and to analyze the risk factors for AMD. Methods A population-based cross-sectional epidemiologic study was conducted involving 14,440 individuals. We assessed the prevalence of dry and wet AMD in diabetic and non-diabetic subjects and analyzed the risk factors for AMD. Results The prevalence of wet AMD in diabetic and non-diabetic patients was 0.3% and 0.5%, respectively, and the prevalence of dry AMD was 17% and 16.4%, respectively. The prevalence of wet AMD in healthy, hypertensive, hyperlipidemic, and hypertensive/hyperlipidemic populations was 0.5%, 0.3%, 0.2%, and 0.7%, respectively. The prevalence of dry AMD in healthy, hypertensive, hyperlipidemic, and hypertensive/hyperlipidemic populations was 16.6%, 16.2%, 15.2%, and 17.2%, respectively. Age, sex, body mass index, and use of hypoglycemic drugs or lowering blood pressure drugs were corrected in the risk factor analysis of AMD. Diabetes, diabetes/hypertension, diabetes/hyperlipidemia, and diabetes/hypertension/hyperlipidemia were analyzed. None of the factors analyzed in the current study increased the risk for the onset of AMD. Conclusion There was no significant difference in the prevalence of wet and dry AMD among diabetic and non-diabetic subjects. Similarly, there was no significant difference in the prevalence of wet and dry AMD among subjects with hypertension and hyperlipidemia. Diabetes co-existing with hypertension and hyperlipidemia were not shown to be risk factors for the onset of dry AMD.

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Protein Requirements in Healthy Adults:A Meta-analysis of Nitrogen Balance Studies (1095 day view times: 290)

LI Min, SUN Feng, PIAO Jian Hua, YANG Xiao Guang

2014, 27(8): 606-613. doi: 10.3967/bes2014.093

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Objective The goal of this study was to analyze protein requirements in healthy adults through a meta-analysis of nitrogen balance studies.
Methods A comprehensive search for nitrogen balance studies of healthy adults published up to October 2012 was performed, each study were reviewed, and data were abstracted. The studies were first evaluated for heterogeneity. The average protein requirements were analyzed by using the individual data of each included studies. Study site climate, age, sex, and dietary protein source were compared.
Results Data for 348 subjects were gathered from 28 nitrogen balance studies. The natural logarithm of requirement for 348 individuals had a normal distribution with a mean of 4.66. The estimated average requirement was the exponentiation of the mean of the log requirement, 105.64 mg N/kg·d. No significant differences between adult age, source of dietary protein were observed. But there was significant difference between sex and the climate of the study site (P<0.05).
Conclusion The estimated average requirement and recommended nutrient intake of the healthy adult population was 105.64 mg N/kg·d (0.66 g high quality protein/kg·d) and 132.05 mg N/kg·d (0.83 g high quality protein/kg·d), respectively.

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Protective Effect of Angiotensin (1-7) on Silicotic Fibrosis in Rats (1095 day view times: 290)

ZHANG Bo Nan, XU Hong, GAO Xue Min, ZHANG Gui Zhen, ZHANG Xin, YANG Fang

2019, 32(6): 419-426. doi: 10.3967/bes2019.057

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Objective Silicosis, caused by inhalation of silica dust, is the most serious occupational disease in China and the aim of present study was to explore the protective effect of Ang (1-7) on silicotic fibrosis and myofibroblast differentiation induced by Ang Ⅱ. Methods HOPE-MED 8050 exposure control apparatus was used to establish the rat silicosis model. Pathological changes and collagen deposition of the lung tissue were examined by H.E. and VG staining, respectively. The localizations of ACE2 and α-smooth muscle actin (α-SMA) in the lung were detected by immunohistochemistry. Expression levels of collagen type Ⅰ, α-SMA, ACE2, and Mas in the lung tissue and fibroblasts were examined by western blot. Levels of ACE2, Ang (1-7), and Ang Ⅱ in serum were determined by ELISA. Co-localization of ACE2 and α-SMA in fibroblasts was detected by immunofluorescence. Results Ang (1-7) induced pathological changes and enhanced collagen deposition in vivo. Ang (1-7) decreased the expressions of collagen type Ⅰ and α-SMA and increased the expressions of ACE2 and Mas in the silicotic rat lung tissue and fibroblasts stimulated by Ang Ⅱ. Ang (1-7) increased the levels of ACE2 and Ang (1-7) and decreased the level of Ang Ⅱ in silicotic rat serum. A779 enhanced the protective effect of Ang (1-7) in fibroblasts stimulated by Ang Ⅱ. Conclusion Ang (1-7) exerted protective effect on silicotic fibrosis and myofibroblast differentiation induced by Ang Ⅱ by regulating ACE2-Ang (1-7)-Mas axis.

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Overexpression of Protein Phosphatase 2 Regulatory Subunit B"Alpha Promotes Glycolysis by Regulating Hexokinase 1 in Hepatocellular Carcinoma (1095 day view times: 289)

JIAO Ning, JI Wan Sheng, ZHANG Biao, SHANG Yu Kui, ZHANG Yu Chen, YU Wei Qun, JIN Hai Long, LI Chao, ZHANG Cheng Ying, YAN Cheng, YUE Wen, ZHANG Qing

2022, 35(7): 622-632. doi: 10.3967/bes2022.082

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Objective To investigate the regulatory relationship of Protein Phosphatase 2 Regulatory Subunit B"Alpha (PPP2R3A) and hexokinase 1 (HK1) in glycolysis of hepatocellular carcinoma (HCC). Methods In HepG2 and Huh7 cells, PPP2R3A expression was silenced by small interfering RNA (siRNA) and overexpression by plasmid transfection. The PPP2R3A-related genes were searched by RNA sequencing. Glycolysis levels were measured by glucose uptake and lactate production. QRT-PCR, ELISA, western blot and immunofluorescence assay were performed to detect the changes of PPP2R3A and HK1. Cell proliferation, migration and invasion assay were used to study the roles of HK1 regulation by PPP2R3A. Results RNA sequencing data revealed that PPP2R3A siRNA significantly downregulated the expression of HK1. PPP2R3A gene overexpression promotes, while gene silencing suppresses, the level of HK1 and glycolysis in HCC cells. In HCC tissue samples, PPP2R3A and HK1 were colocalized in the cytoplasm, and their expression showed a positive correlation. HK1 inhibition abrogated the promotion of glycolysis, proliferation, migration and invasion by PPP2R3A overexpression in liver cancer cells. Conclusion Our findings showed the correlation of PPP2R3A and HK1 in the glycolysis of HCC, which reveals a new mechanism for the oncogenic roles of PPP2R3A in cancer.

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Contents (1095 day view times: 289)

2023, 36(7): 1-2.

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Data Resource Profile: A Protocol of China National Diabetic Chronic Complications Study (1095 day view times: 286)

HOU Xu Hong, WANG Li Min, CHEN Si Yu, LIANG Ye Bei, ZHANG Mei, HUANG Zheng Jing, CHEN Hong Li, WU Jing Zhu, WU Jing, JIA Wei Ping

2022, 35(7): 633-640. doi: 10.3967/bes2022.078

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Health Effect of Forest Bathing Trip on Elderly Patients with Chronic Obstructive Pulmonary Disease (1095 day view times: 282)

JIA Bing Bing, YANG Zhou Xin, MAO Gen Xiang, LYU Yuan Dong, WEN Xiao Lin, XU Wei Hong, LYU XIAO Ling

2016, 29(3): 212-218. doi: 10.3967/bes2016.026

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Correlation between Anxiety, Depression, and Sleep Quality in College Students (1095 day view times: 281)

ZHANG Yu Tong, HUANG Tao, ZHOU Fang, HUANG Ao Di, JI Xiao Qi, HE Lu, GENG Qiang, WANG Jia, MEI Can, XU Yu Jia, YANG Ze Long, ZHAN Jian Bo, CHENG Jing

2022, 35(7): 648-651. doi: 10.3967/bes2022.084

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Associations of Sarcopenia, Handgrip Strength and Calf Circumference with Cognitive Impairment among Chinese Older Adults (1095 day view times: 280)

WU Bing, LYU Yue Bin, CAO Zhao Jin, WEI Yuan, SHI Wan Ying, GAO Xiang, ZHOU Jin Hui, KRAUS Virginia Byers, ZHAO Feng, CHEN Xin, LU Feng, ZHANG Ming Yuan, LIU Ying Chun, TAN Qi Yue, SONG Shi Xun, QU Ying Li, ZHENG Xu Lin, SHEN Chong, MAO Chen, SHI Xiao Ming

2021, 34(11): 859-870. doi: 10.3967/bes2021.119

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Objective To evaluate the associations of sarcopenia, handgrip strength and calf circumference with cognitive impairment among Chinese older adults. Methods Totally 2,525 older adults were recruited from the Healthy Aging and Biomarkers Cohort Study. Cognitive impairment was assessed by the Chinese Mini-Mental State Examination. Handgrip strength was calculated from the means of the right and left hand values. Calf circumference was measured at the site of maximum circumference of the non-dominant leg. The formula developed by Ishii was used to define sarcopenia. Multiple logistic regression was performed to evaluate the associations of sarcopenia, handgrip strength, and calf circumference with cognitive impairment. Results The prevalence of cognitive impairment was 34.36%. The adjusted odds ratio (OR) for cognitive impairment in individuals with sarcopenia was 2.55 [95% confidence interval (95% CI): 1.86−3.50]. Compared with individuals in the first quartile (Q₁) of calf circumference, the adjusted ORs in the second, third, and fourth quartiles (Q₂, Q₃, and Q₄) were 0.75 (95% CI: 0.58−0.96), 0.59 (95% CI: 0.44−0.79), and 0.62 (95% CI: 0.45−0.8), respectively. Compared with individuals in Q₁ of handgrip strength, the adjusted ORs for Q₂, Q₃, and Q₄ were 0.49 (95% CI: 0.38−0.62), 0.31 (95% CI: 0.23−0.41), and 0.30 (95% CI: 0.21−0.44), respectively. Conclusion Sarcopenia, identified by low handgrip strength and low calf circumference, was positively associated with cognitive impairment.

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Supplementation of Fermented Barley Extracts with Lactobacillus Plantarum dy-1 Inhibits Obesity via a UCP1-dependent Mechanism (1095 day view times: 280)

XIAO Xiang, BAI Juan, LI Ming Song, ZHANG Jia Yan, SUN Xin Juan, DONG Ying

2019, 32(8): 578-591. doi: 10.3967/bes2019.076

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Objective We aimed to explore how fermented barley extracts with Lactobacillus plantarum dy-1 (LFBE) affected the browning in adipocytes and obese rats. Methods In vitro, 3T3-L1 cells were induced by LFBE, raw barley extraction (RBE) and polyphenol compounds (PC) from LFBE to evaluate the adipocyte differentiation. In vivo, obese SD rats induced by high fat diet (HFD) were randomly divided into three groups treated with oral gavage:(a) normal control diet with distilled water, (b) HFD with distilled water, (c) HFD with 800 mg LFBE/kg body weight (bw). Results In vitro, LFBE and the PC in the extraction significantly inhibited adipogenesis and potentiated browning of 3T3-L1 preadipocytes, rather than RBE. In vivo, we observed remarkable decreases in the body weight, serum lipid levels, white adipose tissue (WAT) weights and cell sizes of brown adipose tissues (BAT) in the LFBE group after 10 weeks. LFBE group could gain more mass of interscapular BAT (IBAT) and promote the dehydrogenase activity in the mitochondria. And LFBE may potentiate process of the IBAT thermogenesis and epididymis adipose tissue (EAT) browning via activating the uncoupling protein 1 (UCP1)-dependent mechanism to suppress the obesity. Conclusion These results demonstrated that LFBE decreased obesity partly by increasing the BAT mass and the energy expenditure by activating BAT thermogenesis and WAT browning in a UCP1-dependent mechanism.

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TaqMan Real-time RT-PCR Assay for Detecting and Differentiating Japanese Encephalitis Virus (1095 day view times: 279)

SHAO Nan, LI Fan, NIE Kai, FU Shi Hong, ZHANG Wei Jia, HE Ying, LEI Wen Wen, WANG Qian Ying, LIANG Guo Dong, CAO Yu Xi, WANG Huan Yu

2018, 31(3): 208-214. doi: 10.3967/bes2018.026

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Objective To detect Japanese encephalitis virus (JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction (RT-PCR) detection system was developed. Methods By aligning the full-length sequences of JEV (G1-G5), six sets of highly specific TaqMan real-time RT-PCR primers and probes were designed based on the highly conserved NS1, NS2, and M genes of JEV, which included one set for non-specific JEV detection and five sets for the detection of specific JEV genotypes. Twenty batches of mosquito samples were used to evaluate our quantitative PCR assay. Results With the specific assay, no other flavivirus were detected. The lower limits of detection of the system were 1 pfu/mL for JEV titers and 100 RNA copies/μL. The coefficients of variation of this real-time RT-PCR were all < 2.8%. The amplification efficiency of this method was between 90% and 103%. Conclusion A TaqMan real-time RT-PCR detection system was successfully established to detect and differentiate all five JEV genotypes.

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