1.Visual acuity and corrected visual acuity of children and adolescents in Shanghai City
Chinese Journal of School Health 2025;46(1):24-28
		                        		
		                        			Objective:
		                        			To investigate the visual acuity and correction conditions of children and adolescents in Shanghai, so as to provide a scientific basis for developing intervention measures to prevent myopia and protect vision among children and adolescents.
		                        		
		                        			Methods:
		                        			From October to December 2022, a stratified cluster random sampling survey was conducted, involving 47 034 students from 16 municipal districts in Shanghai, covering kindergartens (≥5 years), primary schools, middle schools, general high schools and vocational high schools. According to the Guidelines for Screening Refractive Errors in Primary and Secondary School Students, the Standard Logarithmic Visual acuity Chart was used to examine naked vision and corrected vision of students, and general information was collected. The distribution and severity of visual impairment in different age groups were analyzed, and  χ 2 tests and multivariate Logistic regression were used to explore factors associated with visual impairment.
		                        		
		                        			Results:
		                        			The detection rate of visual impairment among children and adolescents was 76.2%, with a higher rate among females (78.8%) than males ( 73.8 %), higher among Han ethic students ( 76.2 %) than minority students (71.2%), and higher among urban students (76.7%) than suburban students (75.8%), all with statistically significant differences ( χ 2=162.6, 10.4, 5.5,  P <0.05). The rate of visual impairment initially decreased and then increased with age, reaching its lowest at age 7 (53.8%) and peaking at age 17 (89.6%) ( χ 2 trend = 3 467.0 ,  P <0.05). Severe visual impairment accounted for the majority, at 56.6%, and there was a positive correlation between the severity of visual impairment and age among children and adolescents ( r =0.45,  P <0.05). Multivariate Logistic regression showed that age, BMI, gender, ethnicity and urban suburban status were associated with visual impairment ( OR =1.18, 1.01,  1.38 , 0.79, 0.88,  P <0.05). Among those with moderate to severe visual impairment, the rate of spectacle lens usage was 62.8%, yet only  44.8 % of those who used spectacle lens had fully corrected visual acuity. Females (64.9%) had higher spectacle lens usage rates than males (60.6%), and general high school students had the highest spectacle lens usage (83.9%), and there were statistically significant differences in gender and academic stages ( χ 2=57.7, 4 592.8,  P <0.05).
		                        		
		                        			Conclusions
		                        			The rate of spectacle lens usage among students with moderate to severe visual impairment is relatively low, and even after using spectacle lens, some students still do not achieve adequate corrected visual acuity. Efforts should focus on enhancing public awareness of eye health and refractive correction and improving the accessibility of related health services.
		                        		
		                        		
		                        		
		                        	
2.Component Analysis of Anmeidan and Its Mechanism in Regulating ERK1/2/MNK/ELF4E Signaling Pathway to Improve Circadian Rhythm in Insomnia Rats
Yi GAO ; Bo XU ; Jing XIA ; Linlin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):44-53
		                        		
		                        			
		                        			ObjectiveTo identify the main chemical constituents of Anmeidan (AMD) and to explore the mechanism of AMD in regulating the extracellular signal-regulated kinase 1/2 (ERK1/2)/mitogen-activated protein kinase (MAPK)-interacting serine/threonine-protein kinase (MNK)/eukaryotic translation initiation factor 4E (eIF4E) signaling pathway to improve circadian rhythm disturbances in insomnia rats. MethodsThe main chemical constituents of AMD were identified using ultra-high-performance liquid chromatography-linear ion trap-electrostatic orbital trap mass spectrometry (UPLC-LTQ/Orbitrap/MS) in combination with reference standards. Sixty male Sprague-Dawley (SD) rats were randomly divided into control, model, melatonin, and AMD low-, medium-, and high-dose groups, with 10 rats in each group. Except for the control group, all rats were administered p-chlorophenylalanine via intraperitoneal injection to establish an insomnia model. The activity-rest rhythm of rats was assessed using the open field test and circadian rhythm test. Hematoxylin-eosin (HE) staining and Nissl staining were used to observe structural changes in hypothalamic neurons. Immunofluorescence, real-time quantitative polymerase chain reaction (Real-time PCR), and Western blot analysis were employed to detect mRNA and protein expression levels of ERK1/2, MNK, and eIF4E in the hypothalamus. ResultsA total of 50 chemical components, including flavonoids, phenylpropanoids, triterpenoid saponins, alkaloids, and lignans, were identified in AMD. Compared with the control group, the model group exhibited significantly increased total distance traveled, average speed, central area residence time, and cumulative rearing time (P<0.01), as well as prolonged cumulative activity time and total activity time in both light and dark phases (P<0.01). Hypothalamic neurons in the model group were sparsely arranged, reduced in number, and exhibited nuclear disappearance or nucleolar rupture, with a significantly increased apoptosis index (P<0.01). The cytoplasm appeared turbid, Nissl body staining was lighter, and the Nissl body apoptosis index was significantly increased (P<0.01). The mRNA expression levels of ERK1/2, MNK, and eIF4E were significantly decreased (P<0.01), along with a significant reduction in protein expression levels of ERK1/2, phosphorylated ERK1/2 (p-ERK1/2), MNK, phosphorylated MNK (p-MNK), eIF4E, and phosphorylated eIF4E (p-eIF4E) (P<0.01). Compared with the model group, the total distance, average speed, central area residence time and body upright cumulative time of the AMD high-dose group were significantly reduced (P<0.01). The total distance, average speed and body upright cumulative time of the AMD medium-dose group were significantly reduced (P<0.01). The cumulative time of light activity and total time of activity in each dose group of AMD were significantly shortened (P<0.01). The cumulative time of dark activity in the high-dose group of AMD was prolonged (P<0.01). The neurons in the middle and high dose groups of AMD were closely arranged, the number of neurons increased, and the apoptosis index of hypothalamic cells decreased significantly (P<0.05, P<0.01). The cytoplasm of the low, middle and high dose groups of AMD was clear, the color of Nissl body became darker, and the apoptosis index of Nissl body decreased significantly (P<0.01). The expression of ERK1/2, MNK and eIF4E mRNA and protein in the hypothalamus of the middle and high dose groups of AMD increased significantly (P<0.05, P<0.01). ConclusionAMD primarily contains 50 chemical constituents, including flavonoids, phenylpropanoids, and triterpenoid saponins. It exhibits a "synergistic enhancement" effect through multiple components and multiple pathways to improve insomnia. AMD ameliorates circadian rhythm disturbances in p-chlorophenylalanine-induced insomnia rats by upregulating ERK1/2/MNK/eIF4E signaling pathway-related proteins. 
		                        		
		                        		
		                        		
		                        	
3.Effect of Anmeidan on Cognitive Function and Metabolic Profiling in Insomnia Model Rats Based on Untargeted Metabolomics
Feizhou LI ; Bo XU ; Zijing YE ; Lianyu LI ; Andong ZHANG ; Ping WANG ; Linlin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):54-64
		                        		
		                        			
		                        			ObjectiveTo elucidate the potential mechanisms by which the classic prescription Anmeidan alleviates cognitive impairment in insomnia model rats through metabolic profiling. MethodsA total of 60 SD rats were randomly divided into six groups: blank group, model group, low-, medium-, and high-dose Anmeidan groups, and the Suvorexant group, with 10 rats in each group. Except for the blank group, the insomnia model was established in all other groups via intraperitoneal injection of para-chlorophenylalanine. The Suvorexant group was administered Suvorexant solution (30 mg·kg-1·d-1) by gavage, while the low-, medium-, and high-dose Anmeidan groups received Anmeidan decoction (4.55, 9.09, 18.18 g·kg-1·d-1) by gavage. The blank group received an equivalent volume of normal saline. The open field test was used to assess spatial exploration and anxiety/depressive-like behaviors in rats. Serum levels of epidermal growth factor (EGF), brain-derived neurotrophic factor (BDNF), and vasoactive intestinal peptide (VIP) were measured using enzyme-linked immunosorbent assay (ELISA). Untargeted metabolomics was employed to identify differential metabolites in rat serum, and systematic biological methods were applied to analyze the potential targets and pathways of Anmeidan. ResultsCompared to the blank group, the model group exhibited significant reductions in total distance traveled, average speed, number of entries into the central area, time spent in the central area, and frequency of upright events (P<0.01), along with significant decreases in VIP, EGF, and BDNF levels (P<0.05,P<0.01). A total of 100 differential metabolites were identified between the model and blank groups. Compared to the model group, the low-, medium-, and high-dose Anmeidan groups showed significant increases in total distance traveled, average speed, number of entries into the central area, time spent in the central area, and frequency of upright events (P<0.05,P<0.01), as well as a significant increase in VIP levels (P<0.05,P<0.01). Anmeidan significantly reversed abnormal changes in 67 metabolites compared to the model group. A combined analysis identified 134 potential targets of Anmeidan, with network topology analysis suggesting that Caspase-3, B-cell lymphoma 2 (Bcl-2), nuclear transcription factor-κB (NF-κB), interleukin-1β (IL-1β), interleukin-2 (IL-2), matrix metalloproteinase-9 (MMP-9), and Toll-like receptor 4 (TLR4), among others, may serve as key targets of Anmeidan. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed major enriched pathways, including the cyclic adenosine monophosphate (cAMP) signaling pathway, hypoxia inducible factor-1 (HIF-1) signaling pathway, and IL-17 signaling pathway. ConclusionThis study demonstrates that Anmeidan can recalibrate abnormal metabolic profiles in insomnia model rats to mitigate cognitive impairment, with its mechanisms of action potentially involving the regulation of immune-inflammatory responses, energy metabolism, and apoptosis-related pathways. 
		                        		
		                        		
		                        		
		                        	
4.Cost-effectiveness analysis of chemical treatment with drones for Oncomelania hupensis control in marshland and lake areas
Yong CHEN ; Xiaojuan XU ; Daolong WEN ; Bo DAI ; Lan GAO ; Rong ZHANG ; Qingqing HUANG ; Linlin LI ; Fan ZHA ; Liang FANG ; Ping ZHANG ; Shiqing ZHANG ; Chunli CAO
Chinese Journal of Schistosomiasis Control 2024;36(5):502-506
		                        		
		                        			
		                        			 Objective To evaluate the molluscicidal effect and cost of spraying molluscicides with drones against Oncomelania hupensis snails in marshland and lake areas, so as to provide new insights into field snail control in China. Methods A marshland and lake setting measuring approximately 12 000 m2 was selected in Wanzhi District, Wuhu City on June 2023 as the test field, and assigned to four groups, of 3 000 m2 in each group. Environmental cleaning was not conducted in groups A or B, which were given 5% niclosamide ethanolamine salt granules sprayed with knapsack-type sprayers and drones at a dose of 40 g/m2, and environmental cleaning was conducted in groups C and D, which were given 5% niclosamide ethanolamine salt granules sprayed with drones and knapsack-type sprayers at a dose of 40 g/m2, respectively. O. hupensis snails were surveyed before chemical treatment and 1, 3, 5, 7, 14 days post-treatment. The uniformity of chemicals was determined on the day of treatment, and the snail mortality, corrected snail mortality and density of living snails were calculated and compared among groups. The cost of molluscicides, labor fees of environmental cleaning and chemical treatment and cost of equipment were calculated, and the cost for a 1% reduction in the mean density of living snails was calculated 14 days post-treatment. Results The mean densities of living snails and mortality rates of snails were 1.82 to 2.85 snails/0.1 m2 and 1.41% to 2.94% in groups A, B, C and D before chemical treatment, and the mortality and corrected mortality of snails were 55.75%, 49.32%, 85.94% and 87.50%, and 55.00%, 48.47%, 85.70% and 87.29% in groups A, B, C and D 14 days post-treatment. There was a significant difference in the mortality of snails among the four groups 14 days post-treatment (χ2 = 38.735, P < 0.005), and there was a higher snail mortality in Group D than in Group A (χ2 = 16.876, P < 0.005), and higher in Group C than in Group B (χ2 = 20.508, P < 0.005). The density of living snails reduced by 55.00%, 43.94%, 90.43% and 87.14% 14 days post-treatment relative to pre-treatment in groups A, B, C and D, respectively. The test for uniformity of chemicals showed that the mean dose of molluscicides were 57.34, 55.21, 40.19 g/m2 and 32.37 g/m2 in groups A, B, C and D, respectively, and the minimal standard deviation (7.07) and coefficient of variation (0.18) of mean doses were seen in Group C. The costs for chemical treatment were 0.33 Yuan in groups A and B and 1.53 Yuan in groups C and D, respectively. The costs for a 1% reduction in the mean density of living snails were 17.82, 22.47, 50.73 Yuan and 52.56 Yuan in groups A, B, C, and D 14 days post-treatment, respectively. Conclusions The molluscicidal effect and cost of spraying 5% niclosamide ethanolamine salt granules with drones are comparable to manual spraying, and chemical treatment with drones are high in uniformity of molluscicides, time- and labor-saving, and feasible for applications in complex environments, which deserves widespread applications in the field of snail control. 
		                        		
		                        		
		                        		
		                        	
5.Association between parental education level and time spent outdoors among children
Chinese Journal of School Health 2024;45(12):1704-1707
		                        		
		                        			Objective:
		                        			To explore the association between parental education level and time spent outdoors among children, so as to provide the scientific evidence for formulating policies of myopia prevention and control among children.
		                        		
		                        			Methods:
		                        			The study was based on secondary analysis of data from outdoor intervention studies in Shanghai. The follow up period was from March to December 2018. It included control group children ( n =1 117) with complete questionnaire surveys, ocular examinations, and time spent outdoors. Generalized linear regression models and trend tests were used to analyze the effect of parental education level on time spent outdoors among children.
		                        		
		                        			Results:
		                        			The median time spent outdoors was 76.4(59.7, 94.6) minutes. After adjusting for covariates including children s sex and age, generalized linear regression model suggested that there was no statistical significance between father s education level and outdoor activity time ( P >0.05). Compared with children whose mothers had a junior high school education or below, children whose mothers had high school/vocational high school education, college or above had shorter time spent outdoors ( β=-6.64,  -8.84 , P <0.05). Trend tests revealed that time spent outdoors among children decreased with the increase of parental education level ( P trend <0.01).
		                        		
		                        			Conclusions
		                        			The higher the education level of fathers or mothers, the shorter time spent outdoors of children. In addition to highlight outdoor activities at school, myopia prevention and control efforts should be focused on the role of parents in increasing children s outdoor activities.
		                        		
		                        		
		                        		
		                        	
6.Prospective clinical study of effect of neck muscle training on reducing setup error of radiotherapy for nasopharyngeal carcinoma
Lihua ZHANG ; Xiaowei YAO ; Bo LI ; Linlin XU ; Zhaodi XU ; Fei BAI
Chinese Journal of Radiation Oncology 2024;33(10):942-949
		                        		
		                        			
		                        			Objective:To evaluate the effect of neck muscle training on reducing the neck setup error during radiotherapy in patients with nasopharyngeal carcinoma.Methods:Clinical data of nasopharyngeal carcinoma patients who were treated with radiotherapy in the First Affiliated Hospital of Air Force Medical University from February 2021 to October 2022 were selected and analyzed. All subjects were randomly divided into the experimental group ( n=48) and control group ( n=51) at a ratio of 1:1 using a random number table method. In the experimental group, patients received neck muscle training prior to treatment, and those in the control group received conventional treatment without additional interventions. Cone beam computed tomography (CBCT) was performed weekly to measure and analyze the setup errors at the level of the slopes, 4th cervical vertebra (C4) and 7th cervical vertebra (C7). The four-dimensional displacement [left-right (LR), superior-inferior (SI), anterior-posterior (AP), rotation (Rtn)] systematic errors and random errors at each level were calculated, and the planning target volume (PTV) boundary was calculated. The differences at three different levels of slope, C4, C7 (LR, SI, AP axis) were compared between two groups. The correlation of setup errors in each direction was analyzed by Spearman correlation analysis. The changes of cervical curvature, cutaneous toxicity (common terminology criteria for adverse events V3.0) and pain assessment (numerical rating scale) were compared between two groups. Qualitative data between two groups were compared by χ2 test. Quantitative data between two groups were compared by t-test. Results:Baseline features were well balanced in both groups. The setup error in the experimental group was smaller than that in the control group. For the setup error in the AP direction, the setup errors at the levels of slope, C4 and C7 in the experimental group were (0.94±0.88) mm, (1.13±1.03) mm and (1.32±1.22) mm, significantly less than (1.66±1.23) mm, (1.63±1.35) mm and (1.89±1.48) mm in the control group (all P<0.001). In the SI direction, the setup errors at the levels of slope, C4 and C7 in the experimental group were (1.14±0.87) mm, (1.31±0.93) mm and (1.39±0.95) mm, compared with (1.22±0.95) mm, (1.40±1.11) mm and (1.52±1.08) mm in the control group ( P=0.278, 0.272, 0.100). The differences in the AP direction at the level of C4 and C7 in the experimental group were smaller than those in the control group ( P=0.014, 0.005). The required PTV boundary in the AP direction in the experimental group was increased from 1.77 mm at the slope level to 2.98 mm at the level of C7. In the control group, it was increased from 3.02 mm at the slope level to 4.78 mm at the level of C7. Correlation analysis showed that at the C4 and C7 levels, and the setup errors in the SI direction were moderately negatively correlated with those in the AR direction. There were no significant differences in cervical curvature, skin toxicity and pain assessment between two groups. Conclusion:Neck muscle training can reduce the setup error in the AP direction and PTV boundary of radiotherapy in patients with nasopharyngeal carcinoma, which is worthy of clinical promotion.
		                        		
		                        		
		                        		
		                        	
7.Based on probe near-infrared autofluorescence imaging technique of parathyroid gland application in thyroid surgery
Zheng WAN ; Bo XU ; Xiaodong YANG ; Wensong CAI ; Gaosong WU ; Chen LI ; Linlin ZHANG ; Xin MIAO ; Jing YAO ; Bing WANG ; Jianhua FENG ; Wen TIAN
Chinese Journal of Endocrine Surgery 2023;17(4):404-409
		                        		
		                        			
		                        			Objective:To investigate the effectiveness of probe-based near-infrared autofluorescence imaging (NIRAF) of the parathyroid gland.Methods:A total of 71 patients with thyroid cancer eligible for admission from May 4, 2023 to May 26, 2023 were selected, including 42 patients with thyroid cancer enrolled in the Department of Thyroid (hernia), Department of General Surgery, PLA General Hospital, including 29 females and 13 males, with a median age of 41 years, ranging from 21 to 76 years. A total of 29 patients with thyroid cancer were enrolled in the Department of Thyroid Surgery of Guangzhou First People’s Hospital, including 22 females and 7 males. The median age was 42.5 years, ranging from 24 to 72 years. The follow-up period was 1 month.Results:Among them, 196 suspicious parathyroid tissues were identified by the naked eye, and 207 suspicious parathyroid tissues were identified by probe NIRAF technology. Naked eye identification sensitivity, specificity and accuracy were 84.86%, 56% and 81.89%. The sensitivity, specificity and accuracy of parathyroid tissue identification by probe NIRAF were 92.66%, 80.00% and 90.53%, which were better than that of naked eye identification, and had a good coincidence rate with the results of immunocolloidal gold test or intraoperative freezing pathology (Kappa=0.61, P<0.001) . Conclusion:The probe-based NIRAF technique has a good ability to identify parathyroid tissue.
		                        		
		                        		
		                        		
		                        	
8.Effects of sunlike spectrum LED illumination on retinal blood perfusion in children and adolescents: a randomized controlled trial
Chinese Journal of School Health 2022;43(3):338-340
		                        		
		                        			Objective:
		                        			To analyze the effect of sunlike spectrum LED illumination on retinal blood flow perfusion, and to explore the the correlation between sunlike spectrum LED illumination and eye health indicators in children and adolescents.
		                        		
		                        			Methods:
		                        			A randomized control double blind trial was conducted. The ordinary LED table lamp in the control group(11) and the sunlike spectrum LED table lamp in the experimental group(12) had a fitting degree of 87% and 95% with the daylighting spectrum, respectively. Two sample independent t test and multivariable linear regression model were applied to compare the changes of retinal blood perfusion before and after the trial.
		                        		
		                        			Results:
		                        			After near reading for 1 hour, the retinal capillary density in the superficial and deep layers of the subjects in the ordinary LED illumination group decreased (superficial layer: -3.05±2.04 ,  P <0.01; deep layer:  -4.03± 4.94,  P =0.02), no significant decrease was found in the sunlike spectrum LED illumination group (superficial layer:  -0.59± 1.44,  P =0.18; deep layer: -0.49±4.27,  P =0.70). Multivariable regression analysis found that compared with ordinary LED illumination, sunlike spectrum LED illumination could significantly alleviate the decrease in capillary density in the superficial and deep retinal layers, respectively ( β =2.83, 95% CI =1.54-4.12,  P <0.01; β =4.21,95% CI =0.58-7.84,  P =0.02).
		                        		
		                        			Conclusion
		                        			Sunlike spectrum LED illumination can alleviate the decrease in retinal blood perfusion caused by near work among children and adolescents, suggesting that it may delay the onset and development of myopia. Prevention and control of myopia needs to pay attention to the spectral power distribution of artificial illumination.
		                        		
		                        		
		                        		
		                        	
9.Analyze of obesity indicators and effect of fertility preservation treatment in patients with endometrial atypical hyperplasia and early endometrial cancer
Linlin BO ; Yiqin WANG ; Yuanyuan LIU ; Xiaodan LI ; Rong ZHOU ; Jianliu WANG
Chinese Journal of Obstetrics and Gynecology 2022;57(10):767-774
		                        		
		                        			
		                        			Objective:To investigate the clinical features of obesity indicators in patients with endometrial atypical hyperplasia (EAH) and early endometrial cancer (EC) and analyze the relationship between these indexes and effect of fertility preservation therapy.Methods:The clinical data of patients with EAH, EC and endometrial benign lesions treated in Peking University People′s Hospital from January 1, 2018 to June 30, 2021 who required fertility-sparing treatment were collected, and obesity indicators were calculated and analyzed retrospectively.Results:(1) Obesity indicators: the obesity [body mass index (BMI) ≥28 kg/m 2] rate of patients with fertility preservation treatment was 40% (32/80), and abdominal obesity [waist circumference (WC) ≥80 cm] rate was 79% (63/80), and obesity indicators [BMI, WC, waist-hip ratio (WHR), weight height ratio (WHTR), body roundness index (BRI), lipid accumulation index (LAP), visceral adiposity index (VAI)] were higher than those with endometrial benign lesions (all P<0.001). (2) Related factors affecting the efficacy of fertility preservation treatment and their predictive value: EC, higher BMI, WC, WHR, WHTR and BRI were risk factors for lower complete remission rate after nine months′ treatment (all P<0.05). The predictive values of BRI and WHTR combined with pathological type were superior to other indicators [area under the curve (AUC)=0.716; AUC=0.714]. (3) Relation of obesity indicators and glucolipid indicators:BMI, WC, WHR, WHTR, BRI, LAP and VAI were positively correlated with homeostasis model assessment-insulin resistance index, glycosylated hemoglobin, and triacylglycerol (all P<0.05); while VAI was negatively correlated with high density lipoprotein cholesterol ( P<0.001). Conclusions:For patients with EAH and EC treated with fertility preservation therapy, abnormal obesity indexes affect the treatment effect. BRI and WHTR combined pathology have good predictive value for effect of fertility preservation treatment. In clinical practice, appropriate indicators could be selected to evaluate body shape, glucolipid metabolism and predict efficacy.
		                        		
		                        		
		                        		
		                        	
10.Molecular evolutionary analysis reveals Arctic-like rabies viruses evolved and dispersed independently in North and South Asia
Xin YU ; Hongwei ZHU ; Yongheng BO ; Youzhi LI ; Jianlong ZHANG ; Linlin JIANG ; Guozhong CHEN ; Xingxiao ZHANG ; Yongjun WEN
Journal of Veterinary Science 2021;22(1):e5-
		                        		
		                        			 Background:
		                        			Arctic-like (AL) lineages of rabies viruses (RABVs) remains endemic in some Arctic and Asia countries. However, their evolutionary dynamics are largely unappreciated. 
		                        		
		                        			Objectives:
		                        			We attempted to estimate the evolutionary history, geographic origin and spread of the Arctic-related RABVs. 
		                        		
		                        			Methods:
		                        			Full length or partial sequences of the N and G genes were used to infer the evolutionary aspects of AL RABVs by Bayesian evolutionary analysis. 
		                        		
		                        			Results:
		                        			The most recent common ancestor (tMRCA) of the current Arctic and AL RABVs emerged in the 1830s and evolved independently after diversification. Population demographic analysis indicated that the viruses experienced gradual growth followed by a sudden decrease in its population size from the mid-1980s to approximately 2000.Genetic flow patterns among the regions reveal a high geographic correlation in AL RABVs transmission. Discrete phylogeography suggests that the geographic origin of the AL RABVs was in east Russia in approximately the 1830s. The ancestral AL RABV then diversified and immigrated to the countries in Northeast Asia, while the viruses in South Asia were dispersed to the neighboring regions from India. The N and G genes of RABVs in both clades sustained high levels of purifying selection, and the positive selection sites were mainly found on the C-terminus of the G gene. 
		                        		
		                        			Conclusions
		                        			The current AL RABVs circulating in South and North Asia evolved and dispersed independently. 
		                        		
		                        		
		                        		
		                        	
            

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