Modified suture-assisted canaloplasty in Asians with primary open-angle glaucoma: a prospective study with 12-month follow-up.

BACKGROUND: To evaluate the efficacy and safety of modified suture-assisted canaloplasty in Asians with primary open-angle glaucoma (POAG). METHODS: A prospective, consecutive cases study, evaluating a modified canaloplasty by twisted 6/0 suture was performed on Asian POAG patients. Three modifications of this canaloplasty included opening the Schlemm's canal by viscocanalostomy, circumferential probing by a twisted 6/0 suture and loose suture of the superficial scleral flap. The twisted 6/0 suture was selected as a prober based on characteristical analysis of size and contact measurement as well as chemical composition conducted among 5/0, twisted 6/0 polypropylene sutures and the microcatheter. Success criteria were defined as intraocular pressure (IOP) ≤ 21 mmHg, 18 mmHg, 15 mmHg, and ≥ 20% reduction without (complete success) or with medications (qualified success). Efficacy was assessed by the success rate of circumferential catheterization, IOP values, the success rate of the surgery, the number of IOP-lowering medications, best-corrected vision acuity (BCVA), cup-to-disc ratio (C/D), and mean deviation (MD). Safety was evaluated by adverse events. RESULTS: Forty eyes from 40 consecutive patients were included with a mean follow-up of 14.8 ± 3.0 months. Circumferential catheterization was successfully conducted in 36 eyes (90%). Mean IOP decreased from 26.2 ± 6.9 mmHg to 14.5 ± 2.7 mmHg at 12 months postoperatively. While medication numbers were reduced from 3.2 ± 0.6 to 0.5 ± 0.8 at month 12 (both p < 0.001). Qualified success rate was 97.2% [95% confidence interval (CI) 0.92-1.03], 86.1% (95% CI 0.74-0.98) and 66.7% (95% CI 0.51-0.83) at 12 months with three criteria. BCVA, C/D and MD did not show progression at 1-year follow-up (p > 0.05). Age, baseline IOP, and spherical equivalent negatively influenced the success rate significantly (all p < 0.05). Adverse events included hyphema (30.6%), IOP spike > 25 mmHg (8.3%), and peripheral synechia to the trabecular-Descemet's membrane (2.7%). CONCLUSION: Twisted 6/0 suture can be an ideal material for cannulation. Modified suture-assisted canaloplasty is an effective, safe alternative with a cost-efficient feature for patients with POAG, especially in developing countries. TRIAL REGISTRATION: This trial was registered in the Chinese Clinical Trial Registry ( ChiCTR1900028618 , 29/12/2019).

Protection motivation theory in predicting cervical cancer screening participation: A longitudinal study in rural Chinese women.

OBJECTIVE: To examine factors longitudinally associated with cervical cancer screening uptake among rural Chinese women, guided by protection motivation theory (PMT). METHODS: A large sample of women (n = 2408, aged 35-65 years old) was randomly selected from a rural county in China in 2015 and followed up for 2 years. Data for demographic factors, knowledge of cervical cancer screening, screening outcome, and six PMT constructs measured at the baseline in 2015 were used to predict cervical cancer screening participation at the follow-up in 2017 using structural equation model method. RESULTS: Among the 2408 women at the baseline, 1879 (78.03%) participated in the screening services at the follow-up. In addition to significant direct effect of age, social status and baseline screening outcome, and three (perceived severity, fear arousal and response efficacy) of the six PMT subconstructs, four variables (age, social status, knowledge of cervical cancer screening, and baseline screening outcome) at the baseline were indirectly associated with screening participation, mediated by the three significant PMT subconstructs. CONCLUSIONS: Findings of this study indicate that the rate of participating in cervical cancer screening for rural women needs to be further improved. In addition to the commonly reported influential factors, PMT subconstructs play important roles in encouraging rural women in China to participate in cervical cancer screening. These longitudinal findings provided data much needed for future research to develop evidence-based intervention programs to enhance cervical cancer screening among rural women in China.

Viscocanalostomy combined with nearly 360-degree suture trabeculotomy for the treatment of primary congenital glaucoma: a preliminary report of a novel technique for trabeculotomy.

PURPOSE: The aim of this study was to determine the preliminary efficacy and safety of a novel technique for trabeculotomy for the treatment of primary congenital glaucoma (PCG). METHODS: This retrospective interventional study was conducted on patients with PCG. Pliability test was performed among 5/0 and 6/0 polypropylene sutures and the flexible illuminated microcatheter. Viscocanalostomy was first performed and the Schlemm's canal was identified. Then, suture trabeculotomy was completed except the region for viscocanalostomy. The preoperative and follow-up data were recorded and analyzed. Success criteria were defined as intraocular pressure (IOP) ≤ 21 mmHg without (complete success) or with medications (qualified success). RESULTS: 5/0 polypropylene suture was an appropriate option for cannulation of Schlemm's canal. A total of 33 eyes from 23 consecutive patients were included with a mean follow-up of 9.3 ± 3.6 months (range 6-18 months). Circumferential cannulation by suture was successfully performed in 28 eyes (84.8%) of 18 patients. Mean IOP decreased from 33.9 ± 9.4 mmHg preoperatively to 10.5 ± 3.5 mmHg at 6 months (p < 0.001) and 11.3 ± 4.1 mmHg at 9 months (p < 0.001). Complete success rate was 92.9% [95% confidence interval (CI:0.83-1.03)] and 87.5% (CI:0.69-1.06) at 6 months and 9 months, respectively. Hyphema was found in 5 eyes (17.9%), all of which were absorbed within 1 week. No other complications were observed. CONCLUSIONS: Viscocanalostomy combined with nearly 360-degree suture trabeculotomy as a novel technique controls IOP in patients with PCG without any severe complications. It is suitable for extended applications because of accurate identification of Schlemm's canal and low cost.

Comparison of ultrasound cycloplasty and transscleral cyclophotocoagulation for refractory glaucoma in Chinese population.

BACKGROUND: To compare the efficacy and safety of focused ultrasound cycloplasty (UCP) and transscleral cyclophotocoagulation (TSCP) in the treatment of refractory glaucoma in a Chinese population. METHODS: We retrospectively compared twenty-eight eligible patients with refractory glaucoma, who were divided into the UCP group and TSCP group. Patients in these two groups underwent a corresponding procedure from June 2018 to February 2019. The intraocular pressure (IOP), visual acuity, the number of anti-glaucoma agents used and complications were reviewed and compared between groups. Proper statistical methods were selected according to comparison models under IBM SPSS 25 software. RESULTS: After the 12-months follow-up, postoperative IOP and number of anti-glaucoma agents used in the two groups were both reduced than the baseline level, and the differences were statistically significant (P < 0.05). There were no significant differences in IOP, number of anti-glaucoma agents and the best-corrected visual acuity between the two groups at each follow-up time point (P>0.05). In terms of complications, the pain at 1 day after surgery in the UCP group was significantly milder than that in the TSCP group (P < 0.05). And there were no significant differences in other complications between the two groups (P > 0.05). CONCLUSIONS: Both UCP and TSCP are safe and effective methods for the treatment of refractory glaucoma. Nevertheless, pain is less severe after UCP.

Human papillomavirus type 18/16 infection and prevalence among middle-aged and older Chinese rural women: a cross-sectional survey in Wufeng, Hubei Province.

This study aimed to measure the prevalence and potential factors related to HPV 16/18 infection among middle-aged and older Chinese rural women. The study was conducted among women aged 35-65 years in rural villages within Wufeng in Hubei Province. Data were collected using a pretested questionnaire between July and August 2015. Cervical specimens were collected for HPV DNA detection and typing by using careHPV. Pearson's Chi-square and logistic regression analyses were used to examine associations with HPV positivity. Among 1001 participants, the prevalence of HPV 18/16 positive genotyping was 15.88% (n = 159). Husbands' extramarital sex (adjusted odds ratio [OR] = 15.85, 95% confidence interval [CI] 5.76-43.59), cervicitis (adjusted OR = 9.27, 95% CI 5.06-16.99), condom usage (adjusted OR = 0.16, 95% CI 0.05-0.53), higher average number of live births (adjusted OR = 1.98, 95% CI 1.32-2.96 for two live births; adjusted OR = 3.29, 95% CI 1.39-7.81 for three or more live births) were associated with HPV infection. The prevalence of HPV infection among participating women from rural areas was higher than that in several other areas in China. Our findings can aid efforts to prevent HPV infection to lower the risk of cervical cancer.

Diastereoselective Synthesis of Adjacent P,C-Stereogenic ß-N-Glycosidic Linked α-Aminophosphinates.

The diastereoselective formation of adjacent P,C-stereogenic ß-N-glycosidic linked α-aminophosphinates is developed in high yields via a phospha-Mannich reaction. The reaction was performed by employing (Rp)-O-(-)-menthyl H-phenylphosphinate and O-pivaloylated N-galactosylimine for double stereodifferentiation and BF3·Et2O as a promoter in THF. O-Pivaloylated N-galactosylphenyl imine 2 and (Rp)-O-(-)-menthyl H-phenylphosphinate 1 were converted to N-galactosyl α-aminoalkylphosphinate 3 with ratios of diastereomers up to 20:1. The synthetic method of the conversion provides a rapid access to adjacent P,C-stereogenic chiral α-aminophosphinates.

Hydroxycamptothecin regulates scar formation of the filtration channel under scleral flap by inhibiting the proliferation of scleral fibroblasts.

BACKGROUND: To investigate the inhibitory effect of a hyaluronic acid hydrogel loaded with hydroxycamptothecin (HCPT) on scar formation after filtration surgery in a rabbit model. METHODS: Scleral fibroblasts were isolated and extracted from rabbits' eyes. After treatment with different concentrations of HCPT, cytotoxicity was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and proliferation and extent of apoptosis were analysed using flow cytometry. Hydrogels loaded with different dosages of HCPT were prepared and placed under the scleral flap after the filtration surgery. One day, one week, and two weeks after surgery, follicular, conjunctival, corneal, and anterior chamber inflammation and iris and lens changes were observed. RESULTS: In vitro, compared with cells not treated with HCPT, cells treated with HCPT had decreased survival rate and proliferation, and the apoptosis level increased with increasing HCPT concentrations (p < 0.05). In vivo, the flattening time of filtering blebs in the three groups treated with different dosages of HCPT hydrogel was delayed. The degrees of oedema, inflammation, and bleeding were similar to those observed in the control group. The HCPT hydrogel effectively downregulated the expression of collagen 1 and 3 and tissue inhibitor of metalloproteinase 2 and upregulated the expression of matrix metalloproteinase 2 in a dose-dependent manner. CONCLUSIONS: HCPT significantly inhibited the growth of rabbits' scleral fibroblasts and effectively inhibited scar formation after filtering surgery by accelerating the degradation of extracellular matrix deposition.

Expression of angiotensin converting enzyme 2 mRNA in different SARS-CoV-2 infection states and times.

To measure the expression of angiotensin converting enzyme 2 (ACE2) mRNA in SARS-CoV-2 infection with different infection status and at different stages during infection, we used RT-qP CR to measure the expression of ACE2 mRNA. Measurements were analyzed by two-way repeated measures analysis of variance (RMANOVA). Expression of ACE2 mRNA was downregulated in initial stages of SARS-CoV-2 infection both in the asymptomatic infection (ASY) group and the confirmed cases (CON) group (t=-8.0845, P < 0.0001; t=-8.1904, P < 0.0001, respectively). The expression of ACE2 mRNA in the incubation period of CON group was lower compared with the intinal period of ASY group (F = 6.084, p = 0.016, partialη2 = 0.070). Relative expression of ACE2 mRNA was upregulated at the late stage of SARS-CoV-2 infection in the ASY and CON groups (F = 23.489, p = 0.000, partialη2 = 0.225; F = 46.555, p = 0.000, partialη2 = 0.365, respectively). The relative expression of ACE2 mRNA was down-regulated (mean ± SEM:0.69 ± 0.03) after inoculation with SARSCoV- 2 Spike pseudovirus, and there was a statistical difference (one-way t test, mean diffience =-0.31, 95%CI: -0.37˜-0.24, t=-8.1904, P < 0.0001). The expression of ACE2 mRNA is downregulated in the initial stage of SARS-CoV-2 infection, and then upregulated in the late stage of SARS-CoV-2 infection. The lower expression of ACE2 mRNA during the incubation period can lead to clinical symptoms. Downregulation of ACE2 mRNA was related to the interaction between SARS-CoV-2 S protein and ACE2.Communicated by Ramaswamy H. Sarma.

Comparison of viscocanalostomy plus suture-assisted near-360-degree trabeculotomy and viscocanalostomy plus rigid probe trabeculotomy in primary congenital glaucoma.

PURPOSE: To compare the efficacy and safety of viscocanalostomy plus near-360-degree suture trabeculotomy (VST) with viscocanalostomy plus rigid probe trabeculotomy (VT) in treating primary congenital glaucoma (PCG) over a one-year follow-up. METHODS: This consecutive retrospective study included patients with PCG confirmed within 3 years of age from March 2017 to October 2019. Efficacy was evaluated by comparing the postoperative intraocular pressure (IOP) curve and the success rate at one year after surgery. Safety was assessed by comparing the postoperative complications. The number of anti-glaucoma agents, horizontal corneal diameter (HCD) and cup-to-disc ratio (C/D) of the two surgical methods were also compared. RESULTS: Data of 90 eyes from 61 patients were analysed. The baseline parameters of the two groups were similar. The IOP at 12 months after surgery in the VST group was 12.7 ± 4.8 mmHg, while that in the VT group was 15.8 ± 6.5 mmHg. The IOP at 6, 9 and 12 months postoperatively in the VST group was significantly lower than in the VT group (p < 0.05). Viscocanalostomy plus near-360-degree suture trabeculotomy (VST) remained a significant favourable factor for complete one-year success (93.6% versus 74.4%, p = 0.005) but not qualified one-year success (97.9% versus 88.4%, p = 0.06). The number of anti-glaucoma agents, HCD and C/D were reduced in both groups. Postoperative complications were not significantly different between the two groups. CONCLUSIONS: In children with PCG, VST provides a more durable IOP control than VT over the one-year follow-up, with a similar safety profile.

Genomic Epidemiology and Phylodynamic Analysis of Enterovirus A71 Reveal Its Transmission Dynamics in Asia.

Enterovirus A71 (EV-A71) is one of the main pathogens causing hand, foot, and mouth disease (HFMD) outbreaks in Asian children under 5 years of age. In severe cases, it can cause neurological complications and be life-threatening. In this study, 200 newly sequenced EV-A71 whole-genome sequences were combined with 772 EV-A71 sequences from GenBank for large-scale analysis to investigate global EV-A71 epidemiology, phylogeny, and Bayesian phylodynamic characteristics. Based on the phylogenetic analysis of the EV-A71 3Dpol region, six new evolutionary lineages (lineages B, J, K, O, P, and Q) were found in this study, and the number of evolutionary lineages was expanded from 11 to 17. Temporal dynamics and recombination breakpoint analyses based on genotype C revealed that recombination of nonstructural protein-coding regions, including 3Dpol, is an important reason for the emergence of new lineages. The EV-A71 epidemic in the Asia-Pacific region is complex, and phylogeographic analysis found that Vietnam played a key role in the spread of subgenotypes B5 and C4. The origin of EV-A71 subgenotype C4 in China is East China, which is closely related to the prevalence of subgenotype C4 in the south and throughout China. Selection pressure analysis revealed that, in addition to VP1 amino acid residues VP1-98 and VP1-145, which are associated with EV-A71 pathogenicity, amino acid residues VP1-184 and VP1-249 were also positively selected, and their functions still need to be determined by biology and immunology. This study aimed to provide a solid theoretical basis for EV-A71-related disease surveillance and prevention, antiviral research, and vaccine development through a comprehensive analysis. IMPORTANCE EV-A71 is one of the most important pathogens causing HFMD outbreaks; however, large-scale studies of EV-A71 genomic epidemiology are currently lacking. In this study, 200 new EV-A71 whole-genome sequences were determined. Combining these with 772 EV-A71 whole-genome sequences in the GenBank database, the evolutionary and transmission characteristics of global and Asian EV-A71 were analyzed. Six new evolutionary lineages were identified in this study. We also found that recombination in nonstructural protein-coding regions, including 3Dpol, is an important cause for the emergence of new lineages. The results provided a solid theoretical basis for EV-A71-related disease surveillance and prevention, antiviral research, and vaccine development.

Genetic characterization and molecular epidemiology of Coxsackievirus A12 from mainland China during 2010-2019.

Coxsackievirus A12 (CVA12) is an enterovirus that has been isolated in many countries in recent years. However, studies on CVA12 are limited, and its effective population size, evolutionary dynamics and recombination patterns have not been clarified now. In this study, we described the phylogenetic characteristics of 16 CVA12 strains isolated from pediatric HFMD patients in mainland China from 2010 to 2019. Comparison of the nucleotide sequences and amino acid sequences with the CVA12 prototype strain revealed that the 16 CVA12 strains are identical in 78.8-79% and 94-94.2%, respectively. A phylodynamic analysis based on the 16 full-length VP1 sequences from this study and 21 sequences obtained from GenBank revealed a mean substitution rate of 6.61 × 10-3 substitutions/site/year (95% HPD: 5.16-8.20 × 10-3), dating the time to most recent common ancestor (tMRCA) of CVA12 back to 1946 (95% HPD: 1942-1947). The Bayesian skyline plot showed that the effective population size has experienced twice dynamic fluctuations since 2007. Phylogeographic analysis identified two significant migration pathways, indicating the existence of cross-provincial transmission of CVA12 in mainland China. Recombination analysis revealed two recombination patterns between 16 CVA12 strains and other EV-A, suggesting that there may be extensive genetic exchange between CVA12 and other enteroviruses. In summary, a total of 16 full-length CVA12 strains were reported in this study, providing valuable references for further studies of CVA12 worldwide.

Prevalence and correlates of cigarette smoking among Dulong adults in China: A cross-sectional survey in 2020.

Background: The Dulong people are one of the minorities in China with the lowest population. In recent years, the lifestyle of the Dulong people has also changed drastically due to income growth and urbanization. This study aims to identify cigarette smoking prevalence and potential risk factors among Dulong adults in China. Methods: This study was conducted among 1,018 adults based on the Dulong Health Status Investigation and Evaluation (DHSIE) in Gongshan Dulong and Nu Autonomous County of Yunnan province, Southwest China. A cross-sectional design and face-to-face questionnaire were used to collect cigarette smoking habits and demographic information. Data were weighted by post-stratification weights according to the age and gender composition of Dulong resident. We also analyzed univariate and multivariate unconditional logistic regression to explore current smoking correlates. Results: The weighted prevalence of ever-smoking, currently smoking, and formerly smoking among Dulong adults is 31.3, 27.7, and 3.6%, respectively. The prevalence of ever-smoking and currently smoking among male participants (57.0 and 50.6%) is much higher than that of female participants (4.0 and 3.4%). Nearly 60% of ever-smokers and current smokers smoked more than 20 cigarettes per day, which are higher than former smokers (35.2%). Among current smokers, 33.1% relapsed, and 28.3% intend to quit smoking. By adjusting for potential confounding variables, multiple logistic regression analysis indicated that male participants (OR = 48.982, 95% CI: 25.026-95.869) and current drinkers (OR = 4.450, 95% CI: 2.556-7.746) are more likely to be current smokers. On the contrary, current smokers are also more likely to be exposed to secondhand smoke (OR = 4.269, 95% CI: 2.330-7.820) and have a higher risk of chronic respiratory disease (OR = 4.955, 95% CI: 1.669-14.706). Conclusion: Cigarette smoking is highly prevalent among the Dulong people in Southwest China. An appropriate and effective tobacco control strategy is an urgent need for this population.

Eliciting national and subnational sets of disability weights in mainland China: Findings from the Chinese disability weight measurement study.

Background: The disability weight (DW) quantifies the severity of health states from disease sequela and is a pivotal parameter for disease burden calculation. We conducted a national and subnational DW measurement in China. Methods: In 2020-2021, we conducted a web-based survey to assess DWs for 206 health states in 31 Chinese provinces targeting health workers via professional networks. We fielded questions of paired comparison (PC) and population health equivalence (PHE). The PC data were analysed by probit regression analysis, and the regression results were anchored by results from the PHE responses on the DW scale between 0 (no loss of health) and 1 (health loss equivalent to death). Findings: We used PC responses from 468,541 respondents to estimate DWs of health states. Eight of 11 domains of health had significantly negative coefficients in the regression of the difference between Chinese and Global Burden of Disease (GBD) DWs, suggesting lower DW values for health states with mention of these domains in their lay description. We noted considerable heterogeneity within domains, however. After applying these Chinese DWs to the 2019 GBD estimates for China, total years lived with disability (YLDs) increased by 14·9% to 177 million despite lower estimates for musculoskeletal disorders, cardiovascular diseases, mental disorders, diabetes and chronic kidney disease. The lower estimates of YLDs for these conditions were more than offset by higher estimates of common, low-severity conditions. Interpretation: The differences between the GBD and Chinese DWs suggest that there might be some contextual factors influencing the valuation of health states. While the reduced estimates for mental disorders, alcohol use disorder, and dementia could hint at a culturally different valuation of these conditions in China, the much greater shifts in YLDs from low-severity conditions more likely reflects methodological difficulty to distinguish between health states that vary a little in absolute DW value but a lot in relative terms. Funding: This work was supported by the National Natural Science Foundation of China [grant number 82173626], the National Key Research and Development Program of China [grant numbers 2018YFC1315302], Wuhan Medical Research Program of Joint Fund of Hubei Health Committee [grant number WJ2019H304], and Ningxia Natural Science Foundation Project [grant number 2020AAC03436].

MAU-Net: A Retinal Vessels Segmentation Method.

Detailed extraction of retinal vessel morphology is of great significance in many clinical applications. In this paper, we propose a retinal image segmentation method, called MAU-Net, which is based on the U-net structure and takes advantages of both modulated deformable convolution and dual attention modules to realize vessels segmentation. Specifically, based on the classic U-shaped architecture, our network introduces the Modulated Deformable Convolutional (MDC) block as encoding and decoding unit to model vessels with various shapes and deformations. In addition, in order to obtain better feature presentations, we aggregate the outputs of dual attention modules: the position attention module (PAM) and channel attention module (CAM). On three publicly available datasets: DRIVE, STARE and CHASEDB1, we have achieved superior performance to other algorithms. Quantitative and qualitative experimental results show that our MAU-Net can effectively and accurately accomplish the retinal vessels segmentation task.

Effects and Molecular Mechanism of L-Type Calcium Channel on Fluoride-Induced Kidney Injury.

This study aimed to investigate the role and molecular mechanism of L-type calcium channel (LTCC) on fluoride exposure-induced kidney injury. Subchronic and chronic fluoride exposures were included in the experiment. Each part contained 140 ICR male mice. They were randomly divided into 7 groups: control group, high-fluoride group (NaF 30 mg/L), low-fluoride group (NaF 5 mg/L), high/low-fluoride + agonist (FPL64176) group, high/low-fluoride + inhibitor (nifedipine) group. One week before the end of fluoride exposure, each mouse in the fluoride exposure group was injected intraperitoneally with LTCC agonist (FPL64176) or inhibitor (nifedipine) (5 mg/kg day). The apoptosis of kidney cell was observed by TUNEL, and the protein expression levels of Cav1.2 and CaM, CaMKII, Bcl-2, and Bax were detected by Western blot. Compared with the control group, the protein expression levels of Cav1.2, CaM, and Bax significantly increased, and those of CaMKII and Bcl-2 significantly decreased, the ratio of Bax/Bcl-2 also significantly increased, and the number of apoptotic kidney cells significantly increased in the high/low-fluoride group and in the high/low-fluoride + agonist group. The above indicators and fluoride exposure concentrations showed in time- and dose-dependent changes. Compared with the high/low-fluoride + agonist group, the protein expression level of the molecular in the kidney cells above mentioned was significantly opposite and the number of apoptotic kidney cells significantly decreased in the high/low-fluoride + inhibitor group. In conclusion, LTCC mediates the kidney injury induced by fluoride exposure in mice. Fluoride exposure induced abnormal expression of the Cav1.2 protein, Ca2+ signal transduction pathway, and apoptosis-regulated proteins, which is one of the molecular mechanisms. Nifedipine may be a new and effective anti-fluoride drug.

Genetic recombination in fast-spreading coxsackievirus A6 variants: a potential role in evolution and pathogenicity.

Hand, foot, and mouth disease (HFMD) is a common global epidemic. From 2008 onwards, many HFMD outbreaks caused by coxsackievirus A6 (CV-A6) have been reported worldwide. Since 2013, with a dramatically increasing number of CV-A6-related HFMD cases, CV-A6 has become the predominant HFMD pathogen in mainland China. Phylogenetic analysis based on the VP1 capsid gene revealed that subtype D3 dominated the CV-A6 outbreaks. Here, we performed a large-scale (near) full-length genetic analysis of global and Chinese CV-A6 variants, including 158 newly sequenced samples collected extensively in mainland China between 2010 and 2018. During the global transmission of subtype D3 of CV-A6, the noncapsid gene continued recombining, giving rise to a series of viable recombinant hybrids designated evolutionary lineages, and each lineage displayed internal consistency in both genetic and epidemiological features. The emergence of lineage -A since 2005 has triggered CV-A6 outbreaks worldwide, with a rate of evolution estimated at 4.17 × 10-3 substitutions site- 1 year-1 based on a large number of monophyletic open reading frame sequences, and created a series of lineages chronologically through varied noncapsid recombination events. In mainland China, lineage -A has generated another two novel widespread lineages (-J and -L) through recombination within the enterovirus A gene pool, with robust estimates of occurrence time. Lineage -A, -J, and -L infections presented dissimilar clinical manifestations, indicating that the conservation of the CV-A6 capsid gene resulted in high transmissibility, but the lineage-specific noncapsid gene might influence pathogenicity. Potentially important amino acid substitutions were further predicted among CV-A6 variants. The evolutionary phenomenon of noncapsid polymorphism within the same subtype observed in CV-A6 was uncommon in other leading HFMD pathogens; such frequent recombination happened in fast-spreading CV-A6, indicating that the recovery of deleterious genomes may still be ongoing within CV-A6 quasispecies. CV-A6-related HFMD outbreaks have caused a significant public health burden and pose a great threat to children's health; therefore, further surveillance is greatly needed to understand the full genetic diversity of CV-A6 in mainland China.

Genomic epidemiology of coxsackievirus A16 in mainland of China, 2000-18.

Hand, foot, and mouth disease (HFMD), which is a frequently reported and concerning disease worldwide, is a severe burden on societies globally, especially in the countries of East and Southeast Asia. Coxsackievirus A16 (CV-A16) is one of the most important causes of HFMD and a severe threat to human health, especially in children under 5 years of age. To investigate the epidemiological characteristics, spread dynamics, recombinant forms (RFs), and other features of CV-A16, we leveraged the continuous surveillance data of CV-A16-related HFMD cases collected over an 18-year period. With the advent of the EV-A71 vaccine since 2016, which targeted the EV-A71-related HFMD cases, EV-A71-related HFMD cases decreased dramatically, whereas the CV-A16-related HFMD cases showed an upward trend from 2017 to October 2019. The CV-A16 strains observed in this study were genetically related and widely distributed in the mainland of China. Our results show that three clusters (B1a-B1c) existed in the mainland of China and that the cluster of B1b dominates the diffusion of CV-A16 in China. We found that eastern China played a decisive role in seeding the diffusion of CV-A16 in China, with a more complex and variant transmission trend. Although EV-A71 vaccine was launched in China in 2016, it did not affect the genetic diversity of CV-A16, and its genetic diversity did not decline, which confirmed the epidemiological surveillance trend of CV-A16. Two discontinuous clusters (2000-13 and 2014-18) were observed in the full-length genome and arranged along the time gradient, which revealed the reason why the relative genetic diversity of CV-A16 increased and experienced more complex fluctuation model after 2014. In addition, the switch from RFs B (RF-B) and RF-C co-circulation to RF-D contributes to the prevalence of B1b cluster in China after 2008. The correlation between genotype and RFs partially explained the current prevalence of B1b. This study provides unprecedented full-length genomic sequences of CV-A16 in China, with a wider geographic distribution and a long-term time scale. The study presents valuable information about CV-A16, aimed at developing effective control strategies, as well as a call for a more robust surveillance system, especially in the Asia-Pacific region.

Effects of drinking water fluorosis on L-type calcium channel of hippocampal neurons in mice.

The study aimed to investigate the effects of drinking water fluorosis on L-type calcium channels (LTCCs) in mouse hippocampal neurons. A total of 60 newly weaned ICR male mice were randomly divided into control, low fluoride and high fluoride groups. After 3 and 6 months of exposure to fluoride, the patch clamp technique was used to detect the peak and relative values (I/Imax), steady-state activation curve ratio (G/Gmax), decay time constant, and tail current time constant of LTCCs currents in hippocampal CA1 region of mouse brain slices. Fluoride greatly reduced the serum and urinary calcium concentrations in mice, and the chronic fluorosis has a greater impact than subchronic fluorosis. The peak value of LTCCs current in pyramidal neurons of hippocampal CA1 area was significant and increased with the prolonged exposure time. The relative values of current and steady-state coefficients were changed greatly. The decay and tail current time increased significantly. High fluorine concentration indicates great peak value and open time of LTCCs opening. LTCCs are sensitive to fluoride exposure. The activation voltage of calcium channels induced by fluoride exposure is decreased, the opening time of calcium channels is prolonged, and the calcium influx per unit time increased, thereby overloading calcium concentration in neurons and this may be an explanation for intracellular calcium overload caused by fluoride. The imbalance of calcium metabolism caused by fluorosis may be a pathogenesis of brain injury induced by fluoride. Furthermore, the risk of brain damage from low-fluorine exposure cannot be ignored.

Fundus Image Enhancement Method Based on CycleGAN.

In this paper, we propose a retinal image enhancement method, called Cycle-CBAM, which is based on CycleGAN to realize the migration from poor quality fundus images to good quality fundus images. It does not require paired training set any more, that is critical since it is quite difficult to obtain paired medical images. In order to solve the degeneration of texture and detail caused by training unpaired images, we enhance the CycleGAN by adopting the Convolutional Block Attention Module (CBAM). To verify the enhancement effect of our method, we not only analyzed the enhanced fundus image quantitatively and qualitatively, but also introduced a diabetic retinopathy (DR) classification module to evaluate the DR level of the fundus images before and after enhancement. The experiments show that our method of integrating CBAM into CycleGAN has superior performance than CycleGAN both in quantitative and qualitative results.

Emerging recombination of the C2 sub-genotype of HFMD-associated CV-A4 is persistently and extensively circulating in China.

Sporadic outbreaks caused by coxsackievirus A4 (CV-A4) have been reported worldwide. To further elucidate the detailed genetic characteristics and evolutionary recombination events of CV-A4, virus samples from nationwide hand, foot and mouth disease (HFMD) surveillance, encompassing 27 out of the 31 provinces in China, were investigated. Comprehensive and systematic phylogenetic analyses were performed by using 29 complete genomes, 142 complete CV-A4 VP1 sequences. Four genotypes (A, B, C and D) and five sub-genotypes (C1-C5) were re-identified based on the complete VP1 sequences. C2 is the predominant sub-genotype of CV-A4 associated with HFMD and has evolved into 3 clusters. Cluster 1 is a major cluster that has been persistently and extensively circulating in China since 2006 and has been associated with all severe cases. All the sequences showed high homology with the CV-A4 prototype in the P1 region, while higher identities with CV-A5, CV-14 and CV-16 in the P2 and P3 regions. Recombination analysis revealed that C2 had two specific genetic recombination patterns with other EV-A prototypes in the 5'-UTR and 3D region compared with C5. These recombination patterns might be associated with the increased transmissibility of C2 viruses, which were obtained due to their persistent and extensive circulation in populations.