Objective:To characterize the molecular evolution of coxsackievirus A10 (CVA10) strains in Anhui Province.Methods:The nucleic acids of CVA10 isolates in Anhui Province were extracted for whole-genome PCR amplification. One-generation sequencing was performed and the complete whole-genome sequences of 10 isolates were obtained. Nucleotide and amino acid sequence similarity analysis of CVA10 isolates and reference strains was performed using MegAlign in DNAStar software. MEGA 11.0 was used to classify the genotypes of CVA10 isolates and representative strains based on phylogenetic analysis of the VP1 region, and the average evolutionary differences between genotypes were calculated. BioEdit 7.2 was used to calculate the entropy of amino acid substitution in the P1-P3 region of the isolates and analyze the amino acid non-synonymous substitution sites. Recombination signals associated with the Anhui isolates were detected using RDP4 and further verified using Simplot 3.5. DnaSp6 software was selected to analyze the selection pressure of CVA10 isolates and prototype strains.Results:Based on the VP1 region, CVA10 isolates and CVA10 representative strains were categorized into A-F genotypes, and most of the CVA10 prevalent in mainland China belonged to the F genotype, with some isolates in Anhui Province being more closely related to the Yunnan isolates. The average rate of evolutionary difference in nucleotides among genotypes ranged from 18.70% to 33.70%. The isolates shared 17 non-synonymous amino acid mutation sites in the VP1 region, and amino acid substitutions in the VP1, 3A and 3C regions might affect the pathogenicity of the strains. The isolates frequently recombined with a variety of other EVA strains in the 5′UTR, 2C, 3C, 3D, and 3′UTR regions. Selection pressure analysis of the isolates showed that the isolate genes were affected by negative selection pressure.Conclusions:Genetic evolutionary analysis of CVA10 suggests that mutations and recombination with other types of EVA strains are prevalent, affecting the molecular epidemiological trend of CVA10, and that molecular surveillance of CVA10 strains in Anhui Province should continue to be strengthened.

Objective:To characterize the molecular evolution of coxsackievirus A10 (CVA10) strains in Anhui Province.Methods:The nucleic acids of CVA10 isolates in Anhui Province were extracted for whole-genome PCR amplification. One-generation sequencing was performed and the complete whole-genome sequences of 10 isolates were obtained. Nucleotide and amino acid sequence similarity analysis of CVA10 isolates and reference strains was performed using MegAlign in DNAStar software. MEGA 11.0 was used to classify the genotypes of CVA10 isolates and representative strains based on phylogenetic analysis of the VP1 region, and the average evolutionary differences between genotypes were calculated. BioEdit 7.2 was used to calculate the entropy of amino acid substitution in the P1-P3 region of the isolates and analyze the amino acid non-synonymous substitution sites. Recombination signals associated with the Anhui isolates were detected using RDP4 and further verified using Simplot 3.5. DnaSp6 software was selected to analyze the selection pressure of CVA10 isolates and prototype strains.Results:Based on the VP1 region, CVA10 isolates and CVA10 representative strains were categorized into A-F genotypes, and most of the CVA10 prevalent in mainland China belonged to the F genotype, with some isolates in Anhui Province being more closely related to the Yunnan isolates. The average rate of evolutionary difference in nucleotides among genotypes ranged from 18.70% to 33.70%. The isolates shared 17 non-synonymous amino acid mutation sites in the VP1 region, and amino acid substitutions in the VP1, 3A and 3C regions might affect the pathogenicity of the strains. The isolates frequently recombined with a variety of other EVA strains in the 5′UTR, 2C, 3C, 3D, and 3′UTR regions. Selection pressure analysis of the isolates showed that the isolate genes were affected by negative selection pressure.Conclusions:Genetic evolutionary analysis of CVA10 suggests that mutations and recombination with other types of EVA strains are prevalent, affecting the molecular epidemiological trend of CVA10, and that molecular surveillance of CVA10 strains in Anhui Province should continue to be strengthened.

Objective:To understand the genome-wide sequence variation and molecular evolution of coxsackievirus A4 (CV-A4) strain in Anhui province, so as to provide a theoretical basis for the pathogenic monitoring and scientific prevention and treatment of hand, foot and mouth disease in the future.Methods:Five CVA4 isolates of 2020 were sequenced by first-generation sequencing method. MEGA11.0 was used to construct a phylogenetic tree based on VP1 region for 5 CV-A4 isolates, 32 CV-A4 strains and Enterovirus A71(EV-A71) prototype strain BrCr, and the isolates and enterovirus A (EV-A) prototype strains based on P1, P2 and P3 regions respectively, and DNAStar was used for amino acid sequence comparison in VP1 region. BioEdit7.2 was used for amino acid displacement entropy analysis and amino acid site entropy mapping. SimPlot3.5 and RDP4 were used for recombination analysis of CV-A4 isolate and EV-A prototype representative strains, and DnaSP6 software was used for selection pressure analysis of isolates and reference representative strains.Results:The phylogenetic tree showed that the isolates belonged to the C2 subtype, which belonged to the same clade as the CV-A4 strain circulating in Chinese mainland, and the amino acid sequence homology of the C2 subtype branch was 97.3%-100%, and the isolates had 6 amino acid variation sites compared with the prototype. The selection pressure analysis showed that the CV-A4 strain of the C2 subtype was affected by negative selection pressure, and there were 25 mutagenic sites in the amino acid sequence in the coding region of the displacement entropy analysis, accounting for 1.14%, and the evolution of the strain mainly depended on recombination. Recombination analysis showed that the isolates recombined with a variety of EV-A prototype strains in the P2 and P3 regions, and the recombination section with the CV-A5 prototype strain was longer, especially in the 3A-3C section, and P1 was a relatively conserved region.Conclusions:CV-A4 has frequent recombination events with other EV-A prototype strains in P2 and P3, and the molecular evolution of CV-A4 in Anhui should continue to be closely monitored.

Objective : To analyze the epidemiological characteristics and pathogen spectrum of hand，foot mouth disease (HFMD) in Anhui province from 2015 to 2022，and to provide scientific evidence for prevention and control measures of HFMD． Methods : The surveillance data of hand，foot and mouth disease in Anhui province from 2015 to 2022 were analyzed by descriptive epidemiology. Real-time PCR was used to detect and classify HFMD samples. Results : A total of 650 590 HFMD cases were reported in Anhui province from 2015 to 2022，including 1 406 se- vere cases and 17 deaths．The annual reported incidence was 131. 45 /100 000．The epidemic features of“low incidence in odd years and high incidence in even years”were presented from 2015 to 2019．The incidence showed a continuous decline from 2020 to 2022．The monthly distribution showed the characteristics of bimodal epidemic，and the main peak was not obvious in 2020．Hefei，Fuyang，Bozhou，Chuzhou and Suzhou ranked the top five cities in terms of cumulative incidence．The age of onset was mainly distributed in children aged 5 years and below，accounting for 89. 26% of the total cases．The male to female ratio was 1. 48 ∶ 1．A total of 28 657 laboratory-confirmed cases had been reported from 2015 to 2022．EV71 cases accounted for 10. 57% ，Cox A16 cases accounted for 24. 90% ，and other enterovirus cases accounted for 64. 53%．The dominant pathogens showed dynamic changes in different years．Since 2018，the proportion of EV71 decreased significantly，and the proportion of other enteroviruses gradually increased to become the dominant pathogens．Among other enteroviruses，Cox A6 strain was dominant (80. 48% ) ． Conclusion This study suggests that the prevention and control of HFMD in Anhui province should be paid more attention from April to July and from October to December．The focus areas are the cities in northern Anhui and Hefei where the floating population is large．The focus of prevention and control is on children aged 5 years and below．Other enteroviruses have become the dominant pathogens of hand-foot-mouth disease in Anhui province，Cox A6 strain is dominant.

Metabolomics, which mainly studies the metabolite components of organisms, tissues, cells and their dynamic changes, is an emerging omics technology following genomics and proteomics. Metabolites are the final products of cellular regulation, and the concentration of metabolites is considered to be the ultimate response of a biological system to genetic or environmental changes. Secondary metabolites with chemical diversity are widely present in living organisms, thus accurate quantification of secondary metabolites through appropriate analytical platforms is an important task of metabolomics. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the most commonly used method for the detection of metabolites, providing a basis for the wide application of plant secondary metabolites. This review summarizes the advances of using LC-MS/MS techniques for the detection of phytohormone, folic acid, flavonoids and other secondary metabolites.
Chromatography, Liquid/methods* ; Tandem Mass Spectrometry/methods* ; Metabolomics/methods* ; Plants ; Proteomics

Objective:To investigate the effect of hyperbaric oxygen (HBO) on the prevention of chemotherapy-induced peripheral neuropathic pain (CIPNP), and to observe its mechanism by targeting spinal cannabinoid receptors (CBRs).Methods:A total of 75 male Sprague-Dawley(SD) rats were randomly divided into 5 groups (15 rats in each group), i. e. blank control group, CIPNP control group, CIPNP+ HBO group, CIPNP+ HBO+ AM630 group, and CIPNP+ HBO+ AM251 group. The model rats with CIPNP were established by injecting paclitaxel (i.p.). Each group with HBO intervention received the HBO treatment on the second day after each of the 5 times of paclitaxel injection. The CIPNP+ HBO+ AM630 and CIPNP+ HBO+ AM251 groups were administered with AM630 (an antagonist of CBR2) and AM251 (an antagonist of CBR1) respectively before each HBO treatment. The behavioral test was carried out to measure the mechanical withdrawal threshold (MWT) of rats by von fery filaments before the experiment and every 7 days during the experiment. The expressions of CBR1 and CBR2 were tested by Western blotting. The expression of glial fibrillary acidic protein (GFAP) was tested by immunohistochemistry (ICH) and Western blotting. And the expressions of inflammatory cytokines in the spinal cord, i. e. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), were detected by enzyme-linked immunosorbent assay (ELISA).Results:Compared with that of the blank control group, the MWT of the CIPNP control group was significantly decreased ( P<0.01), and the difference was most significant [(15.46±2.83) g vs. (4.33±3.53) g; P<0.01] especially on the 21st day of the experiment. The expressions of spinal GFAP, IL-1β, and TNF-α were significantly increased, and the differences were statistically significant ( P<0.05). Compared with those of the CIPNP control group, the MWT and spinal CBR2 of the CIPNP+ HBO group were significantly increased ( P<0.05), the GFAP, IL-1β, and TNF-α in the spinal cord were significantly decreased ( P<0.05), and the above-mentioned effects could be blocked by intraperitoneal injection of AM630, while there was no such reverse effect after intraperitoneal injection of AM251. Conclusion:HBO can prevent paclitaxel-induced CIPNP, and its mechanism may be related to the activation of spinal CBR2 and then the blocking of the activation of GFAP and the expression of inflammatory cytokines in the spinal cord.

Objective:To investigate the effect of hyperbaric oxygen (HBO) on the prevention of chemotherapy-induced peripheral neuropathic pain (CIPNP), and to observe its mechanism by targeting spinal cannabinoid receptors (CBRs).Methods:A total of 75 male Sprague-Dawley(SD) rats were randomly divided into 5 groups (15 rats in each group), i. e. blank control group, CIPNP control group, CIPNP+ HBO group, CIPNP+ HBO+ AM630 group, and CIPNP+ HBO+ AM251 group. The model rats with CIPNP were established by injecting paclitaxel (i.p.). Each group with HBO intervention received the HBO treatment on the second day after each of the 5 times of paclitaxel injection. The CIPNP+ HBO+ AM630 and CIPNP+ HBO+ AM251 groups were administered with AM630 (an antagonist of CBR2) and AM251 (an antagonist of CBR1) respectively before each HBO treatment. The behavioral test was carried out to measure the mechanical withdrawal threshold (MWT) of rats by von fery filaments before the experiment and every 7 days during the experiment. The expressions of CBR1 and CBR2 were tested by Western blotting. The expression of glial fibrillary acidic protein (GFAP) was tested by immunohistochemistry (ICH) and Western blotting. And the expressions of inflammatory cytokines in the spinal cord, i. e. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), were detected by enzyme-linked immunosorbent assay (ELISA).Results:Compared with that of the blank control group, the MWT of the CIPNP control group was significantly decreased ( P<0.01), and the difference was most significant [(15.46±2.83) g vs. (4.33±3.53) g; P<0.01] especially on the 21st day of the experiment. The expressions of spinal GFAP, IL-1β, and TNF-α were significantly increased, and the differences were statistically significant ( P<0.05). Compared with those of the CIPNP control group, the MWT and spinal CBR2 of the CIPNP+ HBO group were significantly increased ( P<0.05), the GFAP, IL-1β, and TNF-α in the spinal cord were significantly decreased ( P<0.05), and the above-mentioned effects could be blocked by intraperitoneal injection of AM630, while there was no such reverse effect after intraperitoneal injection of AM251. Conclusion:HBO can prevent paclitaxel-induced CIPNP, and its mechanism may be related to the activation of spinal CBR2 and then the blocking of the activation of GFAP and the expression of inflammatory cytokines in the spinal cord.

Objective:To examine 2019 novel coronavirus (2019-nCoV) RNA in clinical specimens of COVID-19 patients in Anhui province, and provide evidence for laboratory diagnosis of COVID-19 and risk assessment of clinical specimens.Methods:ORF1ab gene and N gene of 2019-nCoV were detected by real-time fluorescence RT-PCR in 466 clinical specimens of 197 COVID-19 cases. Chi-square test was used to analyze the differences in positive rates of specimens with clinical classification and time of onset.Results:The positive rates of 2019-nCoV in throat swab, sputum, serum, blood sample were 88.83%, 94.67%, 6.78% and 5.08%. The positive rate for 2019-nCoV RNA in throat swabs and sputum differed significantly ( χ2=8.994, P=0.003) in common cases during 7 days after illness onset. Conclusions:The positive rate of RNA in sputum was higher than throat swabs. 2019-nCoV RNA was detected in serum and blood specimens of COVID-19 cases. There was a risk of serum and blood specimens for transmission of COVID-19.

Objective: To analyze the genotype diversity and phylogenetic characterization of norovirus(NoV) in patients with diarrhea from Anhui province. Methods: NoV positive fecal specimens from sentinel hospitals were collected from January, 2016 to December, 2017. The samples were detected by Real-time fluorescent quantitative PCR. Positive samples were of randomly selected and amplified by RT-PCR and the products were sequenced. Phylogenetic tree was constructed by the Neighbor-Joining method based on partial VP1 gene regions of NoV to perform phylogenetic analysis. Results: A total of 263 NoV positive samples were genotyped, of which 239 belonged to genogroup II, 24 belonged to genogroup I. Fifty-five positive samples were successfully sequenced. There were 6 NoV GII genotypes, which included GII.2, 3, 4/Sydney_2012, 13, 17 and 21, while NV GII.17 and GII.4 were the dominant genotypes from 2016 to 2017. The predominant genotype was GII.4/Sydney 2012 (47.27%, 26/55), followed by GII.17 (23.64%, 13/55) and GII.2 (14.55%, 8/55). Phylogenetic tree showed that 26 strains belonged to genotype GII.4/Sydney 2012, NoV. The nucleotide homology among the 26 VP1 genes was 97.8% to 100%. Analysis of the partial VP1 genes of 26 strains showed that it shared the highest homology of 98.9% with the strain of GII.4Sydney2012 (GenBank ID: KU720515). However, the prevailing genotype in the Anhui province has shifted on two separate occasions, the GII.17 strain was dominant in 2016, and the GII.4/Sydney 2012 strain was dominant in 2017. Conclusions NoV GII was the major pathogen causing sporadic diarrhea in Anhui province during from 2016 to 2017, the genotypes are widely distributed, and shifted into the two predominant strains.

OBJECTIVE:To evaluate economic effectiveness of 3 kinds of antimicrobial drugs for acute lithiasic cholecystitis,and to provide reference for rational use of antimicrobial drugs in biliary disease surgery.METHODS:A multicenter prospective randomized controlled study was conducted in 493 clinical cases of acute lithiasic cholecystitis.Cost-effectiveness analysis of cefoperazone/sulbactam sodium(therapy A n=180),cefuroxime(therapy B n=148) and levofloxacin(therapy C n=165) for acute lithiasic cholecystitis were carried out.RESULTS:Effective rates of three kinds of therapeutic regimens 95.56%,73.65%,84.24%,respectively.CONCLUSION:Incremental cost-effectiveness analysis of three kinds of therapeutic regimens shows therapy A is economical and the best choice for acute lithiasic cholecystitis compared with group B and C.