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.

ObjectiveTo investigate the effect of Tangzhi pills on the improvement of insulin resistance （IR） in the liver with type 2 diabetes （T2DM） by regulating phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway based on differential genes and its possible molecular mechanism. MethodT2DM rat models were prepared by high fat （HFD） diet combined with streptozotocin （STZ） intraperitoneal injection. The experiment was divided into blank group， model group， metformin hydrochloride group （0.18 g·kg^-1）， Tangzhi pills high （1.08 g·kg^-1）， medium （0.54 g·kg^-1） and low （0.27 g·kg^-1） dose groups. Rat serum， liver， and pancreatic tissue were collected， and the pathological tissue of the liver and pancreas was observed using hematoxylin-eosin （HE） staining. The fasting blood glucose level （FBG） was detected， and oral glucose tolerance （OGTT） tests were conducted. Enzyme-linked immunosorbent assay （ELISA） was used to detect fasting serum insulin （FINS） and glycated hemoglobin （GHb） levels in rats. IR homeostasis model index （HOMA-IR）， β cellular homeostasis index （HOMA-β）， and insulin sensitivity index （ISI） were calculated. Biochemical methods were used to determine the levels of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein （LDL-C）， and high-density lipoprotein （HDL-C） in rat serum. Transcriptomics obtained differentially expressed mRNA from liver tissue and enriched differentially expressed pathways. Real-time reverse transcriptase polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of cyclic adenylate responsive element binding protein 3-like protein 2 antibody （CREB3l2）， B-lymphocyte tumor 2 （Bcl-2）， Toll-like receptor 2 （TLR2）， cyclin-dependent kinase inhibitor 1A （CDNK1A）， and DNA damage induced transcription factor 4-like protein （DDIT4） in liver tissue. Western blot was used to detect the protein expression of phosphorylated phosphatidylinositol 3-kinase （p-PI3K）， phosphorylated protein kinase B （p-Akt）， glucose transporter 4 （GLUT4）， insulin receptor （INSR）， and insulin receptor substrate 2 （IRS2）. ResultThe pharmacodynamic experiment results showed that compared with model group， Tangzhi pills groups repaired liver and pancreatic tissue to varying degrees， reduced blood sugar （P<0.01）， and promoted a decrease in serum FINS， GHb， and HOMA-IR （P<0.05， P<0.01）. In addition， HOMA-β and ISI increased （P<0.05， P<0.01）. The levels of TC， TG， and LDL-C decreased （P<0.05， P<0.01）， while the levels of HDL-C increased （P<0.05， P<0.01）. The transcriptomics experimental results confirmed that the PI3K/Akt signaling pathway was significantly expressed in both the blank group and model group， as well as in the high-dose Tangzhi pills group and model group. CDNK1A， DDIT4， CREB3l2， Bcl-2， and TLR2 were significantly differentially expressed mRNA during TG intervention in T2DM. Compared with the model group， the protein expression of p-PI3K， p-Akt， GLUT4， INSR， and IRS2 increased in all Tangzhi pills groups （P<0.01）. The mRNA expression of CREB3l2， Bcl-2， and TLR2 increased （P<0.01）， while that of CDNK1A and DDIT4 decreased （P<0.01）. ConclusionTangzhi pills may regulate the PI3K/Akt signaling pathway based on the differential mRNA expression of CREB3l2， Bcl-2， TLR2， CDNK1A， and DDIT4， thereby improving IR in the liver with T2DM.

Diabetic nephropathy(DN)is one of the most serious microangiopathies in diabetes mellitus and the leading cause of death in patients with end-stage renal disease.Ferroptosis,as a mode of programmed cell death,is mainly manifested by excessive accumulation of intracellular lipid peroxides and iron.Ferroptosis is involved in a series of pathological processes such as damage to DN renal podocytes,mesangial cells,and renal tubular epithelial cells.Chinese materia medica has the characteristics of significant therapeutic effects and minimal adverse reactions in the treatment of diseases,and has been widely used in the prevention and treatment of DN.This article summarized the key factors regulating ferroptosis in DN,as well as the active components and TCM formulas targeting the inhibition of ferroptosis in the prevention and treatment of DN,providing reference for the development of DN targeted drugs.

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.

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 explore the value and disadvantage of micro-mirror in the intracranial aneurysm surgery. Methods: Micro-mirror was used to assist microsurgical clipping to 36 intracranial aneurysms in 31 cases, of which 3 were carotid-ophthalmic artery aneurysms, 3 anterior choroidal artery aneurysm, 11 were posterior communicating artery aneurysms, 7 were middle cerebral artery aneurysms, 10 anterior communicating artery or anterior cerebral artery aneurysms, and the others were a posterior cerebral artery aneurysm and a posterior inferior cerebellar artery aneurysm. The micro-mirror was used before and after clipping to observe the anatomic features of necks hidden behind and medial to aneurysms, to visualize surrounding neurovascular structures, and to verify the optimal clipping position. Intraoperative indocyanine green fluorescein angiography confirmed the success of sufficient clipping. Results: All aneurysms were clipped successfully. The parent arteries were occluded temporarily in 26 cases, and 9 aneurysms ruptured during the operation. Postoperative follow-up lasted from 2 weeks to 1 year. After operations digital subtraction angiography (DSA) were repeated in 10 cases, computed tomography angiography(CTA) in 21 cases, and no insufficient clipping or occlusion of parent arteries were revealed. Conclusions Micro-mirror assisting microsurgical clipping to the intracranial aneurysm is conducive to accurate clipping, and is a convenient and practical operation.

Objective To understand the infection status and genetic characteristics of Norovirus from foodborne diseases in sentinel hospital of Ma′anshan city. Methods The 911 stool samples and epidemiological data of the patients with foodborne disease were collected from three hospitals of Ma′anshan city during January 2015 to June 2018. The GⅠ and GⅡ Norovirus were detected by real?time reverse transcription PCR. Some of the positive specimens were amplified by conventional reverse transcription PCR, and the PCR products were sequenced for sequence alignment and phylogenetic analysis. Results The positive rate of Norovirus was 14.7% (134/911), in which 7 strains were GⅠ, 124 strains were GⅡ and 3 strains were mixed infection. Norovirus can be detected throughout the year, with high positive rate from December to April of the next year [24.4%(20/82)-45.3%(24/53)]. The 78 males (15.5%) and 55 females (13.7%) were positive for Norovirus (χ2=0.58, P=0.448). There was no significant difference in different age groups (χ2=9.55, P=0.089). A total of 79 strains were successfully sequenced, 4 strains were GⅠ group (5.1%), 75 strains were GⅡ group (94.9%). The predominant strains were GⅡ. 17 and GⅡ. 4, aud the number were 35 and 15 respectively. The predominant strains were different in different years. The main strain was GⅡ.17 in 2015 (30, 68.2%),GⅡ.4 in 2016 (5/9) and 2017 (8/16),but GⅡ.3 in 2018 (3/6). Conclusion Norovirus diarrhea was popular in Ma′anshan city throughout the year,especially in winter and spring.Theprevalent strain was GⅡ,genotypes were diversified distribution,the dominant strains were GⅡ.17 and GⅡ.4. The predominant strains were different in different years.

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 explore the clinical effects of antibiotic bone cement in the treatment of diabetic foot ulcers. Methods: According to the treatment methods, 18 patients with diabetic foot ulcers (11 males and 7 females, aged 53-79 years), who were conformed to the study criteria and admitted to our hospital from January 2016 to January 2017, were enrolled in traditional group; 18 patients with diabetic foot ulcers (11 males and 7 females, aged 55-80 years), who were conformed to the study criteria and admitted to our hospital from February 2017 to February 2018, were enrolled in bone cement group. Wounds of patients in traditional group were treated with vacuum sealing drainage after conventional debridement. Wounds of patients in bone cement group were covered with antibiotic bone cement after conventional debridement. The number of patients with positive bacterial culture in wound exudate in the 2 groups on admission and 3, 6, 9, and 15 days after surgery, the length of hospital stay, the number of operation, and the wound complete healing time were retrospectively recorded. Data were processed with Fisher′s exact probability test and independent sample t test. Results: Compared with (29±10) d and (4.6±1.2) times of patients in traditional group, the length of hospital stay [(9±3) d] of patients was obviously shortened, the number of operation [(1.3±0.6) times] of patients was obviously reduced, the number of patients with positive bacterial culture in wound exudate at each time point post surgery was obviously reduced (t=8.177, 9.896, P<0.05 or P<0.01) in bone cement group. There were no statistically significant differences in the number of patients with positive bacterial culture in wound exudate on admission and wound complete healing time between patients in the 2 groups (t=0.175, P>0.05). Conclusions The antibiotic bone cement treatment of diabetic foot ulcers can reduce the number of patients with positive bacterial culture in wound exudate and the number of operation, as well as shorten the length of hospital stay.

Objective: To understand the infection status and genetic characteristics of Norovirus from foodborne diseases in sentinel hospital of Ma′anshan city. Methods: The 911 stool samples and epidemiological data of the patients with foodborne disease were collected from three hospitals of Ma′anshan city during January 2015 to June 2018. The G Ⅰ and G Ⅱ Norovirus were detected by real-time reverse transcription PCR. Some of the positive specimens were amplified by conventional reverse transcription PCR, and the PCR products were sequenced for sequence alignment and phylogenetic analysis. Results: The positive rate of Norovirus was 14.7% (134/911), in which 7 strains were G Ⅰ, 124 strains were G Ⅱ and 3 strains were mixed infection. Norovirus can be detected throughout the year, with high positive rate from December to April of the next year [24.4%(20/82)-45.3%(24/53)]. The 78 males (15.5%) and 55 females (13.7%) were positive for Norovirus (χ²=0.58,P=0.448). There was no significant difference in different age groups (χ²=9.55, P=0.089). A total of 79 strains were successfully sequenced, 4 strains were G Ⅰ group(5.1%), 75 strains were G Ⅱ group (94.9%). The predominant strains were GⅡ.17 and GⅡ.4, aud the number were 35 and 15 respectively. The predominant strains were different in different years. The main strain was GⅡ.17 in 2015 (30, 68.2%),GⅡ.4 in 2016 (5/9) and 2017 (8/16), but GⅡ.3 in 2018 (3/6). Conclusion Norovirus diarrhea was popular in Ma′anshan city throughout the year,especially in winter and spring. The prevalent strain was GⅡ,genotypes were diversified distribution,the dominant strains were GⅡ.17 and GⅡ.4. The predominant strains were different in different years.