Abstract: Objective To analyze the genetic evolution and molecular characteristics of the H5N6 avian influenza virus A/Changsha/1/2022(H5N6) in a human infection case in Changsha City, providing insights for the prevention and control of H5N6 avian influenza transmission to humans. Methods The sample underwent whole-genome sequencing on a third-generation sequencing platform. Reference sequences were downloaded from the NCBI and GISAID databases for comparison. The phylogenetic tree was constructed and key amino acid mutation sites were analyzed using MEGA7 software. Results The sequence analysis results revealed that the strain studied belongs to the H5 subtype Clade 2.3.4.4b branch. The PB1 gene shared 99.69% homology with the H5N6 virus A/duck/Hunan/S40199/2021(H5N6) from Hunan Province; PB2 had 98.58% homology with A/Whooper swan/Sanmenxia/Y36/2020(H5N8); and other sequences showed high homology with the strain A/Guangdong/1/2021(H5N6) discovered in Guangzhou in 2021. These results indicate that an infection with a recombinant H5N6 subtype avian influenza virus caused the case. The amino acid composition of the cleavage site was RERRRKR↓GLF, consistent with high-pathogenicity avian influenza characteristics. The HA sequence showed no mutations at the Q226L and G228S sites, indicating the preservation of characteristics binding to avian receptors. However, mutations at the S127P, S137A, T160A, T192R, and A267T sites increased the virus's affinity for humans. A deletion found in positions 59-70 of the NA protein stalk implies enhanced viral virulence in mammals. Mutations were noted in the internal genes; S622G in PB1, K389R and V598T in PB2, N409S in PA, N30D and T215A in M1, and P42S in NS1, which may enhance the virulence of avian influenza virus in infecting mice. Conclusions The H5N6 virus strain detected in a human case in Changsha City in this study is highly pathogenic, prone to bind to avian receptors, but numerous mutations at key amino acid sites facilitate infection in humans. Continuous monitoring and research on H5N6 avian influenza virus should be strengthened

OBJECTIVE To provide reference for improving the rational use of antimicrobial drugs in primary township medical institutions. METHODS Based on the county prescription pre-review center， a team led by anti-infective clinical pharmacists constructed the management mode for the use of antimicrobial drugs in county-level medical communities with clinical pharmacists as the main team by finding out the main problems in the use of antimicrobial drugs in primary township medical institutions， providing feedback on the problems， organizing relevant training for the problems， improving the customization rules of the prescription pre-review software， implementing the automatic interception and pharmacist online prescription review and other measures. Data on the use of antimicrobial drugs were collected and compared in the 15 primary township medical institutions between January-June in 2022 （before the implementation of the mode） and January-June in 2023 （after the implementation of the mode）. RESULTS Compared with before the implementation of the mode， the utilization rate of antimicrobial drugs in outpatients of primary township medical institutions decreased from 24.97% before the implementation of the mode to 19.39% after the implementation of the mode； the utilization rate of antimicrobial injection in outpatients decreased from 66.10% to 46.80%； the utilization rate of intravenous drip of antimicrobial drugs in outpatients decreased from 52.33% to 40.35%； the rates of combined use of antimicrobial drugs in outpatients decreased from 12.70% to 8.19%； the reasonable rate of antimicrobial prescribing in outpatients increased from 55.28% to 73.93%. After the implementation of the mode， the proportion of antimicrobial prescriptions for each diagnosis was basically the same as before； the defined daily dose system （DDDs） and proportion of a few antimicrobial drugs changed compared with before according to the anatomical therapeutic chemical classification of drugs， among which DDDs of lincomycin， gentamicin and other drugs declined significantly； DDDs of antimicrobial drugs for each classification was basically the same as before according to AWaRe classification. CONCLUSIONS The management mode of the use of antimicrobial drugs led by anti-infective clinical pharmacists is constructed in the prescription pre-reviewing center of county-level medical communities， which can effectively improve the rational use of antimicrobial drugs in the primary township medical institutions.

Nowadays,the prevalence of nonalcoholic fatty liver disease(NAFLD)is constantly rising in China and globally,and its incidence rate is increasing year by year,which has seriously affected human life and health.Lipophagy is molecular chaperone-mediated autophagy and has the functions of promoting lipolysis,maintaining the lipid homeostasis of hepatocytes,and alleviating hepatocyte fatty degeneration.Lipophagy has three main processes of lipid droplet catabolism,lipid droplet autophagy,and fatty acid β-oxidation,which are regulated by key genes,receptors,and enzymes.Currently,important advances have been achieved for the intervention methods of traditional Chinese medicine,Western medicine,diet,and exercise in the research on lipophagy,which provides new perspectives for the prevention and treatment strategies for NAFLD.

Emerging SARS-CoV-2 variants have made COVID-19 convalescents susceptible to re-infection and have raised concern about the efficacy of inactivated vaccination in neutralization against emerging variants and antigen-specific B cell response. To this end, a study on a long-term cohort of 208 participants who have recovered from COVID-19 was conducted, and the participants were followed up at 3.3 (Visit 1), 9.2 (Visit 2), and 18.5 (Visit 3) months after SARS-CoV-2 infection. They were classified into three groups (no-vaccination (n = 54), one-dose (n = 62), and two-dose (n = 92) groups) on the basis of the administration of inactivated vaccination. The neutralizing antibody (NAb) titers against the wild-type virus continued to decrease in the no-vaccination group, but they rose significantly in the one-dose and two-dose groups, with the highest NAb titers being observed in the two-dose group at Visit 3. The NAb titers against the Delta variant for the no-vaccination, one-dose, and two-dose groups decreased by 3.3, 1.9, and 2.3 folds relative to the wild-type virus, respectively, and those against the Omicron variant decreased by 7.0, 4.0, and 3.8 folds, respectively. Similarly, the responses of SARS-CoV-2 RBD-specific B cells and memory B cells were boosted by the second vaccine dose. Results showed that the convalescents benefited from the administration of the inactivated vaccine (one or two doses), which enhanced neutralization against highly mutated SARS-CoV-2 variants and memory B cell responses. Two doses of inactivated vaccine among COVID-19 convalescents are therefore recommended for the prevention of the COVID-19 pandemic, and vaccination guidelines and policies need to be updated.

Liver transplant patients require lifelong immunotherapy， and if they are infected by SARS-Cov-2， their immune function will face double whammy. This report described the integrated approach of traditional Chinese and western medicine in the treatment of a liver transplant patient with severe COVID-19. The treatment was involved with western medicine such as antiviral， immunosuppressive， focusing on maintaining immune balance. Traditional Chinese medicine was given based on the differentiation of syndromes， targeting at the core pathogenesis and using methods such as promoting qi circulation， clearing heat and resolving dampness， draining lung and relieving panting. Following the treatment， the patient exhibited notable improvement in clinical symptoms and liver function， leading to the effective cessation of disease progression and a shortened recovery period.

This paper reported a case of severe COVID-19 in the recovery stage with acute lymphoblastic leukemia treated by integrated traditional Chinese and western medicine， with the intention of shedding light on the clinical diagnosis and treatment of similar conditions. The patient， who had acute lymphoblastic leukemia， developed COVID-19 infection during the bone marrow suppression period after chemotherapy. Treatment with western medicine was mainly anti-infection， symptomatic management， and supportive care. During the recovery stage， considering the patient's chemotherapy history and disease progression， the overall syndrome was identified as deficiency of both qi and yin and binding of phlegm and blood. Based on the “state-target” combined treatment strategy， herbal prescriptions were selected and modified to address the “deficiency state”， “disease target”， and “symptom target”. In addition to western medicine， the patient was administered with Shengmai Powder （生脉散） and Compound Zhebei Granules （复方浙贝颗粒） in its modifications to boost qi， nourish yin， and reinforce healthy qi， nourish and cool the blood， ultimately achieving satisfactory therapeutic effects.

Objective:To investigate the impact of diabetes mellitus (DM) and stroke on long-term outcomes in patients with acute coronary syndrome (ACS) undergoing percutaneous coronary intervention (PCI).Methods:Total 411 consecutive ACS patients undergoing PCI at the Ninth People′s Hospital of Zhengzhou between December 2014 and June 2018 were recruited, including 319 males and 92 females with a mean age of (64.7±10.1) years. These patients were divided into 4 groups according to the presence or absence of history of diabetes or stroke: non-DM non-stroke group ( n=192) , DM group ( n=140), stroke group ( n=41), and DM+stroke group ( n=38). The data of baseline demographic characteristics, clinical feature, coronary angiographic findings, and cardiovascular adverse events during long-term follow-up were obtained. Kaplan-Meier curves were used to investigate the long-term clinical outcomes among groups. Results:The mean interval of follow-up was (24.1±13.8) months. Patients with DM+stroke had the highest rates of non-fetal myocardial infarction (χ 2=24.932) , non-fetal stroke (χ 2=9.434) , hospitalization due to heart failure/angina (χ 2=69.290) , revascularization (χ 2=22.918) , cardiovascular death(χ 2=13.473)and all-cause death(χ 2=17.724)as well as hard endpoint events (the sum of non-fetal myocardial infarction, non-fetal stroke, and all-cause death) (χ 2=30.268)and combined major adverse cardiovascular events (MACE) (the sum of hard endpoint events, hospitalization due to heart failure/angina, and revascularization) (χ 2=119.556)among 4 groups(all P<0.01). In Kaplan-Meier survival analysis, the cumulative ratio of freedom from all-cause death decreased significantly in DM+stroke group compared with no DM no stroke group ( HR=17.121, 95 %CI: 2.527-115.934, P<0.01), but no statistical difference was observed in the cumulative ratio of freedom from all-cause death between DM+stroke group and DM group or stroke group respectively ( HR=3.178, 95 %CI: 0.744-13.582; HR=1.383, 95 %CI: 0.374-5.118; all P>0.05) . Meanwhile, patients with DM+stroke presented significantly lower cumulated ratio free from combined MACE than patients with non-DM non-stroke ( HR=5.423, 95 %CI:2.941-10.036, P<0.01), and the cumulated ratio free from combined MACE also decreased significantly in DM+stroke group as compared to DM group or stroke group respectively ( HR=1.859,95 %CI: 1.167-2.962; HR=1.991,95 %CI: 1.178-3.364; all P<0.01) . Conclusions:ACS patients with combined history of DM and stroke have a worse long-term outcomes after PCI than those with DM alone or stroke alone or without DM or stroke. DM and stroke seemed to have an additive effect on decrease in the cumulative ratio free from combined MACE in ACS patients following PCI.

OBJECTIVE: To determine the types and frequency of deafness-related variants among 7875 newborns from Dongying area of Shandong Province. METHODS: One hundred loci of 18 common deafness genes were subjected to semiconductor sequencing. Variant site, frequency and distribution of the variants were analyzed. RESULTS: In total 552 deafness gene variants were detected among the 7875 newborns, which yielded a detection rate of 7.01%. Among these, common variant sites for GJB2, SLC26A4 and GJB3 genes were c.235delC, IVS7-2A>G and c.538C>T, respectively. The variant frequencies of matrilinear inheritance deafness genes MT-CO1, MT-RNR1, MT-TL1 and MT-TS1 were 0.38%, 0.25%, 0.1% and 0.01%, respectively. Four newborns were diagnosed with deafness, among which one had unilateral hearing loss. Analysis of the proportions of neonatal deafness-related variants in five counties of Dongying showed that the highest variant rate for the SLC26A4 gene compared with GJB2 was in Lijin county (51.76% vs. 40%), while the lowest was in Hekou county (30.77% vs. 56.41%). CONCLUSION The carrier rate of deafness-related variants in Dongying area is higher than other regions of China, which may be attributed to the increased types and variant sites covered by the semiconductor sequencing method compared with the chip method and time-of-flight mass spectrometry. Due to geographical and population aggregation factors, the proportion of deafness variants in the five counties of Dongying differed significantly. Above results may provide a guide for the prevention of congenital deafness in Dongying area.

Objective: To establish and evaluate a microfluidic chip platform for the rapid diagnosis of post-neurosurgical bacterial infection. Methods: The pathogens isolated from patients with post-neurosurgical bacterial infection in Beijing Tiantan Hospital Affiliated to Capital Medical University during 2007 and 2016 and the epidemiological data from China drug resistance monitoring network CHINET were analyzed retrospectively. Based on the retrospective data and the molecular epidemiological information of drug-resistant bacteria reported in the literature, target pathogens and drug resistance gene parameters were selected. The microbial identification parameters from 10 different bacteria, including Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus epidermidis, Enterobacter cloacae, Staphylococcus aureus, Escherichia coli, Enterococcus faecium, Enterococcus faecalis, Stenotrophomonas maltophilia and Pseudomonas aeruginosa, and the parameters of 15 drug resistance genes, including mecA, vanA, vanB, aacC1, aadA1, bla CTX-M-1 , bla CTX-M-9 , bla GES-1 , bla OXA-23 , bla OXA-24 , bla OXA-58 , bla OXA-66 , bla KPC-2 , bla IMP-4 and bla VIM-2 , were selected for designing a microfluidic chip platform. Using MAIDI-TOF MS for bacterial identification, multiplex PCR for the detection of drug resistance genes, micro-broth dilution method for the detection of drug resistance phenotypes and ESBLs screening test as reference methods, 13 known bacteria were used to evaluate the preliminary performance of the established microfluidic chip platform, and 108 cerebrospinal fluid bacterial culture positive specimens were used to evaluate the clinical application value of the microfluidic chip platform. Results: The identification rates of 13 known strains and the coincidence rate of drug resistance genes were 100%. The coincidence rate of identification results for 108 cerebrospinal fluid bacterial culture positive specimens between the microfluidic chip platform and the MALDI-TOF MS method was as high as 94.44%. The coincidence rates of drug resistance phenotype of carbapenems, oxacillin, vancomycin, ESBLs and genotype between the microfluidic chip platform and the micro-broth dilution method or ESBLs screening test were above 90%. Conclusion The established microfluidic chip platform is fast and accurate, and has application value in microbial identification and the prediction of drug resistance, which may be used as an important supplementary method in the diagnosis of post-neurosurgical bacterial infection.

Objective To investigate the diagnostic value of procalcitonin ( PCT) and lactate in cerebrospinal fluid (CSF) combined with conventional biomarkers for post-neurosurgical bacterial meningitis (PBM).Methods Clinical data of 213 patients with post-neurosurgical meningitis admitted in Beijing Tiantan Hospital, Capital Medical University from March 2017 to December 2017 were retrospectively analyzed, including 85 cases of PBM and 128 cases of post-neurosurgical aseptic meningitis ( PAM).The diagnostic value of CSF procalcitonin , lactate and other 12 conventional biomarkers for PBM was analyzed by multivariate logistic regression.A prediction algorithm was generated and its diagnostic value for PBM was assessed with receiver operating characteristic curve (ROC).Results The univariate analysis showed that CSF cell count, CSF leukocyte count , CSF protein concentration , CSF glucose concentration, CSF glucose／blood glucose ratio, CSF PCT and CSF lactate were significantly associated with PBM.Multivariate logistic regression analysis showed that CSF PCT , CSF lactate, CSF protein concentration and CSF glucose ／blood glucose ratio were independent predictive factors for PBM.The predictive algorithm score ＝4.315 ×CSF PCT+0.822×CSF Lactate+0.009×CSF protein concentration -5.480×CSF glucose／blood glucose ratio-3.074.The predictive algorithm has the largest area under the ROC curve ( AUC ＝0.947), and the sensitivity and specificity of the predictive algorithm score were 90.60% and 85.10%, respectively.The positive predictive value , negative predictive value and the accurate rate of the algorithm in diagnosis of PBM were 84.06%, 94.44% and 90.40%, respectively.Conclusion The predictive algorithm based on the combination of CSF PCT and CSF lactate with CSF protein concentration and CSF glucose ／blood glucose ratio has a good diagnostic value for PBM.It can shorten the diagnosis time of PBM and improve the clinical outcomes.