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 observe the impact of mild hypothermia (MH)on the reactive oxygen species (ROS)and expression of cacpase-3mRNA and light chain 3 (LC3,a subunit of immunoglobulin)in hippocampus nerve cells of rats after cardiopulmonary resuscitation (CPR).Methods A total of 65 healthy male Sprague Dawley (SD)adult rats were randomly (random number)divided into 2 groups:blank control group (n =5)and CPR group (n =60).Cardiac arrest (CA)was induced in rats of CPR group by asphyxia.The survival rats after CPR were randomly (random number)divided into 2 groups:normothermia CPR group (NT)and hypothermia CPR group (HT).Homeothermia of 37 ℃ was maintained in NT group after restoration of spontaneous circulation (ROSC),and hypothermal intervention to 32 ℃ was carried out in HT group for 4 hours immediately after ROSC.Both NT group and HT group were then randomly divided to 2 subgroups 12 hours and 24 hours after ROSC (NT-12,NT-24,HT-12,HT-24 subgroups).During observation,the neurological deficit (NDS)of rats was scored,then the bilateral hippocampi were obtained from rats'head,and monoplast suspension of fresh hippocampus tissue was made immediately to determine the level of intracellular ROS by flow cytometry.Transmission electron microscope was used to observe the ultrastructure changes of cellular nucleus and mitochondria.Reverse transcription-polymerase chain reaction (RT-PCR)was used to determine the expression of caspase-3mRNA and Western-blotting (WB)was used to determine the level of LC3 in frozen hippocampus tissue.Measured data were analyzed with paired sample T test and One-Way ANOVA.Results Of 60 rats with CA,44 were successfully resuscitated (73%)and 33 survived until the end of the experiment (55%).The NDSs of rats in NT and HT groups were significantly reduced in comparison with BC group (F=8.107,P<0.05),while the NDSs of rats in HT-12 subgroup and HT-24 subgroup were significantly increased in comparison with NDSs of rats in NT-12 subgroup and NT-24 subgroup,respectively (t=9.692,P<0.01;t=14.374,P<0.01 ).The ROS in hippocampus nerve cells of rats in NT group and HT group were significantly increased compared to BC group (F=16.824,P<0.05 ),whereas the ROS in HT-12 and HT-24 subgroups were significantly reduced compared to ROS in NT-12 and NT-24 subgroups,respectively (t =9.836,P<0.01;t =7.499,P<0.01).The expressions of caspase-3 mRNA in hippocampus nerve cells of rats in NT and HT groups were significantly increased compared to BC group (F=24.527,P<0.05),while the expressions of caspase-3 mRNA in rats of HT-12 and HT-24 subgroups were significantly reduced compared to NT-12 and NT-24 subgroups,respectively (t =6.935,P <0.01;t =4.317,P <0.01 ).The level of LC3B-II/I in hippocampus nerve cells of rats in NT and HT groups were significantly increased compared to BC group (F=6.584,P<0.05),while the levels of LC3B-II/I in rats of HT-12 and HT-24 subgroups were significantly reduced compared to NT-12 and NT-24 subgroups,respectively (t=10.836,P<0.001;t=2.653,P=0.02).Ultrastructure damage of nucleus and mitochondria in NT group was more evident compared to BC group,and eumorphism of nucleus and mitochondria were maintained in rats of HT group compared to NT group.Conclusions The mild hypothermia reduced the injury of nerve cells and improved the neurological function of rats survived from cardiac arrest likely by reducing ROS production of nerve cells and inhibition the expression of caspase-3mRNA and lowering the level of LC3 leading to reducing cellular apoptosis and massive autophagy in rats survived from cardiac arrest after CPR.

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.

Marked by the Third Plenary Session of the Eleventh Central Committee of the CPC, Shanghai health care started a journey of reform and opening up as with other sectors. This essay reviews various stages Shanghai health care experienced and the achievements for the last 30 years since reform and opening up, and analyzes the challenges Shanghai health care faces currently. On this basis, the essay summaries the experiences and lessons learned from Shanghai health care reform and development.

Under the general requirements of China's national health reform, the basic framework of Shanghai's health reform was constructed at five different aspects the long term vision, goal, strategies, measures, supportive reform activities. The general thought of Shanghai's health reform can be concluded into two aspects: first, establishing a basic healthcare system of covering both urban and rural residents universally; second, enhancing the foundation of enabling sustainable health development.

Objective:to apply Fuzhengkangai AIDS particles combined therapy with effective antiretroviral therapy (HAART) at the beginning of the HIV/AIDS (human immunodeficiency virus/acquired immunodeficiency syndrome) patients,observe its clinical curative effect and improve the related adverse reactions.Methods:a multicenter,randomized,double-blind,placebo-controlled clinical research method,divided into experimental group and the control group (n =57),experimental group to "Fuzhengkangai particles" joint efficient antiretroviral therapy (HAART) antiviral treatment,control group to "Fuzhengkangai particles" placebo (5％ low dose) joint efficient antiretroviral therapy (HAART) antiviral treatment,treatment for 3 months.Two groups of patients with simultaneous liver and kidney function,blood routine,CD4 + T lymphocyte count,the observation of TCM symptoms and signs,survival quality of integral.Results:after 12 weeks of treatment,the control group and experimental group,the experimental group significantly elevated levels of peripheral blood RBC,lower levels of CR,reduce the signs and symptoms (total score,fatigue,stay,and spontaneous perspiration)integral,higher levels of CD4 + T lymphocyte count and survival quality score,the difference had statistical significance (p ＜ 0.05).Conclusion:Fuzhengkangai particles joint efficient antiretroviral therapy (HAART) antiviral treatment,can improve HAART after initial treatment of the patient's clinical symptoms,blood toxicity,the quality of survival,reduce the antiviral treatment of adverse reactions,combination drug alone can significantly increase the CD4 + T lymphocytes,so as to improve clinical curative effect.

Objective To evaluate the regulatory effect of Diwu Yanggan(DWYG)Decoction on lysoglycerophospholipids(Lyso-GPLs)in circulating exosomes in a mouse model of nonalcoholic fatty liver disease(NAFLD).Methods Circulating exosomes isolated from mouse serum by size exclusion chromatography were morphologically characterized using transmission electron microscope and examined for surface markers CD9,CD63 and TSG101 using Western blotting.Twenty-four male Kunming mice were randomized into 3 groups for normal feeding(control,n=8)or high-fat diet feeding for 1 week to induce NAFLD,after which the latter mice were given DWYG decoction(treatment group,n=8)or normal saline(model group,n=8)by gavage for 4 weeks.After the last treatment,blood samples were collected from the mice for testing serum TC,HDL-C,LDL-C,ALT and AST levels and isolating circulating exosomes.Using multivariate statistical analysis based on targeted metabolomics strategy,the potential biomarkers for Lyso-GPLs in the exosomes were screened.Results The isolated exosomes about 100 nm in size had a typical saucer-like structure with distinct double-layer membranes and a mean particle size of 137.5 nm and expressed the specific surface marker proteins CD9,CD63 and TSG101.The mouse models of NAFLD had significantly increased serum levels of TC,HDL-C,LDL-C and AST and lowered serum ALT level.A total of 43 Lyso-GPLs with significant reduction after DWYG Decoction treatment were identified in NAFLD mice.Conclusion DWYG Decoction can regulate Lyso-GPLs in circulating exosomes in NAFLD mice,which provides a new clue for studying the therapeutic mechanism of DWYG Decoction for liver disease.

Objective To evaluate the regulatory effect of Diwu Yanggan(DWYG)Decoction on lysoglycerophospholipids(Lyso-GPLs)in circulating exosomes in a mouse model of nonalcoholic fatty liver disease(NAFLD).Methods Circulating exosomes isolated from mouse serum by size exclusion chromatography were morphologically characterized using transmission electron microscope and examined for surface markers CD9,CD63 and TSG101 using Western blotting.Twenty-four male Kunming mice were randomized into 3 groups for normal feeding(control,n=8)or high-fat diet feeding for 1 week to induce NAFLD,after which the latter mice were given DWYG decoction(treatment group,n=8)or normal saline(model group,n=8)by gavage for 4 weeks.After the last treatment,blood samples were collected from the mice for testing serum TC,HDL-C,LDL-C,ALT and AST levels and isolating circulating exosomes.Using multivariate statistical analysis based on targeted metabolomics strategy,the potential biomarkers for Lyso-GPLs in the exosomes were screened.Results The isolated exosomes about 100 nm in size had a typical saucer-like structure with distinct double-layer membranes and a mean particle size of 137.5 nm and expressed the specific surface marker proteins CD9,CD63 and TSG101.The mouse models of NAFLD had significantly increased serum levels of TC,HDL-C,LDL-C and AST and lowered serum ALT level.A total of 43 Lyso-GPLs with significant reduction after DWYG Decoction treatment were identified in NAFLD mice.Conclusion DWYG Decoction can regulate Lyso-GPLs in circulating exosomes in NAFLD mice,which provides a new clue for studying the therapeutic mechanism of DWYG Decoction for liver disease.

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.