Since the proposal of the concept of the gut-liver axis,a large number of studies have focused on exploring the connection between gut microbiota and liver disease;however,the idea of using pyroptosis as a hub to explore the intrinsic mechanism of gut-liver crosstalk is still in its infancy.This article mainly describes the process by which gut microbiota dysbiosis affects the integrity of mucosal barrier and bile acid metabolism,induces pyroptosis,and thereby affects the development and progression of liver diseases,and it also concludes that gut microbiota dysbiosis affects liver diseases by inducing NLRP3/AIM2/Caspase-1-dependent,Caspase-4/11/GSDMD-dependent,and Caspase-3/GSDME-dependent pyroptosis.In summary,this study aims to provide new ideas and targets for the future diagnosis and treatment of liver diseases by establishing the connection between pyroptosis and intestinal-liver immune crosstalk.

Biochemical analysis is of great significance in many fields,such as disease diagnosis,environmental pollution source identification,food safety,etc.Immunoassay based on the reaction of antigen and antibody has the characteristics of high speci-ficity and diversity.Compared with other analytical methods such as liquid chromatography,which requires a large and expensive in-strument,immunodetection has the advantages of simplicity and good selectivity and is thus widely used in sample testing.Quench-body(Q-body)is a novel fluorescent immunosensor based on antigen-antibody reaction.It is constructed by labeling antibody fragment with fluorescent dye.Tryptophan in antibody leads to electron transfer and quenching of fluorescent,when it specifically binds with an-tigen,its fluorescence intensity recovers and presents concentration dependence.Therefore,the antigen concentration can be detected by detecting the change of fluorescence intensity of the Q-body.Q-body immunoassay is widely used in the rapid detection of environ-mental pollutants and disease markers due to its simple operation and high sensitivity,which is of great significance for environmental detection and disease diagnosis.In this paper,the principle,construction method,detection technology development and domestic and foreign research progress of Q-body are comprehensively discussed,and the future application of this technology is prospected.

In order to achieve the purpose of rapid detection of duck astrovirus type 1(DAstV-1),specific primers were designed based on the conservative region of ORF1a which belonged to DAstV-1(WF1202 strain).A real-time fluorescent quantitative PCR(qPCR)detective method for DAstV-1 was established.Clinical samples were detected by the qPCR method and the positive samples were used for virus isolation and identification.Results showed that the detection limit of the established method was 4.64×103 copies/μL,which was 10 times higher than the normal RT-PCR method.In addition,no cross-reactions were found with other common infectious disease pathogens in poultry,indicating that the qPCR method had good specificity.What's more,the coef-ficient of variations(Cv)in intra-and inter-assays were 0.85％-2.85％and 0.21％-2.94％,re-spectively,both less than 3％,indicating that the qPCR method had a good repeatability.Using this method,35 tissue samples from different duck farms in 10 provinces from 2020 to 2022 were detected for DAstV-1.Results showed that the positive rate was 25.71％(9/35),and the coinci-dence rate was 94.29％when compared with the normal RT-PCR method.A positive sample ran-domly taken for the virus isolation through duck embryo passage,and the allantoic fluid was col-lected and then was verified by the qPCR method and inoculated with 1-day-old healthy ducklings for the animal regression experiment.The infected ducklings suffered from transient disease but did not die.The liver tissues were all positive with DAstV-1 when detected by qPCR.Meanwhile,autopsy showed that there were slight changes in the livers,and the histopathological observation showed that the liver cells were steatosis.These findings indicated that the isolated DAstV-1 strain had weak pathogenicity and might be a low virulent strain.To sum up,the qPCR detection method of DAstV-1 was successfully established in this work,and could provide technical support for clini-cal diagnosis,isolation and identification,and molecular epidemiology monitoring of DAstV-1.

In order to establish a rapid,specific and sensitive detection method for Streptococcus equi subspecies zooepidemicus(SEZ)and to understand the infection status of SEZ in horses ente-ring China,specific primers were designed and synthesized based on the conserved gene comB of standard strain SEZ(ATCC 43079)in this work.Then,the pMD19-T-comB recombinant plasmid was constructed and used as a standard positive template.After that,the fluorescence-based quantitative PCR(qPCR)detection method based on SYBR Green Ⅰ dye was established.Totally,477 equine entry serum samples from 6 countries,including Netherlands,Belgium,Japan,Germa-ny,Argentina and New Zealand,during 2018 to 2023,were randomly selected and detected for SEZ by the qPCR method.Results showed that the established qPCR method had specific amplification for only SEZ,which illustrated a good specificity.Sensitivity test of the method showed that the limited detection amount was 4.58 X101 copies/μL.And the repeatability test showed that the coef-ficient of variation of intra-batch repeatability was less than 0.5％,while the inter-batch repeat-ability was less than 3.0％,which indicated good repeatability and high stability.Retrospective a-nalysis showed that totally 11 of 477 positive samples were detected,with a relatively low positive rate of 2.31％(11/477).Among them,all the 40 samples from Netherlands in 2018 were negative(0/40).In the samples of 2019,one positive was detected from Belgium(1/20),while all other 36 samples which form Japan and Germany were negative.In the samples of 2021,three samples(3/34)from Japan and one sample(1/20)from Argentina were positive,and all the other 40 samples from the Netherlands were negative.In the samples of 2022,76 samples from Netherlands were all negative.While in the 2023,5(5/126)of 126 samples from Netherlands and one(1/88)of 88 from New Zealand were found positive with SEZ.To summarize,The SYBR Green Ⅰ qPCR method for the diagnosis of SEZ was successfully established,and it could provide necessary technical support for the rapid quarantine of China's entry-exit and port departments,as well as the epidemiological investigation of the disease.

In order to understand the prevalence of Akabane disease(AKAD)in Guizhou Province and the molecular characteristics of the isolates,the whole-genome sequence of a strain of Akabane virus(AKAV)from a bovine AKAD-positive sample was determined and analyzed.The genotype and genetic variation of the strain were also explored.Based on the conserved S sequence,a fluores-cence quantitative PCR(qPCR)detection method was established and applied for the investigation of AKAV infection status in four large-scale beef cattle farms of Guizhou.Results showed that the S,M and L fragments of the bovine strain were highly homologous to the Tianjin strain(TJ2016/China/2016)and the Australian strain(JaLAB39/Australia/1959),where they were in the same evolutionary branch and belonged to genotype Ⅱ.Sensitivity assay found that the lowest detection limit was 2.5 X 101 copies/μL.Specificity assay showed the established method detected only AKAV with no amplification on bovine bluetongue virus(BLUV),Pasteurella multocida(PM),bovine infectious rhinotracheitis virus(IBRV)and bovine Mycoplasma bovis.The variation coefficients of inter-and intra batches in the repeatability test were both lower than 2.26％.These findings illus-trated that the established qPCR method had high sensitivity,good specificity and repeatability.A total of 298 serum samples from 4 large-scale beef cattle farms in Qianxi City and Huangping County of Guizhou Province were collected and tested for AKAV by the method.Out of 298 sam-ples,25 positive samples(25/298)were detected as positive with a positive rate of 8.39％.In sum-mary,this work provided the reference data for a deep understanding of the molecular prevalence of AKAV in Guizhou Province and laid foundation for the prevention and control of AKAD.

OBJECTIVE To provide reference for rational use of cyclin-dependent kinase 4/6 （CDK4/6） inhibitors. METHODS Retrieved from Web of Science， PubMed， SpringerLink， CNKI， Wanfang Data and VIP database， and so on， the literature about lung toxicity related to CDK4/6 inhibitors were collected and analyzed statistically with Excel 2013 software. RESULTS A total of 12 literature which met the inclusion and exclusion criteria were included； 13 patients were involved， among which 3 cases were from the United States， 3 from Japan， 2 from India， and 1 from Israel， Spain， France， Australia and Saudi Arabia respectively； all patients were female， aged between 43-89 years， of whom 8 were treated with palbocicilib， 3 with abemacilib， and 2 with ribociclib. The lung toxicity of patients after medication occurred from 1 week to 15 months； the majority of patients were hospitalized with the symptom such as difficulty breathing， chest tightness， shortness of breath， dry cough， etc. The lung toxicity mainly manifested as interstitial lung disease， eosinophilic pneumonia， mediastinal and pulmonary granulomatous reaction， drug-induced pneumonia， diffuse alveolar damage， organizing pneumonia and so on. The shortest treatment duration was 3 weeks， and the longest was 6 months. The treatment measures included drug withdrawal， intravenous use of antibiotics， intravenous use of systemic steroids， oxygen inhalation， and so on； after treatment， 8 patients improved or recovered， and 5 patients died due to deterioration. One patient developed lung toxicity again after reuse of such drugs and must stop drugs permanently. CONCLUSIONS Lung toxicity related to CDK4/6 inhibitors possibly cause mortality. It is necessary to make early judgment， stop the drug in time， and give patients systemic steroids， oxygen inhalation and other treatment measures as soon as possible.

Objective:To evaluate the immunogenicity of a quadrivalent influenza virus subunit vaccine in a healthy population aged 3-64 years.Methods:Healthy people aged 3-64 years old were selected as the study subjects, and a randomized, blind, positive controlled, non-inferiority test was adopted. The subjects were randomly inoculated with one dose of the corresponding experimental vaccine or control vaccine in a ratio of 1∶1. Blood samples were collected from all subjects before and at 28 d after immunization, and hemagglutination inhibition (HI) test was used to measure the levels of antibodies against H1N1, H3N2, B/Victoria (BV) and B/Yamagata (BY) in serum. The geometric mean titers (GMT), geometric mean increase (GMI), positive conversion rates and protection rates of antibodies against the four types of viruses were analyzed.Results:A total of 2 157 subjects aged 3-64 years were included, including 1 074 in the experimental group and 1 083 in the control group. There were no significant differences in the GMT or protection rates of antibodies against H1N1, H3N2, BV or BY before immunization between the two groups ( P>0.05), and the two groups were balanced at baseline. After full immunization, the GMI of antibodies to H1N1, H3N2, BV and BY in the experimental group was 11.16, 17.77, 9.61 and 15.13, respectively; the positive conversion rates were 84.08% (903/1 074), 92.46% (993/1 074), 86.03% (924/1 074) and 91.71% (985/1 074), respectively; the protection rates were 96.74% (1 039/1 074), 97.58% (1 048/1 074), 88.08% (946/1 074) and 94.97% (1 020/1 074), respectively. In the experimental group, the GMT of each antibody increased by more than 2.5 times after immunization; the lower limit of the 95%CI of the positive conversion rate was higher than 40%, and the lower limit of the 95%CI of the protection rate was higher than 70%. The lower limit of the 95%CI of the difference in the positive conversion rate of each antibody between the experimental group and the control group was >-10%, and the lower limit of the 95%CI for GMT (experimental group)/GMT (control group) was over 2/3. Conclusions:The experimental vaccine had good immunogenicity and was non-inferior to the control vaccine in the population aged 3-64 years.

Objective:To understand the health literacy of young and middle-aged stroke patients and analyze the influencing factors.Methods:A total of 270 young and middle-aged stroke patients treated in ICU of Neurology Department of the First Affiliated Hospital of Chongqing Medical University from November 2020 to May 2022 were selected as the research objects by the convenient sampling method. The General Data Questionnaire, National Institutes of Health Stroke Scale and Stroke Patient Health Literacy Scale were used to investigate the patients. Binomial Logistic regression analysis was used to explore the influencing factors of health literacy in young and middle-aged stroke patients. A total of 270 questionnaires were sent out in this study, and 266 were effectively received, with the effective recovery of 98.52% (266/270) .Results:The total score of the health literacy scale for young and middle-aged stroke patients was (73.44±6.73) , and the patients with health literacy accounted for 33.46% (89/266) . The score rates of the three dimensions in Stroke Patient Health Literacy Scale from high to low were basic skills, healthy lifestyle and behavior and basic knowledge and concept. Binomial Logistic regression analysis showed that gender, educational level, monthly family income and self-rating doctor-patient communication were the influencing factors of health literacy in young and middle-aged stroke patients ( P<0.01) . Conclusions:The health literacy of young and middle-aged stroke patients needs to be improved. The health literacy level of young and middle-aged male stroke patients with low education level, low family monthly income and poor or average self-rated doctor-patient communication needs more attention and intervention. Health education and health promotion should be done according to the characteristics of young and middle-aged stroke patients.

Objective:To investigate the expression levels and clinical significance of glioma-associated oncogene homolog 1 (GLI1) and sonic hedgehog signaling molecule (Shh) in the malignant transformation of ovarian endometriosis (EM).Methods:The expressions of GLI1 and Shh were detected by real-time reverse transcription (RT)-polymerase chain reaction (PCR) and EnVision method in 50 cases of ovarian EM tissues, 35 cases of atypical endometriosis (aEM) and 50 cases of endometriosis-associated ovarian cancer (EAOC). The expression differences of two molecular markers in the malignant transformation of ovarian EM were compared, and the relationships between two molecular markers and the clinicopathological features and prognosis of EAOC were analyzed.Results:(1) RT-PCR showed that the expression levels of GLI1 mRNA in EM, aEM and EAOC group were 1.77±0.40, 3.54±0.44, and 7.80±0.24, respectively. The expression levels of Shh mRNA were 0.95±0.21, 3.14±0.35, and 5.41±0.31, respectively. GLI1 and Shh mRNA in EAOC group were significantly higher than those in EM and aEM group (all P<0.01), and there were statistically significant differences between EM and aEM group (all P<0.01). The percentages of GLI1 in ovarian EM, aEM and EAOC were 32% (16/50), 57% (20/35), and 66% (33/50), respectively, meanwhile, the positive expression rates of Shh were 20% (10/50), 49% (17/35), and 54% (27/50), respectively (all P<0.01). GLI1 mRNA expression was positively correlated with Shh mRNA expression in EAOC tissues ( r=0.721, P<0.01). The expressions of GLI1 protein were proportionated to Shh protein in EAOC tissues ( r=0.608, P=0.001). (2) The expression of GLI1 was significantly related to the International Federation of Gynecology and Obstetrics (FIGO) stage, cancer antigen 125 (CA 125) levels, lymph node metastasis, and Platinum resistance in EAOC patients (all P<0.05). The expression of Shh were related to FIGO stage and lymph node metastasis in EAOC patients (all P<0.05). Logistic regression analysis showed that GLI1 expression was an independent risk factor for poor prognosis in EAOC patients ( P<0.05). Kaplan-meier survival analysis showed that the overall survival rate of EAOC patients with high GLI1 expression and low GLI1 expression was 12.1% and 35.3%, respectively, with statistical significance ( χ2=10.73, P<0.01). The overall survival rate of EAOC patients with high and low expression of Shh protein was 11.1% and 30.4%, in which there was statistically significant difference ( χ2=3.96, P=0.047). Conclusion:GLI1 and Shh are highly associated with the malignant transformation of ovarian EM, which may play a role in promoting malignant degeneration of ovarian EM, and the high expression of GLI1 and Shh indicates a poor prognosis in EAOC patients.

Objective:To determine whether insulin resistance is associated with all-cause mortality in subjects without diabetes.Methods:A total of 505 participants without diabetes, 198 with normal glucose tolerance (NGT) and 307 with impaired glucose tolerance (IGT), were recruited from the Daqing Diabetes Study. The participants were followed up for 30 years. They were stratified into three groups (tertiles) according to baseline homeostasis model assessment of insulin resistance(HOMA-IR) levels, as the HOMA-IR 0, the HOMA-IR 1 and the HOMA-IR 2 groups, to assess the predictive effect of insulin resistance on risk of all-cause mortality.Results:During the 30-year follow-up, 52, 56 and 78 participants died across the three HOMA-IR groups, respectively. The corresponding mortality per 1 000 person-years (95 %CI) were 12.12 (9.56-15.01), 13.10 (10.46-16.03) and 19.91 (16.73-23.15), respectively. Participants in the HOMA-IR 2 group had a significantly higher risk of death than those in the HOMA-IR 0 group after adjustment of age, sex and smoking status ( HR=1.97,95 %CI 1.38-2.81, P<0.001). Cox analyses showed that a one standard deviation increase in HOMA-IR was associated with a 22% increase in the mortality after adjustment of potential confounders ( HR=1.22, 95 %CI 1.08-1.39, P=0.002). Conclusions:Insulin resistance is associated with increased risk of all-cause death in Chinese people without diabetes, suggesting that improving insulin resistance could be beneficial for people without diabetic in reducing risk of long-term all-cause mortality.