Objective This study conducted molecular biological identification of the viruses carried by Culex pipiens pallens specimens collected in Shunyi District, Beijing in 2023, and observed the changes in the infection rate of the viruses carried by Cx. pipiens pallens at different collection times. Methods Cx. pipiens pallens were collected using carbon dioxide mosquito traps. The mosquito samples were ground in batches and analyzed by molecular biology technologies. The virus infection rate at different collection times was analyzed statistically. Results 17 strains of Culex pipiens pallens densovirus(CppDNV)were identified from Cx. pipiens pallens samples collected in Shunyi District, Beijing, in 2023. The nucleotide sequence analysis of the virus genome coding region showed that CppDNV was a single-stranded DNA virus with a total length of 3 335 nt, encoding 2 non-structural proteins(NS1, NS2)and 1 capsid protein(VP). The nucleotide(amino acid)sequence lengths of the three proteins were 2 376 nt(791 aa),1 092 nt(363 aa)and 1 071 nt(356 aa), respectively. Phylogenetic analysis showed that CppDNV was located in genus Brevihamaparvovirus. Statistical analysis showed no significant difference in infection rates across collection times(χ2=4.429, P=0.194). Conclusions CppDNV was identified in Cx. pipiens pallens in Beijing, and it was stably maintained in this natural population.

Objective:To establish a mouse model of liver cancer resistant to PD-1 monoclonal antibody and analyze the changes in its metabolomics to explore the potential mechanism of drug resistance.Methods:BALB/c mice were randomly divided into control and treatment groups after being loaded with tumor,and a normal group was additionally set up.The normal and control groups were injected with saline,and the treatment group was injected with PD-1 monoclonal antibody,after which the mice in the treatment group were screened for drug resistant and response groups.Observed the drug-resistant situation,body mass,tumor growth and survival rate of mice in each group,calculate the spleen index.The pathological features of tumor tissues were observed by HE staining method.Serum metabolites were detected by non-targeted metabolomics.Finally,a bivariate Pearson correlation analysis was conducted between the differential serum metabolites and tumor size.Results:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established,and the drug resistance rate of the mice was 50％.Compared with the normal and response groups,mice in the resistant group showed an increase in body weight,a significant increase in tumor volume,a decrease in survival rate,and a significant increase in splenic index.There was less lymphocyte infiltration in the tumor tissue.Metabolomics analysis showed that the serum levels of glutamic acid and aspartic acid increased and malic acid decreased in the resistant mice compared with the response group,and these changes were closely related to the arginine biosynthesis pathway.Conclusions:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established.The changes in its peripheral serum metabolomics mainly involve arginine metabolism and the related changes of aspartate,malate and glutamate.

Objective:To analyze the clinical features, imaging features and diagnosis and treatment process of a female pelvic epithelioid inflammatory myofibroblastic sarcoma (EIMS) with peritoneal metastasis, so as to improve the clinical understanding and diagnostic ability of the disease and avoid misdiagnosis and missed diagnosis.Methods:The clinical data of a female patient with pelvic EIMS combined with peritoneal metastasis in the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed.Results:The triad examination of the patient involved an irregular solid mass in the pelvic cavity about 8.0 cm×9.0 cm in size. Laboratory examination revealed human epididymal protein 4(HE4)154.00 pmol/L. The ultrasonography showed multiple low-echo masses in deep pelvic cavity with unclear boundary and uneven internal echo, and color doppler flow imaging (CDFI) showed abundant internal blood flow signals. Enhanced CT showed uneven and obvious enhancement of the lesion. Pathological examination showed the infiltration of inflammatory cells in the mucous interstitial background. The tumor cells were round and epithelioid, with large nuclei, deep staining and obvious nucleolus. Immunohistochemistry showed anaplastic lymphoma kinase (ALK) positive, and molecular pathology fluorescence in situ hybridization showed ALK gene amplification (positive). Combined with pathological, immunohistochemical and genetic tests, EIMS was diagnosed. Conclusions:EIMS should be considered when there are single or multiple solid or cystic nodules or masses in the pelvic cavity with obvious enhancement, invasive growth and peritoneal implantation metastasis, and the correct diagnosis can be made according to the pathological findings, immunohistochemistry and genetic test results.

Objective To analyze the epidemiological and clinical characteristics of scrub typhus in Liuyang People's Hospital Hunan Province for better diagnosis,treatment,prevention and control of scrub typhus.Methods A retrospective study was conducted on 159 confirmed cases of scrub typhus.The demographic data of patients,clinical manifestations of scrub typhus,laboratory tests,and chest CT findings,complications,treatment,and outcomes were analyzed.Results The 159 patients with scrub typhus included 142 males and 17 females.The average age of patients was(53.8±11.9)(18-82)years old.The peak incidence of scrub typhus was in the period from July to September(87.42％).The common clinical manifestations included fever(97.48％),headache(41.51％),and fatigue(23.90％).The most common signs were eschar(92.45％)on the lower limbs(25.16％),scrotum(15.72％),and buttocks(11.32％).Laboratory test results upon admission showed decreased platelet count(64.15％),elevated creatinine(38.36％),elevated AST(90.57％),ALT(80.50％),and LDH(90.57％).Pulmonary imaging study revealed pulmonary inflammation(22.64％),pleural effusion(13.21％),and pericardial effusion(6.29％).The common complications included liver impairment(69.81％)and decreased platelet count(50.94％),as well as other complications such as renal dysfunction,myocardial injury,sepsis,respiratory failure,and disseminated intravascular coagulation(DIC).Doxycycline-based therapies resulted in cure rate of 98.11％(156/159).Three patients died due to severe complications.Conclusions The peak incidence of scrub typhus was in the period from July to September in Hunan Province.Eschar is a key feature for clinical diagnosis.Scrub typhus can affect multiple organ systems,leading to various systemic complications.The cure rate is high with doxycycline treatment,but some patients may die from severe complications due to delayed medical treatment.

Objective:To establish a mouse model of liver cancer resistant to PD-1 monoclonal antibody and analyze the changes in its metabolomics to explore the potential mechanism of drug resistance.Methods:BALB/c mice were randomly divided into control and treatment groups after being loaded with tumor,and a normal group was additionally set up.The normal and control groups were injected with saline,and the treatment group was injected with PD-1 monoclonal antibody,after which the mice in the treatment group were screened for drug resistant and response groups.Observed the drug-resistant situation,body mass,tumor growth and survival rate of mice in each group,calculate the spleen index.The pathological features of tumor tissues were observed by HE staining method.Serum metabolites were detected by non-targeted metabolomics.Finally,a bivariate Pearson correlation analysis was conducted between the differential serum metabolites and tumor size.Results:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established,and the drug resistance rate of the mice was 50％.Compared with the normal and response groups,mice in the resistant group showed an increase in body weight,a significant increase in tumor volume,a decrease in survival rate,and a significant increase in splenic index.There was less lymphocyte infiltration in the tumor tissue.Metabolomics analysis showed that the serum levels of glutamic acid and aspartic acid increased and malic acid decreased in the resistant mice compared with the response group,and these changes were closely related to the arginine biosynthesis pathway.Conclusions:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established.The changes in its peripheral serum metabolomics mainly involve arginine metabolism and the related changes of aspartate,malate and glutamate.

Objective:To analyze the clinical features, imaging features and diagnosis and treatment process of a female pelvic epithelioid inflammatory myofibroblastic sarcoma (EIMS) with peritoneal metastasis, so as to improve the clinical understanding and diagnostic ability of the disease and avoid misdiagnosis and missed diagnosis.Methods:The clinical data of a female patient with pelvic EIMS combined with peritoneal metastasis in the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed.Results:The triad examination of the patient involved an irregular solid mass in the pelvic cavity about 8.0 cm×9.0 cm in size. Laboratory examination revealed human epididymal protein 4(HE4)154.00 pmol/L. The ultrasonography showed multiple low-echo masses in deep pelvic cavity with unclear boundary and uneven internal echo, and color doppler flow imaging (CDFI) showed abundant internal blood flow signals. Enhanced CT showed uneven and obvious enhancement of the lesion. Pathological examination showed the infiltration of inflammatory cells in the mucous interstitial background. The tumor cells were round and epithelioid, with large nuclei, deep staining and obvious nucleolus. Immunohistochemistry showed anaplastic lymphoma kinase (ALK) positive, and molecular pathology fluorescence in situ hybridization showed ALK gene amplification (positive). Combined with pathological, immunohistochemical and genetic tests, EIMS was diagnosed. Conclusions:EIMS should be considered when there are single or multiple solid or cystic nodules or masses in the pelvic cavity with obvious enhancement, invasive growth and peritoneal implantation metastasis, and the correct diagnosis can be made according to the pathological findings, immunohistochemistry and genetic test results.

Objective To analyze the epidemiological and clinical characteristics of scrub typhus in Liuyang People's Hospital Hunan Province for better diagnosis,treatment,prevention and control of scrub typhus.Methods A retrospective study was conducted on 159 confirmed cases of scrub typhus.The demographic data of patients,clinical manifestations of scrub typhus,laboratory tests,and chest CT findings,complications,treatment,and outcomes were analyzed.Results The 159 patients with scrub typhus included 142 males and 17 females.The average age of patients was(53.8±11.9)(18-82)years old.The peak incidence of scrub typhus was in the period from July to September(87.42％).The common clinical manifestations included fever(97.48％),headache(41.51％),and fatigue(23.90％).The most common signs were eschar(92.45％)on the lower limbs(25.16％),scrotum(15.72％),and buttocks(11.32％).Laboratory test results upon admission showed decreased platelet count(64.15％),elevated creatinine(38.36％),elevated AST(90.57％),ALT(80.50％),and LDH(90.57％).Pulmonary imaging study revealed pulmonary inflammation(22.64％),pleural effusion(13.21％),and pericardial effusion(6.29％).The common complications included liver impairment(69.81％)and decreased platelet count(50.94％),as well as other complications such as renal dysfunction,myocardial injury,sepsis,respiratory failure,and disseminated intravascular coagulation(DIC).Doxycycline-based therapies resulted in cure rate of 98.11％(156/159).Three patients died due to severe complications.Conclusions The peak incidence of scrub typhus was in the period from July to September in Hunan Province.Eschar is a key feature for clinical diagnosis.Scrub typhus can affect multiple organ systems,leading to various systemic complications.The cure rate is high with doxycycline treatment,but some patients may die from severe complications due to delayed medical treatment.

This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

The Animal Molecular Biology course is a crucial and foundational course for both Animal Medicine and Animal Science majors.Apart from teaching fundamental principles of molecular biology,the course provides updated applications of these principles in the field of animal science research.Im-portantly,it plays a fundamental role in cultivating students'research capabilities.With the rise of over-whelming information and their optimal utilization,the demand for integrating digital education with tradi-tional teaching methods is increasing.Based on the five years of teaching practice,this paper summarizes four highlights of the course:the construction of teaching resource,the restructuring of teaching syllabus,the adjustment of classroom teaching hours,and the improvement of assessment methodology.It focuses on Electronic-Learning"E(E-Learning)",offline classroom intensive teaching"C(Classroom)",and post-class extension to construct a blended teaching model that integrates Electronic-Learning and Class-room teaching,namely the"E+C"blended teaching model.Offline classroom teaching emphasizes the combination of theory and knowledge systems,while online Electronic-Learning mainly focuses on popular science and interesting aspects to stimulate students' interests and enthusiasm in learning.Over five years of practice,the"E+C"blended model has been proven to exert a good teaching effect.Students have reported significant gains from the course,with tightly connected and strongly complementary class-room teaching and E-Learning,which greatly aids in mastering professional knowledge.It also cultivates intrinsic motivation for learning and enhances the sense of accomplishment in acquiring knowledge,sig-nificantly improving teaching effectiveness.

Parkinson's disease(PD)is one of the most common neurodegenerative disorders.Recent evidence implicates pyroptosis as one of the pathogenic mechanisms in central nervous system disorders,although its specific mechanisms remain unclear.In this study,SH-SY5Y cells were transfected with py-roptosis-related proteins GSDME full-length(GSDME-F)or GSDME-N terminal(GSDME-N)plasmids revealed that GSDME-N significantly reduced mitochondrial membrane potential(P＜0.0001).To inves-tigate the mechanism by which GSDME mediates mitochondrial dysfunction,Western blotting analysis demonstrated that transfection with GSDME-N plasmids significantly increased BAX expression and en-hanced its translocation to mitochondria in both HEK 293T and SH-SY5Y cells(P＜0.05).SH-SY5Y cells treated with varying concentrations of rotenone(ROT)exhibited GSDME cleavage,elevated BAX expression(P＜0.05),increased mitochondrial BAX aggregation(P＜0.05),and reduced mitochondrial membrane potential(P＜0.01),as confirmed by Western blotting and JC-1 staining.Concurrently,MTT assays assessing cell viability and lactate dehydrogenase(LDH)release assays indicated that ROT in-duced these processes prior to pyroptosis.Furthermore,in a ROT-induced mouse PD model,ROT trig-gered GSDME cleavage,enhanced BAX expression,caused dopaminergic neuronal damage,and induced motor deficits.In summary,this study demonstrates that GSDME-N exacerbates mitochondrial damage and increases cytotoxicity by upregulating BAX expression and facilitating its mitochondrial translocation.This study provides novel insights into the role of GSDME in PD pathogenesis and suggests potential avenues for therapeutic intervention.