Objective To investigate the genetic diversity of sandfly populations in endemic areas of visceral leishmaniasis in China, so as to provide references insights into management of visceral leishmaniasis and the vector sandflies. MethodsSixteen sampling sites were selected from main endemic foci of visceral leishmaniasis in China from June to September 2024, including Shanxi Province, Shaanxi Province, Henan Province, Gansu Province, Sichuan Province, and Xinjiang Uygur Autonomous Region. Sandflies were captured using light traps and manual aspirators from sheep pens, chicken coops, cave dwellings, bovinesheds, and pig pens at each sampling site. A single sandfly sample was washed in phosphate-buffered saline (PBS), and genomic DNA was extracted from sandfly samples. Cytochrome oxidase subunit 1 (COI) gene was amplified using PCR assay with universal primers, and analyzed and retrieved with the nucleotide sequence analysis tool (BLAST) software, and the sequence of COI gene was aligned with the ClustalX 1.83 and MEGA 7.0 software. The base composition and variation site of the COI gene sequence were analyzed using the software MEGA 7.0, and the number of haplotypes, total number of segregating sites, haplotype diversity, nucleotide diversity, and average nucleotide differences were calculated in the COI gene sequence using the software DnaSP 5.10, followed by Tajima’s D test for neutrality. Haplotypes were screened using the software DnaSP 5.10, and the haplotype network map of sandfly samples was plotted using the software Network 5.0. MEGA 7.0 software was employed for gene sequence editing and alignment, and calculation of genetic distances among sandfly species sampled from different regions, and a phylogenetic tree was built with a neighbor-joining method. Results A total of 466 sandflies were captured from 16 sampling sites in China from June to September 2024, and 430 gene sequences were yielded following PCR amplification and sequencing of the COI gene, with 652 to 688 bp in the length of amplification fragments. The captured sandfly samples were characterized as Phlebotomus chinensis, Sergentomyia squamirostris, Se. koloshanensis, Ph. sichuanensis, and Ph. longiductus following the COI gene sequence alignment in BLAST. A total of 251 haplotypes were identified in the 430 gene sequences from sandfly samples (50.5%), and the average haplotype diversity, nucleotide diversity and average number of nucleotide difference were 0.885, 0.257 and 160.761, respectively. The Tajima’s D values were -0.92 for sandfly populations from Yangquan City, Shanxi Province and -1.73 for sandfly populations from Sanmenxia City, Henan Province, and were all more than 0 for sandfly populations from other sampling sites. Haplotype analysis identified 50 haplotypes, which were classified into two haplogroups. Heplogroup 1 included 29 haplotypes, which had a high homology, and heplogroup 2 included 21 haplotypes. The average genetic distance was 0.000 to 0.604 among sandfly samples from different sampling sites, and phylogenetic analysis revealed that the five sandfly species were clustered into distinct clades, all with 100% clade confidence. Conclusions There is a high genetic polymorphism in the COI gene from five sandfly populations in main endemic foci of visceral leishmaniasis in China, and COI gene may serve as a marker gene for analysis of the genetic structure of sandfly populations.

Objective To analyze and evaluate the implementation effect of tuberculosis prevention and control program in Wuhan, and to provide reference for scientific formulation of tuberculosis prevention and control measures. Methods Using the National Tuberculosis Information Management System, descriptive statistical analysis was carried out on the medical record information of pulmonary tuberculosis patients registered in Wuhan , 2016 - 2021. Results A total of 34 937 cases of pulmonary tuberculosis were registered in Wuhan , with an average annual incidence rate of 49.85/100 000. The incidence rate showed a downward trend year by year, with a statistically significant difference in 2016—2021 (χ²_trend = 708.387, P<0.001). The patients mainly came from referrals, accounting for 71.86%, and the proportion of referrals varied significantly among different years (χ²=355.541, P<0.001). The diagnosis type was mainly pathogenic negative, accounting for 49.12%. The proportion of pathogenic negative had statistically significant difference among different years (χ²=1 354.830, P<0.001). The proportion of patients cured and completed the course of treatment reached 93.98%, with statistically significant differences in the proportions among different years (cured, χ²=1 080.252, P<0.001; completed the treatment course, χ²= 933.655, P<0.001). The sputum examination rate of newly diagnosed patients in each year reached over 90%, and the overall completion rate reached over 95%. The proportion of positive pathogens showed an increasing trend year by year. Conclusion The overall epidemic situation of tuberculosis in Wuhan is declining year by year, and tuberculosis prevention and control work has achieved remarkable results. Active screening in key areas and populations should be strengthened, and prevention and control strategies should be formulated by emphasizing the key and difficult points.

Objective To investigate the value of derived neutrophil to lymphocyte ratio(dNLR)as a pre-dictive biomarker for immunotherapy response in advanced non-small cell lung cancer(NSCLC).Methods A total of 92 patients with advanced NSCLC who received anti-programmed cell death receptor(PD-1)combined therapy in the hospital from August 2018 to December 2019 were selected as the research objects.Peripheral blood samples were collected within 24 h before immunotherapy,complete blood cell count was measured,and dNLR was calculated.Patients with advanced NSCLC were treated with PD-1 inhibitors or combination regi-mens,and the response to immunotherapy was evaluated by objective response rate(ORR)and disease control rate(DCR).The receiver operating characteristic(ROC)curve was used to analyze the predictive value of dN-LR for the diagnosis and response to immunotherapy in advanced NSCLC.Multivariate Logistic regression model was used to analyze the relationship between dNLR and immunotherapy response in advanced NSCLC.Kaplan-Meier survival curve and Log-Rank test were used to analyze the overall survival(OS),progression-free survival(PFS)and disease-specific survival(DSS)of the low dNLR group and the high dNLR group.Re-sults The ORR and DCR of advanced NSCLC patients after immunotherapy were 32.61％and 65.22％,re-spectively,and the PFS and OS were 17.0(8.5,25.5)and 24.0(12.7,36.1)months,respectively.The dNLR of DCR group was lower than that of non-DCR group(P＜0.001).The dNLR of ORR group was lower than that of non-ORR group(P＜0.001).The area under the curve of dNLR for predicting DCR or ORR was 0.897(95％CI 0.829-0.965)and 0.874(95％CI 0.795-0.953),respectively.Multivariate Logistic regres-sion analysis showed that dNLR≥2.28 increased the risk of non-response to immunotherapy,and this inde-pendent relationship still existed after further adjustment for objective confounding factors(P＜0.05).Sur-vival curve results showed that patients with high dNLR had significantly shorter PFS,OS,and DSS(P＜0.05).Multivariate Cox regression analysis showed that high dNLR was an independent factor affecting the poor prognosis of patients with advanced NSCLC(P＜0.05).Conclusion High dNLR advanced NSCLC pa-tients are more difficult to benefit from immune therapy,and prognosis is worse.dNLR is promising as a pre-dictive biomarker for immunotherapy response in advanced NSCLC.

Objective To analyze the diagnosis status of negative etiological pulmonary tuberculosis test re-sults in Wuhan,and to provide scientific basis for improving the diagnosis strategy of etiological negative pul-monary tuberculosis.Methods From January 1 to February 28,2019,a total of 241 patients with negative eti-ological tuberculosis who were registered,reported and treated in 1 municipal and 2 district-level designated hospitals were selected.The medical record data,chest imaging examination and laboratory examination re-sults of the selected patients were reviewed and extracted,and the quality of etiological examination and ima-ging examination of patients with negative etiological pulmonary tuberculosis were analyzed.Results Among the 241 patients,88.8％(214/241)of the patients met the diagnostic criteria for negative etiological pulmona-ry tuberculosis,and 96.3％(232/241)of the patients had chest imaging examinations that were consistent with the original diagnostic results.Patients received sputum smear examination,sputum culture,and molecu-lar biology test accounted for 97.9％(236/241),73.9％(178/241)and 65.6％(158/241),respectively.Patients received anti-tuberculosis antibody test,tuberculin skin test,y-interferon release and diagnostic anti-infection treatment accounted for 54.8％(132/241),46.5％(112/241),26.1％(63/241),and 53.1％(128/241),respec-tively.The sputum culture detection rate of urban area was higher than those of central and remote urban are-as,the rate of central urban area was higher than that of remote urban areas,and the molecular biology detec-tion rate of urban area was higher than those of central and remote urban area,with statistical significance(P＜0.001).The detection rate of anti-tuberculosis antibody of urban area was lower than that of central ur-ban area,and the differences were statistically significant(P＜0.001).The rate of diagnostic anti-infective therapy of central urban area was higher than that of urban area and the remote urban area,and the rate in ur-ban area was higher than that of remote urban area,and the differences were statistically significant(P＜0.001).Conclusion It is necessary to further standardize the diagnosis of negative etiological pulmonany tu-berculosis of designated tuberculosis hospitals.The proportion of diagnostic anti-infection treatment and auxil-iary examination at the municipal level needs to be increased,and the frequency and items of laboratory etio-logical examination at the district level need to be increased.

Objective:To develop and evaluate a nomogram prediction model for the 3-month mortality risk of patients with sepsis-associated acute kidney injury (S-AKI).Methods:Based on the American Medical Information Mart for Intensive Care-Ⅳ (MIMIC-Ⅳ), clinical data of S-AKI patients from 2008 to 2021 were collected.Initially, 58 relevant predictive factors were included, with all-cause mortality within 3 months as the outcome event. The data were divided into training and testing sets at a 7∶3 ratio. In the training set, univariate Logistic regression analysis was used for preliminary variable screening. Multicollinearity analysis, Lasso regression, and random forest algorithm were employed for variable selection, combined with the clinical application value of variables, to establish a multivariable Logistic regression model, visualized using a nomogram. In the testing set, the predictive value of the model was evaluated through internal validation. The receiver operator characteristic curve (ROC curve) was drawn, and the area under the curve (AUC) was calculated to evaluate the discrimination of nomogram model and Oxford acute severity of illness score (OASIS), sequential organ failure assessment (SOFA), and systemic inflammatory response syndrome score (SIRS). The calibration curve was used to evaluate the calibration, and decision curve analysis (DCA) was performed to assess the net benefit at different probability thresholds.Results:Based on the survival status at 3 months after diagnosis, patients were divided into 7?768 (68.54%) survivors and 3?566 (31.46%) death. In the training set, after multiple screenings, 7 variables were finally included in the nomogram model: Logistic organ dysfunction system (LODS), Charlson comorbidity index, urine output, international normalized ratio (INR), respiratory support mode, blood urea nitrogen, and age. Internal validation in the testing set showed that the AUC of nomogram model was 0.81 [95% confidence interval (95% CI) was 0.80-0.82], higher than the OASIS score's 0.70 (95% CI was 0.69-0.71) and significantly higher than the SOFA score's 0.57 (95% CI was 0.56-0.58) and SIRS score's 0.56 (95% CI was 0.55-0.57), indicating good discrimination. The calibration curve demonstrated that the nomogram model's calibration was better than the OASIS, SOFA, and SIRS scores. The DCA curve suggested that the nomogram model's clinical net benefit was better than the OASIS, SOFA, and SIRS scores at different probability thresholds. Conclusions:A nomogram prediction model for the 3-month mortality risk of S-AKI patients, based on clinical big data from MIMIC-Ⅳ and including seven variables, demonstrates good discriminative ability and calibration, providing an effective new tool for assessing the prognosis of S-AKI patients.

Objective To construct an evaluation index system for the capability of comprehensive control of mountain-type zoonotic visceral leishmaniasis based on the One Health concept, so as to provide insights into the control and elimination of mountain-type zoonotic visceral leishmaniasis using the One Health approach. Methods A preliminary evaluation index system was constructed based on literature review, panel discussions and field surveys. Thirty-three experts were selected from 7 provincial disease control and prevention centers in Beijing Municipality, Hebei Province, Shanxi Province, Henan Province, Sichuan Province, Shaanxi Province and Gansu Province where mountain-type zoonotic visceral leishmaniasis was endemic, and two rounds of expert consultations were conducted to screen the indicators. The positive coefficient, degree of concentration, degree of coordination, and authority of the experts were calculated, and the normalized weights of each index were calculated with the analytic hierarchy process. Results The response rates of questionnaires during two rounds of expert consultation were both 100.00% (33/33), and the authority coefficients of the experts were 0.86 and 0.88, respectively. The coefficients of coordination among experts on the rationality, importance, and operability of the indicators were 0.392, 0.437, 0.258, and 0.364, 0.335, 0.263, respectively (all P values < 0.05). Following screening, the final evaluation index system included 3 primary indicators, 17 secondary indicators, and 50 tertiary indicators. The normalized weights of primary indicators “external environment”, “internal support” and “comprehensive control” were 16.98%, 38.73% and 44.29%, respectively. Among the secondary indicators of the primary indicator “external environment”, the highest weight was seen for natural environment (66.67%), and among the secondary indicators of the primary indicator “internal support”, the lowest weight was seen for the scientific research for visceral leishmaniasis control (8.26%), while other indicators had weights of 12.42% to 13.38%. Among the secondary indicators of the primary indicator “comprehensive control”, the weight was 16.67% for each indicator. Conclusions An evaluation index system has been constructed for the capability of comprehensive control of mountain-type zoonotic visceral leishmaniasis based on the One Health concept. In addition to assessment of the effect of conventional mountain-type zoonotic visceral leishmaniasis control measures, this index system integrates the importance of top-level design, organizational management, and implementation of control measures, and includes indicators related to multi-sectoral cooperation.

Phage antibody display technology is currently the most widely used in vitro antibody screening technology,which uses bacteriophages as a vector,and inserts exogenous antibody library genes into phage capsid protein genes,and expresses the capsid protein on the phage surface while also displays the antibody protein.Antibody drugs play an important role in tumor immunity and microbial immunity due to their targeting advantages,which is also an important driving force for them to become a hot spot in the field of pharmaceutical research and development.Therefore,this article reviews the background,basic principles,antibody library types and antibody fragment types of phage display technology,and looks forward to the latest progress and application prospects of fully human antibodies.

Objective: To determine the knowledge, attitudes and practices (KAP) regarding tuberculosis prevention and control among college students in Wuhan, China, while analyzing the factors affecting tuberculosis prevention behavior. Methods: Referring to the National KAP Questionnaire for Tuberculosis Prevention and Control among College Students, which was designed by China Center for Disease Control and Prevention, 22 colleges and universities in Wuhan were selected using a judgment sampling method, and a questionnaire survey was carried out through the online questionnaire platform. The formation of tuberculosis prevention and control behavior among this population was analyzed by single factor and multi factor Logistic regression. Results: The total awareness rate of KAP regarding tuberculosis prevention and control among college students in Wuhan was 79.69%; the awareness rate of core knowledge was 79.28%, and the passing rate was 92.46%. The correct attitude holding rate was 89.69%, while the passing rate was 90.56%. In respect to correct behavior, the formation rate was 72.11%, with a passing rate of 96.62%. The multivariate Logistic regression analysis showed that the following factors were more likely to promote tuberculosis prevention and control behaviors that met the target: female college students ( OR =1.86); college students whose father had a primary school education level, junior high school and equivalent, high school and equivalent, and junior high school students ( OR =2.94, 3.05, 3.17, 3.24 ); no history of tuberculosis ( OR =3.32); the passing knowledge of core knowledge ( OR =9.91), and the passing attitude holding score ( OR =7.35)( P <0.05). Conclusion The overall awareness rate of KAP regarding tuberculosis prevention and control among college students in Wuhan is acceptable,the mainly influenced factors are gender, history of tuberculosis, awareness rate of core knowledge about prevention and control, attitudes and cognition, and parents educational background.

Objective:To investigate the role of septum incision technique in laparoscopic transcystic common bile duct exploration (LTCBDE).Methods:The data of 22 patients with choledocholithiasis undergoing LTCBDE by septum (a membrane-like wall of the cystic duct running parallel to the common hepatic duct) incision technique at the Suzhou Ninth Hospital Affiliated to Soochow University from April 2020 to March 2023 were retrospectively analyzed, including 13 males and 9 females, aged (54.4±20.3) years. The operative time, postoperative hospital stay, postoperative complications (bile leakage, abdominal hemorrhage, abdominal infection, etc.) and follow-up data were collected and analyzed.Results:All 22 patients (including two patients undergoing common bile duct exploration through the residual cystic duct) underwent the exploration of common hepatic duct, left and right hepatic duct openings, and secondary branch openings successfully. It was confirmed using this procedure that there were no obstruction of the common hepatic duct and intrahepatic bile ducts, or residual stones. The operation time was (79.6±23.2) min and the postoperative hospital stay was (6.3±1.7) d. No complications such as biliary leakage, abdominal hemorrhage or infection occurred after surgery. No cases of biliary stricture or residual stones were observed during short-term postoperative follow-ups.Conclusion:The septum incision technique improved the visualization of hilar bile duct in LTCBDE, which could be a safe and effective procedure to facilitate the LTCBDE and increase its success rate.

Objective: To analyze the epidemiological characteristics of pulmonary tuberculosis among students in Wuhan from 2011 to 2020, and to provide a basis for the scientific development of interventions and strategies. Methods: Epidemiological distribution, time of onset and treatment, as well as treatment outcomes of student tuberculosis in Wuhan from 2011 to 2020 registered in the national tuberculosis information management system were described. Results: During 2011-2020, 4 337 student tuberculosis patients in Wuhan were registered. The average annual incidence rate was 22.44/10 million, and the annual decreasing rate of 7.86% The incidence of male and female patients was 1.76:1, and the incidence rate of male was higher than that of female( χ 2=184.18, P <0.01). Most of patients aged 19-22 years old, accounting for 47.89%; Tuberculosis reports were highest during March to May, and September to November, and lowest during January to February, and July to August. Student patients were mainly concentrated in Hongshan District, Jiangxia District and Wuchang District, where schools were more distributed in Hongshan District, Dongxihu District, Wuchang District and Xinzhou District. The median duration from tuberculosis onset to treatment was 9(3, 21) days, which varied significantly by region, age, nationality, and patient residence ( Z =-9.25, 47.14, 9.88,43.96, P <0.01). The treatment and outcome of student tuberculosis patients were varied significantly by year and nationality( P <0.05). Conclusion The incidence of student tuberculosis in Wuhan City showed a slow downward trend. Most of student tuberculosis are college and high school students. Time and place of case detection are relatively fixed. The time of treatment and the outcome of treatment vary significantly. Tuberculosis prevention and control strategies should be formulated according to the local conditions according to the tuberculosis distribution characteristics, as well as enhancing surveillance, health promotion, active discovery and supervision management of tuberculosis in school settings.