Cats, owing to their physiological and immunological similarities with humans, have become increasingly valuable as model animals in virology research, drug development, and vaccine evaluation. They are irreplaceable in studies of feline immunodeficiency virus, feline coronavirus, and other related pathogens. However, cats are temperamentally sensitive, exhibit strong stress responses, and possess well-developed nervous systems as well as sharp claws and teeth. Consequently, the biosafety risks associated with infectious experiments using cats in animal biosafety level 2 laboratory (ABSL-2) are significantly higher than those encountered with conventional rodents. Drawing on long-term ABSL-2 operational experience, this article systematically reviews the entire workflow of infectious experiments in laboratory cats — from animal selection, pre-entry preparation, reception and quarantine, housing management, to infectious experimental procedures and incident response — identifying and addressing critical risk points at each stage. For strain selection, SPF-grade shorthair cats with defined genetic backgrounds and docile temperaments are recommended; sex and age should be scientifically matched to experimental objectives. During pre-entry preparation, emphasis is placed on dual-credential personnel management, health surveillance, standardized disinfection of environments and cages, feed and water standards, and robust record-keeping. During reception and quarantine, standardized protocols are established for transport control, appearance inspection, isolation quarantine, pathogen exclusion, and positive-reinforcement training. During infectious experimentation, a "three-fixed" husbandry principle is clearly implemented: dedicated caretakers, fixed feeding/cleaning times, and fixed cage positions. Disinfectant selection, autoclaving of waste, and daily veterinary rounds are rigorously enforced. Operational risk control includes detailed measures for graded personal protection, animal anesthesia and restraint, zoned operation within biosafety cabinets, and disposal of experimental waste. Contingency plans are formulated to address animal death, escape, personnel exposure, and spills of infectious materials. This study provides a reproducible and scalable technical pathway and operational standard for conducting infectious experiments in laboratory cats in ABSL-2 laboratories, offering a reference for other facilities undertaking similar work.

ObjectiveTo prepare rabbit anti-porcine inhibin polypeptide-keyhole limpet hemocyanin(KLH) conjugated polyclonal antibody and evaluate its effect on superovulation in C57BL/6J mice. MethodsNew Zealand white rabbits were immunized with a synthesized porcine inhibin polypeptide conjugated with KLH to produce anti-inhibin serum (AIS, i.e., inhibin polyclonal antibody). Female C57BL/6J mice received intraperitoneal injections of purified AIS in combination with pregnant mare serum gonadotropin (PMSG), followed by human chorionic gonadotropin (hCG) after 48 hours to induce superovulation. Oocytes obtained from superovulation were collected and counted 15 hours post-hCG administration, and the number of 2-cell embryos was assessed 24 hours after in vitro fertilization. ResultsAIS prepared by immunizing New Zealand White rabbits with KLH-conjugated porcine inhibin polypeptide was subjected to titer determination by indirect ELISA, showing titers reaching 1∶ 512 000. SDS-polyacrylamide gel electrophoresis analysis of ammonium sulfate-purified AIS revealed distinct 50 kDa and 25 kDa bands corresponding to the theoretical molecular weights of IgG antibody heavy and light chains, confirming successful production of porcine inhibin polyclonal antibody. Compared with conventional superovulation methods, AIS diluted 10-fold combined with PMSG significantly increased the number of oocytes obtained from superovulation in mice (P<0.05) by approximately 1.5-fold. ConclusionPorcine inhibin polyclonal antibody, as an improved superovulation reagent, can improve superovulation efficiency in C57BL/6J mice, and shows promising prospects for future applications.

Acute-on-chronic liver failure （ACLF） refers to severe liver dysfunction that occurs on the basis of chronic liver diseases， and it is characterized by rapid disease progression， poor prognosis， and high mortality rate. In recent years， inflammatory markers have become a research hotspot due to their significant role in assessing the prognosis of ACLF. This article reviews the advances in the application of inflammatory markers in assessing the prognosis of ACLF， such as systemic immune inflammatory index， lymphocyte-white blood cell ratio， neutrophil-lymphocyte ratio， and C-reactive protein， and discusses their clinical value and future research directions， in order to provide a theoretical basis for the early intervention and prognosis management of ACLF patients.

ObjectiveTo improve the accuracy of discrete element method in simulating the processing of traditional Chinese medicine（TCM） powder system under low shear conditions. MethodsIn this study， extract powders of Tongsaimai tablets and Qige granules were used as the research objects， the angle of repose（AOR） and effective angle of internal friction of the two materials were determined by AOR test method and shear cell test method. Based on the Hertz-Mindlin with JKR V2 contact model and particle scaling theory， taking the particle-particle restitution coefficient（A）， particle-particle static friction coefficient（B）， particle-particle rolling friction coefficient（C）， particle-steel restitution coefficient（D）， particle-steel static friction coefficient（E）， particle-steel rolling friction coefficient（F） and Johnson-Kendall-Roberts（JKR） surface energy（G） as test factors， the simulated contact parameters of Tongsaimai tablets extract powder were first calibrated with a single reference value using AOR as the reference value， and then the simulated contact parameters of Tongsaimai tablets extract powder as well as Qige granules extract powder were co-calibrated with AOR and effective angle of internal friction as the joint reference value， respectively. Then， Plackett-Burman design was used to screen the critical contact parameters that have a significant effect on the simulated reference value， and the steepest ascent design was used to determine the optimal range of the critical contact parameters， finally， the regression model between the critical contact parameters and the simulated reference values was established through the design of the response surface test， and the critical contact parameters were calibrated based on the regression model and the desirability function approach. ResultsThe optimal combination of discrete elemental contact parameters A-G for Tongsaimai tablets extract powder under a single reference value was 0.100， 0.718， 0.616， 0.100， 0.400， 0.250 and 0.075 J·m^-2， which was validated to have relative errors of 0.10% and -8.64% for the simulated AOR and the simulated effective angle of internal friction， respectively. And the optimal combination of discrete elemental contact parameters A-G for Tongsaimai tablets extract powder at the joint reference values was 0.100， 0.682， 0.598， 0.100， 0.521， 0.294 and 0.075 J·m^-2， which was verified to have relative errors of 0.10% and -0.18% for the simulated AOR and the simulated effective angle of internal friction， respectively. The optimal combination of discrete elemental contact parameters A-G for Qige granules extract powder at the joint reference values was 0.150， 0.370， 0.330， 0.150， 0.500， 0.500 and 0.100 J·m^-2， which was verified to have relative errors of 2.70% and -1.30% for the simulated AOR and the simulated effective angle of internal friction， respectively. Compared with the single reference value method， the joint calibration method not only increased the number of the critical contact parameters for characterizing particle-device interactions， but also was more accurate and reliable. ConclusionCompared with the results of single reference value calibration， the results obtained by the method of joint calibration of discrete element simulation contact parameters with AOR and effective angle of internal friction as the reference values are more accurate， which can provide more accurate and reliable simulation physical property data for the simulation experiments of TCM extract powder under low shear process conditions.

Background::Computer-aided detection (CAD) software has been introduced to automatically interpret digital chest X-rays. This study aimed to evaluate the performance of CAD software (JF CXR-1 v3.0, which was developed by a domestic Hi-tech enterprise) in tuberculosis (TB) case finding in China.Methods::In 2019, we conducted an internal evaluation of the performance of JF CXR-1 v3.0 by reading standard images annotated by a panel of experts. In 2020, using the reading results of chest X-rays by a panel of experts as the reference standard, we conducted an on-site prospective study to evaluate the performance of JF CXR-1 v3.0 and local radiologists in TB case finding in 13 township health centers in Zhongmu County, Henan Province.Results::Internal assessment results based on 277 standard images showed that JF CXR-1 v3.0 had a sensitivity of 85.94% (95% confidence interval [CI]: 77.42%, 94.45%) and a specificity of 74.65% (95% CI: 68.81%, 80.49%) to distinguish active TB from other imaging conditions. In the on-site evaluation phase, images from 3705 outpatients who underwent chest X-ray detection were read by JF CXR-1 v3.0 and local radiologists in parallel. The imaging diagnosis of local radiologists for active TB had a sensitivity of 32.89% (95% CI: 22.33%, 43.46%) and a specificity of 99.28% (95% CI: 99.01%, 99.56%), while JF CXR-1 v3.0 showed a significantly higher sensitivity of 92.11% (95% CI: 86.04%, 98.17%) ( p < 0.05) and maintained high specificity at 94.54% (95% CI: 93.81%, 95.28%). Conclusions::CAD software could play a positive role in improving the TB case finding capability of township health centers.

Objective:To analyze the effects of fertilization methods, quality of inner cell mass (ICM) and trophectoderm (TE) on the sex ratio at birth (SRB) during single blastocyst transfer cycles.Methods:5 367 single blastocyst transfer cycles in in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) of patients with successful pregnancy and delivery in Reproductive Medical Center of Zhanjiang Jiuhe Hospital from January 2010 to December 2022 were included in this retrospective cohort study. According to different types of embryos, the cycles were divided into fresh embryo group ( n=1 487, total population; n=1 258, ≤35 years old) and frozen-thawed embryo group ( n=3 880, total population; n=3 199, ≤35 years old). According to different fertilization methods, the cycles were divided into IVF group ( n=4 424, total population; n=3 667, ≤35 years old) and ICSI group ( n=943, total population; n=780, ≤35 years old). Chi-square test was used to analyze the differences of SRB among all the patients and the patients aged ≤35 years who underwent different embryo types and fertilization methods, and logistic regression was used to analyze the effects of ICM and TE ratings on SRB in each group of blastocysts. Results:There were 917 male infants (61.7%, 917/1 487) in the fresh embryo transfer group and 2 317 male infants (59.7%, 2 317/3 880) in frozen-thawed embryo transfer group. The SRB of the fresh embryo transfer group (160.9∶100) was slightly higher than that in frozen-thawed embryo transfer group (148.2∶100), but the difference was not statistically significant ( P=0.201). There were 2 732 male infants (61.8%, 2 732/4 424) in IVF group and 502 male infants (53.2%, 502/943) in ICSI group. The SRB of IVF group (161.5∶100) was significantly higher than that of ICSI group (113.8∶100), and the difference was statistically significant ( P<0.001). In female patients aged ≤35 years, the effects of different embryo transfer methods and different fertilization methods on SRB were consistent with the results of total population. Univariate logistic regression analysis showed that the SRB of patients with ICSI was lower than that of patients with IVF, except for 3-6AA. There were significant differences in SRB between ICSI cycles patients (88.8∶100; 72.8∶100; 156.1∶100) and IVF cycles patients (130.7∶100; 124.8∶100; 206.3∶100) when the blastocyst grade was 3-6BB ( OR=0.679, 95% CI: 0.516-0.896, P=0.006), 3-6AB( OR=0.583, 95% CI: 0.421-0.809, P=0.001) and 3-6BA OR=0.757, 95% CI: 0.585-0.979, P=0.034). Taking 3-6BB blastocysts as reference, the SRB of 3-6AA and 3-6BA blastocysts were higher (IVF: OR=1.527, 95% CI: 1.258-1.854, P<0.001; OR=1.579,95% CI: 1.341-1.859, P<0.001; ICSI: OR=2.566, 95% CI: 1.661-3.966, P<0.001; OR=1.758, 95% CI: 1.250-2.472, P=0.001). Compared with 3-6BA, the SRB of 3-6BC blastocysts was lower (IVF: OR=0.621, 95% CI: 0.447-0.862, P=0.004; ICSI: OR=0.442, 95% CI: 0.238-0.818, P=0.009). Taking 3-6AB as reference, the SRB of 3-6AA blastocysts was higher (IVF: OR=1.600, 95% CI: 1.307-1.958, P<0.001; ICSI: OR=3.130, 95% CI: 1.964-4.987, P<0.001). Conclusion:The SRB of fresh embryo group is slightly higher than that of frozen-thawed group in single blastocyst transfer cycles. Different fertilization methods can affect SRB, and the SRB in IVF is significantly higher than that in ICSI group. The quality of blastocysts can affect SRB, and blastocysts with higher quality TE have significantly increased SRB, while ICM quality have no significant effect on SRB.

Panic disorder has the characteristics of early onset, late detection, high comorbidity rate and low treatment rate, which have a lasting impact on the physical and mental health of children and adolescents. Early intervention can reduce the risk of various mental disorders in adulthood. Cognitive-behavioral therapy(CBT), as a routine social psychological treatment, is also recommended as a first-line intervention for the treatment of panic disorder in children and adolescents. However, due to diagnostic and application difficulties, CBT has not received sufficient attention in the field of panic disorder in children and adolescents. Through a systematic review of the application of CBT in children and adolescents with panic disorder, it found that panic control therapy(PCT)and intensified CBT(iCBT) therapy are the main methods for treating panic disorder in children and adolescents, both of which can effectively alleviate the symptoms of panic disorder in children and adolescents.Cognitive restructuring, exposure, respiratory training and parental involvement are effective components of CBT treatment for panic disorder.In the future, it is necessary to increase attention to underage panic disorder, improve CBT cultural adaptation, use diverse treatment methods, objectively evaluate standards, and optimize research design, to promote the improvement of treatment effectiveness and the development of clinical research.

Objective:To observe the efficacy of thrombectomy combined with ezetimibe and atorvastatin in patients with acute ischemic stroke (AIS) and the serum levels of absent in melanoma 2 (AIM2) and leukotriene B4 (LTB4).Methods:A prospective study method was used to select 60 patients with AIS thrombectomy treated in Marina Bay Central Hospital of Dongguan City from June 2021 to December 2022, and they were divided into two groups according to random number table method, with 30 cases in each group. The control group was given atorvastatin treatment, and the study group was combined with ezetimibe treatment on the basis of the control group treatment. Blood routine indexes, blood lipid indexes and serum levels of AIM2 and LTB4 were compared between the two groups at admission, discharge and follow-up for 3 months. The National Institutes of Health Stroke Scale (NIHSS) was used to assess the degree of neurological impairment of the patients, and the modified Rankin Scale (mRS) was used to assess the patients' ability to live independently. The occurrence of adverse reactions in the two groups was observed and compared.Results:The neutrophil to lymphocyte ratio (NLR) and neutrophil (N) in the study group were lower than those in the control group : 1.71 ± 0.05 vs. 2.04 ± 0.31, 0.261 ± 0.012 vs. 0.403 ± 0.021 at follow-up for 3 months, there were statistical differences ( P<0.05). The levels of low density lipoprotein cholesterol (LDL-C), total cholesterol (TC) and triacylglycerol (TG) in the study group were lower than those in the control group at follow-up for 3 months:(1.59 ± 0.48) mmol/L vs. (2.09 ± 0.69) mmol/L, (2.92 ± 0.44) mmol/L vs. (3.57 ± 0.71) mmol/L, (1.19 ± 0.24) mmol/L vs. (1.37 ± 0.21) mmol/L, the level of high density lipoprotein cholesterol (HDL-C) was higher than that in the control group : (1.24 ± 0.19) mmol/L vs. (1.06 ± 0.21) mmol/L, there were statistical differences ( P<0.05). The levels of AIM2 and LTB4 in the study group were lower than those in the control group at discharge and follow-up for 3 months: (945.20 ± 123.60) ng/L vs. (1 094.40 ± 227.10) ng/L, (948.90 ± 135.80) ng/L vs. (1 236.90 ± 125.70) ng/L, (115.80 ± 20.60) ng/L vs. (129.70 ± 31.50) ng/L, (116.40 ± 21.10) ng/L vs. (130.30 ± 32.60) ng/L, there were statistical differences (P<0.05). The NHISS scores in the study group was lower than that in the control group at follow-up for 3 months: 2.00(1.00, 3.00) scores vs. 4.00(3.00, 5.00) scores, there was statistical difference ( P<0.05). After follow-up for 3 months, the mRS scores and incidence of adverse reactions between the two groups had no statistical differences ( P<0.05). Conclusions:The combination of atorvastatin and ezetimibe in the early stage of AIS thrombectomy can significantly reduce the levels of NLR, serum AIM2 and LTB4, improve blood lipid levels, reduce the degree of neurological impairment, and improve the ability of patients to carry out activities of daily living, with high safety.

Objective To explore the efficacy of the self-formulated Zhuzhang Formula in trea-ting the spleen deficiency with dysfunction of transportation syndrome of idiopathic short stature(ISS)and its effects on the growth rate,disease-related indicators,and bone metabolism indicators in affect-ed children.Methods A total of 80 children with ISS were enrolled as study subjects.They were randomly assigned to control group and observation group using random number table method,with 40 cases in each group.The control group received conventional western medicine treatment,while the observation group was administered self-formulated Zhuzhang Formula in addition to the treatment of the control group.Comparisons were made between the two groups in terms of clinical efficacy,TCM syndrome scores,growth parameters(monthly average height increment,annual growth rate),levels of disease-related indicators[serum insulin-like growth factor-1(IGF-1),bone morphogenetic pro-tein-2(BMP-2),insulin-like growth factor binding protein-3(IGFBP-3)],levels of bone metabo-lism indicators[bone alkaline phosphatase(BAP),type Ⅰ procollagen N-terminal propeptide(PⅠNP),type Ⅰ collagen cross-linked C-terminal telopeptide(β-CTX)],and the incidence of adverse reac-tions.Results The overall effective rate in the observation group was 97.50％,which was significantly higher than the 80.00％ in the control group(P＜0.05).After 3 and 6 months of intervention,the scores for symptoms such as short stature,sallow complexion,loss of appetite,weak voice and shortness of breath,limb weakness,and loose stools,as well as the total TCM syndrome scores,were lower in the observation group than those in the control group,with statistically significant differences(P＜0.05).After 6 months of intervention,the monthly average height increment and annual growth rate were higher in the observation group than those in the control group,and the differences were statistically significant(P＜0.05).After 3 and 6 months of intervention,the levels of IGF-1,BMP-2,IGFBP-3,BAP,and PⅠNP were higher in the observation group than those in the control group,while the level of β-CTX was lower,and the differences were statistically significant(P＜0.05).There was no statistically significant difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion The self-formulated Zhuchang Formula demon-strates remarkable efficacy in treating the spleen deficiency with dysfunction of transportation syndrome of ISS.It can alleviate clinical symptoms by regulating the levels of IGF-1,BMP-2,IGFBP-3,and bone metabolism indicators,thereby promoting the growth and development of affected children,and exhibits high safety.

Objective:To establish and evaluate a second-tier screening method for neonatal congenital adrenal hyperplasia (CAH) and develop appropriate screening interpretation criteria.Methods:We employed liquid chromatography-tandem mass spectrometry to simultaneously detect five steroid hormones in dried blood spots: 17α-hydroxyprogesterone (17α-OHP), androstenedione (A4), 11-deoxycortisol (11-DOC), 21-deoxycortisol (21-DOC), and cortisol (F), calculating (17α-OHP+A4)/F and (17α-OHP+21-DOC)/F ratios for second-tier CAH screening. The study utilized 429 residual dried blood spot samples from neonates (0-7 days) who completed first-tier screening between January 2020 and March 2024 in Guangzhou Women and Children's Medical Center, Guangzhou Medical University, including first-tier negatives ( n=369), confirmed false positives ( n=50), and CYP21A2-confirmed 21-hydroxylase deficiency patients ( n=10). Mann-Whitney U and Kruskal-Wallis tests analyzed steroid concentration variations across gestational ages and birth weights in all negative samples, with reference intervals established via P2.5- P97.5 percentiles and screening cutoffs set at population P97.5. Receiver operating characteristic (ROC) curve analysis identified optimal interpretation indicators among steroid hormone profiles, with second-tier screening performance evaluated by comparing sensitivity and specificity across different steroid hormone indicators to establish the optimal diagnostic criteria. Results:The five steroid hormones demonstrated intra-assay precision with coefficient of variation (CV) of 9.8%-14.2% and inter-assay precision with CV of 4.7%-14.4% across three different concentration levels of quality control materials. Accuracy ranged from 98.5% to 110.0% and the lower limits of quantification were 0.25 ng/ml for 17α-OHP, 0.05 ng/ml for A4/11-DOC, 0.31 ng/ml for 21-DOC, and 0.1 ng/ml for F. Stratification by gestational age categorized 17α-OHP into ≤31, 32-34, and ≥35 weeks; A4 into ≤31, 32-36, and ≥37 weeks; and 11-DOC into ≤31 and ≥32 weeks, while the remaining indicators were not stratified. When grouped by birth weight (low/normal), all measured parameters except 21-DOC showed statistically significant differences between groups (all P<0.05). Established reference intervals included 17α-OHP: 0.53-7.82 ng/ml (≤31 weeks), <0.25-3.60 ng/ml (32-34 weeks), <0.25-1.64 ng/ml (≥35 weeks); A4: 0.12-2.36 ng/ml (≤31 weeks), <0.05-1.45 ng/ml (32-36 weeks), 0.17-0.95 ng/ml (≥37 weeks); 11-DOC: 0.43-4.04 ng/ml (≤31 weeks), 0.08-1.46 ng/ml (≥32 weeks); F: 1.70-83.70 ng/ml; 21-DOC: <0.31-0.69 ng/ml; (17α-OHP+A4)/F: 0.01-0.74; and (17α-OHP+21-DOC)/F: 0.01-0.69. Comprehensive comparison of CAH second-tier screening performance demonstrated that interpretation based on elevated 17α-OHP accompanied by either elevated 21-DOC or elevated ratios [(17α-OHP+A4)/F or (17α-OHP+21-DOC)/F] achieved 100% sensitivity, 96% specificity, and a 96% reduction in false-positive rate. Conclusion:The application of liquid chromatography-tandem mass spectrometry for multi-steroid hormone profiling in second-tier neonatal CAH screening, utilizing gestational age-specific 17α-OHP cutoffs combined with elevated 21-DOC or ratio criteria, demonstrated 100% screening sensitivity while substantially reducing false-positive rates from primary screening, though further validation with expanded sample sizes remains necessary.