Objective To study the changes in peripheral blood T lymphocytes in patients with ankylosing spondylitis and their correlation with the disease's severity and prognosis.Methods We selected 120 patients with ankylosing spondylitis treated between January 2020 and March 2023 as the research group and 120 healthy people who had medical examinations in the same period as the health group.We detected the changes in CD4+,CD8+and CD4+/CD8+values of peripheral blood T lymphocyte subsets with flow cytometry,compared the differences in T lymphocyte subsets between the two groups,and analyzed the correlation with the disease severity of the patients.All the 120 patients with ankylosing spondylitis were followed up for 6 months after treatment to assess their prognosis.General information and T lymphocyte sub-groups CD4+,CD8+level,CD4+/CD8+value changes were compared among patients with different prognosis.We analyzed the value of T lymphocyte sub-groups in predicting the prognosis of patients with ankylosing spondylitis.Results In the research group CD4+and CD4+/CD8+were lower but CD8 1 was higher than those in the healthy group(P＜0.05).CD4+and CD4+/CD8 were lower but CD8+was higher in patients with advanced ankylosing spondylitis than in early and mid-term patients(P＜0.05).The ROC curve analysis showed that the AUC of CD4+,CD8+,and CD4+/CD8+combined diagnosis of ankylosing spondylitis patients was 0.878,with higher diagnostic sensitivity than that of the single diagnosis(P＜0.05).In the poor prognosis group,CD8+was higher than that in the excellent prognosis group,but CD4+and CD4+/CD8 value were lower than the latter(P＜0.05).The results of Pearson test showed that CD4+and CD4+/CD8+were negatively correlated with the prognosis of patients with ankylosing spondylitis(r=-0.568,-0.656,P＜0.001).CD8+was positively correlated with the prognosis of patients with ankylosing spondylitis(r=0.623,P＜0.001).ROC curve analysis showed that the AUC of the combined diagnosis of CD4+,CD8+and CD4+/CD8+for ankylosing spondylitis patients was 0.910,and the diagnostic sensitivity was higher than that of single diagnosis(P＜0.05).Conclusion The abnormal levels of peripheral blood T lymphocyte subsets in patients with ankylosing spondylitis are closely related to the severity and prognosis of the disease,and can be used as a reference indicator for diagnosing the severity and prognosis of ankylosing spondylitis.

Background::Hyperglycemia frequently induces apoptosis in endothelial cells and ultimately contributes to microvascular dysfunction in patients with diabetes mellitus (DM). Previous research reported that the expression of integrins as well as their ligands was elevated in the diseased vessels of DM patients. However, the association between integrins and hyperglycemia-induced cell death is still unclear. This research was designed to investigate the role played by integrin subunit β5 (ITGB5) in hyperglycemia-induced endothelial cell apoptosis.Methods::We used leptin receptor knockout (Lepr-KO) ( db/ db) mice as spontaneous diabetes animal model. Selective deletion of ITGB5 in endothelial cell was achieved by injecting vascular targeted adeno-associated virus via tail vein. Besides, we also applied small interfering RNA in vitro to study the mechanism of ITGB5 in regulating high glucose-induced cell apoptosis. Results::ITGB5 and its ligand, fibronectin, were both upregulated after exposure to high glucose in vivo and in vitro. ITGB5 knockdown alleviated hyperglycemia-induced vascular endothelial cell apoptosis and microvascular rarefaction in vivo. In vitro analysis revealed that knockdown of either ITGB5 or fibronectin ameliorated high glucose-induced apoptosis in human umbilical vascular endothelial cells (HUVECs). In addition, knockdown of ITGB5 inhibited fibronectin-induced HUVEC apoptosis, which indicated that the fibronectin-ITGB5 interaction participated in high glucose-induced endothelial cell apoptosis. By using RNA-sequencing technology and bioinformatic analysis, we identified Forkhead Box Protein O1 (FoxO1) as an important downstream target regulated by ITGB5. Moreover, we demonstrated that the excessive macroautophagy induced by high glucose can contribute to HUVEC apoptosis, which was regulated by the ITGB5-FoxO1 axis. Conclusion::The study revealed that high glucose-induced endothelial cell apoptosis was positively regulated by ITGB5, which suggested that ITGB5 could potentially be used to predict and treat DM-related vascular complications.

Objective This study aims to analyze the proteomic characteristics of peripheral blood in patients with lung cancer coexistent with pulmonary tuberculosis(LC-PTB),identify the differential proteins compared with lung cancer(LC)patients,and conduct functional analysis on these proteins.Methods The study included 8 LC-PTB patients and 10 LC patients.The LC patients were newly diagnosed and confirmed by pathology and did not receive any anti-tumor treatment before,while the PTB patients were Mycobacterium tuberculosis positive at the time of sampling.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was applied to perform proteomic mass spectrometry to assess the differential proteins,and then functional analysis was conducted via bioinformatics.Results A total of 5,185 proteins were detected between two groups.Through differential expression screening,190 proteins(58 upregulated and 132 downregulated)were identified to be differentially expressed.Subcellular localization analysis revealed that the differential proteins were mainly concentrated in the cytoplasm,nucleus,and extracellular matrix.KEGG pathway and GO analysis showed the roles of differential proteins in biological processes including immune response,metabolism,and secretion regulation.Protein interaction network analysis highlighted the importance of SORT1,SAR1B,RPS6KB1,VWF,SHC1,SRPRB,CTSD,TARDBP,RPLP0,PSMA2,RPS6,XPO1,PRKACB,and HLA-DRB1 in LC-PTB.Additionally,the expression changes in proteins like ADA2,MAP3K1,and GLS2 might be closely associated with the development of LC-PTB.Conclusions The proteomic profile comprehensively described the proteomic characteristics of LC-PTB and identified numerous differentially expressed proteins,which could provide further clues for research on biological mechanism of LC-PTB.

As the incidence of male infertility has been increasing during recent years, it is urgent to reveal the pathogenesis of male infertility, as well as to develop the new drugs for treatment of male infertility, in order to solve the declining birth rate and aging problems. The construction and application of male infertile animal models is critical for drug development, which plays an important role in accurately evaluating the efficacy and mechanism of infertility treatment. A suitable infertility model not only can reduce the repeated drug efficacy evaluations, reduce animal usage and the cost of new drug development, but also has important reference value for subsequent clinical trial research. Male infertility laboratory animal models can be constructed through chemical, physical, endocrine, environmental estrogen, gene modification, and immune methods. This article mainly introduces the existing male infertility animal models available for drug development, and briefly introduces the application progress of each model to provide reference for the male infertility drug researchers.

In recent years, with the rapid development of life sciences, the use of laboratory fish in toxicology, genetics, developmental biology and medicine has increased dramatically, and they have gradually become important new model organisms. At the same time, the welfare of laboratory fish has also received increasing attention. Although the research level of experimental fish welfare is still in a relatively early stage compared to terrestrial experimental animals, developed regions such as Europe and America have established corresponding legal frameworks to safeguard the welfare of laboratory fish in research. This article elucidates the current developmental status of laboratory fish welfare, discusses the rationale behind the imperative to prioritize and enhance their welfare, deeply investigates factors influencing their welfare from the feeding stage and experimental stage. Moreover, it explores strategies for augmenting welfare standards, with the overarching aim of propelling the continual improvement of laboratory fish welfare in our country.

ObjectiveTo observe the fine structure of the trunk kidney in zebrafish, and to identify its secreted exosomes. MethodsThe microstructure and ultrastructure of the trunk kidney in zebrafish were observed by light microscopy and electron microscopy, and the particle size of exosomes was detected by nanoparticle tracking analysis (NTA). ResultsThe trunk kidney was close and parallel to the spine in adult zebrafish. The nephron consisted of renal tubules and renal corpuscles. The renal tubules could be further divided into three types: proximal convoluted tubules, distal convoluted tubules, and cervical segments. The renal corpuscles were composed of glomerulus and renal capsules. The periodic acid-Schiff (PAS) staining results revealed that there were abundant glycogen granules in the proximal convoluted tubules, with brush-like outline in the apical surface of epithelial cells. Under transmission electron microscopy (TEM), there were exosomes distributed in the lumen of renal tubules, with numerous late endosomes and few number of multivesicular bodies (MVBs) in the cytoplasm of the epithelial cells concentrating on the apical side. Meanwhile, MVBs were also distributed in the apical regions of the renal tubules and the podocytes of the renal glomeruli. Immunohistochemical staining results showed that CD9, CD63 and TSG101 were strongly expressed in the lumen surface of the renal tubules, but weakly expressed in the corpuscles and lumen. NTA and TEM results showed that the exosomes isolated from zebrafish trunk kidney were saucer-like outline, and the particle size mode was 144.4 nm, which was consistent with the characteristics of morphological futures of exosome. ConclusionThe zebrafish somatic kidney has the typical structure of the mammalian kidney and is the urinary organ in the body. The renal tubules have the ability to secrete exosomes, and their formation is a process of releasing poly-vesicles to the free surface of epithelial cells into the extracellular space. This study laid a morphological foundation for further study of exosomes in urinary function in aquatic experimental animals as well as the development and application of related models.

Objective To compare the differences of bacterial distribution of intestinal flora in Microtus fortis living under laboratory feeding and wild survival conditions. Methods The 16S rDNA-V4-V5 region of bacteria in the ileocecal contents from Microtus fortis raised in lab and captured in wild were measured by high-throughput sequencing. The number of operational taxonomic units(OTUs)were sorted and calculated,and the species abundance and distribution and difference were analyzed. Results The rarefaction curves indicated that adequate sampling was achieved. At the phylum level,the distribution of intestinal flora between two groups was similar. The experimental group had a unique phylum, Lentisphaerae. The wild type group had 3 unique phylums,Fusobacteria,Thaumarchaeota and an unclassified phylum. At the genus level, the kind of intestinal flora in the wild type group was more abundant than the experimental group. Ruminococcus is the largest differential genus. Conclusions The microbial community structure and differences of Microtus fortis living under different conditions are obtained. It may further enrich the basic biology data of Microtus fortis.

Objective To more intuitively understand the quality control for laboratory animals and further achie-ving a more scientific and reasonable management of laboratory animals, the infection index as evaluation criteria was intro-duced. Then the best way to calculate infection index was explored in order to more scientifically reflect the infection status of laboratory animals. Methods Infection index, also called the degree of infection, is a qualitative indicator of monito-ring laboratory animal quality. After arranging, analyzing, processing and gathering the data from laboratory animal quality monitoring, the index reflects synthetically the pathogen infection status or trend of a particularly investigated experimental animal population or the development of certain experimental animals. Results In general, the pathogen infection index of mice was slightly decreased, while the pathogen infection index of rats roughly increased year by year. In comparing infec-tion index by different pathogens, the parasite infection index of mice was found to be higher than bacteria and virus infec-tion indexes, while the bacteria infection index of rats was higher than parasite infection index and virus ones. Conclusions The infection index model intuitively reflects the quality control status of laboratory animals. The analysis also reveals that the parasite monitoring of the mice and the bacteria detection of rat needs to be reinforcement. In addition, the index of infection reveals that the pathogen infection of mice is well under control, while that of rats tends to be more serious year by year.

Objective To investigate the possible protective effect of adenoviral vector expressing interleukin-1β (IL-1β) small hairpin RNA (shRNA) on spinal cord injury (SCI) and its mechanism in rats.Methods Forty-eight adult male Sprague-Dawley rats were randomly assigned to 4 groups including the Sham, the Vehicle,the Ad-GFP and the Ad-shIL-1β groups.SCI was induced by epidural compression.Motor function of hind limbs was evaluated by Basso-Beattie-Bresnahan (BBB) score, the expressions of green fluorescence in injured spinal cord tissue were observed by fluorescence microscope.Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence were also performed.Results The expressions of green fluorescence in injured spinal cord tissue were observed in the Ad-GFP and Ad-shIL-1β groups one day after SCI.Significant functional improvement was observed in the Ad-shIL-1β group (8.17 ± 1.17, 10.17 ± 0.98 and 11.33 ± 0.82, respectively) compared to the Vehicle (4.00 ± 0.89, 5.67 ± 1.03 and 6.17 ± 1.17, respectively) and Ad-GFP (3.83 ± 0.98, 5.33 ± 1.21 and 5.67 ± 1.03, respectively) groups at 7, 14 and 21 days after SCI (P ＜ 0.05).Rats in the Ad-shIL-1β group had less neuronal loss 21 days after SCI.In addition, IL-1β downregulation significantly decreased IL-1β, tumor necrosis factor-or (TNF-α) and IL-6 levels (138.83 ± 7.96,143.38 ± 10.20 and 120.43 ± 9.79 in Ad-shIL-1β group;169.33 ± 11.45, 172.33 ± 8.26 and 163.00 ± 9.57 in Vehicle group;172.83 ± 10.85,167.48 ± 8.19 and 159.48 ± 10.98 in Ad-GFP group, respectively) one day after SCI (P ＜ 0.05).Conclusion This study demonstrated that the IL-1β downregulation may have potential therapeutic benefits for improving the outcomes after SCI.

Objective To explore application value of combined detection of procalcitonin(PCT ) and high‐sensitivity C‐reactive protein (hs‐CRP) in clinical diagnosis of infectious diseases .Methods 162 cases of hospitalized patients with infectious diseases from June 2012 to December 2014 were enrolled in this study and divided into bacterial infection group(92 cases)and viral infection group (70 cases) according to the results of isolation and culture of pathogen and virus antibody detection .Other 100 cases of healthy individuals were selected as healthy control .The serum levels of PCT and hs‐CRP were detected by using immunochroma‐tography and immunofluorescence assay respectively and were statistically analysed .Results The serum levels of PCT and hs‐CRP in the bacterial infection group were higher than those in the healthy control group ,the differences were statistically significant (P<0 .05) .The serum levels of PCT and hs‐CRP in the viral infection group were higher than those in the healthy control group , but only the difference of hs‐CRP between the two groups was statistically significant(P<0 .05) .In patients with bacterial infection and viral infection ,the positive rates of combined detection of hs‐CRP and PCT were higher than single detection of hs‐CRP or PCT ,the differences were statistically significant(P<0 .05) .Conclusion PCT and hs‐CRP could be important serum markers of bacterial infections ,combined detection of the two markers have clinical significance for diagnosing infectious diseases and assessing its prognosis .