[The role of Treg/Thl7 and calprotectin in ulcerative colitis rat model].

OBJECTIVE: To investigate the roles of helper T cells (Th), regulatory T cells (Treg) and calprotectin in the pathogenesis of ulcerative colitis (UC) in the rat model. METHODS: Thirty rats were randomly divided into normal control (NC) group and UC model (UC) group. The rats of UC group were replicated using the mixed solution of 2, 4,6 trinitrobenzene sulfonic acid (TNBS) / ethanol through the intestinal tract. The rats of NC group were given NaCI solution enema. Three weeks after UC modeling, the morphology changes of colon tissue were observed. The serum levels interleukin (IL)-1ß, IL-10, IL-17, IL-23 and calprotectin (CP) were detected by enzyme-linked immunosorbent assay. The changes of Treg in peripheral blood were measured by flow cytometry. The expressions of IL-17, CP, and transcriptional regulatory proteins of decision Treg cell differentiation and function (FoxP3) in colon tissue were detected by Western blot and immunohistochemistry. RESULTS: UC group was observed with obvious colon tissue injury and inflammatory cells infiltration in intestinal tissue, compared with NC group, the expression of IL-1ß,IL-17,IL-23,CP in serum were increased, while the expression of IL-10, the number of Treg were decreased in UC group (P<0. 05 or P(<0. 01). Compared with NC group, the expression of IL-17, CP in colon tissue were significantly increased in UC group (P(<0.05), and the expression of FoxP3 was increased (P(0. 05). CONCLUSION: Expressions of calprotectin, IL-17 of intestinal tissue, serum are increased in rat of ulcerative colitis, which to lead to the imbalance of Treg/Thl7 cells, and elevate inflammatory responses. These factors have promote the occurrence and development of ulcerative colitis.

Exploring the regulatory role of Linc00893 in asthenozoospermia: Insights into sperm motility and SSC viability.

The role of long intergenic noncoding RNA 00893 (Linc00893) in asthenozoospermia (AS) and its impact on sperm motility remains unclear The present study explored the effect of Linc00893 on AS, specifically its effect on sperm motility and its relationship with spermatogonial stem cell (SSC) vitality and myosin heavy chain 9 (MYH9) protein expression. Linc00893 expression was analyzed in semen samples using reverse transcription­quantitative PCR, revealing a significant downregulation in samples from individuals with AS compared with those from healthy subjects. This downregulation was found to be negatively correlated with parameters of sperm motility. To further understand the role of Linc00893, small interfering RNA was used to knockdown its expression in SSCs. This knockdown led to a marked decrease in cell vitality and an increase in apoptosis. Notably, Linc00893 knockdown was shown to inhibit MYH9 expression by competitively binding with microRNA­107, a finding verified by dual­luciferase reporter and RNA immunoprecipitation assays. Furthermore, using the GSE160749 dataset from the Gene Expression Omnibus database, it was revealed that MYH9 protein expression was downregulated in AS samples. Subsequently, lentiviral vectors were constructed to induce overexpression of MYH9, which in turn reduced SSC apoptosis and counteracted the apoptosis triggered by Linc00893 knockdown. In conclusion, the present study identified the role of Linc00893 in AS, particularly its regulatory impact on sperm motility, SSC vitality and MYH9 expression. These findings may provide information on the potential regulatory mechanisms in AS development, and identify Linc00893 and MYH9 as possible targets for diagnosing and treating AS­related disorders.

Identification of potential biomarkers and pathways for asthenozoospermia by bioinformatics analysis and experiments.

Background: Asthenozoospermia, a type of male infertility, is primarily caused by dysfunctional sperm mitochondria. Despite previous bioinformatics analysis identifying potential key lncRNAs, miRNAs, hub genes, and pathways associated with asthenospermia, there is still a need to explore additional molecular mechanisms and potential biomarkers for this condition. Methods: We integrated data from Gene Expression Omnibus (GEO) (GSE22331, GSE34514, and GSE160749) and performed bioinformatics analysis to identify differentially expressed genes (DEGs) between normozoospermia and asthenozoospermia. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were conducted to gain insights into biological processes and signaling pathways. Weighted Gene Co-expression Network Analysis (WGCNA) identified gene modules associated with asthenozoospermia. Expression levels of key genes were assessed using datasets and experimental data. Gene Set Enrichment Analysis (GSEA) and correlation analysis identified pathways associated with the hub gene and explore the relationship between the ZNF764 and COQ9 and mitochondrial autophagy-related genes. Competitive endogenous RNA (ceRNA) networks were constructed, and in vitro experiments using exosome samples were conducted to validate this finding. Results: COQ9 was identified as a marker gene in asthenozoospermia, involved in autophagy, ATP-dependent chromatin remodeling, endocytosis, and cell cycle, etc. The ceRNA regulatory network (LINC00893/miR-125a-5p/COQ9) was constructed, and PCR demonstrated that LINC00893 and COQ9 were downregulated in asthenozoospermia, while miR-125a-5p and m6A methylation level of LINC00893 were upregulated in asthenozoospermia compared to normozoospermic individuals. Conclusion: The ceRNA regulatory network (LINC00893/miR-125a-5p/COQ9) likely plays a crucial role in the mechanism of asthenozoospermia. However, further functional experiments are needed to fully understand its significance.

Significance of elevated levels of collagen type IV and hyaluronic acid in gastric juice and serum in gastric cancer and precancerous lesion.

BACKGROUND AND AIMS: Collagen type IV and hyaluronic acid (HA) are the major components of basement membrane and extracellular matrix, respectively. Cathepsin D is an aspartyl lysosomal protease involved in the degradation of the basement membrane and extracellular matrix. The aim of this study is to investigate the clinical significance of collagen type IV and hyaluronic acid in gastric juice and serum in diagnosis of gastric cancer and the degrading effect of cathepsin D on collagen type IV and HA. METHODS: Fifty gastric cancer patients were enrolled in our study compared with 41 patients with precancerous lesion and 30 control subjects. Collagen type IV and HA in gastric juice and serum were analyzed by radioimmunoassay. Expression of cathepsin D and collagen type IV in tissue were analyzed by immunohistochemical staining with monoclonal antibodies. RESULTS: The contents of collagen type IV and HA in gastric juice and HA in serum were significantly higher in patients with gastric cancer than those in patients with precancerous lesion and control group (p < 0.05, p < 0.0001). Gastric cancer patients with lymph node metastasis had a higher level of collagen type IV and HA in gastric juice than those in patients without metastasis (p = 0.049, p = 0.043). The expression of cathepsin D had significantly increased in patients with gastric cancer compared to the control group (p < 0.0001). The continuous expression of collagen type IV in basement membrane in gastric cancer group was lower than that in the precancerous lesion group and control group (p < 0.0001). CONCLUSIONS: The analysis of collagen type IV and HA in gastric juice and serum may provide a simple aid in diagnosing gastric cancer and evaluating whether metastasis is occurring or not.

[Analysis of Genetic Screening in Couples of Reproductive Age for Thalassemia in Lingshui Li Autonomous County of Hainan Province].

OBJECTIVE: To investigate the genetic carrier rate of thalassemia and its gene mutation types as well as the distribution characteristics among the people in Lingshui Li autonomous county of Hainan province, so as to provide the basis for making the prevention programs of thalassemia in administrative departments. METHODS: Samples were collected from couples undergoing premarital and pregestational screenings, in which the positive ones in preliminary screening were further tested by genetic diagnoses and the genotypes were analyzed. RESULTS: The rate of thalassemia gene carriers was 19.41% （274/1412） of the couples of childbearing age in Lingshui Li autonomous County of Hainan Province. In these carriers,α-thalassemia accounted for 83.21%（228/274）, ß-thalassemia for 8.03%（22/274）, and both α-and ß-thalassemia gene accounted for 8.76% （28/274）. CONCLUSION: The carrying rate of thalassemia gene in population Lingshui Li autonomous county of Hainan province is high, and its distribution has geographical characteristics,the major type is α-thalassemia. Blood screening and genetic diagnosis of thalassemia should be strengthened, and corresponding measures should be taken to reduce its gene frequency.

[Expression and correlation analysis between inflammatory cytokines and calprotectin in the rat model of ulcerative colitis].

OBJECTIVE: To observe the changes of inflammatory cytokines and calprotectin (CP) in the rat model of ulcerative colitis (UC) and investigate the mechanism of the inflammatory response. METHODS: Twenty rats were randomly divided into normal control (NC) group and model (MC) group, with 10 rats in each group. The rats of MC group were given through the intestinal tract the mixed solution of 2, 4, 6 trinitrobenzene sulfonic acid (TNBS)/ethanol to induce UC. The rats of NC group were given normal saline instead. Three weeks after modeling, the changes of colon tissue morphology were observed with HE staining. The levels of interferon gamma (INF-γ), interleukin (IL)-4, IL-10, IL-12 and CP in serum were detected by ELISA. The expressions of INF-γ, IL-4, CP of colon tissue were detected by Western blotting. RESULTS: Indicators of inflammatory activity were significantly elevated, and colon tissue injury was seen in MC group. Compared with the NC group, the expressions of INF-γ, IL-12 and CP of serum increased in the MC group (P<0.05 or P<0.01). The expressions of IL-4 and IL-10 in the MC group were lower than those in the NC group (P<0.01). Compared with the NC group, the expressions of INF-γ and CP of colon tissue significantly increased (P<0.01 or P<0.05), and the expression of IL-4 decreased in the MC group (P<0.05). Correlation analysis showed that there was a positive correlation between CP and IFN-γ, and a negative correlation between CP and IL-4 (P<0.05). CONCLUSION: Increased CP in UC rats can promote the expression of inflammatory factors, lead to the imbalance of Th1/Th2 cells, and enhance inflammatory responses. These factors promote the occurrence and development of UC.

Emodin promoted pancreatic claudin-5 and occludin expression in experimental acute pancreatitis rats.

AIM: To investigate the effect of emodin on pancreatic claudin-5 and occludin expression, and pancreatic paracellular permeability in acute pancreatitis (AP). METHODS: Experimental pancreatitis was induced by retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. Emodin was injected via the external jugular vein 0 or 6 h after induction of AP. Rats from sham operation and AP groups were injected with normal saline at the same time. Samples of pancreas were obtained 6 or 12 h after drug administration. Pancreatic morphology was examined with hematoxylin and eosin staining. Pancreatic edema was estimated by measuring tissue water content. Tumor necrosis factor (TNF)-α and interleukin (IL)-6 level were measured by enzyme-linked immunosorbent assay. Pancreatic paracellular permeability was assessed by tissue dye extravasation. Expression of pancreatic claudin-5 and occludin was examined by immunohistology, quantitative real-time reverse transcriptase polymerase chain reaction and western blotting. RESULTS: Pancreatic TNF-α and IL-6 levels, wet/dry ratio, dye extravasation, and histological score were significantly elevated at 3, 6 and 12 h following sodium taurocholate infusion; treatment with emodin prevented these changes at all time points. Immunostaining of claudin-5 and occludin was detected in rat pancreas, which was distributed in pancreatic acinar cells, ductal cells and vascular endothelial cells, respectively. Sodium taurocholate infusion significantly decreased pancreatic claudin-5 and occludin mRNA and protein levels at 3, 6 and 12 h, and that could be promoted by intravenous administration of emodin at all time points. CONCLUSION: These results demonstrate that emodin could promote pancreatic claudin-5 and occludin expression, and reduce pancreatic paracellular permeability.