ObjectiveTo establish a highly specific and sensitive triplex TaqMan quantitative PCR method for the detection of mouse hepatitis virus (MHV), mouse parvovirus (MPV), and reovirus type 3 (Reo-3) in laboratory mice. MethodsRecombinant plasmid standards were constructed using conserved genomic regions of the three target viruses. Specific primers and probes were designed, and a triplex TaqMan quantitative PCR (qPCR) system was optimized through preliminary experiments. Sensitivity was evaluated using 10-fold serial dilutions (10¹-10⁷ copies/μL). Intra- and inter-assay repeatability were assessed via reproducibility experiments. ResultsA strong linear correlation (R ²>0.99) was observed between copy number and Ct value at template concentrations of 10²-10⁷ copies/μL. Intra-assay and inter-assay coefficients of variation (CV) were both below 5%. No cross-reactivity was observed with mouse cytomegalovirus (MCMV), vesicular stomatitis virus (VSV), Sendai virus (SeV), or pneumonia virus of mice (PVM). ConclusionThe triplex TaqMan quantitative PCR method established in this study has high sensitivity, good repeatability, and strong specificity. It enables the rapid, specific, and sensitive detection of MHV, MPV, and Reo-3, allows the simultaneous detection of multiple pathogens in a single tube, and can be applied to the detection of clinical samples from laboratory animals and to epidemiological investigations, thereby providing effective scientific and technological support for the prevention and control of viral infection and the interruption of transmission.

Objective:To observe the effects of acupuncture on intestinal flora and inflammatory factor interleukin(IL)-6 in ulcerative colitis(UC)model rats and to explore the related mechanism of acupuncture in treating UC.Methods:Thirty-two Sprague-Dawley rats were randomly divided into a normal control(NC)group,a model(MO)group,a mesalazine(ME)group,and an acupuncture(AC)group,with 8 rats in each group.Rats,except those in the NC group,were given a 2.5%sodium dextran sulfate solution to establish UC models.After successful modeling,rats in the ME group were treated with mesalazine by gavage twice a day for 7 continuous days;rats in the AC group received acupuncture at bilateral Tianshu(ST25)and Zusanli(ST36)with needles retained for 20 min each time,once a day for 7 consecutive days.After intervention,changes in the intestinal flora diversity,colon tissue damage degree,and inflammatory factor expression in each group of rats were evaluated.Results:The body mass increased slowly in the MO group compared to the NC group.After intervention,the body mass increased more significantly,colon injuries recovered,and intestinal flora diversity improved in the ME and AC groups compared to the MO group(P<0.05).The IL-6 expression level was higher in the MO group compared to the NC group(P<0.05).The IL-6 expression level decreased in the AC group compared to the MO group(P<0.05).Conclusion:Acupuncture at Tianshu(ST25)and Zusanli(ST36)improves body mass loss and colon injuries,regulates intestinal flora diversity,and reduces the expression of inflammatory factor IL-6 in the colon tissue of UC rats,therefore alleviating the disease severity of UC.

Objective To explore the casual relationship between immune cells and chronic pancreatitis(CP)using Mendelian randomization(MR)analysis.Methods The immune cell phenotypes and CP GWAS data used in this study were obtained from public databases,and 731 immune cell phenotypes were included.The bidirectional MR analysis was used to explore the causal relationship between immune cells and CP,and various sensitivity analysis methods were used to verify the heterogeneity and level multiplicity of the results.Results This study identified 33 immune cell phenotypes with a causal relationship with CP,of which 18 were inhibitory factors,and the rest were risk factors.Among the 18 inhibitory factors,CD25 on CD4+in the Treg cell group showed the most significant inhibitory effect.Among the 15 risk factors,CD8br AC in the TBNK cell group,CD8br on TD CD8br in the mature T cell group,and CD39+CD8br％ T cell and CD28 on CD4+in the Treg cell group showed statistical significance.The reverse MR results further confirmed the unidirectionality of the causal relationship.Conclusion Our study revealed the close relationship between immune cells and CP through MR method,highlighting the complex interaction pattern between the immune system and CP.

Objective:To evaluate the causal relationship between autism spectrum disorder (ASD) and chronic pancreatitis (CP) using a bidirectional two-sample Mendelian randomization (MR) analysis.Methods:Based on genome-wide association study (GWAS) data, forward and reverse two-sample MR analyses were conducted to examine the causal relationship between ASD and CP. Single-nucleotide polymorphisms (SNPs) reaching genome-wide significance were selected from the GWAS data as candidate instrumental variables, and the Steiger directionality test was used to confirm the causal direction. The inverse-variance weighted (IVW) method was applied as the primary analysis to estimate the causal effect of the exposure on the outcome. MR-Egger regression and weighted median methods were used as supplementary sensitivity analyses to assess the robustness of the results. Pleiotropy and heterogeneity tests were performed to evaluate the reliability of the findings.Results:The forward MR analysis ultimately identified 29 SNPs. The IVW analysis indicated that ASD had a significant positive causal effect on CP risk ( OR=1.197, 95% CI 1.047-1.368, P=0.008), with no evidence of horizontal pleiotropy or significant heterogeneity. In the reverse MR analysis, 17 SNPs were included; the IVW analysis did not detect a significant causal effect of CP on ASD ( OR=0.990, 95% CI 0.935-1.047, P=0.717), also with no evidence of horizontal pleiotropy or significant heterogeneity. Causal effect estimates from MR-Egger regression and weighted median analyses were generally consistent with those of IVW. Conclusions:These findings indicate that, at the level of genetic susceptibility, ASD is a risk factor for CP.

Objective:To evaluate the causal relationship between autism spectrum disorder (ASD) and chronic pancreatitis (CP) using a bidirectional two-sample Mendelian randomization (MR) analysis.Methods:Based on genome-wide association study (GWAS) data, forward and reverse two-sample MR analyses were conducted to examine the causal relationship between ASD and CP. Single-nucleotide polymorphisms (SNPs) reaching genome-wide significance were selected from the GWAS data as candidate instrumental variables, and the Steiger directionality test was used to confirm the causal direction. The inverse-variance weighted (IVW) method was applied as the primary analysis to estimate the causal effect of the exposure on the outcome. MR-Egger regression and weighted median methods were used as supplementary sensitivity analyses to assess the robustness of the results. Pleiotropy and heterogeneity tests were performed to evaluate the reliability of the findings.Results:The forward MR analysis ultimately identified 29 SNPs. The IVW analysis indicated that ASD had a significant positive causal effect on CP risk ( OR=1.197, 95% CI 1.047-1.368, P=0.008), with no evidence of horizontal pleiotropy or significant heterogeneity. In the reverse MR analysis, 17 SNPs were included; the IVW analysis did not detect a significant causal effect of CP on ASD ( OR=0.990, 95% CI 0.935-1.047, P=0.717), also with no evidence of horizontal pleiotropy or significant heterogeneity. Causal effect estimates from MR-Egger regression and weighted median analyses were generally consistent with those of IVW. Conclusions:These findings indicate that, at the level of genetic susceptibility, ASD is a risk factor for CP.

Objective To explore the casual relationship between immune cells and chronic pancreatitis(CP)using Mendelian randomization(MR)analysis.Methods The immune cell phenotypes and CP GWAS data used in this study were obtained from public databases,and 731 immune cell phenotypes were included.The bidirectional MR analysis was used to explore the causal relationship between immune cells and CP,and various sensitivity analysis methods were used to verify the heterogeneity and level multiplicity of the results.Results This study identified 33 immune cell phenotypes with a causal relationship with CP,of which 18 were inhibitory factors,and the rest were risk factors.Among the 18 inhibitory factors,CD25 on CD4+in the Treg cell group showed the most significant inhibitory effect.Among the 15 risk factors,CD8br AC in the TBNK cell group,CD8br on TD CD8br in the mature T cell group,and CD39+CD8br％ T cell and CD28 on CD4+in the Treg cell group showed statistical significance.The reverse MR results further confirmed the unidirectionality of the causal relationship.Conclusion Our study revealed the close relationship between immune cells and CP through MR method,highlighting the complex interaction pattern between the immune system and CP.

Objective:To observe the effects of acupuncture on intestinal flora and inflammatory factor interleukin(IL)-6 in ulcerative colitis(UC)model rats and to explore the related mechanism of acupuncture in treating UC.Methods:Thirty-two Sprague-Dawley rats were randomly divided into a normal control(NC)group,a model(MO)group,a mesalazine(ME)group,and an acupuncture(AC)group,with 8 rats in each group.Rats,except those in the NC group,were given a 2.5%sodium dextran sulfate solution to establish UC models.After successful modeling,rats in the ME group were treated with mesalazine by gavage twice a day for 7 continuous days;rats in the AC group received acupuncture at bilateral Tianshu(ST25)and Zusanli(ST36)with needles retained for 20 min each time,once a day for 7 consecutive days.After intervention,changes in the intestinal flora diversity,colon tissue damage degree,and inflammatory factor expression in each group of rats were evaluated.Results:The body mass increased slowly in the MO group compared to the NC group.After intervention,the body mass increased more significantly,colon injuries recovered,and intestinal flora diversity improved in the ME and AC groups compared to the MO group(P<0.05).The IL-6 expression level was higher in the MO group compared to the NC group(P<0.05).The IL-6 expression level decreased in the AC group compared to the MO group(P<0.05).Conclusion:Acupuncture at Tianshu(ST25)and Zusanli(ST36)improves body mass loss and colon injuries,regulates intestinal flora diversity,and reduces the expression of inflammatory factor IL-6 in the colon tissue of UC rats,therefore alleviating the disease severity of UC.

Based on introducing the total price adjustments in pilot cities and analyzing the existing problems,it further analyzes the objectives of the total price adjustment allocation of medical service items,the characteristics of various types of medical service items and the possible impact of price adjustment,concludes that the priority of the total price adjustment allocation should be as follows:new items,special tasks,complex items,general items,and medical services for special needs.It also combines the practical experience of the pilot cities to establish the total price adjustment allocation mechanism,and provides opinions on the total price adjustment allocation before the dynamic adjustment of medical service prices in the future.

【Objective】 To explore the causal relationship between education level and pancreatitis risk through Mendelian randomization. 【Methods】 A two-sample Mendelian randomization analysis was conducted using genome-wide association study (GWAS) summary data. The GWAS data for education level and pancreatitis were obtained from SSGAC database and the FinnGen database (version R9). Causal relationship between education level and pancreatitis was explored using the inverse variance weighted (IVW), MR-Egger, and weighted median methods. Heterogeneity and directional pleiotropy were evaluated using Cochran’s Q test and funnel plots. 【Results】 Totally 604 SNPs associated with education level were included. The results provided evidence that there was negative relationship between education level and pancreatitis risk. For acute pancreatitis, OR=0.52, 95% CI: 0.44-0.62, P=2.43×10^-14 while for chronic pancreatitis, OR=0.51, 95% CI: 0.41-0.64, P=7.20×10^-9. Results from MR-Egger and weighted median analyses obtained the same results. The results of sensitivity analysis indicated that this study did not violate the basic assumptions of Mendelian randomization. 【Conclusion】 There is a causal relationship between education level and the occurrence of pancreatitis. The educational level is negatively correlated with the risk of pancreatitis.

AIM: To investigate the relationship between objective ocular torsion and near stereopsis in patients with primary superior oblique overaction(PSOOA).METHODS: Retrospective study. A total of 59 strabismus patients with PSOOA who underwent strabismus surgery at Renmin Hospital of Wuhan University between January 2019 and November 2023 were collected. Based on the collected fundus photographs and the position of the fovea relative to the horizontal line through the optic disc, the eyes were categorized as incyclotorsion, excyclotorsion, or no cyclotorsion. Additionally, the fovea-disc angle(FDA)and the relationship between objective ocular torsion status, FDA, and near stereopsis function in the patients were further measured and analyzed.RESULTS: Totally 59 patients(92 eyes)showed superior oblique overaction. There were no cases of excyclotorsion, 32 cases with no cyclotorsion, and 27 cases with incyclotorsion. The total binocular FDA was significantly smaller in the no-cyclotorsion group compared with the incyclotorsion group(2.83°±2.89° vs 16.12°±5.74°, P<0.001). The preservation rates of near stereopsis were 66% and 15% in the no-cyclotorsion and incyclotorsion groups, respectively, with a significant statistical difference(P<0.001), and the preservation rates of fine near stereopsis were 38% and 11% in the no-cyclotorsion and incyclotorsion groups, respectively, with a significant statistical difference(P=0.02). Among all patients, near stereopsis was correlated with total binocular FDA(r=-0.526, P<0.001), with the strongest correlation observed with the FDA of the incyclotorsion(r=-0.546, P<0.001). In the incyclotorsion group, there was no correlation between near stereopsis and total binocular FDA(r=-0.366, P=0.060), with a negative correlation between near stereopsis and the FDA of both the incyclotorsion and the overaction(r=-0.424, P=0.028; r=-0.485, P=0.010). In the no-cyclotorsion group, near stereopsis was not correlated with total binocular FDA, incyclotorsion FDA, or overaction FDA.CONCLUSION:PSOOA patients with incyclotorsion have a lower preservation rate of near stereopsis than those without cyclotorsion. Near stereopsis function of patients with PSOOA is negatively correlated with total binocular FDA, especially the greater the FDA of the incyclotorsion and overaction, the worse the near-stereoscopic function.