BackgroundIndividuals with chronic multimorbidity exhibit a high prevalence of sleep disturbances， which impair their physical and psychological well-being and impede chronic disease management. Although sleep quality is closely influencing with psychological functioning， empirical evidence regarding the longitudinal trajectories of sleep quality and their influencing factors in this population remains insufficient. ObjectiveTo investigate the longitudinal trajectories of sleep quality and associated factors among patients with chronic multimorbidity from hospitalization through 6 months post-discharge， aiming to facilitate the early identification of high-risk individuals and the development of personalized sleep interventions. MethodsA total of 228 inpatients with chronic multimorbidity， primarily with cardiovascular and metabolic chronic conditions， were enrolled via random sampling at The Third People's Hospital of Fuyang from January 2023 to September 2024. Baseline assessments were conducted during hospitalization using the general socio-demographic questionnaire， the Sleep Dysfunction Rating Scale （SDRS）， the Ruminative Response Scale （RRS）， and the Hospital Anxiety and Depression Scale （HADS）. Sleep quality was further evaluated using the SDRS at 1-， 3-， and 6-month post-discharge. Latent class growth model was employed to delineate trajectories of sleep quality. Multivariate logistic regression analysis was subsequently performed to identify influencing factors of sleep quality. ResultsA total of 201 patients with chronic diseases were included.Based on four predefined assessment time points （baseline， 1 month， 3 months， 6 months post-discharge）， the latent class growth model identified three distinct trajectories of sleep quality： persistent high sleep disturbance （n=72， 35.82%）， moderate sleep disturbance-progression （n=73， 36.32%）， and low sleep disturbance （n=56， 27.86%）. Multivariate logistic regression analysis revealed that， compared with the low sleep disturbance group， multiple factors increased the odds of membership in the persistent high sleep disturbance group： per capita monthly household income of <3 000 yuan （β=13.131， P<0.01） or 3 000–5 000 yuan （β=5.913， P<0.05）， aged 45–65 years （β=9.536， P<0.05）， presence of 3 （β=7.792， P<0.05） or >3 comorbid chronic conditions （β=6.626， P<0.05）， as well as higher RRS （β=0.334， P<0.01） and HADS score （β=1.628， P<0.01）. Additionally， patients aged 45–65 years （β=2.777， P<0.05）， with 3 （β=3.802， P<0.01） or >3 comorbid chronic conditions （β=2.463， P<0.01）， and with higher RRS （β=0.111， P<0.01） and HDAS scores （β=0.350， P<0.05） also demonstrated significantly increased odds of belonging to the moderate sleep disturbance–progression group. ConclusionThere is significant population heterogeneity in sleep quality trajectories among patients with chronic multimorbidity， with age， per capita household monthly income， number of comorbidities， rumination， and anxiety/depression symptoms being identify as the primary influencing factors.

To establish a TaqMan-based multiplex RT-qPCR method for the identification of Japa-nese encephalitis virus(JEV),Porcine reproductive and respiratory syndrome virus(PRRSV),and Classical swine fever virus(CSFV),this study designed and synthesized three pairs of specific primers and probes based on the conserved sequences of JEV E,PRRSV ORF6,and CSFV E2 a-vailable in the NCBI GenBank.By optimizing the reaction system and protocol,a multiplex RT-qPCR method for detecting these three viruses was developed and applied to the detection of clini-cal samples.The results showed that the established TaqMan multiplex RT-qPCR specifically am-plified the gene fragments of JEV,PRRSV,and CSFV,and did not amplify other non-target genes,indicating good specificity of the method.Intra-assay and inter-assay repeatability tests showed that the coefficient of variation(Cv)values were all below 3％,demonstrating that the method has ex-cellent repeatability.Sensitivity tests revealed that the minimum detectable amount for the recom-binant plasmids of the three viruses was 100 copies/pL.Using the established method,a total of 969 samples,including blood,aborted fetuses,semen,and deceased pigs,from 26 pig farms in Guizhou Province were tested.The detection rates were 34.3％(332/969)for JEV,28.3％(274/969)for PRRSV,and 19.8％(192/969)for CSFV.The co-infection rates were 10.1％(98/969)for JEV and PRRSV,12.1％(117/969)for JEV and CSFV,and 14.6％(141/969)for CSFV and PRRSV.Additionally,the triple co-infection rate of JEV,PRRSV,and CSFV was 7.9％(77/969).These results indicate that the TaqMan multiplex RT-qPCR method developed in this study is ef-fective for detecting these three viruses in pig farms,providing technical support for identifying vi-ral causes of reproductive disorders.

Objective To investigate serum exosomal miRNA spectrum in patients with gestational diabetes mellitus(GDM)and eval-uate its clinical value in the diagnosis of GDM complicated with premature delivery.Methods Serum samples of pregnant women with GDM registered and delivered in our hospital were collected and divided into the premature delivery group and term labor group based on pregnancy outcomes,with 22 cases in each group.Serum exosomal miRNAs were sequenced,and the differentially expressed miRNAs were further verified by fluorescence quantitative PCR.The receiver operating characteristic(ROC)curve was drew according to the verified expression level of miRNAs,and the value of exosomal miRNAs in the diagnosis of GDM complicated with premature de-livery was analyzed.The potential functions of candidate miRNAs were predicted using the Kyoto Encyclopedia of Genes and Genomes(KEGG).Results A total of 94 differentially expressed miRNAs,including 50 up-regulated and 44 down-regulated,were identified in the premature delivery group and term labor group.The verification results of fluorescence quantitative PCR showed that 7 miRNAs had significant difference between the two groups(P<0.05).Moreover,the expression trend was consistent with the sequencing results.The analysis results of the ROC curve showed that the seven miRNAs had good diagnostic efficacy for GDM combined with premature delivery.The areas under the ROC curve(AUCROC)of hsa-miR-16-5p,hsa-miR-25-3p,hsa-miR-30b-5p,and hsa-miR-92a-3p were all more than 0.7.Their sensitivity and specificity were 0.375 and 1.000,0.563 and 0.941,0.563 and 0.824,and 0.765 and 0.647,respectively.The binary logistic regression analysis showed that the AUCROC of the combination of hsa-miR-16-5p,hsa-miR-25-3p,hsa-miR-30b-5p,and hsa-miR-92a-3p for the diagnosis of GDM complicated with premature delivery increased to 0.982,and that its sensi-tivity and specificity were both more than 0.850.These candidate miRNAs were related to the ubiquitin-mediated proteolysis pathway,actin cytoskeleton regulatory pathway,mTOR signaling pathway,and P53 signaling pathway.Conclusion Serum exosomal miRNAs in GDM patients complicated with premature delivery have significant difference,which may be served as potential diagnostic markers.

Objective:To analyze Hepatitis B virus(HBV)drug resistance mutations in patients with chronic hepatitis B(CHB)infection who have undergone long-term monotherapy with Entecavir(ETV)and those receiving combination therapy with ETV and Lamivudine(LAM), and to explore the related factors affecting HBV drug resistance mutations.Methods:The study retrospectively analyzed patients with CHB, compensated cirrhosis, decompensated cirrhosis, and liver cancer who received long-term nucleotide analogue antiviral therapy at the Fifth Medical Center of PLA General Hospital from August 2012 to August 2019.The patients were divided into an ETV monotherapy group and a combined LAM+ETV therapy group.Chi-square tests, independent sample t-tests, and Wilcoxon rank-sum tests were used to compare the clinical baseline characteristics and HBV drug resistance mutation features between the two therapy groups.A multivariate logistic regression model was used to analyze the factors related to HBV drug resistance mutations. Results:A total of 533 patients were enrolled in this study, 357 in the ETV monotherapy group and 176 in the LAM+ETV group. The ETV monotherapy group had 122 (34.17%) patients with resistance mutations, while the LAM+ETV group had 126 (71.59%).In general, the difference in gene mutation rate between the two therapy groups was statistically significant( χ2=66.337, P<0.001). The median age and alanine aminotransferase levels of patients with drug resistance mutations in the two therapy groups were higher than those in the non-mutation group[( t=-4.743, P<0.001)/( Z=-4.809, P<0.001), ( Z=-2.667, P=0.007)/( Z=-2.001, P=0.045)].Age( OR=1.044, 95% CI:1.023-1.066), compensated cirrhosis( OR=2.163, 95% CI:1.193-3.922), liver cancer( OR=4.017, 95% CI:2.170-7.436) and the treatment regimen( OR=6.075, 95% CI:3.889-9.489) were associated with drug resistance gene mutations( P<0.001).The mutation rates in different stages of chronic liver disease(CHB, cirrhosis, and liver cancer)showed statistically significant( χ2=41.038, P<0.001; χ2=15.894, P<0.001).The overall mutation rates of ETV-related genes in the two therapy groups were 25.49% and 32.39%, respectively.Additionally, 10 mutation sites and 38 variant combinations were identified, containing five common combinations being rtL180M, rtM204V, rtS202G;rtL180M, rtM204V, rtT184A; rtL180M, rtM204V, rtT184L;rtM204I and rtL180M, rtM204V. Conclusion:In CHB patients undergoing long-term therapy, the rate of HBV resistance mutations is higher in those receiving ETV and LAM combination therapy than in those receiving ETV monotherapy.Monitoring older patients and those with cirrhosis or liver cancer is especially important for preventing resistance mutations.

Objective:To investigate the dosimetric impact of dwell position spacing in the design of three-dimensional (3D) interstitial brachytherapy plans for cervical cancer, and to provide a reference for selecting dwell spacing in clinical planning.Methods:A total of 15 patients with cervical cancer who underwent 3D interstitial brachytherapy at Tianjin Medical University Cancer Institute & Hospital between March 2022 and March 2024 were selected using simple random sampling. For each patient, 10 brachytherapy plans were generated with different dwell position spacings set at 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 mm, respectively. Key parameters among different dwell spacings compared included D 90%, V 100%, V 200%, and V 300% for the high-risk clinical target volume (HRCTV); D 90% for the intermediate-risk clinical target volume (IRCTV); D 2 cm3 for organs at risk (OARs) (bladder, small intestine, colon, and rectum); and the total dwell time. Statistical analyses were performed using repeated measurement ANOVA or the Friedman test. Results:Among different dwell spacings, there were no statistically significant differences in HRCTV D 90%, HRCTV V 100%, bladder D 2 cm3, and rectum D 2 cm3 among different dwell spacings ( P=0.075, 0.061, 0.480, 0.639). All plans with dwell spacings ≤ 3 mm met clinical dose requirements. When the dwell spacing was set to 1 mm, HRCTV V 200% and V 300% had the smallest mean values, while IRCTV D 90% and total dwell time had the largest mean values; all differences were statistically significant ( P<0.05). When the dwell spacing was ≥6 mm, an increase in spacing led to a decrease in mean small intestine D 2 cm3, and total dwell time, but an increase in HRCTV V 200% and a decrease in IRCTV D 90%, with statistically significant differences compared to spacings of 1-4 mm ( P<0.05). When the dwell spacing was ≥8 mm, the median colon D 2 cm3 decreased, with statistically significant differences compared to spacings of 1-3 mm ( P<0.05). Conclusions:For 3D interstitial brachytherapy planning in cervical cancer, dwell position spacings ≤ 3 mm can meet clinical dose requirements, with 1 mm providing optimal target coverage. Spacings ≥6 mm / ≥8 mm can reduce radiation dose to the small intestine and colon, respectively, while also shortening dwell time.

Objective To investigate the effects of perioperative blood glucose(BG)levels on postoperative acute kidney injury(AKI)and rehabilitation in diabetic patients with coronary heart disease after coronary artery bypass grafting(CABG).Methods Randomly select data from 142 patients who underwent CABG treatment at The Fuwai Cardiovascular Hospital in Qingdao from January to December 2022,and compare the BG indicators,AKI incidence,cardiac function improvement and quality of life between good control group(n=57)and poor control group(n=85).Results The operation time,ventilator use time,postoperative ICU time,hospital stay time and various BG indexes in the good control group were significantly lower than those in the poor control group(P<0.05 or P<0.01).27 patients(19.01%)developed AKI after surgery,and the incidence of AKI in the good control group was significantly lower than that in the poor control group(10.53%vs 24.71%,P<0.05).After treatment,the cardiac function indexes in both groups were significantly improved compared with those before surgery,and the good control had significantly better cardiac function and quality of life than the poor control(P<0.05).Conclusions The incidence of AKI after CABG in patients with diabetes and coronary heart disease was 19.01%.Good perioperative BG level control can reduce the risk of postoperative AKI and help early postoperative recovery.

Objective:To analyze Hepatitis B virus(HBV)drug resistance mutations in patients with chronic hepatitis B(CHB)infection who have undergone long-term monotherapy with Entecavir(ETV)and those receiving combination therapy with ETV and Lamivudine(LAM), and to explore the related factors affecting HBV drug resistance mutations.Methods:The study retrospectively analyzed patients with CHB, compensated cirrhosis, decompensated cirrhosis, and liver cancer who received long-term nucleotide analogue antiviral therapy at the Fifth Medical Center of PLA General Hospital from August 2012 to August 2019.The patients were divided into an ETV monotherapy group and a combined LAM+ETV therapy group.Chi-square tests, independent sample t-tests, and Wilcoxon rank-sum tests were used to compare the clinical baseline characteristics and HBV drug resistance mutation features between the two therapy groups.A multivariate logistic regression model was used to analyze the factors related to HBV drug resistance mutations. Results:A total of 533 patients were enrolled in this study, 357 in the ETV monotherapy group and 176 in the LAM+ETV group. The ETV monotherapy group had 122 (34.17%) patients with resistance mutations, while the LAM+ETV group had 126 (71.59%).In general, the difference in gene mutation rate between the two therapy groups was statistically significant( χ2=66.337, P<0.001). The median age and alanine aminotransferase levels of patients with drug resistance mutations in the two therapy groups were higher than those in the non-mutation group[( t=-4.743, P<0.001)/( Z=-4.809, P<0.001), ( Z=-2.667, P=0.007)/( Z=-2.001, P=0.045)].Age( OR=1.044, 95% CI:1.023-1.066), compensated cirrhosis( OR=2.163, 95% CI:1.193-3.922), liver cancer( OR=4.017, 95% CI:2.170-7.436) and the treatment regimen( OR=6.075, 95% CI:3.889-9.489) were associated with drug resistance gene mutations( P<0.001).The mutation rates in different stages of chronic liver disease(CHB, cirrhosis, and liver cancer)showed statistically significant( χ2=41.038, P<0.001; χ2=15.894, P<0.001).The overall mutation rates of ETV-related genes in the two therapy groups were 25.49% and 32.39%, respectively.Additionally, 10 mutation sites and 38 variant combinations were identified, containing five common combinations being rtL180M, rtM204V, rtS202G;rtL180M, rtM204V, rtT184A; rtL180M, rtM204V, rtT184L;rtM204I and rtL180M, rtM204V. Conclusion:In CHB patients undergoing long-term therapy, the rate of HBV resistance mutations is higher in those receiving ETV and LAM combination therapy than in those receiving ETV monotherapy.Monitoring older patients and those with cirrhosis or liver cancer is especially important for preventing resistance mutations.

Objective To explore the mechanism of Bushen Zhupai Decoction in treating ovarian fibrosis in polycystic ovary syndrome(PCOS)model rats based on IL-17/TRAF6/NF-κB signaling pathway.Methods Totally 60 SPF grade female SD rats were randomly divided into blank group,model group,Yousiyue group and TCM low-,medium-and high-dosage groups,with 10 rats in each group.The PCOS model was replicated by gavage of 1 mg/kg letrozole for 21 days.The Yousiyue group was given a 10 mg/kg solution of drospirenone ethinylestradiol tablets by gavage,TCM low-,medium-and high-dosage groups were given 8.82,17.64 and 35.28 g/kg of Bushen Zhupai Decoction by gavage,once a day for 28 consecutive days.HE and Masson staining were used to observe the morphology of ovarian tissue and ovarian fibrosis,the contents of serum IL-17,IL-18 and IL-1β were detected by ELISA,the mRNA expression of IL-17,TRAF6,NF-κB p65 and TGF-β1 in ovarian tissue were determined by RT-qPCR,the protein expression of TRAF6,NF-κB p65,TGF-β1,α-SMA,E-cadherin and the positive expression of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue were detected by Western blot and immunohistochemical staining respectively.Results Compared with the blank group,the body mass,ovarian mass and ovarian index significantly increased of rats in model group,with follicular cystic dilation,thinning of granulosa cell layer,and significant increase in fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly increased(P<0.01),the expressions of IL-17,TRAF6,NF-κB p65,TGF-β1 mRNA and IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA proteins significantly increased(P<0.01,P<0.05),the expression of E-cadherin protein significantly decreased(P<0.01).Compared with the model group,the ovarian index of rats in Yousiyue group and TCM medium-and high-dosage groups significantly decreased(P<0.01,P<0.05),the rats in TCM high-dosage group showed dominant follicles,increased thickness of granulosa cell layer,and significantly reduced fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly decreased in Yousiyue group and TCM low-,medium-and high-dosage groups(P<0.01),the mRNA expressions of IL-17,TRAF6,NF-κB p65,TGF-β1,and the protein expressions of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue significantly decreaed in TCM high-dosage group(P<0.01,P<0.05),the expression of E-cadherin protein significantly increased(P<0.01).Conclusion Bushen Zhupai Decoction can decrease the body mass,improve the ovarian index,decrease the expressions of inflammatory factors and improve the status of ovarian fibrosis in PCOS rats.The mechanism may be related to the regulation of IL-17/TRAF6/NF-κB signaling pathway.

Objective To explore the mechanism of Bushen Zhupai Decoction in treating ovarian fibrosis in polycystic ovary syndrome(PCOS)model rats based on IL-17/TRAF6/NF-κB signaling pathway.Methods Totally 60 SPF grade female SD rats were randomly divided into blank group,model group,Yousiyue group and TCM low-,medium-and high-dosage groups,with 10 rats in each group.The PCOS model was replicated by gavage of 1 mg/kg letrozole for 21 days.The Yousiyue group was given a 10 mg/kg solution of drospirenone ethinylestradiol tablets by gavage,TCM low-,medium-and high-dosage groups were given 8.82,17.64 and 35.28 g/kg of Bushen Zhupai Decoction by gavage,once a day for 28 consecutive days.HE and Masson staining were used to observe the morphology of ovarian tissue and ovarian fibrosis,the contents of serum IL-17,IL-18 and IL-1β were detected by ELISA,the mRNA expression of IL-17,TRAF6,NF-κB p65 and TGF-β1 in ovarian tissue were determined by RT-qPCR,the protein expression of TRAF6,NF-κB p65,TGF-β1,α-SMA,E-cadherin and the positive expression of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue were detected by Western blot and immunohistochemical staining respectively.Results Compared with the blank group,the body mass,ovarian mass and ovarian index significantly increased of rats in model group,with follicular cystic dilation,thinning of granulosa cell layer,and significant increase in fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly increased(P<0.01),the expressions of IL-17,TRAF6,NF-κB p65,TGF-β1 mRNA and IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA proteins significantly increased(P<0.01,P<0.05),the expression of E-cadherin protein significantly decreased(P<0.01).Compared with the model group,the ovarian index of rats in Yousiyue group and TCM medium-and high-dosage groups significantly decreased(P<0.01,P<0.05),the rats in TCM high-dosage group showed dominant follicles,increased thickness of granulosa cell layer,and significantly reduced fibrosis positive range(P<0.01),the serum contents of IL-17,IL-18 and IL-1β significantly decreased in Yousiyue group and TCM low-,medium-and high-dosage groups(P<0.01),the mRNA expressions of IL-17,TRAF6,NF-κB p65,TGF-β1,and the protein expressions of IL-17RA,TRAF6,NF-κB p65,TGF-β1 and α-SMA in ovarian tissue significantly decreaed in TCM high-dosage group(P<0.01,P<0.05),the expression of E-cadherin protein significantly increased(P<0.01).Conclusion Bushen Zhupai Decoction can decrease the body mass,improve the ovarian index,decrease the expressions of inflammatory factors and improve the status of ovarian fibrosis in PCOS rats.The mechanism may be related to the regulation of IL-17/TRAF6/NF-κB signaling pathway.

Objective:To explore the improvement effect of Tiaojing Zhuyun formula combined with Dai meridian moxibustion on endometrial fibrosis in rats with intrauterine adhesions(IUA)by regulating M2 type macrophage polarization.Methods:SD rats were selected to establish an IUA model using a dual injury method of mechanical curettage and lipopolysaccharide infection,which were randomly separated into model group,Tiaojing Zhuyun formula group,Dai meridian moxibustion group,Tiaojing Zhuyun formula+Dai meridian moxibustion group and Bujiale group,with 10 rats in each group,another 10 rats were regarded as a sham operation(control)group,after grouping and treatment,the number of embryo implantation of rats in each group after cage closure was detected;HE staining was applied to detect pathological morphology of uterine tissue in rats in different groups,and their endometrial thickness and glandular number were compared;Masson staining was applied to detect endometrial fibrosis in rats of each group,and the collagen volume fraction(CVF)was compared;ELISA was applied to detect serum levels of profibrotic factor TGF-β1,pro-inflammatory factors IL-1β and IL-6,and levels of procollagen type Ⅰ carboxy-terminal peptide(PⅠNP)and procollagen Ⅲ N-terminal peptide(PⅢNP)of rats in each group;immunofluorescence staining was applied to detect expressions of M2 type macrophage markers CD163 and CD68 in endometrium of rats in each group.Results:Compared with control group,model group showed obvious pathological damage to uterine tissue,thickness of endometrium was thinner,the number of glands and embryo implantation were obviously reduced(P<0.05),endometrial CVF,levels of serum PⅠNP,P Ⅲ NP,TGF-β1,IL-1β and IL-6,and the number of CD163 and CD68 positive cells in endometrium were increased(P<0.05);compared with model group,pathological damage to uterine tissue of rats in Tiaojing Zhuyun formula group,Dai meridian moxibustion group,Tiaojing Zhuyun formula+Dai meridian moxibustion group and Bujiale group was reduced,thickness of endometrium,the number of glands and embryo implantation,and the numbers of CD163 and CD68 positive cells in endometrium were increased(P<0.05),endometrial CVF and levels of serum PⅠNP,P Ⅲ NP,TGF-β1,IL-1β and IL-6 were reduced(P<0.05);compared with Tiaojing Zhuyun formula group and Dai meridian moxibustion group,pathological damage to uterine tissue of rats in Tiaojing Zhuyun formula+Dai meridian moxibustion group and Bujiale group was alleviated,thickness of endometrium,the number of glands and embryo implantation,and the number of CD163 and CD68 positive cells in endometrium were increased(P<0.05),endometrial CVF and levels of serum PⅠNP,P Ⅲ NP,TGF-β1,IL-1β and IL-6 were reduced(P<0.05);there was no obvious change in all indicators between Tiaojing Zhuyun formula+Dai meridian moxibustion group and Bujiale group(P>0.05).Conclusion:Combination of Tiaojing Zhuyun formula and Dai meridian moxibustion can promote the polarization of M2 type macrophages,thereby inhibiting inflammation and endometrial fibrosis in IUA rats,and improving their symptoms of uterine adhesions.