ObjectiveTo investigate the mechanism through which miR‑21 improves chronic renal fibrosis in mice via targeted modulation of the phosphatase and tensin homolog （PTEN）/protein kinase B （AKT）/mammalian target of rapamycin （mTOR） pathway. MethodsThirty‑two chronic kidney disease model mice were randomly divided into four groups （n=8 each group）： model group， miR‑21 overexpression group， miR‑21 inhibition group， and miR‑21 inhibition + MK‑2206 group. Eight healthy mice were included as the control group. The miR‑21 overexpression， miR‑21 inhibition， and miR‑21 inhibition + MK‑2206 groups received tail‑vein injections of lentivirus （50 μL， 1×10⁸ TU per mouse） once weekly for three weeks. The control and model groups were injected with an equal volume of empty vector （LV‑NC）. The miR‑21 inhibition + MK‑2206 group additionally received gavage of the AKT/mTOR pathway inhibitor MK‑2206 （480 mg/kg） once weekly for three weeks. The expressions of miR‑21， 24 h urinary protein， serum creatinine （Scr）， blood urea nitrogen （BUN）， and renal tissue levels of collagen Ⅰ， collagen Ⅲ， α‑smooth muscle actin （α‑SMA）， and PTEN protein， as well as p‑AKT/AKT and p‑mTOR/mTOR ratios， were compared among groups. HE staining was used to observe pathological changes in renal tissue， and Masson staining was used to observe the degree of renal fibrosis. A dual‑luciferase assay was performed to verify the targeting relationship between miR‑21 and PTEN. ResultsCompared with the model group， miR‑21 expression in renal tissue increased in the miR‑21 overexpression group （P<0.05） and decreased in the miR‑21 inhibition group （P<0.05）. Compared with the model group， the miR‑21 overexpression group showed increased 24 h urinary protein， Scr， BUN， and renal tissue expression of collagen Ⅰ， collagen Ⅲ， and α‑SMA （all P<0.05）， while these indicators decreased in the miR‑21 inhibition group （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group exhibited lower 24‑h urinary protein， Scr， BUN， and renal tissue expression of Collagen Ⅰ， Collagen Ⅲ， and α‑SMA （all P<0.05）. Compared with the model group， the miR‑21 overexpression group showed decreased PTEN protein expression （P<0.05） and increased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， while the miR‑21 inhibition group showed increased PTEN expression （P<0.05） and decreased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group had lower p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， with no significant difference in PTEN protein expression. HE and Masson staining showed normal kidney structure and almost no fibrosis in the control group. The model group exhibited glomerular enlargement， capillary loop adhesion， and focal fibrosis. The miR-21 overexpression group showed severe destruction of glomerular structure， accompanied by extensive fibrosis and renal tubular atrophy. The pathological changes and degree of fibrosis were alleviated in the miR-21 inhibition group. The miR-21 inhibition + MK-2206 group showed only mild pathological changes and mild fibrosis， with the interstitium being largely normal. Compared with PTEN-WT + NC mimics 1， the relative luciferase activity in the PTEN-WT + miR-21 mimics group decreased （P<0.001）. There was no statistically significant difference in relative luciferase activity between PTEN-WT + NC mimics group and PTEN-MUT + miR-21 mimics group. ConclusionmiR‑21 may improve renal function indicators and alleviate renal fibrosis in chronic kidney disease mice via targeted modulation of PTEN and subsequently inhibiting the AKT/mTOR pathway.

Objective: To evaluate the diagnostic performance of magnetic resonance spectroscopy (MRS) in discriminating fumarate hydratase-deficient (FH-d) uterine leiomyomas (ULs) from FH-preserved ULs. Materials and Methods: This study consisted of three stages, with independent cohorts recruited for each stage: 1) sample-size estimation was retrospectively performed on UL specimens (diameter ≥3 cm; age, 20–40 years) from our database with immunohistochemistry (IHC) for 2-succinocysteine (2-SC) as the reference, without genetic testing, 2) MRS sequence optimization in confirmed FH germline mutation participants with ultrasound-detected ULs (diameter ≥3 cm), without IHC analysis, and 3) prospective diagnostic test accuracy was evaluated in consecutive participants with ultrasound-detected ULs (diameter ≥3 cm;age, 20–40 years), using IHC for 2-SC for determining the FH status and subsequent genetic testing in those with positive 2-SC results to identify whether FH mutations were germline or somatic in origin. The choline and fumarate peaks in MRS were classified as positive, negative, or technical failure (TF). TFs were analyzed separately and excluded from the primary diagnostic accuracy calculations. T1-, T2-, and diffusion-weighted images were interpreted as hyperintense or hypointense. The enhancement rate and apparent diffusion coefficient were also acquired. Diagnostic performance was compared between MRS and various magnetic resonance imaging (MRI) features. Results: The optimal MRS parameters for the fumarate peak were echo time (TE) = 140 ms and an average of 256. Among the 360 prospective participants, 37 were confirmed to have FH-dULs. MRS showed positive fumarate peaks in 35 of 37 FH-dULs.After excluding six TFs, the positive fumarate peak on MRS showed 94.6% (35/37) sensitivity, 99.7% (316/317) specificity, and 99.2% (351/354) accuracy, all of which were significantly superior to those of other MRI features (P ≤ 0.002). Conclusion A positive fumarate peak on MRS may be a useful imaging biomarker for diagnosing FH-dULs.

Objective To investigate the prevalence of thyroid nodules in the physical examination population in Mianyang region and analyze its association with metabolic syndrome. Methods A retrospective study was conducted on 9 978 individuals who underwent health examinations at our hospital from January 2024 to May 2025. Thyroid examinations were performed using color Doppler ultrasound to analyze the prevalence of thyroid nodules in this population. Clinical data of all subjects were collected, and logistic regression analysis was employed to assess the association between metabolic syndrome and the risk of thyroid nodule development. Results The prevalence of thyroid nodules in the physical examination population of Mianyang region was 17.98% (1 794/9 978). The logistic regression results showed that after adjusting for gender, age, BMI, occupation, consumption of non-iodized salt, staying up late, daily sleep duration, anxiety, and depression, metabolic syndrome (OR=6.593, 95% CI: 3.961-10.975) was associated with thyroid nodules (P<0.05). Conclusion The prevalence of thyroid nodules among the physical examination population in the Mianyang area is 17.98%, and metabolic syndrome remains associated with the risk of thyroid nodules after effectively controlling for confounding factors.

Objective: To assess the status of sexual behaviors, HIV/AIDS knowledge awareness, and novel synthetic drug use among school attending young students in Guangdong Province, so as to provide evidence base for formulating targeted intervention strategies. Methods: From September to December 2024, a convenience sampling method was employed to conduct an anonymous online survey among 41 119 students from 20 higher education institutions and vocational colleges in Guangdong Province. Chi square test was utilized for rate comparisons and univariate analysis, and a multivariate Logistic regression model was applied to analyze factors associated with novel synthetic drug use. Results: The self reported rate of sexual behavior was 5.2%. Among sexually active students, the rates of unprotected sexual intercourse, having temporary sexual partners, and having commercial sexual partners were 32.4%, 34.5%, and 26.3%, respectively. The awareness rate of HIV related knowledge was 75.7%. The overall use rate of novel psychoactive substances(NPS) was 1.0%, and among sexually active students it was 6.5%. Multivariate Logistic regression analysis showed that students with sexual experience ( OR =4.15) and those with HIV testing history ( OR =9.38) had higher risks of NPS use; among sexually active students, those with unprotected sexual intercourse ( OR =6.83) and those with commercial sexual partners ( OR = 4.68 ) had higher risks of NPS use (all P <0.05). Conclusions The HIV/AIDS knowledge level among surveyed students needs further improvement, and high risk sexual behaviors are cross linked with novel synthetic drug use. It is recommended to carry out integrated health education for all students and implement comprehensive targeted interventions for sexually active students.

Objective:To investigate the pulmonary function in patients with Parkinson's disease(PD)and the association between pulmonary function and motor symptoms.Methods:A total of 104 PD patients were included in the study,and a portable spirometer was used to assess pulmonary function.According to the results of pulmonary function,the patients were divided into ventilation dys-function group with 37 patients and non-ventilation dysfunction with 67 patients,and clinical data were compared between the two groups.Univariate and multivariate logistic regression analyses were used to investigate the factors associated with abnormal pulmonary function,and a Spearman correlation analysis was used to investigate the correlation between pulmonary function parameters and motor symptoms.Results:There were significant differences in the scores of bradykinesia and postural gait between the two groups(P<0.05),and the logistic regression analysis showed a positive correlation between pulmonary ventilation dysfunction and postural gait score(regression coefficient β=0.171,95%CI=1.015-1.386,P=0.032).There were significant differences between the patients with different stages of PD in the pulmonary function parameters of the percentage of forced vital capacity in predicted value(t=2.143,P=0.034),the percentage of forced expiratory volume in 1 second(t=2.118,P=0.037),the percentage of predicted peak expiratory flow(t=2.002,P=0.048),maximum expiratory pressure as a percentage of predicted value(t=2.412,P=0.018),and maximum inspiratory pressure as a percentage of predicted value(t=2.119,P=0.036).Conclusion:The pulmonary function parameters of PD patients are lower than expected values,and the incidence rate of pulmonary ventilation dysfunction is 35.58%,with the main manifestation of restrictive ventilatory dysfunction.There is a correlation between pulmonary function and motor symptoms in PD patients.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

Objective·To investigate the mechanisms by which different subtypes of G protein-coupled receptor kinases(GRKs)regulate the biased signaling transduction mediated by the muscarinic acetylcholine receptor 1(M1 receptor),focusing on their molecular effects in modulating the binding of the M1 receptor to the downstream heterotrimeric G protein(Gαq-Gβ1-Gγ2)andβ-arrestin 2(βarr2).Methods·By establishing a highly sensitive protein interaction detection system based on bioluminescence resonance energy transfer(BRET),six M1 receptor agonists/allosteric modulators were selected to measure the dynamic interactions between the M1 receptor and four GRK subtypes(GRK2/3/5/6),βarr2,and the G protein under stimulation.All BRET data were statistically quantified using the area under the curve(AUC)of the time-response curves.First,concentration-effect curves were established by treatment with gradient concentrations of agonists/allosteric modulators and AUC fitting,to comprehensively analyze the differences in efficacy between each agonist/allosteric modulator and the endogenous agonist acetylcholine chloride(ACh)in promoting the interactions of M1 receptor with GRK3/5,βarr2,and the G protein;next,GRKs were divided into two groups based on subtypes:GRK2/3 and GRK5/6.The maximum AUC values for the interaction between the M1 receptor and the two GRK groups under high concentrations were calculated respectively,to further evaluate the regulatory propensity of different types of GRKs on the binding strength of the M1 receptor to βarr2 or the G protein.Results·All six agonists/allosteric modulators effectively induced the association of the M1 receptor with GRK3,while simultaneousey inducing dissociation of the M1 receptor from GRK5.The allosteric modulator BQCA not only activated the M1 receptor alone and triggered its binding to downstream signaling proteins,but also,when co-treated with ACh,caused a significant leftward shift of the concentration-effect curves in the M1-G protein and M1-βarr2 systems,suggesting that its potentiation effect on ACh was mainly achieved by reducing the half-maximal effective concentration.A moderate positive correlation was observed between the maximum AUC values of M1-βarr2 and M1-G protein interactions induced by the seven groups of drug treatments(r=0.722,P=0.067).Further analysis showed that the ratio of the maximum AUC for M1-GRK2/3 interaction to that for M1-GRK5/6 interaction was also positively correlated with the ratio of the maximum AUC for M1-βarr2 interaction to that for M1-G protein interaction(r=0.760,P=0.047).Conclusion·The M1 receptor may be pre-coupled with GRK5/6 under basal conditions,and they dissociate upon receptor activation,suggesting that GRK5/6 may be involved in M1 receptor inactivation or signal reprogramming.The relative efficiency of the M1 receptor's interaction with different GRK subtypes determines its preference for downstream signaling pathways.

Objective·To investigate the mechanisms by which different subtypes of G protein-coupled receptor kinases(GRKs)regulate the biased signaling transduction mediated by the muscarinic acetylcholine receptor 1(M1 receptor),focusing on their molecular effects in modulating the binding of the M1 receptor to the downstream heterotrimeric G protein(Gαq-Gβ1-Gγ2)andβ-arrestin 2(βarr2).Methods·By establishing a highly sensitive protein interaction detection system based on bioluminescence resonance energy transfer(BRET),six M1 receptor agonists/allosteric modulators were selected to measure the dynamic interactions between the M1 receptor and four GRK subtypes(GRK2/3/5/6),βarr2,and the G protein under stimulation.All BRET data were statistically quantified using the area under the curve(AUC)of the time-response curves.First,concentration-effect curves were established by treatment with gradient concentrations of agonists/allosteric modulators and AUC fitting,to comprehensively analyze the differences in efficacy between each agonist/allosteric modulator and the endogenous agonist acetylcholine chloride(ACh)in promoting the interactions of M1 receptor with GRK3/5,βarr2,and the G protein;next,GRKs were divided into two groups based on subtypes:GRK2/3 and GRK5/6.The maximum AUC values for the interaction between the M1 receptor and the two GRK groups under high concentrations were calculated respectively,to further evaluate the regulatory propensity of different types of GRKs on the binding strength of the M1 receptor to βarr2 or the G protein.Results·All six agonists/allosteric modulators effectively induced the association of the M1 receptor with GRK3,while simultaneousey inducing dissociation of the M1 receptor from GRK5.The allosteric modulator BQCA not only activated the M1 receptor alone and triggered its binding to downstream signaling proteins,but also,when co-treated with ACh,caused a significant leftward shift of the concentration-effect curves in the M1-G protein and M1-βarr2 systems,suggesting that its potentiation effect on ACh was mainly achieved by reducing the half-maximal effective concentration.A moderate positive correlation was observed between the maximum AUC values of M1-βarr2 and M1-G protein interactions induced by the seven groups of drug treatments(r=0.722,P=0.067).Further analysis showed that the ratio of the maximum AUC for M1-GRK2/3 interaction to that for M1-GRK5/6 interaction was also positively correlated with the ratio of the maximum AUC for M1-βarr2 interaction to that for M1-G protein interaction(r=0.760,P=0.047).Conclusion·The M1 receptor may be pre-coupled with GRK5/6 under basal conditions,and they dissociate upon receptor activation,suggesting that GRK5/6 may be involved in M1 receptor inactivation or signal reprogramming.The relative efficiency of the M1 receptor's interaction with different GRK subtypes determines its preference for downstream signaling pathways.

Objective To explore the diagnostic value of epicardial adipose tissue(EAT)thickness combined with serum disintegrin metalloprotease 10(ADAM10)and stromal interaction molecule 1(STIM1)in coronary heart disease(CHD).Methods A total of 119 CHD patients were included as the study group,and 100 non-CHD patients in the same period were selected as the control group.The clinical data of the patients were collected.Serum ADAM10 and STIM1 levels were detected by enzyme-linked immunosorbent assay(ELISA).Color Doppler ultrasonography was applied to detect EAT thickness.Multivariate Logistic regression was used to analyze influencing factors of CHD occurrence.The diagnostic value of EAT thickness combined with serum ADAM10 and STIM1 levels in the diagnosis of CHD was analyzed by subject work characteristics(ROC)curves.Results The levels of triglycerides,low-density lipoprotein cholesterol(LDL-C),EAT thickness,serum ADAM10 and STIM1 were higher in the study group than those in the control group(P＜0.05).Logistic regression analysis showed that elevated EAT thickness,serum ADAM10,STIM1,triglyceride and LDL-C levels were independent risk factors for the occurrence of CHD(P＜0.05).ROC curve analysis showed that the area under the curve of EAT thickness combined with serum ADAM10 and STIM1 for the diagnosis of CHD[0.864(95%CI:0.811-0.907)]was higher than that of their respective individual diagnoses[0.801(95%CI:0.741-0.851),0.750(95%CI:0.687-0.806),respectively,0.799(95%CI:0.740-0.850)],and the sensitivity and specificity of the combined diagnosis were 82.35%and 83.00%,respectively.Conclusion EAT thickness,serum ADAM10 and STIM1 levels are obviously elevated in CHD patients,and the combination of the three tests has a high diagnostic value for CHD.