ObjectiveTo evaluate the detection specificity for clinical samples and the detection capability for standard substances of five commercially available multiplex polymerase chain reaction (PCR) nucleic acid detection kits (hereinafter referred to as the kits) for respiratory pathogens, and to provide a reference for selecting appropriate detection kits for multi-pathogen nucleic acid testing of respiratory infections. MethodsA total of 60 respiratory pathogen-positive clinical samples with known redults were selected and tested using the five kits (labeled as A, B, C, D, and E). The detection rates and Kappa coefficients were calculated to evaluate the consistency between the results from these kits and those from single-pathogen PCR kits. According to the limit of detection (LOD) provided by the kits, standard substances of respiratory pathogens (including 12 types such as influenza virus, Mycoplasma pneumoniae, and Bordetella pertussis) were diluted to four concentrations (250, 500, 1 000, and 2 000 copies·mL⁻¹). All five kits were used for detection to evaluate their respective detection capabilities. ResultsCompared with the results from single-pathogen PCR kits, the five tested kits demonstrated good consistency (all Kappa >0.80). Among them, Kit A had the highest detection rate (100.00%), followed by Kits C and E (98.33%), and then Kits B and D (95.00%). All five kits showed a relatively low false negative rate (FNR) for samples with a cycle threshold (Ct) value ≤35 (≤2.38%). However, for samples with Ct values>35, the FNR increased accordingly(average FNR=6.67%, P=0.029). Kit C exhibited the highest detection sensitivity for the tested standard substances (average LOD: 458.33 copies·mL⁻¹), followed by Kit D, then Kits A/E, and finally Kit B. ConclusionThe five multiplex PCR kits showed good consistency with single-pathogen detection results, but each had its own performance emphasis. Kit A, with the highest detection rate and high throughput, is suitable for targeted viral screening. Kit B, covering the broadest pathogen spectrum (including fungi/bacteria), is suitable for comprehensive respiratory pathogen screening. Kits C, D and E, are applicable for rapid detection. It is important to note that the detection efficacy of all kits decreases for low viral load samples with Ct values >35. In practical application, selection should be based on specific screening objectives, throughput requirements, and sample types.

Objective To investigate the association between gut microbiota metabolite,trimethy-lamine-N-oxide(TMAO),and total imaging burden in diabetic patients with cerebral small vessel disease(CSVD).Methods A prospective study was conducted on 112 elderly diabetic patients with CSVD admitted in our hospital from June 2022 to May 2024.According to the total imaging burden score,they were divided into a high burden group(burden score≥2,57 cases)and a low burden group(burden score＜2,55 cases).High-performance liquid chromatography with online electrospray ionization tandem mass spectrometry was applied to detect the plasma level of TMAO.Then based on the tertile of plasma TMAO level,the patients were also assigned into low(＜2.44 μmol/L,38 cases),median(2.44 μmol/L≤TMAO＜5.18 μmol/L,37 cases)and high TMAO(≥5.18 μmol/L,37 cases)groups.ROC curve analysis was used to assess the predictive value of plasma TMAO level for high imaging burden in diabetic patients with CSVD.Binary logistic regression analysis was employed to analyze the correlation between plasma TMAO level and high imaging burden.Results The high burden group exhibited significantly higher plasma TMAO level than the low burden group(P=0.002).The AUC value of plasma TMAO level in predicting high imaging burden was 0.669(95％CI:0.569-0.769,P=0.002).The percentage of high imaging burden was 34.2％,54.1％and 64.9％,respectively among the low,median and high TMAO groups,with significant differences among them(Chi-square=7.270,P=0.026).Binary logistic regression analysis indicated the correlation between TMAO and high imaging burden(OR=1.178,95％CI:1.019-1.364,P=0.027).Conclusion In elderly diabetic patients with CSVD,plasma TMAO level is closely associated with high imaging burden,with higher TMAO level,higher risk for high imaging burden.

Objective To investigate the mechanism by which Senegenin(SEN)alleviates microglial inflammatory response through the nuclear factor erythroid 2-related factor 2(Nrf2)/NOD-like receptor protein 3(NLRP3)pathway.Methods BV2 mouse microglia cells were randomly divided into control group,model group,SEN group and MCC950 group.Cells in control group were not treated,and cells in model group were added with 1 μg/ml lipopolysaccharide(LPS);Cells in SEN group were added with 1 μg/ml LPS+4 μmol/L SEN,and cells in MCC950 group were added with 1 μg/ml LPS+10 μmol/L MCC950 for 24 hours.CCK-8 method was used to detect the effect of different concentrations of SEN on the viability of BV2 cells.Griess method was used to determine the release amount of nitric oxide(NO)in the supernatant.Real-time fluorescent quantitative PCR was used to determine the mRNA expression levels of NLRP3,lymphocyte apoptosis-associated spect-like protein containing a CARD(ASC),caspase-1,interleukin(IL)-1β and IL-18 mRNA.Immunofluorescence staining was used to detect the expression levels of ASC,IL-1β,Nrf2 and heme oxygenase-1(HO-1).Western blotting was used to detect the expression levels of NLRP3,caspase-1,ASC,IL-1β,IL-18,Nrf2,HO-1,nuclear factor kappa B(NF-κB)and inducible nitric oxide synthase(iNOS).Results The results of CCK-8 method showed that there was no significant difference in the viability of BV2 cells treated with 2～20 μmol/L SEN compared with control group(P>0.05).Compared with control group,the viability of BV2 cells in model group decreased significantly(P<0.05).Compared with model group,the viability of BV2 cells in 4 μmol/L SEN group was significantly restored(P<0.05).Compared with control group,the results of Griess method showed that the release amount of NO in cells of model group increased significantly(P<0.05);the results of real-time PCR showed that the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA in cells of model group increased significantly(P<0.05);the results of Western blotting showed that the protein expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 proteins in cells of model group increased significantly(P<0.05),and the immunofluorescence staining results showed that the expression levels of iNOS and NF-κB protein in cells of model group increased,and the expression levels of Nrf2 and HO-1 decreased,with statistically significant differences(P<0.05).Compared with model group,the release amount of NO in cells of SEN group and MCC950 group decreased,and the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA and proteins decreased,with statistically significant differences(P<0.05);in the SEN group,the expression levels of iNOS and NF-κB decreased,and immunofluorescence staining showed that Nrf2 was translocated into the nucleus,and the expression levels of Nrf2 and HO-1 proteins increased significantly,with statistically significant differences(P<0.05).Conclusions SEN could alleviate the inflammatory response of mouse microglia cells induced by LPS and inhibit the activation and expression of NLRP3 inflammasome,with an effect comparable to that of the inflammasome inhibitor MCC950.The mechanism may be related to the regulation of the expression of upstream factors Nrf2 and HO-1.

Objective To investigate the diagnostic value of electromyographic(EMG)tremor indicators for Parkinson's disease(PD)using the Logistic regression model.Methods A total of 65 patients with PD(PD group)and 39 patients with essential tremor(ET)(ET group)were enrolled and underwent EMG tremor analysis.General information,disease-related data,and EMG tremor characteristics were compared between the two groups.Multivariate Logistic regression analysis was performed to screen for independent influencing factors of PD,and receiver operating characteristic(ROC)curves were plotted.The area under the curve(AUC)was used to evaluate the diagnostic value of EMG tremor indicators for PD.Results Compared with the ET group,the PD group had a higher proportion of patients with unilateral onset and those with tremor spectrum frequency≥2 times,and a lower proportion of patients with a family history of tremor(P＜0.05).The tremor peak frequencies in the resting,postural,and weight-bearing(1 000 g)states were lower in the PD group than in the ET group(P＜0.05).There were statistically significant differences in the tremor rhythm patterns between the two groups in the resting and weight-bearing states(P＜0.05),with the PD group dominated by alternating contraction patterns and the ET group by synchronous contraction pat-terns.Multivariate Logistic regression analysis revealed that the tremor peak frequency in the weight-bearing state,the tremor rhythm pattern in the resting state,and the frequency of tremor spectrum were independent influencing factors of PD(P＜0.05).The ROC curves showed that the AUCs of the tremor peak frequency in the weight-bearing state,the tremor rhythm pattern in the resting state,and the frequency of tremor spectrum for diagnosing PD were 0.886,0.750,and 0.779,respec-tively.The combination of these three indicators yielded the highest AUC(0.936)for diagnosing PD,with a sensitivity of 81.54％and a specificity of 94.87％.Conclusion The tremor peak fre-quency in the weight-bearing state,the tremor rhythm pattern in the resting state,and the frequency of tremor spectrum provided by EMG tremor analysis can serve as clinical indicators for early diagno-sis of PD,and their combined use offers higher diagnostic value,which can be used to differentiate PD from ET.

The molecular mechanisms underlying the develop-ment and metastasis of prostate cancer remain elusive.This comprehensive review delves into the intricate role of cofilin,an actin-binding protein,in the pathogenesis and progression of prostate cancer.Cofilin is a significant protein in cytoskeletal dynamics,and any dysregulation may result in the morphological changes in normal cells and the invasion and metastasis of tumor cells.Research has revealed that the activity of cofilin is regula-ted by various mechanisms,including phosphorylation/dephos-phorylation and interactions with other molecules.Moreover,this review discusses promising therapeutic interventions,such as co-filin inhibitors and gene therapy,which have demonstrated effica-cy in preclinical models.The challenge of clinically preventing the transition to castration-resistant prostate cancer and tumor metastasis is widely recognized,necessitating the development of precise drug treatments and biomarker identification.As a key regulatory protein,cofilin provides a more comprehensive refer-ence for the prevention and treatment of prostate diseases.

Objective:To explore the influencing factors of swallowing function after stroke based on the semi-quantita-tive assessment of the videofluoroscopic dysphagia scale(VDS),to provide a basis for clinical screening and treatment.Method:A retrospective survey method was conducted on 120 stroke patients who met the inclusion criteria.Clinical characteristics were collected,including gender,age,type of stroke,lesion location,disease duration,food consistency,nasogastric tube placement,tracheostomy,and comorbidities.All patients underwent a video-fluoroscopic swallowing study(VFSS),and swallowing function was assessed using the videofluoroscopic dys-phagia scale(VDS).Univariate and multivariate linear regression analyses were conducted to examine the ef-fects of clinical characteristics on oral phase,pharyngeal phase,and global swallowing function.Result:Univariate analysis revealed that subcortical lesions in the middle cerebral artery(MCA)region(ex-cluding the basal ganglia)(P＜0.001),basal ganglia lesions(P=0.040),and medullary lesions(P＜0.001)were significantly associated with global swallowing function after stroke.Multivariate linear regression analysis indi-cated that medullary lesions(P＜0.001),subcortical lesions in the MCA region(excluding the basal ganglia)(P＜0.001),and MCA(including basal ganglia)lesions(P=0.019)were risk factors for global swallowing function after stroke.Regarding food consistency,consumption of thickened liquids(P=0.018)and pureed foods(P=0.006)were protective factors for global swallowing function.Subgroup analysis of global swallow-ing function revealed that MCA(including basal ganglia)lesions(P=0.003),female gender(P＜0.001)and tra-cheotomy(P＜0.001)were risk factors for oral phase swallowing function after stroke.Compared with patients without a gastrostomy tube,those with a gastrostomy tubes(P=0.034)had less effect on swallowing function in the oral phase.Compared with hemorrhagic stroke,ischemic stroke(P＜0.001)had a less impact on the oral phase swallowing function.Medullary lesions(P＜0.001)and subcortical lesions in the MCA region(ex-cluding the basal ganglia)(P＜0.001)were risk factors for pharyngeal phase swallowing function.For food con-sistency,the consumption of thickened liquids(P=0.005)and pureed foods(P=0.002)were protective factors for pharyngeal phase swallowing function after stroke.Conclusion:Identifying clinical characteristics in the assessment of swallowing function in stroke patients helps to determine the type of dysphagia and the key points of treatment.This approach may facilitate the de-velopment and implementation of more targeted rehabilitation treatment plans,and accelerate the recovery of swallowing function and feeding ability in stroke patients.

Objective To analyze the changes of intestinal microbiota in patients with nonalcoholic fatty liver disease(NAFLD)and healthy controls so as to explore the relationship between intestinal microbiota and NAFLD.Methods A total of 353 samples from the National Center for Biotechnology Information were included,including 110 healthy controls and 243 NAFLD patients(127 NASH and 116 NAFLD).Macrogenome sequencing data were extracted to obtain data on intergroup flora diversity changes and analyze the potential correlation between alterations in gut microbes and NAFLD.Results Alpha diversity analysis showed no significant differences in Invsimpson index(P=0.57)or Simpson index(P=0.57)between NAFLD patients and healthy controls,while there was a trend of decreasing Shannon index(P=0.084),but with no significant difference.β-diversity analysis showed significant differences between the two groups(P=0.002).Species composition analysis showed that at the genus level,NAFLD patients and healthy controls had the same species types,with Bacteroidetes and Firmicutes being the dominant phyla in both groups.However,compared to Control group,NAFLD group had significantly lower Firmicutes,Synergistetes,and Candidatus_Melainabacteria.At the species level,there were significant differences between NAFLD group and Control group for 10 bacterial species,including Bacteroides_uniformis and Faecalbacterium_prausnitzil.Intergroup difference analysis showed that Phocaeicola dorei,Parabacteroides_distasonis,Bilophila_wadsworthia,and Dysosmobacter_welbionis were significantly enriched in NAFLD patients(LDA score＞2.0,P＜0.05),while Prevotella_stercorea,Faecalibacterium prausnitzii,and others were negatively associated with NAFLD.A disease detection model was constructed based on metagenomic data,and 26 intestinal microorganisms related to NAFLD were selected,including Pyramidobacter piscolens and Citrobacter freundii,etc.The diagnostic accuracy of NAFLD was judged by ROC curve discriminant model,and the AUC of the ROC curve was 0.743 8,indicating that the model has a certain accuracy in diagnosing NAFLD.Conclusion This study revealed differences in gut microbial diversity between NAFLD patients and healthy controls and identified key bacterial groups associated with NAFLD.In addition,the disease detection model constructed based on metagenomic data provides a new method and idea for the diagnosis of NAFLD,which is conducive to further study of the disease and the early diagnosis and treatment of clinical patients.

Objective:To observe the effect of Guilu Taohong Formula on semen quality in varicocele(VC)models of rats,and to explore its possible mechanism.Methods:Forty-eight male SD rats were randomly divided into four groups(sham group,model group,Guilu Taohong Formula group and L-carnitine group).After the establishment of models,the rats were treated with intra-gastric administration for eight consecutive weeks.The general condition of the rats was observed.After the gavage,the testicular and epididymal indices were calculated.Semen quality was assessed using an automatic semen analyzer.Apoptosis of testicular cells was assessed by TUNEL staining.And the expression levels of B-cell lymphocytoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and cys-teine aspartate protease-3(caspase-3)in testicular tissue were detected by Western blot.Results:Compared with the sham group,testicular index,epididymal index,sperm concentration,the percentage of progressive motility of sperm(PR％)and the expression level of Bcl-2 decreased in model group(P＜0.01).An increased apoptosis rate of spermatogenic cells and the expression levels of Bax and caspase-3 proteins were observed in model group as well(P＜0.01).Compared with the model group,the testicular index,epidid-ymal index,sperm concentration,PR％and the expression level of Bcl-2 in Guilu Taohong Formula group increased significantly(P＜0.05,P＜0.01).A decreased apoptosis rate of spermatogenic cells and the expression levels of Bax and caspase-3 proteins were de-tected in Guilu Taohong Formula group as well(P＜0.01).Similarly,the L-carnitine group showed increased testicular index,epidid-ymal index,sperm concentration,PR％and the expression level of Bcl-2 protein(P＜0.05,P＜0.01),where showed decreased ap-optosis rate of spermatogenic cells and the expression levels of Bax and caspase-3 proteins compared with model group(P＜0.01,P＜0.05).Conclusion:Guilu Taohong Formula improves semen quality in VC model rats and reduces the apoptosis rate of spermato-genic cells in testicular tissue,which may be related to the promotion of Bcl-2 protein expression and the inhibition of Bax and caspase-3 protein expression levels.

OBJECTIVE: This study aimed to explore the association between body mass index (BMI) and mortality based on the 10-year population-based multicenter prospective study. METHODS: A general population-based multicenter prospective study was conducted at four sites in rural China between 2013 and 2023. Multivariate Cox proportional hazards models and restricted cubic spline analyses were used to assess the association between BMI and mortality. Stratified analyses were performed based on the individual characteristics of the participants. RESULTS: Overall, 19,107 participants with a sum of 163,095 person-years were included and 1,910 participants died. The underweight (< 18.5 kg/m ²) presented an increase in all-cause mortality (adjusted hazards ratio [ aHR] = 2.00, 95% confidence interval [ CI]: 1.66-2.41), while overweight (≥ 24.0 to < 28.0 kg/m ²) and obesity (≥ 28.0 kg/m ²) presented a decrease with an aHR of 0.61 (95% CI: 0.52-0.73) and 0.51 (95% CI: 0.37-0.70), respectively. Overweight ( aHR = 0.76, 95% CI: 0.67-0.86) and mild obesity ( aHR = 0.72, 95% CI: 0.59-0.87) had a positive impact on mortality in people older than 60 years. All-cause mortality decreased rapidly until reaching a BMI of 25.7 kg/m ² ( aHR = 0.95, 95% CI: 0.92-0.98) and increased slightly above that value, indicating a U-shaped association. The beneficial impact of being overweight on mortality was robust in most subgroups and sensitivity analyses. CONCLUSION This study provides additional evidence that overweight and mild obesity may be inversely related to the risk of death in individuals older than 60 years. Therefore, it is essential to consider age differences when formulating health and weight management strategies.
Humans ; Body Mass Index ; China/epidemiology* ; Male ; Female ; Middle Aged ; Prospective Studies ; Rural Population/statistics & numerical data* ; Aged ; Follow-Up Studies ; Adult ; Mortality ; Cause of Death ; Obesity/mortality* ; Overweight/mortality*

Objective:To explore the influencing factors of swallowing function after stroke based on the semi-quantita-tive assessment of the videofluoroscopic dysphagia scale(VDS),to provide a basis for clinical screening and treatment.Method:A retrospective survey method was conducted on 120 stroke patients who met the inclusion criteria.Clinical characteristics were collected,including gender,age,type of stroke,lesion location,disease duration,food consistency,nasogastric tube placement,tracheostomy,and comorbidities.All patients underwent a video-fluoroscopic swallowing study(VFSS),and swallowing function was assessed using the videofluoroscopic dys-phagia scale(VDS).Univariate and multivariate linear regression analyses were conducted to examine the ef-fects of clinical characteristics on oral phase,pharyngeal phase,and global swallowing function.Result:Univariate analysis revealed that subcortical lesions in the middle cerebral artery(MCA)region(ex-cluding the basal ganglia)(P＜0.001),basal ganglia lesions(P=0.040),and medullary lesions(P＜0.001)were significantly associated with global swallowing function after stroke.Multivariate linear regression analysis indi-cated that medullary lesions(P＜0.001),subcortical lesions in the MCA region(excluding the basal ganglia)(P＜0.001),and MCA(including basal ganglia)lesions(P=0.019)were risk factors for global swallowing function after stroke.Regarding food consistency,consumption of thickened liquids(P=0.018)and pureed foods(P=0.006)were protective factors for global swallowing function.Subgroup analysis of global swallow-ing function revealed that MCA(including basal ganglia)lesions(P=0.003),female gender(P＜0.001)and tra-cheotomy(P＜0.001)were risk factors for oral phase swallowing function after stroke.Compared with patients without a gastrostomy tube,those with a gastrostomy tubes(P=0.034)had less effect on swallowing function in the oral phase.Compared with hemorrhagic stroke,ischemic stroke(P＜0.001)had a less impact on the oral phase swallowing function.Medullary lesions(P＜0.001)and subcortical lesions in the MCA region(ex-cluding the basal ganglia)(P＜0.001)were risk factors for pharyngeal phase swallowing function.For food con-sistency,the consumption of thickened liquids(P=0.005)and pureed foods(P=0.002)were protective factors for pharyngeal phase swallowing function after stroke.Conclusion:Identifying clinical characteristics in the assessment of swallowing function in stroke patients helps to determine the type of dysphagia and the key points of treatment.This approach may facilitate the de-velopment and implementation of more targeted rehabilitation treatment plans,and accelerate the recovery of swallowing function and feeding ability in stroke patients.