The diagnostic value of Mycobacterium tuberculosis(MTB)complex and rifampicin resistance test kits(fluorescence PCR method)in detecting for MTB complex and rifampicin resistancein sputum samples was evaluated.A total of 271 patients with suspected tuberculosis were prospectively and consecutively enrolled at Hunan Chest Hospital between April 1,2024,and November 30,2024.Of these,229 patients were confirmed to have tuberculosis,whereas 42 patients were not-tuberculosis samples were col-lected from all patients and subjected to fluorescence PCR,Xpert MTB/RIF(abbreviated as Xpert),and MGIT 960 culture and drug sensitivity testing.Clinical diagnosis and MTB culture results served as reference standards for TB diagnosis,whereas phenotypic drug susceptibility testing and Xpert served as reference standards to for assessment of rifampicin resistance.The sensitivity,specificity,positive predictive value,and negative predictive value of the fluorescence PCR method were analyzed.Kappa tests were performed to analyze the concordance between detection techniques.With clinical diagnosis as the gold standard,the sensitivity and specificity of the fluorescence PCR method for the diagnosis of TB were 65.1%(149/229)and 97.6%(41/42),and the consistency test for the fluo-rescence PCR and Xpert methods showed high consistency(Kappa value=0.993).With the MGIT 960 liquid culture as the reference standard,the positive detection rate of the fluorescence PCR method for the detection of patients with positive cultures was 91.9%(102/111,95%CI:85.2%-96.2%),and the positive detection rate for 147 patients with sputum culture-negative TB was 27.9%(41/147,95%CI:21.3%-35.6%).With the phenotypic drug sensitivity test as the gold standard,the sensitivity and specificity of the fluo-rescence PCR method for the detection of rifampicin resistance were 100.0%(31/31)and 96.6%(28/29)respectively,and the consis-tency between tests was high(Kappa value=0.967).With Xpert as the gold standard,the sensitivity and specificity of fluorescence PCR for the detection of rifampicin resistance were 95.8%(46/48)and 99.0%(99/100),respectively,and the consistency between tests was high(Kappa value=0.953).Finally,samples with rifampicin resistance detected with the fluorescence PCR method had a clear rpoB gene mutation type according to one-generation sequencing.In conclusion,the fluorescence PCR method showed high sen-sitivity in detecting MTB complex groups and rifampicin resistance,and had high concordance with Xpert.Therefore,this technique can serve a rapid test for TB diagnosis to increase the rate of positive TB pathology and detection of rifampicin resistance.This method is particularly suitable for use in lower-income countries and economically disadvantaged grassroots communities.

Objective To explore the therapeutic effect of orthopedic robot assisted percutaneous cannulated screw internal fixation in the treatment of Tilt pelvic fractures.Methods A retrospective analysis was made on clinical data of 21 cases of Tilt pelvic fractures treated with"Tianji"orthopedic surgical robot combined with 3D C-arm fluoroscopy assisted percutaneous cannulated screw internal fixation from January 2020 to October 2023.The Tile classification included 11 cases of type B2.1,3 cases of type B2.2,2 cases of type B2.3,and 5 cases of type B3.Results The percutaneous insertion of cannulated screw was safe in all the 21 cases,with a total of 71 screws inserted,including 21 S1 screws,21 S2 screws,21 pubic branch screws(14 anterograde screws and 7 retrograde screws),4 LC-Ⅱ screws,and 4 modified LC-Ⅱ screws.The insertion time of a single screw was(31.1±10.8)min,the fluoroscopy time of a single screw was(5.9±1.7)min,the surgical time was(100.7±16.2)min,and the bleeding volume was(11.2±4.2)ml.According to the Matta criteria,8 cases had excellent fracture reduction quality,11 cases were good,and 2 cases were fair,with an excellent and good rate of 90.5％(19/21).According to the Gras classification,the quality of screw placement was excellent in 65 cases and good in 6 cases,with an excellent and good rate of 100％(71/71).The 21 cases were followed up for 4-18(mean,10.7±8.4)months.The bone healing was achieved in all the cases,with a fracture healing time of(12.7±1.8)weeks.At the last follow-up,according to the Majeed scoring system,7 cases were rated as excellent pelvic function,11 cases as good,and 3 cases as fair,with an excellent and good rate of 85.7％(18/21).Two cases of lateral femoral cutaneous nerve injury and two cases of intermuscular vein thrombosis were cured.Conclusion The use of orthopedic robot assisted percutaneous cannulated screw fixation for the treatment of Tilt pelvic fractures has satisfactory therapeutic effects,with accurate screw placement and no need for adjustment.

The diagnostic value of Mycobacterium tuberculosis(MTB)complex and rifampicin resistance test kits(fluorescence PCR method)in detecting for MTB complex and rifampicin resistancein sputum samples was evaluated.A total of 271 patients with suspected tuberculosis were prospectively and consecutively enrolled at Hunan Chest Hospital between April 1,2024,and November 30,2024.Of these,229 patients were confirmed to have tuberculosis,whereas 42 patients were not-tuberculosis samples were col-lected from all patients and subjected to fluorescence PCR,Xpert MTB/RIF(abbreviated as Xpert),and MGIT 960 culture and drug sensitivity testing.Clinical diagnosis and MTB culture results served as reference standards for TB diagnosis,whereas phenotypic drug susceptibility testing and Xpert served as reference standards to for assessment of rifampicin resistance.The sensitivity,specificity,positive predictive value,and negative predictive value of the fluorescence PCR method were analyzed.Kappa tests were performed to analyze the concordance between detection techniques.With clinical diagnosis as the gold standard,the sensitivity and specificity of the fluorescence PCR method for the diagnosis of TB were 65.1%(149/229)and 97.6%(41/42),and the consistency test for the fluo-rescence PCR and Xpert methods showed high consistency(Kappa value=0.993).With the MGIT 960 liquid culture as the reference standard,the positive detection rate of the fluorescence PCR method for the detection of patients with positive cultures was 91.9%(102/111,95%CI:85.2%-96.2%),and the positive detection rate for 147 patients with sputum culture-negative TB was 27.9%(41/147,95%CI:21.3%-35.6%).With the phenotypic drug sensitivity test as the gold standard,the sensitivity and specificity of the fluo-rescence PCR method for the detection of rifampicin resistance were 100.0%(31/31)and 96.6%(28/29)respectively,and the consis-tency between tests was high(Kappa value=0.967).With Xpert as the gold standard,the sensitivity and specificity of fluorescence PCR for the detection of rifampicin resistance were 95.8%(46/48)and 99.0%(99/100),respectively,and the consistency between tests was high(Kappa value=0.953).Finally,samples with rifampicin resistance detected with the fluorescence PCR method had a clear rpoB gene mutation type according to one-generation sequencing.In conclusion,the fluorescence PCR method showed high sen-sitivity in detecting MTB complex groups and rifampicin resistance,and had high concordance with Xpert.Therefore,this technique can serve a rapid test for TB diagnosis to increase the rate of positive TB pathology and detection of rifampicin resistance.This method is particularly suitable for use in lower-income countries and economically disadvantaged grassroots communities.

Objective:To systematically review risk prediction models for frailty in maintenance hemodialysis (MHD) patients.Methods:Relevant literature on frailty risk prediction models for MHD patients were retrieved from PubMed, Web of Science, Embase, Cochrane Library, CINHAL, China National Knowledge Infrastructure, WanFang Data, VIP, and China Biology Medicine disc from inception to March 25, 2024. Two researchers independently screened the literature and extracted data. The PROBAST tool was used to assess the methodological quality of the prediction models. Meta-analysis of predictive factors was conducted using Stata 16.0 software.Results:A total of 13 studies were included, comprising 19 frailty risk prediction models for MHD patients. All studies exhibited a high risk of bias. Six studies conducted internal validation, and three studies conducted external validation. The area under the receiver operating characteristic curve ( AUC) of the included prediction models ranged from 0.720 to 0.998. Meta-analysis revealed that age [ OR=1.149, 95% CI (1.070, 1.234), P<0.001], female gender [ OR=4.472, 95% CI (1.799, 11.117), P<0.001], nutritional score [ OR=2.650, 95% CI (1.010, 6.970), P=0.048], comorbidities [ OR=1.990, 95% CI (1.500, 2.650), P<0.001], and serum albumin [ OR=0.830, 95% CI (0.790, 0.880), P<0.001] were significant influencing factors for frailty in MHD patients. Conclusions:Risk prediction models for frailty in MHD patients are still in the research stage. Healthcare professionals should pay close attention to frailty risks in older patients, females, those with malnutrition, comorbidities, and low serum albumin levels, and implement timely interventions.

BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.

Objective:To investigate the molecular mechanism of Xiaoshuantongmai Decoction(XSTMD)targeting JAK2/STAT3 signaling pathway to regulate the function of dendritic cells(DCs)in treatment of deep vein thrombosis(DVT).Methods:After treatment of DVT with XSTMD,expressions of fibrinogen beta chain(FGB)and D-dimer(D2D)protein in plasma of patients with DVT were detected by ELISA,proportion of plasmacytoid dendritic cell(pDC)and conventional dendritic cell(cDC),and expression of HLA-DR protein in peripheral blood mononuclear cells(PBMCs)of patients with DVT were detected by flow cytometry,expressions of CD80 and CD86 mRNA were detected by qRT-PCR,Western blot was used to detect protein levels of JAK2,STAT3 and phosphory-lation(p-JAK2 and p-STAT3)in PBMC of DVT patients and mice.LPS-induced mouse DC2.4 cells were treated with XSTMD drug-containing serum.Western blot was used to determine the protein levels of JAK2,STAT3,p-JAK2 and p-STAT3.ELISA was used to detect protein levels of IL-6,TNF-α,IL-10 and TGF-β1 in cell culture supernatant.Results:After treatment with XSTMD,weight and length of thrombus were significantly reduced in mice with DVT(P<0.001).Compared with before treatment,expressions of FGB and D2D were significantly decreased in plasma of DVT patients(P<0.001),proportion of pDC was significantly increased,while pro-portion of cDC was significantly decreased in PBMC of DVT patients(P<0.01),expression of HLA-DR protein and mRNA levels of CD80 and CD86 were significantly decreased in PBMC of DVT patients(P<0.05,P<0.01,P<0.001)after treatment with XSTMD.Levels of p-JAK2 and p-STAT3 protein were significantly increased in PBMC from DVT patients and mice treated with XSTMD(P<0.05).After treatment with serum containing XSTMD,protein levels of p-JAK2 and p-STAT3 induced by LPS were significantly increased in murine DC2.4 cells(P<0.05).Protein expressions of IL-6 and TNF-α were significantly decreased,while protein expressions of IL-10 and TGF-β1 were significantly increased in cell supernatant(P<0.01,P<0.001).Conclusion:XSTMD effectively treats DVT by pre-cisely regulating the JAK2/STAT3 signaling pathway to promote the differentiation of DCs into pDC and alleviate inflammatory injury.

In recent years, the wearable device market has been developing rapidly. Wearable devices with personalized health management and chronic disease monitoring are widely used in daily life by virtue of their powerful performance and convenience. However, with the popularization of these devices, skin adverse reactions caused by prolonged wearable wear are gradually increasing, among which allergic contact dermatitis (ACD) is the most common. Common allergens such as acrylates, methacrylates, rosin, chromium and nickel are widely present in adhesives and device components and are the main causes of ACD. Understanding the presence of potential sensitizers in wearable devices can help in diagnostic patch testing in users experiencing skin reactions caused by the device. Future innovations in wearable device materials will focus on the adoption of safer bonding technologies and biocompatible materials to reduce the risk of allergy. The introduction of new materials brings both development opportunities and challenges. Educating users on proper device usage, identifying specific allergens, selecting hypoallergenic materials, and optimizing device maintenance are important for reducing the risk of ACD.

Bacterial therapy has attracted increasing attention due to its special mechanism and abundant applications. With the flourishing development of synthetic biology, therapeutic genes have been introduced into engineering bacteria to improve their antitumor efficacy. However, it is difficult to spatiotemporally control the expression of these therapeutic genes at the tumor site in vivo, thereby considerably limiting the application of engineered bacteria in tumor treatment. To resolve this problem, we constructed a temperature-responsive bacterial strain capable of triggering the expression of exogenous genes in a laser-controllable way. Noxa, a pro-apoptotic protein, is chosen to test the expression of exogenous protein and its anti-tumor effect in engineered bacteria upon laser irradiation. Firstly, Noxa was fused to the C-terminus of the bacterial outer membrane protein cytolysin A (ClyA), and then the recombinant gene fragment ClyA-Noxa was inserted into the temperature-sensitive plasmid pBV220 and the recombinant plasmid was transformed into non-pathogenic Escherichia coli MG1655. Thus, we constructed the engineering strain (TRB@Noxa) that could express Noxa on the bacterial surface. TRB@Noxa could target and colonize the tumor tissue without causing notable host toxicity. The bacterial infection triggered thrombosis in the tumor tissue, resulting in the darkness of tumor sites. In a xenograft mouse tumor model, our strategy demonstrated precise tumor targeting and strong tumor inhibition. In conclusion, we successfully constructed a new engineering bacterial strain TRB@Noxa. TRB@Noxa combined with photothermal therapy could arrest tumor growth in the absence of photosensitizers, which represents an appealing method for antitumor therapy in the future.
Escherichia coli/radiation effects* ; Animals ; Humans ; Lasers ; Mice ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Neoplasms/therapy* ; Genetic Engineering ; Cell Line, Tumor ; Escherichia coli Proteins/genetics*

OBJECTIVES: To explore the therapeutic mechanism of Chaihu Shugan (CHSG) Decoction for improving cognitive impairment in rats with epilepsy induced by lithium chloride and pilocarpine. METHODS: Male SD rat models of cognitive impairment model after epilepsy induced by intraperitoneal injection with lithium chloride and pilocarpine were randomly divided into 5 groups (n=12) for treatment with daily gavage of saline, donepezil (90 mg/kg), or CHSG Decoction at 2.5, 5.0, 10, 20 and 40 g/kg for 4 consecutive weeks, with 10 rats with intraperitoneal injection with saline as the blank control group. Morris water maze test was used to evaluate cognitive and behavioral changes of the rats after treatment. The mRNA and protein expressions of ASIC1, NR1, NR2A and NR2B in the hippocampus of rats were detected using RT-qPCR and Western blotting. RESULTS: Compared with those with saline treatment, the rat models treated with CHSG Decoction at 5 and 10 g/kg showed significantly shortened escape latency and prolonged stay in the target quadrant with increased number of platform crossings in Morris water maze test. CHSG Decoction treatment at the two doses significantly increased ASIC1, NR1, NR2A and NR2B protein expressions in the hippocampus of the rat models, and their mRNA expression levels were all increased significantly after the treatment at the doses above 2.5 g/kg. CONCLUSIONS CHSG Decoction can improve cognitive impairment in rats after epilepsy possibly by regulating the expression and channel activity of NMDAR protein and its subunit protein via upregulating ASIC1 to modulate neuronal excitability and synaptic plasticity in the hippocampus.
Animals ; Hippocampus/drug effects* ; Receptors, N-Methyl-D-Aspartate/metabolism* ; Acid Sensing Ion Channels/metabolism* ; Rats, Sprague-Dawley ; Male ; Rats ; Epilepsy/complications* ; Cognitive Dysfunction/drug therapy* ; Drugs, Chinese Herbal/therapeutic use* ; Up-Regulation ; Maze Learning

Objective To explore the therapeutic mechanism of Chaihu Shugan(CHSG)Decoction for improving cognitive impairment in rats with epilepsy induced by lithium chloride and pilocarpine.Methods Male SD rat models of cognitive impairment model after epilepsy induced by intraperitoneal injection with lithium chloride and pilocarpine were randomly divided into 5 groups(n=12)for treatment with daily gavage of saline,donepezil(90 mg/kg),or CHSG Decoction at 2.5,5.0,10,20 and 40 g/kg for 4 consecutive weeks,with 10 rats with intraperitoneal injection with saline as the blank control group.Morris water maze test was used to evaluate cognitive and behavioral changes of the rats after treatment.The mRNA and protein expressions of ASIC1,NR1,NR2A and NR2B in the hippocampus of rats were detected using RT-qPCR and Western blotting.Results Compared with those with saline treatment,the rat models treated with CHSG Decoction at 5 and 10 g/kg showed significantly shortened escape latency and prolonged stay in the target quadrant with increased number of platform crossings in Morris water maze test.CHSG Decoction treatment at the two doses significantly increased ASIC1,NR1,NR2A and NR2B protein expressions in the hippocampus of the rat models,and their mRNA expression levels were all increased significantly after the treatment at the doses above 2.5 g/kg.Conclusion CHSG Decoction can improve cognitive impairment in rats after epilepsy possibly by regulating the expression and channel activity of NMDAR protein and its subunit protein via upregulating ASIC1 to modulate neuronal excitability and synaptic plasticity in the hippocampus.