To evaluate the diagnostic performance of a three-gene (GBP5, DUSP3, and TBP) tuberculosis (TB) score in bacteriologically-negative pulmonary tuberculosis, and to develop and validate a discriminative diagnostic model by integrating inflammatory cytokines (IL-2, IL-5, IL-17, and IFN-γ). A prospective cohort study was conducted, a total of 238 patients admitted to the Affiliated Infectious Disease Hospital of Soochow University from May 2023 to May 2024 were enrolled, including 119 pathogen-negative pulmonary tuberculosis patients and 119 patients with other pulmonary diseases (OPD). The GeneXpert MTB-HR kit was used to detect the three-gene TB scores from residual blood routine samples. The diagnostic performance was assessed using receiver operating characteristic (ROC) curve analysis. Concurrent data on 12 inflammatory cytokines were collected from patients. Potential biomarkers were screened using univariate analysis and multivariate logistic regression, and selected features were incorporated into the construction of four machine learning models: logistic regression, support vector machine (SVM), K-nearest neighbors (KNN), and adaptive boosting (AdaBoost). The samples were randomly split into a training set (85%) and a test set (15%). The models were trained on the training set, and their diagnostic performance was validated using the test set. The predictive ability of each model was evaluated based on ROC curve parameters. The results showed that the three-gene TB score alone yielded an AUC of 0.539 (sensitivity: 50.94%, specificity: 60.50%) in distinguishing pathogen-negative pulmonary tuberculosis from OPD. Four non-col-linear inflammatory factors (IL-2, IL-5, IL-17, and IFN-γ) were selected and combined with the three-gene TB score to construct machine learning models. The AdaBoost model demonstrated the best performance, achieving an AUC of 0.893 (sensitivity: 85.4%, specificity: 73.0%) in the training set and an AUC of 0.873 (sensitivity: 88.2%, specificity: 72.2%) in the test set. In conclusion,the AdaBoost diagnostic model integrating the three-gene TB score with inflammatory factors (IL-2, IL-5, IL-17, and IFN-γ) exhibits superior discriminating performance for pathogen-negative pulmonary tuberculosis compared to OPD, significantly outperforming the three-gene TB score alone.

To evaluate the diagnostic performance of a three-gene (GBP5, DUSP3, and TBP) tuberculosis (TB) score in bacteriologically-negative pulmonary tuberculosis, and to develop and validate a discriminative diagnostic model by integrating inflammatory cytokines (IL-2, IL-5, IL-17, and IFN-γ). A prospective cohort study was conducted, a total of 238 patients admitted to the Affiliated Infectious Disease Hospital of Soochow University from May 2023 to May 2024 were enrolled, including 119 pathogen-negative pulmonary tuberculosis patients and 119 patients with other pulmonary diseases (OPD). The GeneXpert MTB-HR kit was used to detect the three-gene TB scores from residual blood routine samples. The diagnostic performance was assessed using receiver operating characteristic (ROC) curve analysis. Concurrent data on 12 inflammatory cytokines were collected from patients. Potential biomarkers were screened using univariate analysis and multivariate logistic regression, and selected features were incorporated into the construction of four machine learning models: logistic regression, support vector machine (SVM), K-nearest neighbors (KNN), and adaptive boosting (AdaBoost). The samples were randomly split into a training set (85%) and a test set (15%). The models were trained on the training set, and their diagnostic performance was validated using the test set. The predictive ability of each model was evaluated based on ROC curve parameters. The results showed that the three-gene TB score alone yielded an AUC of 0.539 (sensitivity: 50.94%, specificity: 60.50%) in distinguishing pathogen-negative pulmonary tuberculosis from OPD. Four non-col-linear inflammatory factors (IL-2, IL-5, IL-17, and IFN-γ) were selected and combined with the three-gene TB score to construct machine learning models. The AdaBoost model demonstrated the best performance, achieving an AUC of 0.893 (sensitivity: 85.4%, specificity: 73.0%) in the training set and an AUC of 0.873 (sensitivity: 88.2%, specificity: 72.2%) in the test set. In conclusion,the AdaBoost diagnostic model integrating the three-gene TB score with inflammatory factors (IL-2, IL-5, IL-17, and IFN-γ) exhibits superior discriminating performance for pathogen-negative pulmonary tuberculosis compared to OPD, significantly outperforming the three-gene TB score alone.

ObjectiveBased on the experience of traditional quality evaluation， the quality of Atractylodis Macrocephalae Rhizoma（AMR） with different production methods such as direct seeding， transplanting after seedling raising， topping and non-topping， and difference in growth years was compared. MethodVernier caliper was used to measure the trait data of AMR in different production methods. Paraffin sections of AMR with different production methods were made by saffron solid green staining， and the microstructure was observed. The contents of water-soluble and alcohol-soluble extracts in AMR with different production methods were determined according to the 2020 edition of Chinese Pharmacopoeia. The content of water-soluble total polysaccharides in AMR with different production methods was detected by sulfuric acid-anthrone method. Fiber analyzer was used to detect the content of fiber components in AMR with different production methods. The contents of monosaccharides， oligosaccharides and some secondary metabolites in AMR with different production methods were detected by ultra performance liquid chromatography（UPLC）， and the differences of chemical components were compared by multivariate statistical analysis methods such as principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA）. ResultIn terms of traits， the 3-year-old AMR with direct seeding and without topping was close to the high-quality AMR with "phoenix-head and crane-neck， strong sweetness and clear aroma" recorded in ancient materia medica， followed by the 3-year-old AMR with topping after transplanting， while the 2-year-old AMR with topping after transplanting with high market circulation rate was generally fat and strong with mild odor. In the microscopic aspect， the arrangement of xylem vessels and fiber bundles in the 3-year-old samples formed two obvious rings. Compared with the 2-year-old samples cultivated in Bozhou and Zhejiang， the 3-year-old samples without topping after transplanting had more wood fibers. In terms of chemical composition， the contents of 70% ethanol extract， fructose， glucose， sucrose， 1-kestose， atractylenolide Ⅰ， chlorogenic acid， neochlorogenic acid， cryptochlorogenic acid and other components in 3-year-old AMR with direct seeding and without topping were significantly higher than those in the other three samples（P<0.05）. The contents of cellulose， 70% ethanol extract， sucrose， atractylenolide Ⅰ， atractylone and other components in 3-year-old AMR with topping after transplanting were significantly higher than those in the 2-year-old AMR with high market circulation rate（P<0.05）， while the contents of water-soluble extract and water-soluble total polysaccharides in 2-year-old samples with topping after transplanting were significantly higher than those in the 3-year-old AMR with topping after transplanting， direct seeding and without topping（P<0.05）. ConclusionUnder the current mainstream production mode， too much manual intervention makes AMR heavily enriched in polysaccharides and increased the yield， but the accumulation of sweet substances， fragrant substances and fiber substances is insufficient， which affects its quality. The current quality standard of AMR has some shortcomings in guiding the high quality production of it， it is suggested to revise the quality standard of AMR， supplement the quantitative analysis of secondary metabolites， and strengthen the production of imitation wild AMR.

Purpose To explore the clinicopathological features of cholangitis lenta.Methods The clinical data of 12 patients with cholangitis Lenta were collected,HE staining,his-tochemical staining and immunohistochemistry were used to de-tect the clinicopathological changes in 12 cases of cholangitis lenta,and the relevant literature was reviewed.Results All the 12 patients with cholangitis Lenta had acute history and ab-normal liver function,9 cases(75.0％)had abnormal white blood cell count,11 cases(91.7％)had abdominal infection,8 cases(66.7％)had systemic inflammatory response syndrome(SIRS)and 8 cases(66.7％)were diagnosed as sepsis.The common pathological manifestations included spotty necrosis(100％),piecemeal necrosis(100％),confluent necrosis(75.0％),bridging necrosis(66.7％),hepatocyte and capil-lary cholestasis(75.0％),and mild edema in portal area(33.3％).12 cases showed proliferation of bile ductules around the portal area,inspissated bile in dilated bile ductules and no cholestasis in interlobular bile ducts.12 cases had the basis of primary liver disease.2 patients(16.7％)died after being transferred to another hospital for treatment,and 10 patients(83.3％)survived,which lasted for 7 to 55 months.Conclu-sion Cholangitis Lenta shows unique clinical manifestations and morphological features,it is often accompanied by underly-ing diseases.The pathological manifestation of cholangitis Lenta in liver biopsy suggests that patients may have sepsis and/or ab-dominal infection,and therefore its diagnosis should be com-bined with clinical features,laboratory examination and imaging manifestations.

The core competencies of Operating Room specialist nurses are the key to being competent in Operating Room nursing work. However, the core competencies that Operating Room specialist nurses should possess in China are not yet clear, and there is no unified and standardized training model. This paper summarizes the concepts, core competency standards, and training models of Operating Room specialist nurses, explores the existing problems in admission conditions, curriculum arrangement, training content, training methods, and assessment of Operating Room specialist nurses, and puts forward suggestions for the training of Operating Room specialist nurses in China, in order to promote the improvement of core competency of Operating Room specialist nurses and the homogenization and standardization of training models.

Objective To observe the simulation of a finite element model of healthy adult female anal sphincter complex(ASC)established based on MRI.Methods One healthy adult female was prospectively enrolled,and an ASC geometric model was constructed using Mimics 3D reconstruction software based on anal MRI.Then the model was meshed with finite elements,and the parameters of the anal sphincter experiment in vitro and normal human anorectal pressure data were referenced to establish a finite element model,and its physiological characteristics and stress distribution were analyzed.Results After applying boundary loads to the ASC finite element model,under resting-state,the contraction deformation occurred on external anal sphincter,with the maximum displacement of 0.02 mm for anal canal and the smaller maximum displacement for internal anal sphincter,and a significant low stress zone in front of the upper inner surface of anal canal was observed.Under forced state,significant dilation and deformation occurred on anal canal,with the maximum displacement of 4.16 mm for the inner surface of anal sphincter.Under contracted state,the anal canal was compressed,and the maximum displacement of 1.89 mm for the inner surface of anal sphincter.Under both resting-state and contracted state,internal and external anal sphincters wholly moved forward.Conclusion The established ASC finite element model based on MRI could simulate the physiological activities and stress distribution of actual female adult muscle to some extent.

In order to develop a transgenic zebrafish line with green fluorescent protein (enhanced green fluorescent protein, EGFP) expressed specifically in muscle and heart, the recombinant expression vector constructed using the zebrafish ttn.2 gene promoter fragment and EGFP gene coding sequence and the capped mRNA of Tol2 transposase were co-injected into the zebrafish 1-cell stage embryos. The stable genetic Tg (ttn.2: EGFP) transgenic zebrafish line was successfully developed by fluorescence detection, followed by genetic hybridization screening and molecular identification. Fluorescence signals and whole-mount in situ hybridization showed that EGFP expression was located in muscle and heart, the specificity of which was consistent with the expression of ttn.2 mRNA. Inverse PCR showed that EGFP was integrated into chromosomes 4 and 11 of zebrafish in No. 33 transgenic line, while integrated into chromosome 1 in No. 34 transgenic line. The successful construction of this fluorescent transgenic zebrafish line, Tg (ttn.2: EGFP), laid a foundation for the research of muscle and heart development and related diseases. In addition, the transgenic zebrafish lines with strong green fluorescence can also be used as a new ornamental fish.
Animals ; Zebrafish/genetics* ; Animals, Genetically Modified/genetics* ; Green Fluorescent Proteins/metabolism* ; Zebrafish Proteins/genetics* ; Promoter Regions, Genetic

Mitochondrial myopathy is a group of multi-system diseases in which mitochondrial DNA(mtDNA)or nuclear DNA(nDNA)defects lead to structural and functional dysfunction of mitochondria.The clinical manifestations of mitochondrial myopathy are complex and varied,and the testing for mtDNA and nDNA is not widely available,so misdiagnosis or missed diagnosis is common.Chronic progressive external ophthalmoplegia(CPEO)is a common type of mitochondrial myopathy,which is characterized by blepharoptosis.Here we report a 38-year-old female with mitochondrial myopathy presented with chronic numbness and weakness of the limbs,accompanied by blepharoptosis that was recently noticed.Laboratory and head magnetic resonance imaging(MRI)examinations showed no obvious abnormalities.Muscle and nerve biopsies showed characteristic ragged red fibers(RRFs)and large aggregates of denatured mitochondria.Testing for mtDNA and nDNA showed a known mutation c.2857C＞T(p.R953C)and a novel variant c.2391G＞C(p.M797I)in the polymerase gamma(POLG)gene,so the patient was diagnosed as mitochondrial myopathy.Clinicians should pay more attention to long-term unexplained skeletal muscle diseases with recent onset blepharoptosis.Histopathologic examination and genetic testing are of great value in the early diagnosis and therapeutic intervention.

Objective To investigate the application value of pre-implantation genetic testing(PGT)in patients with Turner syndrome.Methods The clinical data,embryonic development,PGT results and pregnancy outcome of 18 patients with Turner syndrome who underwent PGT in the reproductive center of 900th Hospital from January 2016 to June 2023 were retrospectively analyzed.Results All 18 patients had spontaneous puberty development,of which 4 patients had primary ovarian insufficiency(POI).A total of 24 oocyte retrieval cycles were performed in 18 patients,of which 6 patients had no biopsied embryos for 10 cycles.Sixty-one embryos were biopsied and 60 embryos were clearly diagnosed,including 25 with chromosomal abnormalities.Seven patients with mosaic Turner syndrome obtained clinical pregnancies after transplantation,including 4 healthy boys had already been delivered and 3 are in pregnancy.Conclusion There are numerous types of karyotype in Turner syndrome.The clinical phenotypes vary greatly in individuals with Turner syndrome,and prognosis of PGT is significant different.Patients with Turner syndrome who had biopsied embryos can obtain available embryo using PGT,and achieve ideal clinical outcomes.

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins