OBJECTIVE T o establish a simple,rapid,highly sensitive and highly specific visualized detection meth-od for Klebsiella pneumoniae based on recombinase polymerase amplification(RPA)technique combined with the lateral flow dipstick(LFD)technique.METHODS The specific primers targeting the khe gene of K.pneumoniae were designed for RPA,the reaction temperature and reaction time for RPA were optimized,the optimal experi-mental conditions for the LFD were determined based on the colorimetric results.The sensitivity,stability,speci-ficity and clinical practicability of RPA-LFD in detection of the K.pneumoniae were evaluated.RESULTS RPA could achieved the optimal detection result for simplification of K.pneumoniae at 34 ℃ for 10 min.The LFD showed the best color development when Milipore HF 135s was used as the nitrocellulose membrane,phosphate buffered saline containing Tween-20(PBST)was used as the sample developing solution,and the concentrations of streptavidin and secondary antibody were 0.8 mg/ml and 0.1 mg/ml,respectively.The detection limit of the RPA-LFD was 5 CFU/ml for detection of K.pneumoniae,the relative standard deviation of the band color inten-sity was less than 5％in six parallel experiments,and there was no cross reactions with other bacterial strains.In addition,the sensitivity of the RPA-LFD in detection of K.pneumoniae was not affected by the serum compo-nents such as proteins and lipids.CONCLUSION The rapid detection method for K.pneumoniae that is established based RPA-LFD is characterized by the simple operation,high sensitivity and high specificity,and it does not need complicated equipment or strict technical requirement for operators and provides a new technique for early diagno-sis and epidemiological survey of the K.pneumoniae.

BACKGROUND:Three-dimensional(3D)Motion Capture Technology can build accurate,objective,and quantized medical virtual simulation model,which is conducive to clinical learners'precise and in-depth understanding and mastery of various traditional therapies.The virtual simulation model of traditional Chinese medicine based on the 3D Motion Capture Technology has been reported,but such a system of traditional Mongolian medicine therapy has not been reported.OBJECTIVE:To construct an interactive 3D visualization virtual simulation model based on the 3D Motion Capture Technology.METHODS:Motion capture data of the professor of Mongolian Medicine Department were collected using the 3D optical motion capture system(Motion Analysis)and Plantar Force Platform.The 3D motion model of brain vibration therapy was constructed using Motion Builder software,and the role model was constructed using Maya software matched with the action model.Unity3D software was used to build a virtual simulation system of Mongolian medical brain vibration therapy.The system integrated information on 3D animation,kinematic and dynamic parameters of Mongolian medical brain vibration therapy.RESULTS AND CONCLUSION:By using 3D Motion Capture Technology and Computer Simulation Technology to reconstruct the operation of Mongolian medical brain vibration therapy,it can display the posture of the operator and subject and record the key parameters of spatial position and changes of joint motion to obtain kinematic and dynamic parameters.The interactive 3D virtual simulation technology is used to realize the visual presentation of 3D virtual simulation of Mongolian medical brain vibration therapy.It lays a foundation for the standardization,digitization and visualization of Mongolian medical brain vibration therapy.

Objective:To establish the reference interval of 12 types of cytokines(IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,IFN-γ,IFN-α,TNF-α)in adult plasma based on multiple microsphere flow immunofluorescence(MBFFI).Methods:A total of 140 healthy adult patients who were examined at Xuzhou Central Hospital between January 2022 and December 2023 were included in the study.Plasma cytokine levels were detected and reference intervals were established by the flow cytometer and the assay kits produced by Qingdao Raisecare Biotechnology Co.,Ltd and Jiangsu BioPredia Biotechnology Co.,Ltd.Results:All of the cytokines exhibited a non-normal distribution,and there was a discrepancy in the 95％reference interval between the two re-agents.The reference intervals for the 12 cytokine kits produced by Qingdao Raisecare Biotechnology Co.,Ltd.were as follows:IFN-α:<4.91 pg/ml,IL-12 p70:<1.95 pg/ml,IL-5:<12.72 pg/ml,IL-8:<60.68 pg/ml,IL-1β:<27.67 pg/ml,IL-2:<5.01 pg/ml,IL-4:<1.22 pg/ml,IL-6:<6.11 pg/ml,TNF-α:<2.92 pg/ml,IL-17:<10.27 pg/ml,IL-10:<6.88 pg/ml,IFN-γ:<17.68 pg/ml.The reference intervals of the 12 cytokines produced by Jiangsu BioPredia Biotechnology Co.,Ltd.were as follows:IFN-α:<4.05 pg/ml,IL-12 p70:<7.33 pg/ml,IL-5:<7.80 pg/ml,IL-8:<13.24 pg/ml,IL-1β:<19.24 pg/ml,IL-2:<2.42 pg/ml,IL-4:<0.99 pg/ml,IL-6:<2.10 pg/ml,TNF-α:<0.87 pg/ml,IL-17:<1.42 pg/ml,IL-10:<1.10 pg/ml,IFN-γ:<1.34 pg/ml.Conclusion:In this study,the ref-erence range of two reagents for the detection of 12 kinds of cytokines in plasma of healthy adults is established by MBFFI,which pro-vides a valuable reference for the diagnosis and treatment of clinical-related diseases.

Objective:To establish the reference interval of 12 types of cytokines(IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,IFN-γ,IFN-α,TNF-α)in adult plasma based on multiple microsphere flow immunofluorescence(MBFFI).Methods:A total of 140 healthy adult patients who were examined at Xuzhou Central Hospital between January 2022 and December 2023 were included in the study.Plasma cytokine levels were detected and reference intervals were established by the flow cytometer and the assay kits produced by Qingdao Raisecare Biotechnology Co.,Ltd and Jiangsu BioPredia Biotechnology Co.,Ltd.Results:All of the cytokines exhibited a non-normal distribution,and there was a discrepancy in the 95％reference interval between the two re-agents.The reference intervals for the 12 cytokine kits produced by Qingdao Raisecare Biotechnology Co.,Ltd.were as follows:IFN-α:<4.91 pg/ml,IL-12 p70:<1.95 pg/ml,IL-5:<12.72 pg/ml,IL-8:<60.68 pg/ml,IL-1β:<27.67 pg/ml,IL-2:<5.01 pg/ml,IL-4:<1.22 pg/ml,IL-6:<6.11 pg/ml,TNF-α:<2.92 pg/ml,IL-17:<10.27 pg/ml,IL-10:<6.88 pg/ml,IFN-γ:<17.68 pg/ml.The reference intervals of the 12 cytokines produced by Jiangsu BioPredia Biotechnology Co.,Ltd.were as follows:IFN-α:<4.05 pg/ml,IL-12 p70:<7.33 pg/ml,IL-5:<7.80 pg/ml,IL-8:<13.24 pg/ml,IL-1β:<19.24 pg/ml,IL-2:<2.42 pg/ml,IL-4:<0.99 pg/ml,IL-6:<2.10 pg/ml,TNF-α:<0.87 pg/ml,IL-17:<1.42 pg/ml,IL-10:<1.10 pg/ml,IFN-γ:<1.34 pg/ml.Conclusion:In this study,the ref-erence range of two reagents for the detection of 12 kinds of cytokines in plasma of healthy adults is established by MBFFI,which pro-vides a valuable reference for the diagnosis and treatment of clinical-related diseases.

OBJECTIVE T o establish a simple,rapid,highly sensitive and highly specific visualized detection meth-od for Klebsiella pneumoniae based on recombinase polymerase amplification(RPA)technique combined with the lateral flow dipstick(LFD)technique.METHODS The specific primers targeting the khe gene of K.pneumoniae were designed for RPA,the reaction temperature and reaction time for RPA were optimized,the optimal experi-mental conditions for the LFD were determined based on the colorimetric results.The sensitivity,stability,speci-ficity and clinical practicability of RPA-LFD in detection of the K.pneumoniae were evaluated.RESULTS RPA could achieved the optimal detection result for simplification of K.pneumoniae at 34 ℃ for 10 min.The LFD showed the best color development when Milipore HF 135s was used as the nitrocellulose membrane,phosphate buffered saline containing Tween-20(PBST)was used as the sample developing solution,and the concentrations of streptavidin and secondary antibody were 0.8 mg/ml and 0.1 mg/ml,respectively.The detection limit of the RPA-LFD was 5 CFU/ml for detection of K.pneumoniae,the relative standard deviation of the band color inten-sity was less than 5％in six parallel experiments,and there was no cross reactions with other bacterial strains.In addition,the sensitivity of the RPA-LFD in detection of K.pneumoniae was not affected by the serum compo-nents such as proteins and lipids.CONCLUSION The rapid detection method for K.pneumoniae that is established based RPA-LFD is characterized by the simple operation,high sensitivity and high specificity,and it does not need complicated equipment or strict technical requirement for operators and provides a new technique for early diagno-sis and epidemiological survey of the K.pneumoniae.

BACKGROUND:Three-dimensional(3D)Motion Capture Technology can build accurate,objective,and quantized medical virtual simulation model,which is conducive to clinical learners'precise and in-depth understanding and mastery of various traditional therapies.The virtual simulation model of traditional Chinese medicine based on the 3D Motion Capture Technology has been reported,but such a system of traditional Mongolian medicine therapy has not been reported.OBJECTIVE:To construct an interactive 3D visualization virtual simulation model based on the 3D Motion Capture Technology.METHODS:Motion capture data of the professor of Mongolian Medicine Department were collected using the 3D optical motion capture system(Motion Analysis)and Plantar Force Platform.The 3D motion model of brain vibration therapy was constructed using Motion Builder software,and the role model was constructed using Maya software matched with the action model.Unity3D software was used to build a virtual simulation system of Mongolian medical brain vibration therapy.The system integrated information on 3D animation,kinematic and dynamic parameters of Mongolian medical brain vibration therapy.RESULTS AND CONCLUSION:By using 3D Motion Capture Technology and Computer Simulation Technology to reconstruct the operation of Mongolian medical brain vibration therapy,it can display the posture of the operator and subject and record the key parameters of spatial position and changes of joint motion to obtain kinematic and dynamic parameters.The interactive 3D virtual simulation technology is used to realize the visual presentation of 3D virtual simulation of Mongolian medical brain vibration therapy.It lays a foundation for the standardization,digitization and visualization of Mongolian medical brain vibration therapy.

Objective:To investigate the efficacy and safety of dual antiplatelet therapy (DAPT) in patients with minor ischemic stroke (MIS) and tiny unruptured intracranial aneurysm (UIA).Methods:Patients with MIS and tiny UIA admitted to the Department of Neurology, Weifang People's Hospital from October 1, 2022 to February 29, 2024 were included retrospectively. MIS was defined as baseline National Institutes of Health Stroke Scale (NIHSS) score ≤3. Tiny UIA was defined as UIA with a diameter of ≤3 mm. According to the antiplatelet therapy regimen, the patients were divided into an aspirin alone group and an aspirin+clopidogrel DAPT group. The main outcome measure was the clinical outcome at 90 days after onset. The modified Rankin Scale (mRS) score 0-1 was defined as a good outcome and >1 was defined as a poor outcome. Secondary outcome measures included aneurysm rupture, cerebral hemorrhage, and recurrence of cerebral ischemic events. Multivariate logistic regression analysis was used to identify the independent influencing factors for poor outcome. Results:A total of 183 patients with MIS and tiny UIA were included, including 108 males (59.0%), median aged 68 years (interquartile range, 61-73 years). All the UIAs were solitary. The mRS score of all patients before onset was 0; 152 patients (83.1%) had good outcome at 90 days after onset, 31 (16.9%) had poor outcome, and no UIA occurred rupture bleeding. Of the 94 patients (51.4%) who received aspirin monotherapy, 14 patients (14.9%) experienced recurrent cerebral ischemic events during follow-up, and 73 (77.7%) had good outcome. Of the 89 patients (48.6%) who received DAPT, 5 (5.6%) experienced recurrent ischemic events during follow-up, and 79 (88.8%) had good outcome. The recurrence rate of cerebral ischemic events in the aspirin group was significantly higher than that in the DAPT group ( χ2=4.227, P=0.040), while the good outcome rate was significantly lower than that in the DAPT group ( χ2=4.006, P=0.045). Multivariate logistic regression analysis showed that baseline NIHSS score was an independent risk factor for poor outcome (odds ratio 4.597, 95% confidence interval 1.864-11.339; P=0.001), while DAPT was an independent protective factor for good outcome (odds ratio 0.265, 95% confidence interval 0.079-0.892; P=0.032). Conclusion:Compared with aspirin monotherapy, the short-term combination of aspirin and clopidogrel in patients with MIS and tiny UIA may improve the outcome, reduces the recurrence of cerebral ischemic events, and has good safety.

Liquid chromatography-electrospray ionization tandem mass spectrometry(LC-ESI-MS)is a widely utilized technique for in vivo pharmaceutical analysis.Ionization interference within electrospray ion source,occurring between drugs and metabolites,can lead to signal variations,potentially compromising quantitative accuracy.Currently,method validation often overlooks this type of signal interference,which may result in systematic errors in quantitative results without matrix-matched calibration.In this study,we conducted an investigation using ten different groups of drugs and their corresponding me-tabolites across three LC-ESI-MS systems to assess the prevalence of signal interference.Such in-terferences can potentially cause or enhance nonlinearity in the calibration curves of drugs and metabolites,thereby altering the relationship between analyte response and concentration for quanti-fication.Finally,we established an evaluation scheme through a step-by-step dilution assay and employed three resolution methods:chromatographic separation,dilution,and stable labeled isotope internal standards correction.The above strategies were integrated into the method establishment process to improve quantitative accuracy.

Purpose: Prostate cancer (PCa) is an epithelial malignancy that originates in the prostate gland and is generally categorized into low, intermediate, and high-risk groups. The primary diagnostic indicator for PCa is the measurement of serum prostate-specific antigen (PSA) values. However, reliance on PSA levels can result in false positives, leading to unnecessary biopsies and an increased risk of invasive injuries. Therefore, it is imperative to develop an efficient and accurate method for PCa risk stratification. Many recent studies on PCa risk stratification based on clinical data have employed a binary classification, distinguishing between low to intermediate and high risk. In this paper, we propose a novel machine learning (ML) approach utilizing a stacking learning strategy for predicting the tripartite risk stratification of PCa. Methods: Clinical records, featuring attributes selected using the lasso method, were utilized with 5 ML classifiers. The outputs of these classifiers underwent transformation by various nonlinear transformers and were then concatenated with the lasso-selected features, resulting in a set of new features. A stacking learning strategy, integrating different ML classifiers, was developed based on these new features. Results: Our proposed approach demonstrated superior performance, achieving an accuracy of 0.83 and an area under the receiver operating characteristic curve value of 0.88 in a dataset comprising 197 PCa patients with 42 clinical characteristics. Conclusions This study aimed to improve clinicians’ ability to rapidly assess PCa risk stratification while reducing the burden on patients. This was achieved by using artificial intelligence-related technologies as an auxiliary method for diagnosing PCa.

Ectoine is an amino acid derivative and an important natural product in halophilic microorganisms. It plays an important role in protecting cells and stabilizing biological macromolecules, and can be widely used in biomedical fields such as drug preparation adjuvants, organ transplantation and preservation, skin wound repair and cosmetics. Due to the medical value and commercial market demand of ectoine, this article summarized the recent advances in the microbial production of ectoine, including the mutation and breeding of hyper-producing strains, construction of genetically and metabolically engineered strains, optimization of fermentation processes, and extraction and purification processes. The application of multi-omics technologies and computational biology to develop an ectoine producing cell factory was prospected, with the aim to provide a reference for ectoine overproduction.
Amino Acids, Diamino/metabolism* ; Fermentation