This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

OBJECTIVE To observe the etiological characteristics,serum C-reactive protein(CRP),interleukin-10(IL-10),procalcitonin(PCT)and treatment outcomes of the children with pneumonia.METHODS A total of 350 children with pneumonia who were treated in Jinhua People's Hospital from Sep.2021 to Sep.2023 and 30 healthy children matched with age and gender who received physical examination were recruited as the research subjects.The etiological characteristics,serum CRP,IL-6,PCT and treatment outcomes of the children with pneumonia were observed.RESULTS The etiological isolation rate was 74.00％(259/350)among the 350 children,the chil-dren who had viral infection accounted for 39.38％,the children with Mycoplasma pneumoniae(MP)infection 32.82％,the children with bacterial infection 20.08％,the children with mixed infections 7.72％.The viral infec-tion was prevalent in winter and was more common among the children aged between 1 and 3 years old;Myco-plasma pneumoniae infection was more common among the children aged between 3 and 6 years old.The CRP levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(10.53±1.54),(38.11±9.32),(21.07±6.55)and(33.33±8.11)mg/L,respectively,higher than those of the healthy children and the children negative for etiological test.The PCT levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(0.61±0.17),(2.84±0.51),(1.35±0.31)and(2.41±0.46)ng/ml,respectively,higher than those of the healthy children and the children negative for etiological test.The IL-10 levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(23.47±5.12),(26.45±7.15),(25.45±5.03)and(25.29±6.05)pg/ml,respectively,higher than those of the healthy children and the children negative for etiological test(P＜0.05).The levels of CRP and PCT of the bacterial infection group and the mixed infection group were higher than those of the M.pneumoniae infcection group,and the level of above indexes of the M.pneumoniae infcection group were higher than those of the viral infection group(P＜0.05).Among 259 children who were confirmed positive for etiological test,237 had favorable treatment out-comes,and 22 had poor treatment outcomes.The rate of poor treatment outcomes of the children with viral infec-tion was higher than that of the children with M.pneumoniae infcection(x2=5.930,P=0.015);the rate of poor treatment outcomes of the children with mixed infection was higher than that of the children with M.pneumoniae infcection(x2=9.954,P＜0.001).CONCLUSIONS The children with viral infection are dominant among the children with pneumonia,followed by the children with M.pneumoniae infcection.The rate of poor treatment outcomes of the children with viral infection or mixed infection is relatively high,the children with M.pneumoniae infcection generally have favorable treatment outcomes.The children with viral infection have slight increases of peripheral blood CRP and PCT,while the children with bacterial infection and mixed infection have a remarkable rise.

This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

OBJECTIVE To observe the etiological characteristics,serum C-reactive protein(CRP),interleukin-10(IL-10),procalcitonin(PCT)and treatment outcomes of the children with pneumonia.METHODS A total of 350 children with pneumonia who were treated in Jinhua People's Hospital from Sep.2021 to Sep.2023 and 30 healthy children matched with age and gender who received physical examination were recruited as the research subjects.The etiological characteristics,serum CRP,IL-6,PCT and treatment outcomes of the children with pneumonia were observed.RESULTS The etiological isolation rate was 74.00％(259/350)among the 350 children,the chil-dren who had viral infection accounted for 39.38％,the children with Mycoplasma pneumoniae(MP)infection 32.82％,the children with bacterial infection 20.08％,the children with mixed infections 7.72％.The viral infec-tion was prevalent in winter and was more common among the children aged between 1 and 3 years old;Myco-plasma pneumoniae infection was more common among the children aged between 3 and 6 years old.The CRP levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(10.53±1.54),(38.11±9.32),(21.07±6.55)and(33.33±8.11)mg/L,respectively,higher than those of the healthy children and the children negative for etiological test.The PCT levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(0.61±0.17),(2.84±0.51),(1.35±0.31)and(2.41±0.46)ng/ml,respectively,higher than those of the healthy children and the children negative for etiological test.The IL-10 levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(23.47±5.12),(26.45±7.15),(25.45±5.03)and(25.29±6.05)pg/ml,respectively,higher than those of the healthy children and the children negative for etiological test(P＜0.05).The levels of CRP and PCT of the bacterial infection group and the mixed infection group were higher than those of the M.pneumoniae infcection group,and the level of above indexes of the M.pneumoniae infcection group were higher than those of the viral infection group(P＜0.05).Among 259 children who were confirmed positive for etiological test,237 had favorable treatment out-comes,and 22 had poor treatment outcomes.The rate of poor treatment outcomes of the children with viral infec-tion was higher than that of the children with M.pneumoniae infcection(x2=5.930,P=0.015);the rate of poor treatment outcomes of the children with mixed infection was higher than that of the children with M.pneumoniae infcection(x2=9.954,P＜0.001).CONCLUSIONS The children with viral infection are dominant among the children with pneumonia,followed by the children with M.pneumoniae infcection.The rate of poor treatment outcomes of the children with viral infection or mixed infection is relatively high,the children with M.pneumoniae infcection generally have favorable treatment outcomes.The children with viral infection have slight increases of peripheral blood CRP and PCT,while the children with bacterial infection and mixed infection have a remarkable rise.

Objective To observe the clinical effect of Dongjing Tiaoshen Comprehensive Therapy combined with computerized cognitive behavioral therapy for insomnia(CCBT-I)in the treatment of chronic insomnia patients with liver depression and spleen deficiency syndrome.Methods A total of 60 patients with chronic insomnia of liver depression and spleen deficiency type were randomly divided into observation group and control group,30 cases in each group.Both groups of patients were treated with CCBT-I as the basic treatment,the observation group was additionally treated with Dongjing Tiaoshen Comprehensive Therapy,i.e.,mind-regulating acupuncture therapy combined with regular aerobic exercises and Dantian Qigong exercises,and the control group was additionally treated with cranial electrotherapy stimulation.The course of treatment for the two groups covered 4 weeks.The changes of Pittsburgh Sleep Quality Index(PSQI)score,Self-rating Anxiety Scale(SAS)score and Self-rating Depression Scale(SDS)score in the two groups were observed before and after treatment,and the clinical efficacy of the two groups was evaluated after treatment.Results(1)After 4 weeks of treatment,the total effective rate of the observation group was 86.67%(26/30),and that of the control group was 70.00%(21/30).The intergroup comparison(tested by rank sum test)showed that the curative effect of the observation group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).(2)After treatment,the PSQI scores of the two groups were significantly lower than those before treatment(P＜0.05),and the decrease of PSQI scores in the observation group was significantly superior to that in the control group,the difference being statistically significant(P＜0.05).(3)After treatment,the SAS and SDS scores of the two groups were significantly lower than those before treatment(P＜0.05),and the decrease of SAS and SDS scores in the observation group tended to be superior to that in the control group,but there was no significant difference between the two groups(P＞0.05).Conclusion For the treatment of chronic insomnia of liver depression and spleen deficiency type,the combination of Dongjing Tiaoshen Comprehensive Therapy with CCBT-I can significantly improve the sleep quality,relieve anxiety and depression,and improve the quality of life of the patients,which exerts significant efficacy.

Objective:To develop a robotic digestive endoscope system (RDES) and to evaluate its feasibility, safety and control performance by experiments.Methods:The RDES was designed based on the master-slave control system, which consisted of 3 parts: the integrated endoscope, including a knob and button robotic control system integrated with a gastroscope; the robotic mechanical arm system, including the base and arm, as well as the endoscopic advance-retreat control device (force-feedback function was designed) and the endoscopic axial rotation control device; the control console, including a master manipulator and an image monitor. The operator sit far away from the endoscope and controlled the master manipulator to bend the end of the endoscope and to control advance, retract and rotation of the endoscope. The air supply, water supply, suction, figure fixing and motion scaling switching was realized by pressing buttons on the master manipulator. In the endoscopy experiments performed on live pigs, 5 physicians each were in the beginner and advanced groups. Each operator operated RDES and traditional endoscope (2 weeks interval) to perform porcine gastroscopy 6 times, comparing the examination time. In the experiment of endoscopic circle drawing on the inner wall of the simulated stomach model, each operator in the two groups operated RDES 1∶1 motion scaling, 5∶1 motion scaling and ordinary endoscope to complete endoscopic circle drawing 6 times, comparing the completion time, accuracy (i.e. trajectory deviation) and workload.Results:RDES was operated normally with good force feedback function. All porcine in vivo gastroscopies were successful, without mucosal injury, bleeding or perforation. In beginner and advanced groups, the examination time of both RDES and ordinary endoscopy tended to decrease as the number of operations increased, but the decrease in time was greater for operating RDES than for operating ordinary endoscope (beginner group P=0.033; advanced group P=0.023). In the beginner group, the operators operating RDES with 1∶1 motion scaling or 5∶1 motion scaling to complete endoscopic circle drawing had shorter completion time [1.68 (1.40, 2.17) min, 1.73 (1.47, 2.37) min VS 4.13 (2.27, 5.16) min, H=32.506, P<0.001], better trajectory deviation (0.50±0.11 mm, 0.46±0.11 mm VS 0.82±0.26 mm, F=38.999, P<0.001], and less workload [42.00 (30.00, 50.33) points, 43.33 (35.33, 54.00) points VS 52.67 (48.67, 63.33) points, H=20.056, P<0.001] than operating ordinary endoscope. In the advanced group, the operators operating RDES with 1∶1 or 5∶1 motion scaling to complete endoscopic circle drawing had longer completion time than operating ordinary endoscope [1.72 (1.37, 2.53) min, 1.57 (1.25, 2.58) min VS 1.15 (0.86, 1.58) min, H=13.233, P=0.001], but trajectory deviation [0.47 (0.13, 0.57) mm, 0.44 (0.39, 0.58) mm VS 0.52 (0.42, 0.59) mm, H=3.202, P=0.202] and workload (44.62±21.77 points, 41.24±12.57 points VS 44.71±17.92 points, F=0.369, P=0.693) were not different from those of the ordinary endoscope. Conclusion:The RDES enables remote control, greatly reducing the endoscopists' workload. Additionally, it gives full play to the cooperative motion function of the large and small endoscopic knobs, making the control more flexible. Finally, it increases motion scaling switching function to make the control of endoscope more flexible and more accurate. It is also easy for beginners to learn and master, and can shorten the training period. So it can provide the possibility of remote endoscopic control and fully automated robotic endoscope.

Objective To investigate the factors influencing prognosis in patients with acute myocardial infarction complicated with cardiogenic shock undergoing primary percutaneous coronary intervention(PPCI).Methods Patients with acute myocardial infarction complicated with cardiogenic shock who underwent PPCI at our hospital between January 2015 and December 2019 were enrolled.Clinical baseline characteristics,coronary angiography and PCI-related parameters,and mechanical support information were collected.The patients were followed up for one year and divided into survival and death groups based on their survival status within one year.Differences in various factors between the two groups were compared.Results A total of 40 patients were enrolled,including 26 in the survival group and 14 in the death group.There were no differences in baseline data,diagnosis,risk factors,and comorbidities between the two groups.The survival group had a lower heart rate and higher blood pressure trend at admission compared to the death group.Myocardial enzymes were significantly lower in the survival group compared to the death group(median CK peak:496.00(198.25,2 830.00)U/L vs.3 040.00(405.75,5 626.53)U/L,P=0.003;median CK-MB peak:52.65(31.75,219.50)U/L vs.306.00(27.25,489.63)U/L,P=0.006).When comparing coronary angiography and PCI-related indicators between the two groups,the survival group had a higher rate of complete revascularization compared to the control group(53.85％vs.21.43％,P=0.048).The survival group had a higher proportion of extracorporeal membrane oxygenation(ECMO)combined with intra-aortic balloon pump(IABP)support compared to the control group[38.46％vs.7.14％,P=0.034].Conclusions Survival in patients with acute myocardial infarction complicated with cardiogenic shock undergoing PPCI is associated with lower level of myocardial enzymes,ECMO combined with IABP support and complete revascularization.

As artificial intelligence technology rapidly advances, its deployment within the medical sector presents substantial ethical challenges. Consequently, it becomes crucial to create a standardized, transparent, and secure framework for processing medical data. This includes setting the ethical boundaries for medical artificial intelligence and safeguarding both patient rights and data integrity. This consensus governs every facet of medical data handling through artificial intelligence, encompassing data gathering, processing, storage, transmission, utilization, and sharing. Its purpose is to ensure the management of medical data adheres to ethical standards and legal requirements, while safeguarding patient privacy and data security. Concurrently, the principles of compliance with the law, patient privacy respect, patient interest protection, and safety and reliability are underscored. Key issues such as informed consent, data usage, intellectual property protection, conflict of interest, and benefit sharing are examined in depth. The enactment of this expert consensus is intended to foster the profound integration and sustainable advancement of artificial intelligence within the medical domain, while simultaneously ensuring that artificial intelligence adheres strictly to the relevant ethical norms and legal frameworks during the processing of medical data.
Artificial Intelligence/legislation & jurisprudence* ; Humans ; Consensus ; Computer Security/standards* ; Confidentiality/ethics* ; Informed Consent/ethics*

Objective: To characterize the prevalence and genomic epidemiology of Vibrio parahaemolyticus from acute diarrheal patients in Shenzhen City from 2013 to 2021. Methods: Based on the Shenzhen Infectious Diarrhea Surveillance System, acute diarrheal patients were actively monitored in sentinel hospitals from 2013 to 2021. Whole-genome sequencing (WGS) of Vibrio parahaemolyticus isolates was performed, and the genomic population structure, serotypes, virulence genes and multilocus sequence typing were analyzed. Outbreak clusters from 2019 to 2021 were explored based on single-nucleotide polymorphism analysis. Results: A total of 48 623 acute diarrhea cases were monitored in 15 sentinel hospitals from 2013 to 2021, and 1 135 Vibrio parahaemolyticus strains were isolated, with a positive isolation rate of 2.3%. Qualified whole-genome sequencing data of 852 isolates were obtained. Eighty-nine serotypes, 21 known ST types and 5 new ST types were identified by sequence analysis, and 93.2% of strains were detected with toxin profile of tdh+trh-. 8 clonal groups (CGs) were captured, with CG3 as the absolute predominance, followed by CG189. The CG3 group was dominated by O3:K6 serotype and ST3 sequence type, while CG189 group was mainly O4:KUT, O4:K8 serotypes and ST189a and ST189 type. A total of 13 clusters were identified, containing 154 cases. About 30 outbreak clusters with 29 outbreak clusters caused by CG3 strains from 2019 to 2021. Conclusion: Vibrio parahaemolyticus is a major pathogen of acute infectious diarrhea in Shenzhen City, with diverse population structures. CG3 and CG189 have been prevalent and predominant in Shenzhen City for a long time. Scattered outbreaks and persistent sources of contamination ignored by traditional methods could be captured by WGS analysis. Tracing the source of epidemic clone groups and taking precise prevention and control measures are expected to significantly reduce the burden of diarrhea diseases caused by Vibrio parahaemolyticus infection in Shenzhen City.
Humans ; Vibrio parahaemolyticus/genetics* ; Diarrhea/epidemiology* ; Foodborne Diseases/epidemiology* ; Serogroup ; Genomics ; Dysentery ; Vibrio Infections/epidemiology* ; Serotyping

Objective To investigate the effects of glucose and serum deprivation under hypoxia(GSDH)treatment on oxidative stress and apoptosis in rat bone marrow mesenchymal stem cells (BMSCs), so to provide an experimental support for improving the therapeutic efficacy of BMSCs. Methods The cell injury model was established by hypoxia (1% O