An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

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.

To investigate the antimicrobial resistance of Enterococci isolated from 596 anal swabs,feces and environmental samples were collected from pig,cattle,chicken and quail farms in Chang-sha,Hunan Province,Enterococci were isolated and identified by mass spectrometry.The minimum inhibitory concentrations(MICs)of 10 antimicrobial agents were determined by AGAR diffusion method.Whole genome sequencing(WGS)was used to detect the distribution of multilocus se-quence typing(ST),drug resistance genes and virulence genes.A total of 272 strains of Enterococ-cus were isolated(45.6％).The isolates were resistant to cefoxitin(68.9％)and cefotiofur(58.5％),followed by trimethoprim/sulfamethoxazole(52.2％),vancomycin(4.4％),and linezolid(13.6％).In this study,six linezolid highly resistant Enterococci isolates were analyzed by whole genome sequencing to explore the transmission mechanism of linezolid resistance because linezolid is forbidden to be used in aquaculture.ST403(4/6),ST16(1/6)and ST476(1/6)were the most common ST types,which all originated from the same farm.Three oxazolidinone resistance genes(cfr,poxtA and optrA)were found in all 21 strains.One strain(Ecc60)carried all three oxazo-lidinone resistance genes,but none of them were located on the plasmid.tet(M),aph(3')-Ⅲ,and lsa(A)were found in all six Enterococcus isolates.Interestingly,the present study is the first to i-dentify enterococci carrying the optrA gene in quail fecal samples.The analysis of the genetic envi-ronment of oxazolidinone resistance genes showed that the genetic environment of cfr(D),poxtA and optrA of isolates from the same farm was similar.A total of 19 virulence genes were detected in 6 isolates,of which 12 genes(ElrA,SrtA,ace,agg,cCF10,cOB1,cad,camE,ebpA,ebpC,efaAfs,tpx)were carried by all 6 isolates,and the types of virulence genes in strains from the same farm were extremely similar.The results showed that the drug resistance of Enterococci iso-lated from the fish farms in Changsha was serious,and the resistance rate to linezolid,which was prohibited in the fish farms,was high.The oxazolidinone resistance gene loci were accompanied by other resistance genes,especially the florfenicol resistance gene(FexA),which may be related to the abuse of florfenicol in the fish farms.

Objective To investigate the clinical features of patients with China Liver Cancer Staging(CNLC)stage Ⅲhepatocellular carcinoma(HCC)achieving five-year sustained complete remission(CR)after local treatment combined with systemic therapy,as well as potential contributing factors,and to provide a reference for optimizing the treatment of advanced HCC.Methods A retrospective analysis was performed for the clinical data of six patients with CNLC stage Ⅲ HCC who were treated in Department of Interventional Radiology,The First Affiliated Hospital of Soochow University,from January 2016 to December 2019 and achieved five-year sustained CR.Baseline characteristics,treatment modalities,and follow-up data were summarized,and a literature review was performed.Results The six patients had a mean age of 58.3±10.1 years,among whom five had stage Ⅲa HCC and one had stage Ⅲb HCC,and all patients had a history of hepatitis.The mean preoperative MELD score was 8.2±0.8 for the six patients,and there were five patients with Child-Pugh class A liver function and one with Child-Pugh class B liver function.All patients underwent transcatheter arterial chemoembolization,followed by sequential targeted drug therapy after surgery,with sorafenib for four patients and lenvatinib for two patients.Four patients with main portal vein tumor thrombus also received 125I seed implantation,one patient with the single-nodule type underwent radiofrequency ablation,and three patients received immunotherapy with camrelizumab.The median time to AFP normalization was 6 months,the median time from treatment to CR was 5.5 months,and the median follow-up time was 63 months.Conclusion Good liver function at baseline,an early and rapid reduction in AFP,and the combination of local treatment and systemic therapy are key factors for achieving long-term CR in patients with advanced HCC.Multi-center large-scale studies are needed in the future to further explore prognostic factors and optimize treatment regimens.

Objective To investigate the clinical efficacy and safety of facilitated percutaneous coronary intervention(PCI)with half-dose recombinant staphylokinase(r-SAK)in patients with ST-segment elevation myocardial infarction(STEMI)who are expected to undergo PCI within 120 minutes.Methods From October 2021 to August 2022,a total of 200 STEMI patients in eight centers were included and randomly assigned in a 1﹕1 ratio to either r-SAK group or control group.Patients received loading doses of aspirin and ticagrelor and intravenous heparin and were randomized to receive an intravenous bolus of either 5 mg r-SAK or normal saline prior to PCI.The outcomes were set as ST-segment resolution(STR)at 60-90 minutes after PCI,the proportion and transition of pathological Q waves on the 5th day after PCI,and the proportion of high-sensitivity cardiac troponin T(hs-cTnT)peaking within 12 hours of onset.The safety outcome was major bleeding events defined as Bleeding Academic Research Consortium(BARC)≥type 3 bleeding during hospitalization.Results Compared with the control group,the r-SAK group had a higher proportion of STR≥70%within 60-90 minutes after PCI(58.3%vs.40.3%,P=0.009);a lower proportion of pathological Q waves(59.1%vs.74.1%,P=0.040);a lower rate of Q wave progression(14.8%vs.43.2%,P＜0.001);a higher rate of Q wave disappearance(12.5%vs.3.7%,P=0.027);and a higher proportion of hs-cTnT peaking within 12 hours of symptom onset[31/40(77.5%)vs.17/33(51.5%),P=0.027].Regarding the safety outcome,no significant difference in BARC≥type 3 bleeding was found between the two groups during hospitalization(P＞0.05).Conclusions For STEMI patients who were expected to undergo primary PCI within 120 minutes of symptom onset,the facilitated PCI with half-dose r-SAK significantly increased the proportion of STR≥70%at 60-90 minutes after PCI,reduced the formation of pathological Q waves,and shortened the time to peak hs-cTnT,without increasing the risk of bleeding,which should be an alternative reperfusion strategy worthy of further study.

Objective To investigate the clinical features of patients with China Liver Cancer Staging(CNLC)stage Ⅲhepatocellular carcinoma(HCC)achieving five-year sustained complete remission(CR)after local treatment combined with systemic therapy,as well as potential contributing factors,and to provide a reference for optimizing the treatment of advanced HCC.Methods A retrospective analysis was performed for the clinical data of six patients with CNLC stage Ⅲ HCC who were treated in Department of Interventional Radiology,The First Affiliated Hospital of Soochow University,from January 2016 to December 2019 and achieved five-year sustained CR.Baseline characteristics,treatment modalities,and follow-up data were summarized,and a literature review was performed.Results The six patients had a mean age of 58.3±10.1 years,among whom five had stage Ⅲa HCC and one had stage Ⅲb HCC,and all patients had a history of hepatitis.The mean preoperative MELD score was 8.2±0.8 for the six patients,and there were five patients with Child-Pugh class A liver function and one with Child-Pugh class B liver function.All patients underwent transcatheter arterial chemoembolization,followed by sequential targeted drug therapy after surgery,with sorafenib for four patients and lenvatinib for two patients.Four patients with main portal vein tumor thrombus also received 125I seed implantation,one patient with the single-nodule type underwent radiofrequency ablation,and three patients received immunotherapy with camrelizumab.The median time to AFP normalization was 6 months,the median time from treatment to CR was 5.5 months,and the median follow-up time was 63 months.Conclusion Good liver function at baseline,an early and rapid reduction in AFP,and the combination of local treatment and systemic therapy are key factors for achieving long-term CR in patients with advanced HCC.Multi-center large-scale studies are needed in the future to further explore prognostic factors and optimize treatment regimens.

Objective To investigate the clinical efficacy and safety of facilitated percutaneous coronary intervention(PCI)with half-dose recombinant staphylokinase(r-SAK)in patients with ST-segment elevation myocardial infarction(STEMI)who are expected to undergo PCI within 120 minutes.Methods From October 2021 to August 2022,a total of 200 STEMI patients in eight centers were included and randomly assigned in a 1﹕1 ratio to either r-SAK group or control group.Patients received loading doses of aspirin and ticagrelor and intravenous heparin and were randomized to receive an intravenous bolus of either 5 mg r-SAK or normal saline prior to PCI.The outcomes were set as ST-segment resolution(STR)at 60-90 minutes after PCI,the proportion and transition of pathological Q waves on the 5th day after PCI,and the proportion of high-sensitivity cardiac troponin T(hs-cTnT)peaking within 12 hours of onset.The safety outcome was major bleeding events defined as Bleeding Academic Research Consortium(BARC)≥type 3 bleeding during hospitalization.Results Compared with the control group,the r-SAK group had a higher proportion of STR≥70%within 60-90 minutes after PCI(58.3%vs.40.3%,P=0.009);a lower proportion of pathological Q waves(59.1%vs.74.1%,P=0.040);a lower rate of Q wave progression(14.8%vs.43.2%,P＜0.001);a higher rate of Q wave disappearance(12.5%vs.3.7%,P=0.027);and a higher proportion of hs-cTnT peaking within 12 hours of symptom onset[31/40(77.5%)vs.17/33(51.5%),P=0.027].Regarding the safety outcome,no significant difference in BARC≥type 3 bleeding was found between the two groups during hospitalization(P＞0.05).Conclusions For STEMI patients who were expected to undergo primary PCI within 120 minutes of symptom onset,the facilitated PCI with half-dose r-SAK significantly increased the proportion of STR≥70%at 60-90 minutes after PCI,reduced the formation of pathological Q waves,and shortened the time to peak hs-cTnT,without increasing the risk of bleeding,which should be an alternative reperfusion strategy worthy of further study.

Objective To compare the effects of sequential Exoview and Mimics three-dimensional reconstruction with fluorescence method in thoracoscopic pulmonary segmentectomy.Methods The clinical data of 160 patients with lung cancer admitted to our hospital from January 2020 to June 2023 were retrospectively analyzed.Among them,79 patients who underwent thoracoscopic pulmonary segmentectomy with the sequential Exoview three-dimensional reconstruction and fluorescence method before the operation were classified as the Exoview group,and 81 patients who underwent thoracoscopic pulmonary segmentectomy with the sequential Mimics three-dimensional reconstruction and fluorescence method before the operation were classified as the Mimics group.The surgical completion status,the coincidence rate between the number of left and right pulmonary artery branches evaluated before operation and intraoperative findings,reconstruction time,segment display effect,general indicators of operation(operation time,intraoperative blood loss,number of lymph node dissection,thoracic tube placement time,postoperative hospital stay),pulmonary function[forced expiratory volume in one second(FEV1),percentage of forced expiratory volume in one second(FEV1%)]and complications were compared between the two groups.Results All patients in the two groups successfully completed thoracoscopic pulmonary segmentectomy,and indocyanine green was injected once in each group.The operation process was roughly consistent with the preoperative simulation,and no thoracotomy was performed.There was no statistically significant difference in the resection of lung segment between the two groups of patients(P>0.05).The coincidence rate between the number of left and right pulmonary artery branches evaluated before operation and intraoperative findings in the Exoview group was higher than that in the Mimics group(P<0.05).There was no significant difference in the segment display effect between the Exoview group and the Mimics group(P>0.05).The operation time and the reconstruction time in the Exoview group were shorter those that in the Mimics group,and the intraoperative blood loss was less than that in the Mimics group(P<0.05).There was no significant difference in the number of lymph node dissection,the thoracic tube placement time,or the postoperative hospital stay between the two groups(P>0.05).There was no statistically significant difference in FEV1 or FEV1%7 days after surgery compared with those before surgery(P>0.05).The FEV1 and FEV1%of patients in the Mimics group 7 days after surgery were lower than those beforesurgery(P<0.05).There was no statistically significant difference in FEV1 or FEV1%between the Exoview group and the Mimics group before and 7 days after surgery(P>0.05).The total incidence of complications in the Exoview group was 1.27%,compared with 4.94%in the Mimics group,the difference was not statistically significant(P>0.05).Conclusion Both sequential Exoview and Mimics three-dimensional reconstruction with fluorescence method are safe and effective for thoracoscopic pulmonary segmentectomy,while Exoview has more advantages in preoperative assessment of the number of pulmonary artery branches,and it has shorter reconstruction time and operation time,with less impact on lung function.

Objective To compare the effects of sequential Exoview and Mimics three-dimensional reconstruction with fluorescence method in thoracoscopic pulmonary segmentectomy.Methods The clinical data of 160 patients with lung cancer admitted to our hospital from January 2020 to June 2023 were retrospectively analyzed.Among them,79 patients who underwent thoracoscopic pulmonary segmentectomy with the sequential Exoview three-dimensional reconstruction and fluorescence method before the operation were classified as the Exoview group,and 81 patients who underwent thoracoscopic pulmonary segmentectomy with the sequential Mimics three-dimensional reconstruction and fluorescence method before the operation were classified as the Mimics group.The surgical completion status,the coincidence rate between the number of left and right pulmonary artery branches evaluated before operation and intraoperative findings,reconstruction time,segment display effect,general indicators of operation(operation time,intraoperative blood loss,number of lymph node dissection,thoracic tube placement time,postoperative hospital stay),pulmonary function[forced expiratory volume in one second(FEV1),percentage of forced expiratory volume in one second(FEV1%)]and complications were compared between the two groups.Results All patients in the two groups successfully completed thoracoscopic pulmonary segmentectomy,and indocyanine green was injected once in each group.The operation process was roughly consistent with the preoperative simulation,and no thoracotomy was performed.There was no statistically significant difference in the resection of lung segment between the two groups of patients(P>0.05).The coincidence rate between the number of left and right pulmonary artery branches evaluated before operation and intraoperative findings in the Exoview group was higher than that in the Mimics group(P<0.05).There was no significant difference in the segment display effect between the Exoview group and the Mimics group(P>0.05).The operation time and the reconstruction time in the Exoview group were shorter those that in the Mimics group,and the intraoperative blood loss was less than that in the Mimics group(P<0.05).There was no significant difference in the number of lymph node dissection,the thoracic tube placement time,or the postoperative hospital stay between the two groups(P>0.05).There was no statistically significant difference in FEV1 or FEV1%7 days after surgery compared with those before surgery(P>0.05).The FEV1 and FEV1%of patients in the Mimics group 7 days after surgery were lower than those beforesurgery(P<0.05).There was no statistically significant difference in FEV1 or FEV1%between the Exoview group and the Mimics group before and 7 days after surgery(P>0.05).The total incidence of complications in the Exoview group was 1.27%,compared with 4.94%in the Mimics group,the difference was not statistically significant(P>0.05).Conclusion Both sequential Exoview and Mimics three-dimensional reconstruction with fluorescence method are safe and effective for thoracoscopic pulmonary segmentectomy,while Exoview has more advantages in preoperative assessment of the number of pulmonary artery branches,and it has shorter reconstruction time and operation time,with less impact on lung function.