Objective:To compare the enrichment effects of ultrafiltration, polyethylene glycol (PEG) precipitation, aluminum salt precipitation, and anionic membrane adsorption-elution on viruses in drinking water.Methods:Using phage MS2 as the target virus, three different concentrations of drinking water samples were prepared, and the samples were enriched by ultrafiltration 1, ultrafiltration 2, PEG precipitation, aluminum salt precipitation, and anionic membrane adsorption-elution method, respectively. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to quantify MS2 nucleic acid in pre and post concentrated samples and the recovery rates of MS2 in samples with high, medium and low concentrations were compared among the five methods.Results:Comparing the MS2 enrichment recovery rates of individual enrichment method in water samples of different concentrations, ultrafiltration method 1, PEG precipitation method, aluminum salt precipitation method, and membrane adsorption-elution method were not affected by the sample concentration, and the differences of the recovery rates for the three concentration water samples among the four methods were not statistically significant ( P>0.05). The MS2 enrichment recovery rates of the five enrichment methods were significantly different in all concentration samples ( P<0.05). The recovery rates of ultrafiltration method 1 were higher in all three concentration samples, followed by aluminum salt precipitation and anionic membrane adsorption-elution, PEG precipitation were higher in high concentration samples, but lower in low and medium concentration samples, and the recovery rates of ultrafiltration method 2 were the lowest in all three concentration samples. Comparing the Ct values of MS2 in the enriched samples by five methods, the Ct values of ultrafiltration method 1 were the smallest in the three concentration water samples. There was no statistically significant difference in MS2 Ct values among the five enrichment methods in the medium and high concentration water samples ( P>0.05). In low concentration simulated water samples, only the difference of MS2 Ct value between ultrafiltration method 1 and ultrafiltration method 2 was statistically significant ( Z=16.000, P=0.016). Conclusions:Considering the operation simplicity, operation time and virus recovery rate after enrichment, ultrafiltration was the most effective method for virus enrichment in drinking water.

Objective:To evaluate the preservation efficacy of 7 non-inactivating virus preservation solutions.Methods:Equal amounts of H1N1 virus were added to 7 commercially available non-inactivating virus preservation solutions, and the samples were stored at -20 ℃, 4 ℃, 25 ℃ and 37 ℃ for 1 hour, 6 hours, 1 day, 3 days, and 5 days. The viral nucleic acid in each simulated sample under different storage conditions was measured using real-time quantitative PCR. The hemagglutination (HA) titer was determined through viral isolation culture and hemagglutination assay, comparing the differences in viral growth activity across different storage solutions and conditions.Results:Except for solution E, the other solutions effectively protected viral nucleic acid at the 4 storage temperatures. In terms of viral activity, solutions A, B, C, and D effectively maintained viral viability. A and B showing the best performance, E and F showed poorer performance, and G performed the worst.Conclusions:Most non-inactivating virus preservation solutions effectively protect viral nucleic acid, but there are significant differences in their ability to maintain viral viability. To ensure optimal virus preservation, it is recommended that medical institutions evaluate the effectiveness of preservation solutions before use.

Objective:To compare the enrichment effects of ultrafiltration, polyethylene glycol (PEG) precipitation, aluminum salt precipitation, and anionic membrane adsorption-elution on viruses in drinking water.Methods:Using phage MS2 as the target virus, three different concentrations of drinking water samples were prepared, and the samples were enriched by ultrafiltration 1, ultrafiltration 2, PEG precipitation, aluminum salt precipitation, and anionic membrane adsorption-elution method, respectively. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to quantify MS2 nucleic acid in pre and post concentrated samples and the recovery rates of MS2 in samples with high, medium and low concentrations were compared among the five methods.Results:Comparing the MS2 enrichment recovery rates of individual enrichment method in water samples of different concentrations, ultrafiltration method 1, PEG precipitation method, aluminum salt precipitation method, and membrane adsorption-elution method were not affected by the sample concentration, and the differences of the recovery rates for the three concentration water samples among the four methods were not statistically significant ( P>0.05). The MS2 enrichment recovery rates of the five enrichment methods were significantly different in all concentration samples ( P<0.05). The recovery rates of ultrafiltration method 1 were higher in all three concentration samples, followed by aluminum salt precipitation and anionic membrane adsorption-elution, PEG precipitation were higher in high concentration samples, but lower in low and medium concentration samples, and the recovery rates of ultrafiltration method 2 were the lowest in all three concentration samples. Comparing the Ct values of MS2 in the enriched samples by five methods, the Ct values of ultrafiltration method 1 were the smallest in the three concentration water samples. There was no statistically significant difference in MS2 Ct values among the five enrichment methods in the medium and high concentration water samples ( P>0.05). In low concentration simulated water samples, only the difference of MS2 Ct value between ultrafiltration method 1 and ultrafiltration method 2 was statistically significant ( Z=16.000, P=0.016). Conclusions:Considering the operation simplicity, operation time and virus recovery rate after enrichment, ultrafiltration was the most effective method for virus enrichment in drinking water.

Objective:To evaluate the preservation efficacy of 7 non-inactivating virus preservation solutions.Methods:Equal amounts of H1N1 virus were added to 7 commercially available non-inactivating virus preservation solutions, and the samples were stored at -20 ℃, 4 ℃, 25 ℃ and 37 ℃ for 1 hour, 6 hours, 1 day, 3 days, and 5 days. The viral nucleic acid in each simulated sample under different storage conditions was measured using real-time quantitative PCR. The hemagglutination (HA) titer was determined through viral isolation culture and hemagglutination assay, comparing the differences in viral growth activity across different storage solutions and conditions.Results:Except for solution E, the other solutions effectively protected viral nucleic acid at the 4 storage temperatures. In terms of viral activity, solutions A, B, C, and D effectively maintained viral viability. A and B showing the best performance, E and F showed poorer performance, and G performed the worst.Conclusions:Most non-inactivating virus preservation solutions effectively protect viral nucleic acid, but there are significant differences in their ability to maintain viral viability. To ensure optimal virus preservation, it is recommended that medical institutions evaluate the effectiveness of preservation solutions before use.

Objective To investigate the similarities and differences in the pathophysiological responses caused by different infection sites in sepsis patients and to evaluate the clinical significance of the source of infection on the prognosis of patients.Methods A total of 159 patients with a clear source of infection in Foshan Hospital of Traditional Chinese Medicine from January 1,2021 to January 1,2023 were selected and divided into survival group(n=98)and death group(n=61)based on their survival status within 28 days.The independent risk factors affecting the prognosis of sepsis patients were determined by multivariate Logistic regression analysis.The 28-day mortality rates of different infection source groups were compared,and the distribution differences of serum biomarkers in different infection source groups were explored by analysis of variance.Results Among the 159 patients,the frequency of infection source distribution was as follows in descending order:Respiratory tract(62.89％),urinary tract(13.84％),abdomen(13.21％),skin and soft tissue(10.06％).Multivariate Logistic regression analysis showed that age,procalcitonin,sepsis-related organ failure assessment score,and respiratory tract infection source were significant risk factors affecting the 28-day mortality of sepsis patients(P＜0.05).Different infection source groups showed different degrees of differences in inflammation,coagulation,and organ dysfunction,with heterogeneity.Conclusion The differences in the source of infection lead to significant differences in the pathophysiological features(inflammatory response,coagulation activation,and organ dysfunction)and short-term prognosis(28-day mortality)of sepsis.

Objective:To establish a nested PCR method to detect the 2019 novel coronavirus (2019-nCoV), as a supplement to the real-time fluorescent PCR method, and discuss the preliminary application value of this method in clinical diagnosis.Methods:According to the conservative sequences of the 2019-nCoV gene, the nested PCR primers including N gene and S gene, were designed on line. By optimizing the nested PCR reaction systems, the qualitative detection was established by testing N gene and sequencing its PCR product while the preliminary type identification was established by testing S gene and sequencing its PCR product. The sensitivity was evaluated by the gradient dilution of 2019-nCoV positive samples’ nucleic acid and the specificity was evaluated by detecting the human coronavirus OC43, 229E, HKU1, NL63, influenza virus positive samples. The established method was applied to 15 samples with Ct >33 and 15 samples with Ct <33 screened by real-time fluorescent PCR, and the positive amplification result were sequenced and analyzed to verify the result. Results:The established nested PCR method could amplify specific bands of 355 bp N gene fragment and 449 bp S gene fragment. No amplifications occurred in other human coronaviruses samples including 229E、OC43、HKU1、NL63 or in influenza virus samples including H3N2, H1N1(pdm) and B. The minimum detection limit of the N gene fragment could reach Ct value about 37.21. Among the 30 COVID-19 positive samples, the N gene positive coincidence rate detected by nested PCR was 100% (30/30); the S gene positive coincidence rate reached 60% (18/30). 28 samples’ sequences of N gene fragment were completely consistent with 2019-nCoV by BLAST, and the characteristic result of site mutations of 12 samples’ S gene was obtained. Conclusions:A nested PCR method for the specific detection of 2019-nCoV was established, and some characteristic mutations on S gene could be analyzed by sequencing the PCR amplified products. It could be used as a supplement to the real-time fluorescent PCR method.

Objective: To investigate the short-term outcome of patients with acute myocardial infarction complicating cardiogenic shock due to left main disease. Methods: A total of 24 patients with acute myocardial infarction complicating cardiogenic shock due to left main artery disease hospitalized in Fuwai hospital from June 2012 to May 2018 were included. The clinical data were analyzed,and the patients were divided into survivor group (11 cases) and death group (13 cases) according to survival status at 28 days post the diagnosis of shock. The patients were further divided into thrombolysis in myocardial infarction(TIMI) flow grade 0-2 group (11 cases) and TIMI flow grade 3 group (13 cases) according to TIMI flow grade after the procedure. The patients were then divided into non-three-vessel lesions group (14 cases) and three-vessel lesions group (10 cases) according to coronary angiography results. Results: Compared with survivor group, patients in death group presented with lower worst systolic blood pressure within 24 hours after admission (50(48, 70) mmHg (1 mmHg=0.133 kPa) vs. 73(70, 80) mmHg, P<0.01), lower worst diastolic blood pressure with in 24 hours after admission ((33.5±12.4) mmHg vs. (48.9±9.4) mmHg, P<0.01), higher respiratory rates ((27.3±2.5) times/min vs. (21.5±4.0) times/min, P<0.01), less 24 hours urine output ((422±266) ml vs. (1 680±863) ml, P<0.01), lower platelet counts ((161.9±81.9)×10⁹/L vs. (241.6±94.0)×10⁹/L, P=0.03), higher serum creatinine ((250.0±36.8) μmol/L vs. (132.7±34.2) μmol/L, P<0.01), higher alanine aminotransferase (288(76,846) IU/ml vs. 81(42, 109) IU/ml, P=0.04), lower artery pH (7.11±0.17 vs. 7.39±0.09, P<0.01), higher lactic acid ((10.29±3.62) mmol/L vs. (4.21±2.85) mmol/L, P<0.01), higher incidence of invasive ventilation (7/13 vs. 2/11, P=0.02), higher scores of acute physiology and chronic health evaluation (APACHE) Ⅱ (35.4±6.8 vs. 18.7±1.7, P<0.01) and simplified acute physiology score (SAPS) Ⅱ (73.5±17.4 vs. 47.0±4.3, P<0.01), and higher incidence of target vessel TIMI flow grade 0-2 (10/13 vs. 1/11, P<0.01). Kaplan-Meier survival curve analysis showed that survival rate at 28 days post the diagnosis of shock in TIMI flow grade 3 group was higher than that in TIMI flow grade 0-2 group (76.9% vs. 9.1%, log-rank test, P<0.01), and mortality rate was similar at 28 days post the diagnosis of shock between non-three-vessel lesions group and three-vessel lesions group (35.7% vs. 60.0%, log-rank test, P=0.14). Multivariate logistic regression analysis showed that compared with TIMI flow grade 0-2 group, the OR value of death at 28 days post the diagnosis of shock in TIMI flow grade 3 patients with acute myocardial infarction complicating cardiogenic shock due to left main disease was 0.030(95%CI 0.003-0.340, P<0.01). Conclusion Short-term outcomeof patients with acute myocardial infarction complicating cardiogenic shock due to left main disease remains poor, and final flow of TIMI grade 3 is confirmed as independent protective factor of death at 28 days post the diagnosis of shock in these patients.

Objective: To investigate the efficacy of bare metal stent for treating focal coronary artery aneurysm complicating with severe stenosisin single coronary artery. Methods: This retrospective analysis was performed in 7 patients who were diagnosed as local coronary artery aneurysm complicating with severe stenosis(≥70%) in single coronary artery and treated with bare metal stent during the period from December 2012 to June 2015 in Fuwai Hospital. All 7 patients were male with age of (62±11) years old. During the interventional operation, the narrow parts were pre-expanded,and all patients received bare metal stents implantation to cover aneurysms.The clinical and imaging data of patients immediately post procedure and at postoperative follow-up were collected to evaluate the clinical efficacy. Results: There were 5 cases of left anterior descending aneurysms and 2 cases of right coronary artery aneurysms. The diameter of aneurysm was (5.21±1.28)mm, and the length was (13.71±3.88)mm. There was intracranial vortex in coronary arteriography immediately after intervention.Proximalstenosis of coronary artery aneurysm was disappeared,and the distal blood flow was TIMI class 3.There were no signs of aortic dissection and thrombus formation.During 6(6 16) months follow-up, the aneurysms were disappeared,and there were no major adverse cardiovascular events which including myocardial ischemia, acute myocardial infarction, revascularization,bleeding,and death for all patients. Conclusion Initial experience shows that double-layer bare metal stents implantation for patients with localized coronary artery aneurysm complicating with severe stenosis in single vessel is safe and effective.

Objective: To compare predictive value of the current 7 scoring systems and CADILLAC-plus scoring system for risk of short-term deathin patients with acute myocardial infarction complicating cardiogenic shock. Methods: A total of 126 acute myocardial infarction patients complicating cardiogenic shock hospitalized in Fuwai hospital from June 2014 to January 2018 were enrolled in this study, the clinical data were retrospectively analyzed. The patients were divided into survival group(49 cases) and death group(77 cases) according to survival or not at 28 days after diagnosis of cardiogenic shock.The scores of APACHE Ⅱ,APACHE Ⅲ,SAPS Ⅱ,PAMI, TIMI-STEMI,TIMI-NSTEMI,and CADILLAC were calculated within 24 hours in coronary care unit (CCU),and scores of CADILLAC-plus, which is an improved score derived from CADILLAC, was also calculated. The predictive value of the different scoring systems for 28 day smortality of acute myocardial infarction patients complicating cardiogenic shock were compared in this patient cohort. Results: Scores of APACHEⅡ,APACHEⅢ,SAPSⅡ,PAMI,TIMI-STEMI, TIMI-NSTEMI,CADILLAC,and CADILLAC-plus were all significantly higher in death group than in survival group: (28.9±10.2 vs. 21.8±8.3,94.0 (57.0,114.0) vs. 57.0 (45.4,81.5) ,62.0 (46.0,81.0) vs. 47.0 (41.5,60.5) ,7.0 (6.0,9.0) vs. 6.0 (6.0,7.5) ,10.0 (9.0,11.0) vs. 9.0 (8.0,10.0) ,4.0 (3.0,5.0) vs. 3.0 (3.0,4.0) ,10.0 (7.0,12.0) vs. 7.0 (5.0,9.0) ,and 10.0 (8.0,14.0) vs. 7.0 (5.0,10.0) , respectively, all P<0.01).The area under the curve(AUC) of receiver operating characteristic(ROC) curve for predicting 28 days death of APACHE Ⅱ,APACHE Ⅲ,SAPS Ⅱ,PAMI, TIMI-STEMI,TIMI-NSTEMI,and CADILLAC scoring systems were 0.820,0.797,0.785,0.667,0.657,0.711,and 0.821,respectively and cut-off value was 27.5,79.5,66.0,8.5,10.5,3.5,and 8.5, respectively and the sensitivity was 0.766,0.844,0.649,0.494,0.494,0.740,and 0.753, respectively and specificity was 0.816,0.755,0.837,0.204,0.796,0.571,and 0.755,respectively.The AUC of ROC of CADILLAC-plus scoring system was 0.885,cut-off value was 9.5, sensitivity was 0.896,and specificity was 0.735. The predicting efficacy of CADILLAC-plus scoring system was superior to other scoring systems. Conclusion The 7 current scoring systems are all suitable for predicting theshort-term death in acute myocardial infarction patients complicating cardiogenic shock, and the predicting efficacy of the improved CADILLAC-plus score is superior to other scoring systems in this patient cohort.

Objective: To investigate the association between APOB gene R532W polymorphism and the risk of coronary heart disease (CHD) in patients without lipid-lowering treatment and to analyze the interactions between the variation of R532W and different risk factors of CHD. Methods: CHD and non-CHD were diagnosed according to coronary artery angiography (CAG) and/or coronary computed tomography angiogram (CTA) results, as well as clinical features. Blood samples from 771 CHD patients and 772 age- and sex-matched non-CHD controls, who never accepted any lipid-lowering treatments, were collected. R532W was genotyped by HumanExome BeadChip at BGI and strict quality control was made. Firstly, the association between R532W polymorphism and the risk of CHD in 3 genetic models (GA+ AA vs.GG, AA vs. GG+ GA, AA vs. GA vs. GG) after adjusting confounding factors was explored. Then, the interactions between the variation of this loci and risk factors related to CHD were investigated. Results: (1) Total cholesterol (TC) levels were significantly lower in AA genotype than in GA genotype in the total cohort and non-CHD controls, but was similar among the 3 genotypes in CHD patients. (2) R532W GG, GA and AA distribution was 80.7%, 18.2% and 1.2% in CHD patients, and 74.6%, 23.8% and 1.6% in non-CHD controls (P<0.05). (3) R532 polymorphism was related to the incidence of CHD in the dominant model, and A-allele carriers were related to about 35% reduced risk of CHD (OR=0.653, 95% CI 0.502-0.849, P=0.001) after adjusting for confounding factors. (4) R532W polymorphism had positive interactions with hypertension (1.452) and smoke (1.077), while negative interaction with diabetes (0.553) in the occurrence of CHD. Conclusions APOB gene R532W polymorphism is related to TC levels in Chinese north Han population. A-allele carries of R532W loci is linked with reduced risk of CHD in the absence of lipid-lowering treatment. R532W polymorphism has a positive additive interaction with hypertension and smoke, while a negative additive interaction with diabetes mellitus in the occurrence of CHD.