ObjectiveTo investigate the whole genomic characteristics and phylogenetic relationships of clinical isolates of enteroaggregative Escherichia coli (EAEC) in diarrhea outpatients in Pudong New Area, Shanghai. MethodsBased on the diarrheal disease surveillance network in Pudong New Area, Shanghai, whole-genome sequencing was performed on a total of 55 EAEC strains isolated from fecal samples of the diarrhea outpatients from January 2015 to December 2019. The genome analyses based on raw sequencing data encompassed genome size, coding genes, dispersed repeat sequences, genomic islands, and protein coding regions, and pan-genome analyses were conducted simultaneously. Contigs sequences assays were performed to analyze molecular characteristics including serotypes, antibiotic resistance genes, and virulence factors. The phylogenetic clusters and multilocus sequence typing (MLST) were identified, and a phylogenetic tree was constructed. ResultsEAEC exhibited an open pan-genome. The predominant serotype of EAEC in diarrhea outpatients in Pudong New Area was O130:H27, and the carriage rate of β-lactam resistance genes was the highest (67.27%, 37/55). A total of 29 virulence factors and 106 virulence genes were identified, phylogenic group B1 was the predominant group, and clonal group CC31 was the dominant clonal group. The strain distribution was highly heterogeneous. ConclusionThe genomic characteristics of EAEC displayed significant strain polymorphism. It is necessary to develop effective strategies for differential diagnosis and improve detection capabilities for infection with EAEC of different serotypes and genotypes.

Objective:To establish reference values for clot waveform analysis (CWA) and analyze their diagnostic efficacy in distinguishing acquired hemophilia A (AHA) and lupus anticoagulant (LA)-positive patients.Methods:Case-Control Study. A total of 391 healthy individuals(260 males and 131 females) with a mean age of 45.53±14.85 years were enrolled at Nanyang central Hospital between January 6, 2023 and October 10, 2024. Prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT) were measured to establish reference ranges for the CWA parameters, including maximal reaction velocity (Min1), maximal reaction acceleration (Min2), and maximal reaction deceleration (Max2). A total of 158 definitively diagnosed AHA and LA-positive patients (mean age:42.46±14.83 years), including 34 AHA patients and 124 LA-positive patients, were recruited. The Mann Whitney U test was used to analyze the differences in the CWA parameters between the two groups. The diagnostic efficacy of CWA parameters in distinguishing AHA and LA-positive patients was evaluated using the area under the receiver operating characteristic(ROC) curve AUC and the cut-off values were calculated. Results:The reference values for PT-Min1, APTT-Min1, APTT-Min2, APTT-Max2, TT-Min1, TT-Min2, TT-Max2 were 203.41-516.89, 144.63-324.03, 526.46-1 190.03, -404.96±157.22, 159.17±60.34, 272.29-686.99, and -289.47--113.76, respectively. Compared with the CWA parameters in AHA patients, APTT-Max2 was significantly lower in LA-positive patients [-422.74(-577.50, -239.22) vs. -68.87(-92.85,30.28), Z=-7.43, P<0.01], while PT-Min1, APTT-Min1, APTT-Min2, TT-Min1, TT-Min2 were significantly elevated [287.01(188.03, 382.50) vs. 107.45(90.20, 151.39), 972.88(601.20, 1 351.19) vs. 229.10(118.38, 371.67), Z=6.68, 6.69, all P<0.01]. ROC analysis demonstrated the APTT-CWA parameter exhibited high diagnostic efficacy in patients with AHA (AUC>0.900 for both).Additionally, APTT-Min1 and APTT-Max2 were found to be useful in distinguishing between AHA patients and those with LA-positive status accompanied by APTT prolongation (AUC=0.660, 0.700, respectively). Conclusions:Reference values for CWA parameters were successfully established. The APTT-CWA is useful for differentiating between AHA and LA-positive patients and APTT-Max2 demonstrated a good diagnostic value in differentiating AHA patients from those with LA-positive status accompanied by APTT prolongation.

Objective:To establish reference values for clot waveform analysis (CWA) and analyze their diagnostic efficacy in distinguishing acquired hemophilia A (AHA) and lupus anticoagulant (LA)-positive patients.Methods:Case-Control Study. A total of 391 healthy individuals(260 males and 131 females) with a mean age of 45.53±14.85 years were enrolled at Nanyang central Hospital between January 6, 2023 and October 10, 2024. Prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT) were measured to establish reference ranges for the CWA parameters, including maximal reaction velocity (Min1), maximal reaction acceleration (Min2), and maximal reaction deceleration (Max2). A total of 158 definitively diagnosed AHA and LA-positive patients (mean age:42.46±14.83 years), including 34 AHA patients and 124 LA-positive patients, were recruited. The Mann Whitney U test was used to analyze the differences in the CWA parameters between the two groups. The diagnostic efficacy of CWA parameters in distinguishing AHA and LA-positive patients was evaluated using the area under the receiver operating characteristic(ROC) curve AUC and the cut-off values were calculated. Results:The reference values for PT-Min1, APTT-Min1, APTT-Min2, APTT-Max2, TT-Min1, TT-Min2, TT-Max2 were 203.41-516.89, 144.63-324.03, 526.46-1 190.03, -404.96±157.22, 159.17±60.34, 272.29-686.99, and -289.47--113.76, respectively. Compared with the CWA parameters in AHA patients, APTT-Max2 was significantly lower in LA-positive patients [-422.74(-577.50, -239.22) vs. -68.87(-92.85,30.28), Z=-7.43, P<0.01], while PT-Min1, APTT-Min1, APTT-Min2, TT-Min1, TT-Min2 were significantly elevated [287.01(188.03, 382.50) vs. 107.45(90.20, 151.39), 972.88(601.20, 1 351.19) vs. 229.10(118.38, 371.67), Z=6.68, 6.69, all P<0.01]. ROC analysis demonstrated the APTT-CWA parameter exhibited high diagnostic efficacy in patients with AHA (AUC>0.900 for both).Additionally, APTT-Min1 and APTT-Max2 were found to be useful in distinguishing between AHA patients and those with LA-positive status accompanied by APTT prolongation (AUC=0.660, 0.700, respectively). Conclusions:Reference values for CWA parameters were successfully established. The APTT-CWA is useful for differentiating between AHA and LA-positive patients and APTT-Max2 demonstrated a good diagnostic value in differentiating AHA patients from those with LA-positive status accompanied by APTT prolongation.

Objective·To analyze the development trend and current situation of high-level clinical research at Shanghai Jiao Tong University School of Medicine(SJTUSM)from 2015 to 2023,and to summarize the strategies for improving clinical research capability of SJTUSM,aiming to provide reference and inspiration for medical universities to promote high-level clinical research.Methods·Papers published by first authors from SJTUSM or its affiliated hospitals during this period were retrieved in the Web of Science core collection database.General descriptive methods were used to quantify the number of clinical research papers,registered clinical trials,clinical research talent teams,and the current status of clinical research cooperation.Bibliometric methods were used to visualize the international collaboration and topic distribution of retrieved clinical research papers.The differences in the number of clinical research publications,Q1 publications,publications in top clinical medical journals,and full-time researchers between the periods of 2015?2018 and 2019?2023 were compared and analyzed.Results·From 2015 to 2023,a total of 9 468 clinical research papers were published by SJTUSM or its affiliated hospitals as the first author's institution,showing an increasing trend year by year.During this period,the number of Q1 publications of SJTUSM's clinical research,publications in top clinical medical journals,clinical studies registered on the United States Clinical Trial Registry website,clinical studies registered on the Chinese Clinical Trial Registry website,clinical guidelines and expert consensus publications,full-time research team members,and newly added national-level talents all showed an overall upward trend.Tumors,cardiovascular and cerebrovascular diseases,major chronic diseases,and mental illnesses were the main research hotspots.A total of 1 308 papers(13.82%)were international collaborative papers.Developed countries/regions such as the United States(733 papers),Australia(137 papers),and the United Kingdom(99 papers)were closely cooperating with SJTUSM.From 2019 to 2023,there was a significant increase in the number of clinical research papers,Q1 publications,publications in top clinical medical journals,and newly added full-time research team members in affiliated hospitals compared to the period from 2015 to 2018(all P<0.05).Conclusion·SJTUSM has strengthened top-level design,conducted in-depth major diseases,and constructed a systematic,integrated and closed-loop model for enhancing clinical research level along the"platform-talent-methodology"pathway,resulting in a number of high-quality clinical research outcomes.

【Objective】 To investigate the characteristics of HBV serological markers of NAT reactive blood donors under different HBsAg status. 【Methods】 NAT reactive samples, with HBsAg-, HBsAg+ /retest - and HBsAg+ by single reagent were collected from September 2021 to May 2022 in our laboratory. The TMA non-reactive samples were retested by Roche PCR, then HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc were detected by ECLI for statistical analysis. 【Results】 A total of 66 samples were collected, among which 55 were HBsAg-/NAT+. The positive rate of anti-HBc, anti-HBs+ anti-HBc, anti-HBe+ anti-HBc was 87.3% (48/55), 43.6% (24/55) and 45.5% (25/55), respectively. The positive rate of anti-HBs was 10.9% (6/55) and the overall negative rate was 1.8% (1/55). In 7 HBsAg+ initially/retest -/NAT+ samples, the positive rate of anti-HBc was 100%(7/7), and the positive rate of anti-HBe+ anti-HBc was 71.4%(5/7). In 4 HBsAg+ /NAT+ samples by single reagent, the positive rate of HBsAg+ anti-HBs+ anti-HBe+ anti-HBc was 50% (2/4), and positive rate of anti-HBe+ anti-HBc was100% (4/4). Samples, not reactive to TMA discriminatory and anti-HBc negative, were also non-reactive to individual PCR retest. There were significant differences in the positive rates of anti-HBe+ anti-HBc between HBsAg-/NAT+ samples and HBsAg+ /NAT+ (single reagent) samples (P<0.05). 【Conclusion】 Most HBsAg-/NAT+ blood donors were occult hepatitis B virus infection.The anti-HBe+ anti-HBc positive were correlated with HBV infection status. Non-reactivity discriminated by TMA plus anti-HBc negative do not exclude HBV DNA non-reactivity.

BACKGROUND:Cerebral white matter demyelination is outstanding in the images of delayed encephalopathy after acute carbon monoxide (CO) poisoning. Since Nogo-A and Nogo-receptor are expressed in onoligodendrocytes and neurons respectively, we infer that Nogo-A system is involved in brain injury after acute CO poisoning and related to delayed encephalopathy after acute CO poisoning. Endogenous CO is a gaseous messenger, which is the metabolic product of hemeoxygenase. There is no report about the CO effect on Nogo-A system til now. OBIECTIVE: To in vitro culture oligodendrocytes using endogenous CO, inhibit the activity of hemeoxygenase system using zinc protoporphyrin-IX (ZnPPIX) and observe the variation of Nogo-A in oligodendrocytes at mRNA and protein levels. METHODS: Rat oligodendrocytes cultured in vitro were divided into control, CO, ZnPPIX groups. Cels in the CO and ZnPPIX groups were treated with 1% CO directly, In the ZnPPIX group, 10 μmol/L ZnPPIX was added into the culture medium before CO treatment. The expressions of Nogo-A mRNA and protein at 6, 24, 48 hours after culture were compared. Differences in the peak levels of Nogo-A mRNA and protein between CO and ZnPPIX groups were detected using RT-PCR and immunohistochemistry respectively. RESULTS: The expression levels of Nogo-A mRNA and protein were significantly higher in the CO group than the control group and reached the peak at 24 hours of culture. Compared with the CO group, oligodendrocytes cultured with ZnPPIX showed higher expressions of Nogo-A mRNA and protein at 24 hours of culture. These findings suggest that except the influence of hypoxia occurring in CO poisoning, exogenous CO increases the expression of Nogo-A in cultured oligodendrocytes in vitro, and the heme oxygenase system can inhibit the expression of Nogo-A mRNA and protein.

Objective To observe the expression of inducible factor 1(HIF‐1) ,vascular endothelial growth factor(VEGF) and miRNA‐210 and explore the regulatory mechanism after unilateral ureteral occlusion (UUO) .Methods UUO models are made by ligation unilateral ureteral with silk ,and they were divided into sham group and UUO group .All the mice were sacrificed by cer‐vical dislocation after 2 ,5 ,9 and 14 d of UUO .The kidney samples were examined for HIF‐1 mRNA ,VEGF mRNA and miRNA‐210 by quantitative real time reverse transcription polymerase chain reaction (RT‐PCR) ,and HIF‐1 protein by Western blot . Results The expression level of HIF‐1 mRNA was gradually raised after UUO (P<0 .05) ,and compared with the sham group ,the expression level of VEGF mRNA and miRNA‐210 were up regulated after 2 d of UUO and down regulated after 5 ,9 and 14 d of UUO compared with the sham group (P<0 .05);the expression level of HIF‐1 protein increased after UUO compared to the sham group .Conclusion The results of the level of HIF‐1 protein increased and VEGF mRNA and miRNA‐210 were up regulated ,which might be related to the adaptive response of kidney hypoxia and ischemia after UUO .

Objective To compare the bacteriostasis effect of two disinfectants on frequently touched object sur-faces in laminar flow general intensive care unit (GICU),and investigate bacterial contamination on the object sur-faces,so as to provide reference for proper disinfection method and control of infection.Methods Specimens from surfaces of bed rails and bedside tables were taken for bacterial culture before being disinfected.Then object sur-faces around bed units were disinfected with disinfectant containing 500 mg/L of available chloride (routing group) and GammaTM disinfecting wet wipes (control group)respectively,bacteriostasis rate and qualified rate of bacterial count on object surfaces between two kinds of disinfection methods were compared.Results Before routine disinfec-tion,a total of 87 pathogens were isolated from 200 specimens of object surfaces,bacterial contamination rate was 43.50%. Detection rate of gram-positive bacteria was 78.16%% (n =68),the major were Corynebacterium (26.47%,n = 18), Staphylococcus aureus (23.53%,n=16)and Staphylococcus epidermidis (23.53%,n=16);detection rate of gram-nega-tive bacteria was 21.84%(n=19),the major was Acinetobacter baumannii (63.16%,n=12).After a 10-minute disinfec-tion on surfaces,bacteriostasis rate of routine group and control group was (94.89±7.72)% and(96.33 ±12.88)% re-spectively,there was no significant difference between two groups(P >0.05);qualified rates of bacterial count of object surfaces of two groups were both 100%.Conclusion Regular disinfection of object surfaces around bed units,stand-ardization of the manipulation and hand hygiene compliance of medical personnels are simple and effective method of cutting off bacteria dissemination and preventing healthcare-associated infection.

Objective To investigate the screening significance of transcranial Doppler (TCD) for patients with patent foramen ovale ( PFO) induced cerebral embolism and to improve the treatment aiming at the causes.Methods Thirty-six patients of less than 60 years old with unknown-cause cerebral embolism and transient ischemic attack were enrolled.Besides conventional examination,the 24 h Holter,transthoracic echocariiography,carotid artery Doppler ultrasonography (CAU),routine TCD,TCD foaming test,transesophageal echocardiography (TEE),CT angiography (CTA) of head and neck,ultrasonography of vein in bilateral lower limbs were examined in all patients.Results All the patients showed no atrial fibrillation.No structural heart disease and intracardiac occupying lesion were shown in transthoracic echocardiography.No carotid stenosis was shown in carotid artery Doppler ultrasonography.Routine TCD showed no intracranial artery stenosis.No thrombus was found in ultrasonography of deep veins in bilateral lower limbs.CTA of head and neck showed everything was normal.Fourteen patients were observed with right-to-left shunt in heart by TCD foaming test,including 10 cases with microbubble signals(MBS) in 10 s,and 4 cases with MBS after Valsalva maneuver.Among 14 patients,12 patients were proved PFO by TEE.Conclusions PFO is one of the causes of cardiogenic cerebral embolism.TCD is accurate and reliable for the screening of PFO.

Objective To investigate the expression changes of microRNAs and VEGF-NOTCH in renal ischemic injury in mice, and to explore the potential mechanism associated with renal angiogenesis.Method Male Balb/c mice were subjected to a standard renal ischemia to induce acute kidney injury (AKI) after 45 min of bilateral renal artery clamping. Following 4 h, 24 h of reperfusion or sham operation, kindey tissues were collected and subjected to detect the expression changes of microRNAs which relatived with angiogenesis and VEGF, Flk-1, Notch1 mRNA by Quantitative Real-time RT-PCR. Flk-1 protein was detected by Western blotting analysis at 24 h and 72 h following Ischemia/Reperfusion(I/R) injury. The expression of CD31 was examined in tissue sections by immunohistochemistry staining, and the microvessels in ischemic region of each group were counted. Results miRNA-210 and miRNA-92a expression increased significantly, with prominent changes at 4 h and 24 h after reperfusion( P ＜ 0.05 ). VEGF and Flk-1 mRNA expression and Flk-1 protein were increased in renal I/R compared with control group respectively (P＜0.05 ).Immunohistochemistry staining results of CD31 showed a significant increase of microvessels in renal ischemic region. Conclusion This study first reported the changes in miRNAs expression in response to kidney I/R in mouse. our results implied that miRNAs may be involved in targeting VEGF-Notch pathway signaling to regulate angiogenesis after renal I/R injury. It provided novel insights into the angiogenesis mechanism of renal ischemic injury.