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Objective:To establish and evaluate the performance of a stable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of six phytosterols in human plasma.Methods:The methodology was established, evaluated, and applied. A number of 136 healthy subjects were recruited from the Physical Examination Center of Suzhou Municipal Hospital from October 2023 to December 2023, meanwhile plasma phytosterols′ levels were validated in 5 patients with sitosterolemia. Taking deuterium isotope as internal standards, the clear separation was achieved using a Phenomenex C18 (2.1×100 mm, 2.6 μm) column. The mobile phase consisted of water and methanol at a flow rate of 0.4 ml/min with gradient elution, and the column temperature was 25 ℃. The experiments were performed on AB Sciex 6500+triple quadrupole mass spectrometry, and the linearity, limit of quantification, rate of recovery, precision, and stability of the method were evaluated. The testing method was further validated in the sitosterolemia patients.Results:The method had in-run and between-run coefficient of variation ranging 1.8%-12.5% and 2.6%-13.6%, respectively, which accorded with the precision requirements. The rates of recovery were 85.0%-105.1%, which accorded with the accuracy requirements. The samples showed good stability before and after handling stored at 4 ℃ for 72 hours. The plasma levels of sitosterol, campesterol, stigmasterol and cholestanol in sitosterolemia patients were apparently higher compared with those in the healthy group ( P<0.001). Conclusion:LC-MS/MS method for the quantitative measurement of plant sterols in human plasma has been established, which has convenient preprocessing and passed the performance evaluation including linearity, lower limit of quantification, recovery rate, precision and stability, and shown to be suitable for clinical application.

Objective:To quantitatively detect plasma ceramide (Cer) levels using high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyze plasma ceramide levels in homozygous familial hypercholesterolemia (HoFH) patients.Methods:A case-control study was conducted. Plasma samples were collected from 70 healthy individuals who underwent physical examinations at the Health Check-Up Center of Suzhou Municipal Hospital between October 2023 and December 2023, as well as from 34 HoFH patients (including 11 patients with concomitant coronary artery disease) who were treated in the Cardiology Department of the same hospital during the same period. Deuterated isotopes were used as internal standards, and a Phenomenex Kinetex C18 column (2.1×100 mm, 2.6 μm) was employed for separation. An LC-MS/MS method was developed using an AB Sciex 6500+triple quadrupole mass spectrometer for the detection of ceramides, followed by method validation. Additionally, the differences in ceramide levels among healthy individuals, HoFH patients, and HoFH patients with concomitant coronary artery disease were compared. Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic value of various ceramides for HoFH.Results:A method for the quantification of 9 plasma ceramides was established, showing excellent linearity ( r>0.99), precision (1.8%-11.0%), and recovery rates (89.9%-114.7%), meeting the requirements for detection. Compared to the control group, the HoFH group exhibited significantly higher levels of Cer(d18∶1/14∶0), Cer(d18∶1/16∶0), Cer (d18∶1/18∶0), Cer (d18∶1/18∶1), Cer (d18∶1/20∶0), Cer(d18∶1/22∶0), Cer(d18∶1/24∶0), and Cer(d18∶1/24∶1) ( P<0.05). ROC curve analysis revealed that Cer(d18∶1/18∶0) had the highest diagnostic value for HoFH, with an area under the curve (AUC) of 0.83, a cutoff value of 64.6 ng/ml, and sensitivity and specificity of 82.4% and 87.1%, respectively. In HoFH patients complicated with coronary artery disease, the plasma ceramide levels of Cer(d18∶1/18∶0) [(155.7±25.1) ng/ml vs (103.0±11.1) ng/ml, P=0.03] and the ratio of Cer (d18∶1/18∶0) to Cer(d18∶1/24∶0) (0.077±0.012 vs 0.048±0.006, P=0.03) were significantly higher than in HoFH patients without coronary artery disease. Conclusion:A quantitative LC-MS/MS method was developed for detecting ceramides in human plasma, which shows potential diagnostic value for patients with HoFH.

Objective To explore the effect and underlying molecular mechanism of miR-101 on ventricular remod-eling in rats after acute myocardial infarction(AMI).Methods The AMI rat model was established using the left anterior descending coronary artery ligation method.The AMI rats were randomly divided into AMI group,agomir-NC group,miR-101 agomir group and coumermycin A1 group,another 12 rats were selected as sham group with 12 in each.The targeting relationship between miR-101 and JAK2 was analyzed by Target Scan 8.0 database and double luciferase reporter gene assay.The expression of miR-101 in rat myocardium was detected by RT-qPCR.LVESD,LVEDD,LVEF and LVFS were measured by ultrasonography.The level of IL-1β,IL-6 and TNF-α in rats serum was determined by ELISA.The myocardial tissue lesion and fibrosis were detected by HE staining and Mas-son staining.The expression of collagenⅠand TGF-β in rat myocardial tissue was detected by immunohistochemical staining.The expression of E-cadherin,N-cadherin,Vimentin,p-JAK2,JAK2,p-STAT3 and STAT3 proteins was detected by Western blot.Results Compared with AMI group and agomir-NC group,the myocardial tissue lesions and fibrotic area in miR-101 agomir group were significantly decreased(P＜0.05),the level of LVESD,LVEDD,L-1β,IL-6,TNF-α,collagenⅠ,TGF-β,N-cadherin,vimentin,p-JAK2 and p-STAT3 decreased(P＜0.05).The levels of miR-101,LVEF,LVFS and E-cadherin were increased(P＜0.05).Compared with miR-101 agomir group,the myocardial tissue lesions and fibrotic area in coumermycin A1 group significantly increased(P＜0.05),the level of LVESD,LVEDD,L-1β,IL-6,TNF-α,collagenⅠ,TGF-β,N-cadherin,vimentin,p-JAK2 and p-STAT3 was increased(P＜0.05).The level of miR-101,LVEF,LVFS and E-cadherin was decreased(P＜0.05).Conclusions miR-101 inhibits myocardial inflammatory lesions,myocardial fibrosis and epithelial-mesenchymal fransition(EMT)process after AMI with a mechanism targeting at JAK2/STAT3 signaling pathway,thus alleviates ventricular remodeling in rats after AMI.

Objective:To establish and evaluate the performance of a stable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of six phytosterols in human plasma.Methods:The methodology was established, evaluated, and applied. A number of 136 healthy subjects were recruited from the Physical Examination Center of Suzhou Municipal Hospital from October 2023 to December 2023, meanwhile plasma phytosterols′ levels were validated in 5 patients with sitosterolemia. Taking deuterium isotope as internal standards, the clear separation was achieved using a Phenomenex C18 (2.1×100 mm, 2.6 μm) column. The mobile phase consisted of water and methanol at a flow rate of 0.4 ml/min with gradient elution, and the column temperature was 25 ℃. The experiments were performed on AB Sciex 6500+triple quadrupole mass spectrometry, and the linearity, limit of quantification, rate of recovery, precision, and stability of the method were evaluated. The testing method was further validated in the sitosterolemia patients.Results:The method had in-run and between-run coefficient of variation ranging 1.8%-12.5% and 2.6%-13.6%, respectively, which accorded with the precision requirements. The rates of recovery were 85.0%-105.1%, which accorded with the accuracy requirements. The samples showed good stability before and after handling stored at 4 ℃ for 72 hours. The plasma levels of sitosterol, campesterol, stigmasterol and cholestanol in sitosterolemia patients were apparently higher compared with those in the healthy group ( P<0.001). Conclusion:LC-MS/MS method for the quantitative measurement of plant sterols in human plasma has been established, which has convenient preprocessing and passed the performance evaluation including linearity, lower limit of quantification, recovery rate, precision and stability, and shown to be suitable for clinical application.

Objective:To quantitatively detect plasma ceramide (Cer) levels using high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyze plasma ceramide levels in homozygous familial hypercholesterolemia (HoFH) patients.Methods:A case-control study was conducted. Plasma samples were collected from 70 healthy individuals who underwent physical examinations at the Health Check-Up Center of Suzhou Municipal Hospital between October 2023 and December 2023, as well as from 34 HoFH patients (including 11 patients with concomitant coronary artery disease) who were treated in the Cardiology Department of the same hospital during the same period. Deuterated isotopes were used as internal standards, and a Phenomenex Kinetex C18 column (2.1×100 mm, 2.6 μm) was employed for separation. An LC-MS/MS method was developed using an AB Sciex 6500+triple quadrupole mass spectrometer for the detection of ceramides, followed by method validation. Additionally, the differences in ceramide levels among healthy individuals, HoFH patients, and HoFH patients with concomitant coronary artery disease were compared. Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic value of various ceramides for HoFH.Results:A method for the quantification of 9 plasma ceramides was established, showing excellent linearity ( r>0.99), precision (1.8%-11.0%), and recovery rates (89.9%-114.7%), meeting the requirements for detection. Compared to the control group, the HoFH group exhibited significantly higher levels of Cer(d18∶1/14∶0), Cer(d18∶1/16∶0), Cer (d18∶1/18∶0), Cer (d18∶1/18∶1), Cer (d18∶1/20∶0), Cer(d18∶1/22∶0), Cer(d18∶1/24∶0), and Cer(d18∶1/24∶1) ( P<0.05). ROC curve analysis revealed that Cer(d18∶1/18∶0) had the highest diagnostic value for HoFH, with an area under the curve (AUC) of 0.83, a cutoff value of 64.6 ng/ml, and sensitivity and specificity of 82.4% and 87.1%, respectively. In HoFH patients complicated with coronary artery disease, the plasma ceramide levels of Cer(d18∶1/18∶0) [(155.7±25.1) ng/ml vs (103.0±11.1) ng/ml, P=0.03] and the ratio of Cer (d18∶1/18∶0) to Cer(d18∶1/24∶0) (0.077±0.012 vs 0.048±0.006, P=0.03) were significantly higher than in HoFH patients without coronary artery disease. Conclusion:A quantitative LC-MS/MS method was developed for detecting ceramides in human plasma, which shows potential diagnostic value for patients with HoFH.

Objective:To analyze the survival status and death factors of confirmed HIV-infected patients in Hangzhou to provide a basis for the formulation of AIDS prevention and treatment strategies.Methods:A retrospective cohort study was conducted. The data were from the HIV/AIDS Comprehensive Response Information Management System of the Chinese Disease Control and Prevention Information System.Epidemiological characteristics of HIV-infected patients were comparied in Hangzhou City from 2004 to 2023 by using chi-square Test. The survival rate of HIV-infected patients in Hangzhou was calculated by the life table method, the survival curves of different subgroups were described by the Kaplan-Meier method, and the Cox proportional hazard regression model was used to analyze the influencing factors of death risk. The SPSS 26.0 software was used for statistical analysis.Results:Among the 9 457 subjects, the total follow-up time was 58 004.18 person-years, 494 patients died, fatality rate of all-cause cases was 0.85 per 100 person-years.The average survival time was 18.59 (95% CI:18.40-18.78) years. Malignant neoplasms and pneumocystis pneumonia were the first (14.37%,71/494) and second (10.73%, 53/494) causes of death, respectively. Death within 6 months after diagnosis accounted for 42.51% (210/494), and suicide accounted for 4.25% (21/494). Multivariate Cox regression analysis showed that compared with those who received antiviral treatment (ART) within 3 months of diagnosis, those who received ART outside 3 months and those who did not receive ART had a 1.65 (95% CI:1.25-2.19) and 20.68 (95% CI:15.80-27.06) times risk of death, respectively. The HIV-infected patients with high CD4 +T lymphocytes (CD4) counts for the first time had a lower risk of death. The risk of death of patients with baseline CD4 counts of 200-349 cells/μl, 350-499 cells/μl, and ≥500 cells/μl was 0.38 (95% CI:0.29-0.49), 0.26 (95% CI:0.19-0.36), 0.21 (95% CI:0.14-0.31) times higher than that of baseline CD4 counts <200 cells/μl, respectively. Conclusions:The overall survival of the HIV-infected patients was good in Hangzhou from 2004 to 2023. Early detection of HIV infection and timely mobilization to participate in ART was the key to improving the survival rate of patients. At the same time, given the suicide problem of HIV-infected patients, suicide surveillance and depression and anxiety screening of HIV-infected patients should be further strengthened, and targeted psychological intervention policies should be implemented.

Objective:To screen the independent influencing factors of restoration of spontaneous circulation (ROSC) in patients after cardiopulmonary resuscitation (CPR) and establish a predictive model, and explore its clinical value.Methods:A retrospective case control study was conducted. The clinical data of cardiac arrest patients admitted to the emergency department of Tangdu Hospital of Air Force Military Medical University and received CPR from January to July 2023 were analyzed, including general information, blood biochemical indicators, main cause of cardiac arrest, whether it was defibrillation rhythm, duration from admission to CPR, and whether ROSC was achieved. The clinical data between the patients whether achieved ROSC or not were compared. The binary multivariate Logistic regression analysis was used to screen the independent influencing factors of ROSC in in-hospital CPR patients. According to the above influencing factors, the ROSC prediction model was established, and the receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of the model for ROSC.Results:A total of 235 patients who received CPR in the emergency department were enrolled, including 153 cases (65.11%) of in-hospital CPR and 82 cases (34.89%) of out-of-hospital CPR. The ROSC ratio was 30.21% (71/235). Among all patients, the majority were aged 61-80 years [40.43% (95/235)], and cardiogenic disease was the main cause of cardiac arrest [32.77% (77/235)]. Among 153 patients with in-hospital CPR, 89 were non-ROSC and 64 were ROSC with ROSC rate of 41.83%. Compared with the non-ROSC group, the patients in the ROSC group had lower blood lactic acid (Lac), N-terminal pro-brain natriuretic peptide (NT-proBNP), Lac/albumin (Alb) ratio (LAR), and ratio of non-defibrillation rhythm [Lac (mmol/L): 5.50 (2.33, 9.65) vs. 7.10 (3.50, 13.35), NT-proBNP (μg/L): 0.87 (0.20, 8.68) vs. 3.00 (0.58, 20.17), LAR: 0.14 (0.07, 0.29) vs. 0.19 (0.10, 0.43), non-defibrillation rhythm ratio: 68.75% (44/64) vs. 93.26% (83/89)], higher actual base excess (ABE) and Alb [ABE (mmol/L): -3.95 (-12.75, 0.23) vs. -7.50 (-13.50, -3.35), Alb (g/L): 38.13±7.03 vs. 34.09±7.81], and shorter duration from admission to CPR [hours: 3.25 (1.00, 14.00) vs. 8.00 (2.00, 27.50)], the differences were statistically significant (all P < 0.05). Binary multivariate Logistic regression analysis showed that LAR [odds ratio ( OR) = 0.037, 95% confidence interval (95% CI) was 0.005-0.287], non-defibrillation rhythm ( OR = 0.145, 95% CI was 0.049-0.426), and duration from admission to CPR ( OR = 0.984, 95% CI was 0.972-0.997) were independent influencing factors for ROSC in hospitalized CPR patients (all P < 0.05). Based on the above influencing factors, a ROSC prediction model was constructed through regression analysis results. The ROC curve analysis showed that the area under the ROC curve (AUC) for predicting ROSC in in-hospital CPR patients was 0.757 (95% CI was 0.680-0.834), Yoden index was 0.429, sensitivity was 76.6%, and specificity was 66.3%. Conclusions:LAR, non-defibrillation rhythm and duration from admission to CPR were independent influencing factors for ROSC in patients with in-hospital CPR. The ROSC prediction model established based on the above influencing factors has a good predictive value for ROSC of CPR patients in hospital, and can guide clinicians to evaluate the prognosis of patients through relevant indicators as early as possible.

Objective:To analyze the early warning value of laboratory examination on admission of patients with hemorrhagic fever with renal syndrome to critically ill patients.Meetods:In this study, a retrospective case-control study was used to analyze the clinical data and laboratory examination results of patients with hemorrhagic fever with renal syndrome admitted to the emergency department of Tangdu Hospital of Air Force Medical University from January 2021 to January 2022. According to the patient's laboratory indexes and clinical symptoms, the patients were divided into mild, moderate, severe and critical groups. The general data of the two groups were compared, and the independent risk factors of critically ill patients were screened by multi-factor logistic regression analysis, the predictive model of severe HFRS patients was constructed, and the ROC curve was drawn. .Results:Of the 164 patients with HFRS, 50 were in the severe group and 114 in the mild group. The serum levels of WBC, AST, ALT, Cr, BUN, DD and PCT in the severe group were higher than those in the mild group, while the levels of PLT, ALB and PTA in the severe group were lower than those in the mild group. Multiple logistic regression analysis showed that WBC, PLT and PCT were independent influencing factors for the progression of critically ill patients. The predictive model of severe HFRS was established as follows: logit (P) = -0.321 + 0.040 WBC (×10 9/L) -0.045 PLT (×10 9/L) + 0.086 PCT(ng/mL). The early warning ef?cacy of WBC, PLT, And PCT for severe HFRS was further analyzed. The area under the ROC curve (area under curve, AUC) was 0.779, 0.842, 0.862, and the optimal threshold was 10.435×109/L, 41.5 ×109/Land 2.97 ng/mL, respectively. The AUC of joint detection is 0.900, the sensitivity is 88.0%, and the speci?city is 82.5%, which is better than that of a single laboratory. . Conclusions:HFRS laboratory indexes have certain clinical signi?cance for the identi?cation of critically ill patients, in which serum WBC, PLT and PCT indexes are the risk factors of severe HFRS, which provides a theoretical basis for clinical diagnosis, treatment and prognosis of severe HFRS patients.