Objective:To investigate the value of early tight junction protein Claudin-5 levels in predicting the severity of acute pancreatitis (AP).Methods:A prospective observational study was conducted, including patients diagnosed with AP and admitted to the Northern Jiangsu People's Hospital from December 1, 2021 to November 30, 2022. Eligible healthy volunteers were randomly selected to serve as healthy controls during the same period. Patients were classified into mild acute pancreatitis (MAP) group, moderate-severe acute pancreatitis (MSAP) group, and severe acute pancreatitis (SAP) group based on the 2012 Atlanta classification criteria. Patients with SAP were then divided into three subgroups of 1, 3, and 7 days based on the duration of hospitalization. Baseline data, such as gender, age, underlying disease, and probable etiology, was collected from all enrolled individuals. The enzyme-linked immunosorbent assay (ELISA) was employed to detect serum Claudin-5 levels in each cohort of enrollees. Data on additional serologic indicators, including hematocrit (HCT), albumin (Alb), serum Ca 2+, C-reactive protein (CRP), and procalcitonin (PCT) levels, were obtained via the in-hospital test query system in each group of patients with AP. The modified Marshall score (mMarshall), modified CT severity index (mCTSI) score, bedside severity index of severity in acute pancreatitis (BISAP) score, and acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) were recorded for each group of patients with AP. Differences in the above indicators between groups were analyzed and compared. Spearman's correlation method was employed to examine the relationship between Claudin-5 levels and each influential factor. The receiver operator characteristic curve (ROC curve) was plotted to analyze the predictive value of each influencing factor on SAP. Ridge regression was used to screen for independent risk factors for SAP. Results:A total of 109 patients with AP were enrolled, comprising 66 in the MAP group, 15 in the MSAP group, and 28 in the SAP group. Additionally, 27 healthy volunteers were enrolled as the healthy control group. No statistically significant differences were observed in gender and age among the enrolled groups, and no statistically significant differences were identified among the three groups of patients with AP in terms of underlying disease and etiologic composition. As the disease progressed, serum Claudin-5 levels exhibited a notable increase across all AP patient groups, and they were all significantly higher than those in the healthy control group [ng/L: 888.58 (574.52, 1 141.59), 3 749.02 (2 784.93, 5 789.92), 4 667.81 (3 935.21, 7 315.66) vs. 291.13 (250.19, 314.75), all P < 0.05]. Subgroup analyses showed that as the disease duration prolonged, patients in the SAP group exhibited a notable decline in Claudin-5 levels at 3 days post-admission, compared with those at 1 day post-admission [ng/L: 2 052.59 (1 089.43, 4 006.47) vs. 4 667.81 (3 935.21, 7 315.66), P < 0.05]. Spearman correlation analysis showed that serum Claudin-5 levels in patients with AP were significantly positively correlated with CRP, PCT, HCT, and mMarshall, mCTSI, and BISAP scores ( r values were 0.570, 0.525, 0.323, 0.774, 0.670, 0.652, all P < 0.001), and significantly negatively correlated with Alb ( r = -0.394, P < 0.001). A significant trend was observed in patients with AP, with an increase of HCT levels and a decrease of Alb levels as the disease progressed (both P < 0.05). An improvement of aforementioned phenomena was observed in patients with SAP following treatment, indirectly indicating that serum Claudin-5 level was a positive indicator of vascular leakage. ROC curve analysis showed that serum Claudin-5 levels in patients with AP exhibited the highest accuracy for early prediction of SAP, with the area under the ROC curve (AUC) of 0.948. When serum Claudin-5 levels ≥2 997 ng/L, the sensitivity for early screening for SAP was 100% and the specificity was 88.89%. Multifactorial ridge regression analysis showed that serum Claudin-5 level, PCT and APACHEⅡscore could be used as independent risk factors for early prediction of SAP (all P < 0.05). Conclusion:Serum Claudin-5 levels facilitate early prediction of SAP and are strongly associated with inflammatory response and vascular leakage.

Objective To explore the relationship between the early stage HbA1c level and two-year prognosis in the patients with first time onset of acute ischemic stroke(AIS).Methods A total of 513 inpa-tients with first time AIS in this hospital during 2018-2019 were selected as the study subjects.The clinical data,biochemical indicators and discharge situation were collected.The multivariate logistic regression was used to analyze the influencing factors of adverse reactions outcome within 2 years.The Kaplan-Meier survival curve was used to analyze the all-cause mortality and stroke recurrence situation within 2 years.Results The sex,age,TOAST type,,diseases history such as diabetes,arrhythmia and coronary heart disease,medication history such as antidiabetic drugs,lipid-lowering drugs and anticoagulants,BMI,Urea,Crea,ALT,SBP,fasting blood glucose(FBG),total cholesterol(TC),triglyceride(TG),HDL-C,NIHSS score at admission,discharge mode,NIHSS score at discharge,hospitalization duration and hospitalization costs had statistical difference a-mong the patients with different HbA1c levels(P＜0.05).The multivariate logistic regression analysis showed that HbA1c＞7.4％was the independent risk factor for adverse outcome within 2 years(OR=4.470,95％CI:1.105-18.087,P=0.036).The Kaplan-Meier survival curve analysis showed that the survival time within 2 years had statistical difference among the patients with different HbA1c levels(P=0.009).The higher the HbA1c level,the shorter the survival time.Conclusion The high HbA1c level has the influence on the stroke recurrence and all-cause mortality within 2 years in the patients with first onset occurrence of AIS.

Objective: To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected． Methods : CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) ．CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro．The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression，and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ，and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry. Results : CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells，flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P＜0. 01) ． Conclusion In this experiment，CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ，and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.

ObjectiveTo understand the current situation of anxiety level and traditional Chinese medicine （TCM） nursing needs among residents during the COVID-19 epidemic， and to discuss the relationship between them，so as to provide references for maintaining their mental health. MethodsA total of 560 residents were recruited using convenience sampling method， and the selected individuals completed three questionnaires based on online platform， including self-designed general information questionnaire， Generalized Anxiety Disorder Scale （GAD-7）， and TCM nursing needs questionnaire. Then the correlation between anxiety level and TCM nursing needs was analyzed by Spearman correlation test. ResultsA total of 552 valid questionnaires were collected. The results of GAD-7 showed that the total score was （11.04±3.72）， and average score of each item was （1.58±0.53）. There were 328 cases （59.4%） without anxiety symptoms， 180 cases （32.6%） with mild anxiety， 33 cases （6.0%） with moderate anxiety， and 11 cases （2.0%） with severe anxiety. The analysis of TCM nursing needs questionnaire showed that the total score was （21.59±7.92）， and dominated by the needs of medical knowledge （2.90±1.15） and TCM dietary care （2.82±1.13）. The item score and total score of TCM nursing needs questionnaire of residents during the COVID-19 epidemic were positively correlated with GAD-7 total score （r=0.240~0.331， P<0.01）. ConclusionResidents suffer from different levels of anxiety during the COVID-19 epidemic， and TCM nursing needs is on the rise， indicating that the anxiety level of residents is correlated with the needs of TCM nursing.

Immune cell infiltration is of great significance for the diagnosis and prognosis of cancer. In this study, we collected gene expression data of non-small cell lung cancer (NSCLC) and normal tissues included in TCGA database, obtained the proportion of 22 immune cells by CIBERSORT tool, and then evaluated the infiltration of immune cells. Subsequently, based on the proportion of 22 immune cells, a classification model of NSCLC tissues and normal tissues was constructed using machine learning methods. The AUC, sensitivity and specificity of classification model built by random forest algorithm reached 0.987, 0.98 and 0.84, respectively. In addition, the AUC, sensitivity and specificity of classification model of lung adenocarcinoma and lung squamous carcinoma tissues constructed by random forest method 0.827, 0.75 and 0.77, respectively. Finally, we constructed a prognosis model of NSCLC by combining the immunocyte score composed of 8 strongly correlated features of 22 immunocyte features screened by LASSO regression with clinical features. After evaluation and verification, C-index reached 0.71 and the calibration curves of three years and five years were well fitted in the prognosis model, which could accurately predict the degree of prognostic risk. This study aims to provide a new strategy for the diagnosis and prognosis of NSCLC based on the classification model and prognosis model established by immune cell infiltration.
Algorithms ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; physiopathology ; Humans ; Lung Neoplasms ; diagnosis ; physiopathology ; Machine Learning ; Prognosis

Objective:To investigate the clinical characteristic, gene mutations and genotype-phenotype correlation of 21-hydroxylase deficiency (21-OHD) in Hunan.Methods:A total of 48 patients with 21-OHD who were admitted to the Department of Pediatrics, the Second Xiangya Hospital, Central South University from March 2016 to March 2017 were collected.According to the clinical manifestations and biochemical characteristics of the patients, they were divided into salt wasting (SW) and simple virilizing (SV). Sanger sequencing combined with multiplex ligation-dependent probe amplification(MLPA) were used to detect the mutations of CYP21A2 gene.The patients were divided into 3 groups according to their mutations severity: severe mutation group, moderate mutation group and unknown mutation group.Then, the correlation between genotype and phenotype was analyzed. Results:(1) Forty-eight 21-OHD patients included 28 SW cases and 20 SV cases, and the first visiting age of SW was younger than that of SV, and the difference was statistically significant ( U=44.5, P<0.05). The SW cases had high incidence rate of adrenal crisis and the SV patients were liable to advanced bone age and precocious puberty.(2) Forty-four patients were detected abnormal gene mutation and the positive rate of genetic diagnosis was 91.7%.Fourteen mutation types including I2G, Del, I173N, R357W, R484fs(c.1451_1452delGGinsC, c.1450dupC), R483fs, G111Vfs*21, Q319X, c.292+ 1G>A, c.377C>G, E6Cluster, p.H393Q and m. 1647C>T, were found in 88 alleles.The most frequent mutations were I2G(36.4%), I173N(20.4%), and Del(22.7%). p.H393Q and m. 1647C>T were 2 novel mutations.I2G (47.3%) and Del (27.3%) were the most frequent mutations in SW cases, and I173N (48.5%) was the most frequent mutation in SV cases.(3) Severe mutation was in 29 patients, including 26 SW, and moderate mutation was in 13 patients, including 12 SW.The percentage of SW in severe mutation group was 89.7% and SV in moderate mutation was 92.3%. Conclusions:I2G, I173N and Del were the frequent mutations of 21-OHD in Hunan, and the total percen-tage was 79.5%.Genotype of 21-OHD has strong correlation with clinical phenotype, which can effectively predict SW by severe mutation and predict SV by moderate mutation.

Objective:To study the new mechanism of Xuebijing injection improving the function of pulmonary vascular barrier from the perspective of claudin-5 protein.Methods:Acute lung injury (ALI) model was induced by hydrogen sulfide (H 2S) exposure. ① In vivo study: Sprague-Dawley (SD) rats were divided into control group, H 2S exposure group (exposure to 300×10 -6 H 2S for 3 hours), Xuebijing control group (Xuebijing injection 4 mL/kg, twice a day, for 3 days), and Xuebijing intervention group (H 2S exposure after pretreatment of Xuebijing injection) according to random number method, with 6 rats in each group. At different time points (0, 6, 12 and 24 hours) after the model was made successfully, the total protein content in plasma and bronchoalveolar lavage fluid (BALF) of rats were detected respectively, and the pulmonary permeability index (PPI) was calculated (PPI = protein content in BALF/protein content in plasma), lung dry/wet weight ratio (W/D) was detected, and claudin-5 mRNA expression in lung tissue was measured by real time-polymerase chain reaction. ② In vitro test: human pulmonary microvascular endothelial cells (HPMECs) were divided into blank control group, NaHS treatment group (co-incubated with 500 μmol/L NaHS for 12 hours), Xuebijing control group (2 g/L Xuebijing injection for 24 hours), and Xuebijing intervention group (2 g/L Xuebijing injection pre-treated for 24 hours, then co-incubated with 500 μmol/L NaHS for 12 hours). The HPMECs claudin-5 protein expression and monolayer permeability changes were measured at different co-incubation time (1, 3, 6, 12 and 24 hours) by Western Blot and fluoresceinsodium. Results:① In vivo study: compared with the control group, the lung W/D ratio increased significantly at 6 hours and peaked at 12 hours after H 2S exposure in rats (4.67±0.11 vs. 4.26±0.06, P < 0.01). The expression of claudin-5 mRNA in lung tissue was significantly decreased, which was 89% of control group 6 hours after exposure ( P < 0.01). The total protein content in BALF and PPI at 12 hours after exposure were significantly higher than those in the control group [total protein content (mg/L): 262.31±14.24 vs. 33.30±3.09, PPI: (11.72±0.57)×10 -3 vs. (1.21±0.08)×10 -3, both P < 0.01], while the results in Xuebijing intervention group were significantly decreased [total protein content (mg/L): 153.25±7.32 vs. 262.31±14.24, PPI: (5.79±0.23)×10 -3 vs. (11.72±0.57)×10 -3, both P < 0.01]. ② In vitro test: compared with the blank control group, after incubating HPMECs with NaHS, the permeability of monolayer endothelial cells gradually increased, reaching the highest level in 12 hours, about twice of that in the blank control group, while claudin-5 protein expression decreased to the lowest level at 12 hours (claudin-5/β-actin: 0.42±0.03 vs. 1.03±0.05, P < 0.01). After intervention with Xuebijing, the permeability of endothelial cells was significantly improved (fluorescence intensity of fluorescein sodium: 1.46±0.10 vs. 1.89±0.11, P < 0.01), and the decrease of claudin-5 protein was reduced (claudin-5/β-actin: 0.68±0.04 vs. 0.38±0.03, P < 0.01). Conclusion:Xuebijing injection may improve pulmonary vascular barrier function in ALI by upregulating claudin-5 expression.

Objective To investigate the delay of door to signature time in primary percutaneous coronary intervention (PCI) and its influence in patients with ST segment elevation myocardial infarction (STEMI),therefore to provide a scientific basis for further effective shortening the time of primary PCI in patients with STEMI.Methods A total of 226 patients who diagnosed with STEMI and underwent primary PCI at Henan Provincial People's Hospital from June 2016 to December 2017 were enrolled in the study.Observation indicators include:(1) baseline data of patients;(2) time segments in primary PCI:total ischemic time (TIT),door to balloon time (DTBT),door-to-signature time (DTST),signature to balloon time (STBT);(3) the demographic characteristics of the family members who signed informed consent;and (4) the psychological factors and coping strategies of family members before signing informed consent.All data was analyzed using SPSS software (version 22.0).Multiple linear regression analysis was used to analyze the influencing factors of delay of DTST.A P＜0.05 was considered statistically significant.Results In this study,226 patients with STEMI who were first diagnosed in our hospital had a mean age of 55.23±10.80 years,and 181 (80.1％) were male.The median of TIT,DTBT,DTST,STBT were 312 min,166 min,82 min,and 80 min.The ratio of DTST in DTBT and TIT was 50％ and 28.5％,respectively.The multiple linear regression analysis showed that the number of direct family members (P＜0.001),the degree of educational in middle school and below (P=0.010),high school/technical secondary school (P=0.029),families worrying about the high cost of medical care (P=0.020),families consulted each other repeatedly (P=0.022),and consulted the other medical staff(P=0.022) are risk factors of DTST delay,and city residence (P=0.048) is the protection factor of DTST delay.Conclusions The long time of DTS is a reality of the practice of primary PCI in China.The factors that lead to longer DTST include demographic characteristics,psychological factors and coping strategies of family members.The STBT of primary PCI in China should be taken into the value while emphasizing the DTBT.

Objective To compare the efficacy of lung ultrasound and chest X-ray for diagnosis of neonatal transient respiration syndrome (TTN) and respiratory distress syndrome (RDS).Methods Totally 120 infants with respiratory distress underwent lung ultrasound and chest X-ray within 6 h after birth.Taking clinical diagnosis as the standard,the sensitivity,specificity and accuracy of lung ultrasound and X-ray for diagnosis of TTN and RDS were calculated.Results According to clinical diagnosis,there were 75 infants of TTN and 34 of RDS.The sensitivity,specificity and accuracy of lung ultrasound in diagnosis of TTN was 96.00％ (72/75),88.89％ (40/45) and 93.33％ (112/120),respectively,higher than those of chest X-ray (85.33％ [64/75],84.44％ [38/45] and 85.00％ [102/120]).Lung ultrasound showed 85.29％ (29/34) of sensitivity,95.35％ (82/86) of specificity and 92.50％ (111/120) of accuracy for diagnosis of RDS,whereas chest X-ray showed 88.23％ (30/34) of sensitivity,89.53％ (77/86) of specificity and 89.17％ (107/120) of accuracy.Conclusion Lung ultrasound is accurate for the diagnosis of TTN and RDS.

Objective To study the time extended for getting emergency intervention in different modes of transportation and factors influencing the modes of transportation of patients with ST elevation myocardial infarction (STEMI).Methods A total of 564 consecutive patients with STEMI admitted from September 2013 to June 2016 were enrolled in the study.The clinical data about time consumed for getting emergency intervention and modes of transportation were collected.Results According to the mode of transportation,patients were divided into three groups:emergency care system (EMS) transportation group (n =96),self-transportation group (n =206) and referral group in which the patients were sent in from other hospitals (n =262).EMS transportation group had significantly shorter total ischemic time before emergency treatment than self-transportation group (229 rin vs.418 min,P ＜ 0.05) and referral group (229 min vs.512 rin,P ＜ 0.05),and significantly shorter length of pre-hospital time than self-arrival group (55 min vs.110 min;P＜0.05) and referral group (55 min vs.372 min;P＜0,05).The referral group had longer pre-hospital time and the self-transportation group had longer door-to-balloon time,but there was no difference in total ischemic time between the self-arrival and referral group (Z =-1.882,P =0.068).Multivariate logistic regression was used to analyze influence factors in mode of transportation:(1) patients characterized with high school or university education,profession of civil service,and their transportation distance more than 30 km were greater in number than referral group (P ＜ 0.05);(2) patients identified with senior middle school education,staff member of public sectors or company,their transportation distance less than 30 km,and with killip grade above Ⅱ were more likely to have EMS transport (P ＜ 0.05);(3) patients defined as businessmen without taking out new rural cooperative medical insurance,taking up transportation distance less than 80 km,and subjecting to killip grade Ⅰ had a higher proportion of individuals of this kind taking self-transportation (P ＜ 0.05).Conclusion Mode of transportation is an important factor that affects the time extended to get emergency intervention.Education level,occupation,medical insurance type,transportation distance,killip grade are associated with modes of transport.