Objective To investigate the gene transduction efficiency of lentiviral vector in leukemia cells to provide key basis for leukemia gene therapy. Methods A third-generation self-inactivating(SIN) lentiviral vector system based on human immunodeficiency virus type 1(HIV-1) was used to improve transduction efficiency.The transduction efficiency of the HIV-1-based vector was compared directly with the moloney murine leukemia virus(MLV) SIN vector in human leukemia cell line K562.The expression of green fluorescent protein(GFP) in cells was observed by fluorescence microscopy and flow cytometry(FCM) to detect the percentage of gene trasduction cells.Results The GFP expression in K562 cells was observed qualitatively by fluorescence microscopy.At the same gene transduction conditions,the GFP marker gene expression intensity and GFP positive cells in leukemia cells transduced with HIV vectors were significantly higher than those transduced with MLV vectors.Initial transduction efficiencies were almost 100% for the HIV and less than 40% for the MLV vectors. The transduction efficiency had significant difference between HIV vector group and MLV group(P

Objective:To explore the early evaluation value of calprotectin (S100A8/A9) and ischemia modified albumin (IMA) for adult acute appendicitis (AA) and adult non-simple acute appendicitis.Methods:The data of 62 patients with histologically confirmed appendicitis and 57 healthy controls in the physical examination center of our hospital during the same period from May 2018 to October 2019 were collected. According to postoperative pathological data, patients with appendicitis were divided into the simple appendicitis group and the non-simple appendicitis group . The measurement data conforming to normal distribution were expressed as mean ± standard deviation (Mean±SD), and Student's t test was used for comparison between groups. S100A8/A9, IMA, C-reactive protein (CRP), procalcitonin (PCT), and blood routine parameters were compared after grouping.The area under the ROC curve (AUC) was used to evaluate the early efficacy of S100A8/A9 and IMA for acute appendicitis and non-simple acute appendicitis. Results:There were no significant differences in sex, age, platelet count (PLT), and red blood cell count (RBC) between the appendicitis group and healthy control (all P>0.05), while white blood cell count (WBC), CRP, PCT, neutrophils to lymphocytes ratio (NLR), S100A8/A9, and IMA levels in the appendicitis group were higher than those in healthy control (all P<0.05). The AUC of S100A8/A9 (≥366.9 μg/L), IMA (≥0.29), and S100A8/A9 combined with IMA in predicting acute appendicitis were 0.735, 0.891, and 0.913, respectively. There was no significant difference in WBC between the non-simple appendicitis group (21 cases) and the simple appendicitis group (41 cases) ( P>0.05).The levels of CRP, PCT, NLR, S100A8/A9 and IMA in the non-simple appendicitis group were significantly higher than those in the simple appendicitis group ( P<0.05). The AUC of S100A8/A9 (≥532.9 μg /L), IMA (≥0.41) and S100A8/A9 combined with IMA in predicting non-simple acute appendicitis were 0.866, 0.873 and 0.936, respectively. Conclusions:S100A8/A9 and IMA could be used as biomarkers for the diagnosis of AA, which have certain clinical value for the assessment of acute appendicitis and non-simple acute appendicitis.

Objective To investigate the safety and efficacy of a step-up drainage combined with thermostatic lavage in the management of severe acute pancreatitis complicated with infected pancreatic necrosis.Methods From January 2014 to July 2017,100 cases of severe acute pancreatitis complicated with infected pancreatic necrosis admitted to our hospital were prospectively collected.All patients were randomly (random number) assigned into the observation group and control group with 50 cases in each group.The patients in both groups were given a step-up drainage.Moreover,patients in the observation group received lavage under constant temperature (30-35 ℃) while the patients in the control group were at room temperature.The clinical prognosis of the two groups was observed.Results When compared with the control group,patients in the observation group got a lower rate of new onset renal injury (16.00％ vs 34.00％,P=0.038);a lower rate of new onset gastrointestinal dysfunction (12.00％ vs 28.00％,P=0.046);a reduced mechanical ventilation time[(14.82±7.78)vs (18.64±8.12)d,P=0.018];a decreased continuous blood filtration [(12.98±6.20) vs (18.56±10.92)d,P=0.002];and a shorter hospital duration [(43.28±10.72)vs (50.82±12.74) d,P=0.002].No significant difference was found in the APACHE Ⅱ score between the two groups at admission and at the time of puncture [(9.88±2.40) vs (9.46±2.35),P=0.378 and (10.30±3.07vs (10.54±2.96),P=0.692].At 7 and 14 days after the puncture,the APACHE Ⅱ score in the observation group was significantly lower than that in the control group [(9.32±2.49) vs (10.56±2.80),P=0.021 and (6.20±2.54 vs (8.42±2.48),P ＜ 0.01].Conclusions A step-up drainage combined with constant temperature lavage can reduce the incidence of new renal failure and gastrointestinal dysfunction in infected pancreatic necrosis patients and improve the clinical prognosis of the patients.

Objective:To discuss the value of dynamic detection of serum intestinal fatty acid binding protein (I-FABP), heparin binding protein (HBP) and interleukin-1β(IL-1β) in early predicting and evaluating the severity of abdominal compartment syndrome (ACS) in severe acute pancreatitis (SAP) postoperative patients.Methods:The clinical data of 65 SAP patients treated in the Second Hospital of Anhui Medical University from July 2019 to Jan 2021 were retrospective analyzed. According to whether ACS has occurred, the patients were divided into non ACS group (48 cases) and ACS group (17 cases). The serum I-FABP, HBP and IL-1β of the two groups were dynamically monitored. Correlation analysis and receiver operating characteristic (ROC) curve were used to evaluate the efficacy and early prediction value of each observation index in evaluating the severity of SAP patients complicated with ACS.Results:There were no significant differences in age, sex, body mass index (BMI) and pathogenesis between the two groups (all P>0.05). The serum levels of C-reactive protein (CRP), white blood cell (WBC), Acute Physiology and Chronic Health Enquiry (APACHE-Ⅱ) score and intra-abdominal pressure (IAP) in ACS group were significantly higher than those in non ACS group (all P<0.05). The serum levels of I-FABP [(97.41±15.02)ng/ml vs (37.28±18.34)ng/ml, (103.32±18.40)ng/ml vs (56.96±19.12)ng/ml, (85.69±22.94)ng/ml vs (36.88±10.49)ng/ml], HBP [(92.19±14.59)ng/ml vs (24.56±10.96)ng/ml, (106.11±15.03)ng/ml vs (37.17±13.83)ng/ml, (128.11±16.43)ng/ml vs (68.94±15.91)ng/ml] and IL-1β[(15.78±1.44)pg/ml vs (11.26±1.34)pg/ml, (19.34±1.87)pg/ml vs (13.51±2.84)pg/ml, (20.95±1.96)pg/ml vs (16.03±1.04)pg/ml] on 1st, 4th, 7th day in ACS group were continuously and evidently higher than those in non ACS group ( P<0.01). Correlation analysis revealed that I-FABP, HBP and IL-1β were positively correlated with IAP ( r=0.745, 0.793, 0.770) and APACHE Ⅱ score ( r=0.510, 0.489, 0.445) (all P<0.01). ROC curve analysis showed that the AUC of early prediction by I-FABP, HBP and IL-1β on the occurrence of ACS were 0.846, 0.873 and 0.902 respectively, which were higher than the CRP (0.681), WBC (0.765) and APACHE Ⅱ score (0.795), the sensitivity and specificity can be significantly improved to 0.997 and 0.994 by parallel and series tests respectively combined with the three indicators. Conclusions:Dynamic detection of serum I-FABP, HBP and IL-1β has a certain clinical value in evaluating the severity of ACS in SAP patients. At the same time, early detection with serum I-FABP, HBP and IL-1β has high predictive power for ACS in SAP patients and the combined application of three has higher predictive value.

Objective:To analyze the correlation between the severity of acute pancreatitis (AP) and the levels of zonulin, zonula occludens protein-1 (ZO-1), tumor necrosis factor -α (TNF -α) in the peripheral blood of patients with acute pancreatitis (AP), and the value of predicting moderate and severe AP.Methods:The clinical data of 115 AP patients admitted to the Second Affiliated Hospital of Anhui Medical University from June 2020 to January 2022 were retrospectively analyzed. They were divided into mild group (69 cases) and moderate severe group (46 cases). The blood levels of zonulin, ZO-1, and TNF-α were measured for all patients on the 1st, 3rd, and 7th day after admission, and the results of the two group tests were compared. The correlation between zonulin, ZO-1, TNF -α and Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) scores on the 1st day was and the value of various indicators for predicting moderate to severe AP were analyzed.Results:The C-reactive protein (CRP) levels of AP patients in the moderate to severe group were higher than those in the mild group, and the difference was statistically significant (all P<0.05). The levels of zonulin, ZO-1, and TNF -α in AP patients in the moderate to severe group showed an upward trend on the 1st, 3rd, and 7th days after admission. The levels of zonulin, ZO-1, and TNF -α in AP patients in the moderate to severe group were higher than those in the mild group at the same time point, and the differences were statistically significant (all P<0.05). The APACHE Ⅱ score of AP patients on the first day of admission was positively correlated with the levels of zonulin, ZO-1, and TNF -α ( r=0.736, 0.552, 0.621, all P<0.05). Zonulin had the highest area under the curve (AUC) for predicting moderate to severe AP, at 0.892, with an optimal threshold of 2.075 pg/ml. Zonulin had the highest sensitivity, at 0.804, and ZO-1 had the highest specificity, at 0.926. Using zonulin ≥2.075 pg/ml, ZO-1≥399.4 ng/ml, and TNF -α≥40.88 pg/ml as thresholds; the sensitivity and specificity obtained from parallel experiments were 0.976 and 0.710, respectively; The sensitivity and specificity obtained from the series of experiments were 0.326 and 0.999, respectively. Conclusions:There is a correlation between the serum levels of zonulin, ZO-1, and TNF -α in AP patients and the severity of AP. Zonulin, ZO-1, and TNF -α have certain clinical value in predicting moderate to severe AP.

Objective:To investigate the early evaluation potential of serum levels of apolipoprotein B/apolipoprotein A1 (Apo B/A1), microtubule-associated protein 1-light chain 3 (MAP1-LC3) and intercellular adhesion molecule-1 (ICAM-1) in acute pancreatitis (AP) patients.Methods:A total of 413 AP patients who were treated at the Second Affiliated Hospital of Anhui Medical University between January 2019 and August 2020 were enrolled. Serum samples were collected from AP patients within 24 h of admission. Patients were divided into the non-severe acute pancreatitis (Non-SAP, n=315) and severe acute pancreatitis (SAP, n=98) groups according to the severity of the disease. Sixty healthy controls were recruited. The differences of serum Apo B/A1, MAP1-LC3 and ICAM-1 among the three groups were compared by one-way analysis of variance, and the correlation between Apo B/A1, MAP1-LC3 and ICAM-1 and the severity of AP was analyzed by Pearson correlation analysis. Sensitivity and specificity in assessing AP severity were predicted by receiver operating characteristic curve (ROC). Results:The early levels of Apo B/A1, MAP1-LC3 and ICAM-1 were all significantly higher for AP patients than for healthy controls ( P<0.05), and the levels of Apo B/A1, MAP1-LC3 and ICAM-1 in SAP patients were significantly higher than those in non-SAP patients[Apo B/A1: 2.21±1.40 vs. (0.96±0.34); MAP1-LC3: 0.92±0.29 vs. (0.48±0.24) ng/mL and ICAM-1: (235.57±54.50 ) vs. (120.28±61.69)ng/mL; P<0.05]. Pearson correlation analysis showed that levels of Apo B/A1, MAP1-LC3 and ICAM-1 were positively correlated with the first Ranson score after admission ( P<0.05), and ICAM-1 showed the highest degree of correlation with AP severity ( r=0.519). Areas under the receiver operating characteristic curve (AUROC) were 0.769 for Apo B/A1, 0.811 for MAP1-LC3, 0.828 for ICAM-1, and 0.938 for combined detection. Conclusions:Serum levels of Apo B/A1, MAP1-LC3 and ICAM-1 within 24 h after admission are significantly correlated with the severity of AP, which has clinical significance for early prediction of the severity of AP.

Objective:To explore the role and preliminary mechanism of heparin-binding protein (HBP) in the development of acute pancreatitis (AP) through clinical analysis and animal models.Methods:(1) Clinical research: Blood samples were collected from AP patients admitted to the Second Affiliated Hospital of Anhui Medical University from January 1 to December 31, 2021 within 30 min of admission, including 20 patients with severe acute pancreatitis (SAP) and 20 patients with non-severe acute pancreatitis (NSAP). Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of HBP, syndecan-1 and hyaluronic acid (HA). Modified CT severity index (MCTSI), another 20 healthy volunteers were selected as controls (HC). Spearman correlation analysis was used to analyze the correlation between HBP and syndecan-1, HA and MCTSI. Receiver operating characteristic (ROC) curve was used to evaluate HBP to predict AP severity. (2)Animal experiment: The rat model of acute pancreatitis was prepared by intraperitoneal injection of L-arginine. In the normal control group (NC, n=8), the low molecular weight heparin (LMWH) intervention group ( n=8), and the acute pancreatitis group ( AP, n=8), the rats were euthanized 12 h later, and peripheral venous blood was collected to detect the levels of HBP, syndecan-1 and HA. Lung tissue and pancreas tissue were collected to observe the pathological damage, and the polysaccharide coating damage of vascular endothelial cells was observed under a fluoroscopy electron microscope. Results:The level of HBP at admission was significantly higher in the AP group than in the HC group, and the increase in the SAP group was more obvious. Correlation analysis showed that HBP was positively correlated with syndecan-1, HA and MCTSI. Animal studies found that the levels of HBP, syndecan-1 and HA in the AP group were significantly higher than those in the NC group. The pancreatic pathological score showed that the AP group was significantly increased, and the fluoroscopy electron microscope showed that the vascular polysaccharide coating was complete in the NC group, and the structure of the AP group was severely damaged. After LMWH intervention, the structure shedding and damage were significantly reduced, and the difference was statistically significant.Conclusions:HBP can promote the progression of AP, which is related to the destruction of the polysaccharide coating structure of endothelial cells and the increase of vascular permeability caused by HBP.