OBJECTIVETo investigate protective effect of paeonol (Pae) on human umbilical vein endothelial cells (HUVECs)injured by hyperlipidemic serum and explore its possible mechanism.METHODS The injury model was induced by 20％ hyperlipidemic serum incubating HUVECs for 24 h.Pae 124,247 and 495.μmol· L-1 were given followed by administration of hyperlipidemic serum for 24 h.The morphological changes were observed under inverted microscope,cell survival rate was evaluated by MTT method,the nitric oxide (NO) content was measured by nitric acid reductase method and the endothelial nitric oxide synthase (eNOS) mRNA expression was determined by RT-PCR.RESULTSCompared with normal contorl group,most cells in model group split and exfoliated.However,the morphology was tending to normal level after intervention with Pae.Pae significantly improved cell viability(P ＜0.01).Compared with model group,the survival rate increased from (53.0 ±10.1 ) ％ to ( 68.4 ± 9.1 ) ％,( 84.5 ± 6.7 ) ％ and (98.1 ± 7.5 ) ％,respectively.The NO content and eNOS mRNA expression both increased greatly in Pae groups(P ＜ 0.01 ).Compared with model group,content of NO increased from 54 ± 4 to 79 ± 6,115 ± 5 and ( 136 ± 6) μmol · L- 1,respectively.The expression level of eNOSmRNA improved from 0.215 ± 0.060 to 0.451 ± 0.045,0.563 ± 0.013,0.704 ± 0.068,respectively.CONCLUSIONPae exerts protective effect on HUVECs injured by hyperlipidemic serum by increasing eNOS mRNA expression,which might therefore improve the content of NO.

Objective To investigate the inhibitive effects of viral protein r (Vpr) of human immunodeficiency virus 1 ( HIV-1 ) on human colorectal cancer cell line HCT-8,and to find the possible mechanisms.Methods The HCT-8 cells were divided into the control group,adv group and adv-Vpr group.HCT-8 cells were not treated in the control group; HCT-8 cells were treated with Adv or Adv-Vpr at different multiplicity of infection (MOI) in the Adv group or Adv-Vpr group,respectively.Cell proliferation was detected by MTT assay.Cell cycle,apoptosis and mitochondrial membrane potential were detected by flow cytometry.The expression of apoptosisrelated proteins was detected by Western blot.All data were analyzed by using the q test,t test and one-way or two-way analysis of variance.Results The proliferation of HCT-8 cells was significantly inhibited by Vpr.The MTT value of HCT- 8 cells in the Adv-Vpr group was 1.03 ± 0.04,which was significantly lower than 2.46 ± 0.15 in the Adv group and 2.51 ± 0.14 in the control group at 72 hours after Adv-Vpr transfection ( MOI =200) ( F =144.6,P ＜ 0.05).The ratio of HCT-8 cells in the G2/M phase was 37.31％ ± 5.90％ in the Adv-Vpr group,which was significantly higher than 18.30％ ± 6.04％ in the Adv group and 16.66％ ± 3.51％ in the control group ( F =10.08,P ＜ 0.05 ).The ratio of HCT-8 cells with decreased mitochondrial membrane potential was 32.07％ ±5.64％ in the Adv-Vpr group,which was significantly higher than 3.32％ ±0.79％ in the Adv group and 2.76％ ±1.43 ％ in the control group at 48 hours after Adv-Vpr transfection ( MOI =200) ( F =64.45,P ＜ 0.05).The apoptosis rate of HCT-8 cells was 37.62％ ±6.48％ in the Adv-Vpr group,which was significantly higher than 3.44％ ± 1.11％ in the Adv group and 2.93％ ± 1.07％ in the control group at 72 hours after Adv-Vpr transfection ( MOI =200) ( F =122.4,P ＜ 0.05 ).The results of Western blot showed that Vpr induced cleavage and activation of Caspase-9 and Caspase-3 and phosphorylation of Chk1-S345,while the expression levels of Fas,Fas-L,ERK1,ERK2 remained the same at 48 hours after Adv-Vpr treatment ( MOI =200).Conclusions Vpr inhibits the proliferation of the HCT-8 cells in vitro through G2/M phase arrest and apoptosis.Vpr plays its role by activating DNA damaging pathway and initiating mitochondria apoptotic pathway.Vpr is a potential therapeutic agent for colorectal cancer.

Objective To investigate the risk factors of gastrointestinal graft versus host disease(GVHD) following allogeneic hematopoietic stem cell transplantation (allo-HSCT).Method 214 patients receiving allo-HSCT from 2003 to 2012 were enrolled in this study,and assessed on the degree of gastrointestinal GVHD.The effects of the primary diseases status,gender,age,conditioning regimen intensity,donor type,the number of cells positive for the CD34 expression,and the use of anti-lymphocyte immunoglobulin (ALG) in the pretreatment scheme on the occurrence of gastrointestinal GVHD was studied.The responses of different degrees of GVHD to immunotherapy were evaluated.Result Univariate and multivariate analyses revealed that the graft type and the conditioning regimen intensity were the risk factors of gastrointestinal GVHD (P＜0.05).Conclusion Donor type and conditioningregimen intensity may be the main risk factors of gastrointestinal GVHD.

0.05),and there was significant difference in ICA,VP,RI,AT,and the levels of TXB_2 and 6-K-PGF_(1?) of the blood plasma(P

Objective In order to evaluate the possible effects of myeloid-derived suppressor cells (MDSCs) on graft versus host disease (aGVHD) development and clinical outcomes,this study systematically detected the dynamic changes of MDSCs accumulation in patients during the first 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Peripheral blood was obtained from 30 patients and 10 healthy volunteers with heparin anticoagulant tubes for 6 mL.For patients,peripheral blood was collected during the first 100 days after allo-HSCT and MDSCs levels were detected by flow cytometry.For measuring the serum concentrations of IL-6,IL-10,IL-1β,TNF-α,Arg-1,HO-1 and iNOS,samples were analyzed using ELISA kits.Results Patients developing aGVHD were infused with significantly less number of MDSCs [(39.94 ± 8.383) 106/kg]than in those not developing aGVHD [(209.0 ± 57.68) 106/kg,P =0.002 6];Patients developing aGVHD Ⅰ-Ⅱ and patients without aGVHD received significantly greater number of MDSCs [(61.96 ± 13.67) 106/kg and (209.0 ± 57.68) 106/kg] than in those developing aGVHD Ⅲ-Ⅳ [(20.37 ±4.304) 106/kg,P =0.013 9].After allo-HSCT,the mean percentage of MDSCs increased markedly in patients developing aGVHD [(7.725 ± 1.460)％] as compared with those not developing aGVHD [(3.423± 1.044)％,P =0.021 3].The high MDSCs group (＞53.712 × 106/kg) showed more favorable clinical outcomes than in the low MDSCs group (≤53.712 × 106/kg).The 2-year overall survival rate as 100％ in high MDSCs group,and 50％ in low MDSCs group (P =0.001 3).The cumulative incidence of 2-year relapse was 6.250％ and 29.252％ in high MDSCs group and low MDSCs group respectively (P =0.112 3).The cumulative incidence of NRM was significantly lower in high MDSCs group (0％) than in low MDSCs group (49.519％,P=0.001 8).MDSCs frequencies significantly increased in patients developing aGVHD after allo-HSCT.After allo-HSCT,the concentrations of IL-6,IL-10,TNF-α,Arg-1,iNOS and HO-1 were significantly elevated in patients developing aGVHD.Conclusion The number of MDSCs when engraftment may be used as a predictor for the development and severity of aGVHD.MDSCs might be considered as a potential new approach to regulate transplant rejection and achieve long-term survival.

Objective To investigate the effects of resveratrol on inhibiting proliferation and inducing apoptosis of U251 cells. Methods The proliferation activity was detected by MTT assay, and apoptosis of early and late period by method of Annexin V-FITC apoptosis assay, and the cell morphological changes were observed by microscopy after U251 cells being treated with resveratrol. Results Resveratrol could significantly inhibite the proliferation and induce the apoptosis of early and late period of U251 cell in a time-and does-dependent manner. Conclusion Resveratrol can significantly inhibite the proliferation and induce the apoptosis of early and late period of U251 cell.

Objective To clone the full-length of human Gill cDNA and construct the recombinant lentiviral expressing vector pLOX-Gfil for eukaryotic expression,providing a basis for further study on the biological functions of Gfil.Methods Total RNA was isolated from K562 cells,and the full-length Gfil cDNA was amplified by RT-PCR and then ligated with pGEM-T vector after retrieve and purification.The ligation product was transformed into competent cells DH5a.The positive recombinant clones were selected and identified by a complementation,restriction endonuclease digestion.The cloning vector and the lentiviral vector pLOX first digested with BarnH I were ligated and transformed.The enzyme and PCR analyses were performed to confirm the recombinant vector,and then DNA sequence analysis.Results A fragment of 1.2 kb was obtained by RT-PCR.The enzyme and PCR analyses revealed that the correct Gfil cDNA was cloned.The sequence of cloned cDNA was identical to the sequence deposited in GenBank (NM005263).Conclusion Gfil was cloned correctly and the recombinant lentiviral vector pLOX-Gfil for eukaryotic expression was constructed successfully.

Objective To explore brain activity features during the resting state in alcohol dependent individuals,and study the relationship between the brain activity features and alcohol dependent individuals' clinical symptoms.Methods Twenty-four alcohol dependent individuals and 22 healthy control subjects,well matched in gender,age,education and handedness,were enrolled as the alcohol dependent group and control group respectively.AGE 3.0 T MR scanner was used to acquire all the subjects' resting state data. DPARSF software was used to process resting functional MRI data,and then the whole brain fractional amplitudes of low frequency fluctuation (fALFF) data were acquired.Two-sample t test statistical analysis was made to access fALFF difference between the two groups. Results In comparison with the control group,the alcohol dependent group showed reduced fALFF in bilateral medial prefrontal gyrus,right inferior occipital gyrus,left precuneus,left inferior temporal gyrus,and left posterior lobe of cerebellum (0.64-1.69 vs.0.87-1.78,t =- 4.23- - 2.79,P ＜ 0.05 ). fALFF was increased in the alcohol dependent group at the anterior cingulate,bilateral inferior frontal gyrus,right middle frontal gyrus,bilateral insular lobe,bilateral dorsal thalamus ( 0.86-1.82 vs. 0.76-1.58,t =3.56-3.96,P ＜ 0.05 ).Conclusion Alcohol dependent individuals had abnormal activity at the bilateral prefrontal lobe,anterior cingulate,bilateral dorsal thalamus,bilateral insular lobe,left posterior lobe of cerebellum et al,during the resting state,and these abnormal activities might be related with clinical manifestation and pathophysiology.

Objective:To explore the clinical and CT imaging features of immune checkpoint inhibitor-associated pneumonia (CIP) and to improve the early diagnostic ability of CIP.Methods:From June 1, 2020 to October 31, 2021, the clinical data and chest CT images of 2 067 patients with advanced malignant tumor treated with immune checkpoint inhibitor (ICI) in the First Medical Center, Chinese PLA General Hospital were retrospectively analyzed. Patients with CIP were enrolled according to the guidelines for CIP diagnosis, and the incidence, time from the start of medication to the onset of CIP, medication cycle, imaging features, imaging patterns, CT grade and outcomes were analyzed. χ 2 test was used to compare the incidence of CIP in patients with or without basic lung disease. Results:Among 2 067 patients with malignant tumors treated with ICI, 67 patients developed CIP, the incidence of CIP was 3.2%. The incidence of CIP was significantly different between 386 patients with basic lung disease (7.00%, 27/386) and 1 681 patients without basic lung disease (2.4%, 40/1 681) (χ 2=21.32, P<0.001). The time from the start of medication to the onset of CIP was 7-367 d (median 52 days), and the duration of medication was 1-12 cycles (median 2 cycles). The imaging features of CIP presented as ground glass opacities in 54 cases (80.6%), solid nodules in 26 cases (38.8%), consolidations in 25 cases (37.3%) and irregular reticular opacities in 24 cases (35.8%). The main radiologic pattern was organizing pneumonia (OP, 34 cases, 50.7%), and followed by diffuse alveolar damage (DAD) pattern (14 cases, 20.9%). According to CT grading, there were 26 cases in low risk grade, 17 cases in moderate risk grade and 24 cases in high risk grade. Of 43 low-and medium-risk grade cases, 25 were OP pattern, accounting for 58.1%, and among 24 high-risk grade patients, 13 were DAD pattern, accounting for 54.2%. Forty-three of the 52 patients were initially untreated, of which 23 patients progressed, 17 had lesion shrinkage, and 3 had resolution, and relapsed in 8 cases after resolution or drug withdrawal. Conclusions:The imaging manifestations of CIP are mainly ground glass opacities, nodules, consolidations, and irregular reticular opacities. The radiologic patterns are mainly OP and DAD. OP is the most common pattern in low-moderate risk grade CIP and DAD is the most common pattern in high risk grade CIP. Patients with basic lung disease are more likely to get CIP.

OBJECTIVEApplication of real-time shear wave elastography (SWE) measurement of patients with Chronic severe hepatitis B and liver cirrhosis of the liver stiffness, aimed to explore SWE can evaluate the existence of liver cirrhosis patients with esophageal varices (EV) and its severity.

METHODSAccording to the results of gastroscope, 256 cases of patients with chronic liver disease and cirrhosis of the liver can be divided into no EV group,mild EV group,moderate to severe EV group,analysis between groups in patients with liver stiffness, portal vein,spleen vein diameter, the correlation of liver fibrosis indexes and the degree of esophageal varices.Using receives operating characteristic curve (ROC) and area under curve of ROC to evaluate each index prediction ability.

RESULTSCompare the liver stiffiness, portal vein,spleen vein diameter had statistically significant difference in the no EV group, mild EV group,moderate to severe EV group, (F values are respectively 137.86,44.77,73.88, P < 0.05), Patients age, type IV collagen, larninin, hyaluronic acid had no statistically significant difference in the no EV group and mild EV group (P > 0.05) and had statistically significant difference in the other two groups (P < 0.05). Patients with gender, pro-collagen type III N-terminal peptide (PC III NP) had no statistically significant difference in the three groups (P > 0.05). Correlation analysis showed that portal vein, spleen vein diameter, type IV collagen, laminin, hyaluronic acid showed significant positive correlation (P < 0.05),highest correlation was liver stiffness and the degree of esophageal varices, correlation coefficient of 0.689 (P < 0.01). PC III NP and the degree of esophageal varices, liver stiffness showed no correlation (P > 0.05). Liver stiffness area under the ROC curve is 0.923, with a strong ability to predict than the portal vein and splenic vein diameter, LN, IV-C, HA, PCIII NP. Liver stiffness more than 7.55 kPa, diagnose mild EV sensitivity 90.5%, specificity 60%.Liver stiffness more than 18.85 kPa,the sensitivity of the diagnosis of severe EV 82.4%, specificity of 90.5%.

CONCLUSIONSSWE liver stiffness measurement was predicted the existence of the EV and the severity of liver disease patients and effective inspection method, can be used as evaluation of liver disease patients with esophageal varices non-invasive indicator of the initial screening.