Objective:To investigate Helicobactor pylori (H. pylori) infection status and interfamilial transmission pattern in Zhengzhou area. Methods:A cross-sectional study was conducted from September 2020 to march 2021, among 731 individual from 266 families randomly selected from 9 communities of Zhengzhou area. H. pylori infection status was determined by serum antibody tests, and 13C-urea breath test was performed in the previously eradicated population to clarify the current infection status. The individual and familial infection rate, infection status for couples and children and adolescent were analyzed. Results:Among 731 individuals from 266 families, 397 of them were H. pylori positive. The individual infection rate was 54.31% (397/731); among infected individuals 77.83% (307/397) were infected with type Ⅰ strain, 22.67% (90/397) were infected by type Ⅱ strain. Annual household income ( χ2=0.419, 0.410, 0.213, all P>0.05), smoking history (χ 2=0.071, P>0.05), drinking history ( χ2=0.071, P>0.05), dining place ( χ2=0.009, P>0.05), gastrointestinal symptoms ( χ2=0.047, P>0.05), family history of gastric disease ( χ2=0.069, P>0.05), and history of gastric cancer ( χ2=0.004, P>0.05) had no significant differences between H. pylori-positive and -negative groups, but the infection rate in individuals with higher education level was lower ( χ2=4.449, P<0.05). The infection rate was significantly higher in≥18 age groups compared with<18 age groups ( χ2=6.531, 23.362, 20.671, 24.244, 37.948, 14.597 and 5.170, all P<0.05). The familial H. pylori infection rate was 87.59% (233/266), and in 61 families all member were infected (26.18%, 61/233). The positive rate was 23.08% (6/26) in 50 families with children under 18 years when both parents were infected. Among 231 coupled families, both couples were infected in 78 families (33.76%), one couple was infected in 113 families (48.92%), and both couples were not infected in 40 (17.32%). With the increase of marriage time, the infection rate of both spouses increased significantly ( χ2=7.775, 12.662, 15.487, all P<0.05). Conclusions:The distribution of H. pylori infection presents a family cluster pattern, and intrafamilial infection is an important transmission rout of H. pylori. The type I strain of H. pylori is the dominate strain in this area.

Objective To investigate the clinicopathological characteristics and prognosis of well-differentiated rectal neuroendocrine tumor(RNET).Methods A retrospective analysis was conducted using the clinical data from 83 patients with well-differentiated RNET from August 2017 to December 2021,including clinical manifestations,endoscopy,endoscopic treatment,postoperative complications,postoperative pathology,follow-up and prognosis.Pathological results according to the 2019 World Health Organization(WHO)Classification of digestive system tumors,83 patients were divided into G1 stage group(72 cases)and G2 stage group(11 cases);Based on the number of tumors in the patient,83 patients were divided into two groups:single RNET group(77 cases)and multiple RNET group(6 cases),the expressions of chromogranin A(CgA),synapsin(Syn)and CD56 were compared among different groups.Results Based on pathological findings in the group,G1 stage group CgA positive rate was significantly higher than that of G2 stage group,the difference was statistically significant(χ2 = 4.23,P = 0.040);Based on the number of tumors,multiple RNET group CgA positive rate was significantly higher than that of single RNET group,the difference was statistically significant(χ2 = 5.74,P = 0.017).It was no significant difference in Syn and CD56 between the two groups(P＞0.050).Conclusion Well-differentiated RNET has no specific clinical manifestations.It is mostly isolated in G1 stage and single RNET.ESD is safe and has a good prognosis,the positive rate of CgA is higher in G1 stage patients,and the positive rate of CgA is higher in patients with multiple RNET.

To construct cellular senescence model by stimulating primary hepatocytes with hydrogen peroxide (H 2O 2). Primary hepatocytes were transfected with p53 siRNA, progerin siRNA or IGF-1 adenovirus vector. The number of SA-β-Gal stained positive cells and the expression of p53 and progerin were detected. The results showed that p53 siRNA and progerin siRNA had knocked-down the expression of p53 and progerin, and had alleviated the hepatocyte senescence. Transfection of insulin-like growth factor (IGF)-1 adenovirus vector into primary hepatocytes had overexpressed IGF-1, and had alleviated the number of SA-β-Gal-positive cells. The expression of p53 and progerin was down-regulated in the nucleus, while the expression of p53 was up-regulated in the cytoplasm. The co-precipitation and co-localization of p53 and progerin was decreased in the nuclear region of hepatocytes. IGF-1 overexpression can inhibit intranuclear p53 translocation, alleviate the interaction between p53-progerin, and alleviate hepatocyte senescence.

Objective: To explore the gene transduction method of chimeric antigen receptor (CAR) mediated by novel cationic polymer nanocarrier mPEG-P (Asp-AED-g-HFB) (PAEF) and PigyBac transposon system to modify natural killer (NK) cells, providing a new strategy for immunotherapy of cancer cells. Methods: PAEF/DNA (transposase+transposon) complex were prepared. The particle size distribution and surface potential of PAEF/DNA complexes were measured with Nano-ZSE Dynamic Light Scattering System (Malvern Instruments). The DNA encapsulation rate, release and stability of PAEF were evaluated by DNA gel electrophoresis, and then by combiningwithparticlesizeandsurfacepotentialtodeterminethepreferentialN/PratiotoenterNKcells.Thecell cytotoxicity of PAEF/DNA complexes under different N/P ratios was analyzed by CCK-8 cytotoxicity test. Transduction efficiency of NK cells was evaluated by Fluorescence microscopy and Flow cytometry, and the feasibility of PAEF gene transfection vectors was assessed. Results: PAEF could encapsulate DNA to form nano-complexes with the diameter of 100-150 nm, which was suitable to mediate DNA entering into cells. PAEF could completely encapsulate DNA with N/P ratio of 20. In the presence of reducing agent dithiothreitol (DTT), PAEF had a good ability to release DNA. NK-92 cells transfected with PAEF/DNA complex, which was formed at the N/P ratio of 80, attained a significantly higher cell viability than cells of lipofectamine transfection group [(72.50±3.9)% vs (64.03±1.8)%, P<0.05]; Fluorescence microscopic observation showed more fluorescence and higher fluorescence intensity in cells of PAEF/DNA group; Flow cytometry showed the highest transfection efficiency of 83.4%. Conclusions: Nanocarrier PAEF can encapsulate DNA well by electrostatic adsorption, and has good biocompatibility and high efficiency for gene transduction. It provides a good experimental basis for adoptive immunotherapy.

Objective:To evaluate the influence of goal-directed volume management based on cardiac output index (CI), intrathoracic blood volume index (ITBVI) and extravascular lung water index (EVLWI) in patients undergoing off-pump coronary artery bypass surgery.Methods:Forty patients (ASA 2 to 3 grade) undergoing off-pump coronary artery bypass surgery in Lanzhou University Second Hospital from January 2017 to December 2018 were selected. The patients were divided into 2 groups by random digits table method with 20 cases in each group: study group (goal-directed fluid therapy treatment with CI, ITBVI and EVLWI) and control group (conventional fluid therapy). The control group was given central venous pressure (CVP) monitoring rehydration, and the study group was given PiCCO hemodynamic monitoring indicators. The CVP, CI, ITBVI and EVLWI for fluid management were measured. Accurate assessment of volume status of patients was done. The study group received goal-directed fluid therapy based on CVP, CI, ITBVI and EVLWI, with the goal of CI in the 3.0 to 5.0 L/(min·m 2) range, ITBVI in the 800 to 1 000 ml/m 2 range and EVLWI in the 3.0 to 7.0 ml/kg range. The heart rate, mean arterial pressure (MAP), urine volume, central venous oxygen saturation (ScvO 2), lactic acid and renal function were monitored. The ventilator withdrawal time, hospitalization in ICU, length of stay, incidence of acute pulmonary edema, incidence of acute renal failure, mortality of 30 d after surgery were recorded and compared between the two groups. Results:Tissue perfusion and urine volume of the study group was significantly improved compared with that of control group ( P<0.05). ScvO 2 of the study group was higher than that of the routine group ( P<0.05). The concentration of lactic acid of the study group was lower than that of the routine group ( P<0.05). The incidences of acute pulmonary edema, acute renal insufficiency and mortality of the study group were lower than those of the routine group (5.0% vs. 15.0%, 5.0% vs. 10.0% and 5.0% vs. 15.0%), and there were statistical differences ( P<0.05). The length of stay and hospitalization in ICU were both lower than those in the control group ( P<0.01). Conclusions:Goal-directed fluid therapy based on CI, ITBVI and EVLWI can effectively optimize the cardiac preload of patients undergoing off-pump coronary artery bypass surgery, improve cardiac output, ensure microcirculation perfusion, maintain the balance of oxygen supply and demand, and reduce the incidence of complications and mortality.

Objective:To investigate the mechanism of cathepsin G(CatG) in improving the treatment efficacy of T cells in gliomas.Methods:(1) Clinical data of 397 glioma patients in the glioma database were collected, Kaplan-Meier method was used to perform survival analysis, and the correlation between CTSG and β2-microglobulin ( β2M) mRNA expressions in glioma tissues was analyzed. (2) Glioma stem cell (GSC) 387 and GSC3565 were isolated from glioblastoma and differentiated into differentiated glioma cell (DGC) 387 and DGC3565, respectively; GSC387 was divided into CatG group and CatG inhibitor group, and cells in the CatG group and CatG inhibitor group were cultured with 0.1 μg/μL recombinant human leukocyte antigen (HLA)-A*02:01 and HLA-B*15:01 combined with 4 ng/μL CatG or 10 mol/L CatG inhibitor for 10 min, respectively; the expressions of HLA-A*02:01 and HLA-B*15:01 were detected by Thomas bright blue staining, and the protein expressions of major histocompatibility complex (MHC)-I and MHC-DR were detected by Western blotting. (3) GSC387, GSC3565, DGC387, and DGC3565 were divided into 4 groups, including CatG group, CatG inhibitory group, blank antibody group 1 and blank antibody group 2, respectively; 4 ng/μL CatG, 10 μmol/L CatG inhibitor, blank antibody 1 and blank antibody 2 were added into the cells from the 4 groups for 24 h, and the expression of HLA-ABC was detected by flow cytometry. (4) GSC387, GSC3565, DGC387, and DGC3565 were divided into CatG group and CatG inhibitory group, respectively; luciferase assay was used to detect the influence of CatG in the killing effects of T cells and natural killer cells. Results:(1) The survival rate in patients from CTSG mRNA high expression group was significantly higher than that in patients from CTSG mRNA low expression group, and the survival rate in patients from β2M mRNA low expression group was statistically higher than that in patients from β2M mRNA high expression group ( P<0.05); a negative correlation between CTSG mRNA and β2M mRNA expressions was noted in glioma tissues ( r=-9.160, P=0.000). (2) Thomas bright blue staining showed that the expressions of HLA-A*02:01 and HLA-B*15:01 obviously increased in the CatG group as compared with those in the CatG inhibitor group; Western blotting showed that as compared with the CatG inhibitor group, the CatG group had increased MHC-I expression, and decreased expressions of α and β chains of MHC-DR. (3) Flow cytometry showed that the HLA-ABC expressions in GSC387 and GSC3565 of the CatG group were statistically higher than those in the CatG inhibitor group ( P<0.05). (4) Luciferase assay showed that, as compared with the CatG inhibitor group, the CatG group had statistically higher proportion of T cells killing GSCs ( P<0.05). Conclusion:CatG can improve the immunotherapy efficacy in GSCs, mainly by increasing the MHC-I expression on the cell surface.

Objective: To investigate the development of hepatocyte senescence during liver fibrogenesis and to explore the effect and possible mechanism of insulin-like growth factor 1 (IGF-1) on hepatocyte senescence and liver fibrosis. Methods: A total of 42 male Sprague Dawley (SD) rats were selected. Eighteen rats were induced by carbon tetrachloride (CCl₄) to establish the rat model of liver fibrosis. On the day 0, six and 28 after the establishment of the model, six rats were executed respectively to analyze the liver fibrosis and hepatocyte senescence in CCl₄-induced liver fibrosis rat models. Twenty-four rats were divided into control group, CCl₄ group, CCl₄+ lentivirus vector (LV-CTR) group and CCl₄+ LV-IGF-1 group, with six rats in each group.The rats were sacrificed on the 28th day after the establishment of the model. The liver tissues were obtained and the inferior vena cava blood was collected to analyze the effect of IGF-1 overexpression on liver fibrosis and hepatocyte senescence. Analysis variance (ANOVA), least significant difference (LSD) and Dunnett T3 test were performed for statistical analysis. Results: Steatohepatitis on the 6th day and early stage of hepatic fibrosis on the 28th day, which indicated the model was successfully established. The results of the effects of IGF-1 overexpression on hepatic fibrosis and hepatocyte senescence showed that on the 28th day, compared with those of control group, both the score of Ishak liver inflammation and necrosis and the score of Ishak liver fibrosis were increased in the CCl₄ group, CCl₄+ LV-CTR group and CCl₄+ LV-IGF-1 group (0, 14.55±1.94, 15.43±2.19 and 10.29±1.47, respectively; 0, 3.51±0.51, 3.21±0.79 and 1.32±0.40, respectively). The area of liver tissues by Masson staining (0.45±0.40, 5.62±1.08, 6.03±0.65 and 2.88±1.54), SA-β-Gal staining (1.75±0.80, 4.28±1.19, 4.92±1.14, 3.11±0.79), p53 (2.02±0.81, 4.36±1.02, 4.72±0.72 and 3.58±0.70) and progerin (0.72±0.40, 4.52±1.01, 4.01±1.25 and 2.66±0.80) all were increased. The levels of serum IGF-1 all were decreased ((632.00±6.04), (503.00±40.42), (508.00±21.94) and (572.40±5.94) ng/L). However the levels of ALT all were increased ((11.20±5.97), (214.00±73.90), (245.00±76.06) and (30.00±5.00) U/L). The relative expression levels of p53 (0.58±0.06, 1.78±0.18, 1.72±0.10 and 1.23±0.22) and progerin (0.12±0.02, 0.78±0.15, 1.32±0.20 and 0.81±0.16) in the primary hepatocytes were increased. The differences were all statistically significant (F=91.674, 90.778, 32.982, 9.726, 10.640, 17.029, 103.910, 30.059, 64.707 and 97.457, all P<0.05). Compared with those of CCl₄+ LV-CTR group, the score of Ishak liver inflammation and necrosis and the score of Ishak liver fibrosis were decreased in the rats′ liver tissues of CCl₄+ LV-IGF-1 group, the areas of Masson staining, SA-β-Gal staining, p53 and progerin in the liver tissues were decreased, the level of serum IGF-1 was increased, the level of ALT was decreased, and the relative expression levels of p53 and progerin in primary hepatocytes both were decreased. The differences were all statistically significant (all P<0.05, respectively). Conclusions Hepatocyte senescence increases in the process of liver fibrosis induced by CCl₄. Overexpression of IGF-1 may alleviate liver injury, improve hepatocyte senescence and liver fibrogenesis by regulating the nuclear p53/progerin pathway.

[Abstract] Objective:To prepare the third generation CAR-T cells targeting EGFRvⅢ (EGFRvⅢCAR-T) and to detect its specific killing effect against EGFRvⅢ+ U87 cells in vitro and in vivo. Methods: Human CD3+ T cells were transfected with lentiviral EGFRv Ⅲ/3CAR, which was generated by calcium phosphate co-precipitation of three plasmids. The expression of EGFRvⅢ/3CAR in T cells was detected by Western blotting and flow cytometry. In vitro killing effect of EGFRvⅢ/3CAR-T cells on EGFRvⅢ+ U87 cells was detected by 51Cr release assay. The secretion of cytokine IFN-γ of EGFRvⅢ/3CAR-T cells was detected by ELISA. Nude mouse xenograft model was constructed to detect the in vivo cytotoxicity of EGFRvⅢ/3CAR-T cells on xenograft tumor. Results: The EGFRvⅢ/3CAR lentivirus was successfully packaged with an average titer of 5×106 TU/ml. Western blotting showed that a protein band of approximate 58 000 molecular weight was observed in EGFRvⅢ/3CAR-T cells but absent in untransfected T cells. Flow cytometry indicated the average transduction efficiency of EGFRvⅢ/3CAR was 52.3%. 51Cr release assay showed that the specific killing effect of EGFRvⅢ/ 3CAR-T cells was positively correlated with E/T ratio (E∶T=4∶1, 8∶1, 16∶1, 32∶1). ELISA showed that cytokine IFN-γ secretion was (1 836±148.2) pg/ml, which was significantly different from that of NTT and GFP+ T cells (P<0.01). The specific killing activity of EGFRvⅢ/3CAR-T cells and IFN-γ secretion were both dependent on the expression level of EGFRvⅢ in U87 cells. The tumor growth monitoring results showed that the tumor volume of EGFRvⅢ/3CAR-T cell group was significantly different from that of GFP+ T cell group and PBS group around 3 weeks after injection (P<0.01). Conclusion: EGFRvⅢ/3CAR-T cells demonstrated specific antitumor effectagainstEGFRvⅢ+U87cellsbothinvitro and in vivo, providing basis for immunotherapyofgliomainfuture clinical use.

Objective To compare two different regimens of ultrasound-guided Continuous ad-ductor canal block (CACB)for postoperative analgesia and early ambulation after total knee arthro-plasty (TKA).Methods Sixty-seven patients scheduled for unilateral TKA undergoing spinal anes-thesia,13 males and 54 females,aged 18-85 years,BMI 18-30 kg/m2,ASA physical status Ⅰ-Ⅲ, were randomly divided into the continuous infusion group A (n=34)and the intermittent boluses group B (n=33).After the operations,ultrasound-guided CACB were administered and 20 ml of 0.2% ropivacaine was given as the loading dose.From then on,patients in both groups used electronic analgesic pumps containing 240 ml of 0.2% ropivacaine for postoperative analgesia.5 ml/h of 0.2% ropivacaine was continuously infused for 48 hours in the group A.5 ml of 0.2% ropivacaine was automated injected every 60 minutes in the group B.All infusion pumps were setted at a bolus dose of 5 ml,with a lock time of 30 minutes.The total consumptions of analgestic pum solution and dezoine, quadriceps muscle strength, active range of knee flexion, ambulation distance and occurrences of adverse reactions such as nausea and vomiting,dizziness,drowsiness,extravasating and errhysis were recorded at different time points postoperatively.Results The total consumptions of analgestic pum solution at 12,24 h postoperatively of group B were significantly reduced than that of group A (P<0.05).The 48 h total dezoine consumption of group B was significantly reduced than group A (P<0.05).Active range of knee flexion at 24,48 h and ambulation distance at 48,72 h of group B were significantly higher than group A (P<0.05).There was no statistical difference in quadriceps muscle strength between group A and group B.The incidence of nausea and vomiting in group A was significantly higher than that in group B,and there were no statistical difference in other adverse reactions between group A and group B.Conclusion Compared with the continuous infusion group,the intermittent bolus group for CACB after TKA can provide better analgesic effect and de-crease opioid use postoperatively,with little effect on motor nerve,promoting early ambulation.

Objective To evaluate the effect of obstructive jaundice on the accuracy of left ventricular end-diastolic volume (LVEDV) and stroke volume variability (SVV) in monitoring fluid responsiveness.Methods Thirty patients of both sexes,aged 45-60 yr,weighing 55-70 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,with New York Heart Association Ⅰ,scheduled for elective pancreatoduodenectomy,were divided into 2 groups according to the serum total bilirubin levels:A group (serum total bilirubin ≥ 17 μmmol/L,n =16) and B group (serum total bilirubin＜ 17 μmmol/L,n =14).Six percent hydroxyethyl starch 500 ml was infused over 40 min after anesthesia induction.The parameters of VigileoTM such as cardiac output (CO),SVV,systemic vascular resistance (SVR) and pulmonary capillary wedge pressure and indices measured by transesophageal three-dimensional echocardiography such as LVEDV,left ventricular end-systolic volume,CO',left ventricular ejection fraction (LVEF) and ratio of mitral peak velocity of early filling (E) to early diastolic mitral annular velocity (e',E/e'ratio) were recorded before and after fluid loading.Results Compared with that before fluid loading,SVV was significantly decreased in two groups,and CO,LVEDV,CO'and LVEF were significantly increased in group B,and E/e'ratio was significantly increased in group A (P＜0.05).Compared with group B,CO,SVR,CO'and LVEF were significantly decreased,and pulmonary capillary wedge pressure was increased in group A (P＜0.05).Conclusion Obstructive jaundice causes decrease in the accuracy of LVEDV in monitoring fluid responsiveness and no effect on SVV.