Objective To establish the method of combined hepatocyte growth factor (HGF) with epidermal growth factor (EGF) cultured human gallbladder epithelial cells(HGBECs) in vitro .Methods The epithelial layer was peeled away from human gallbladder ,epithelial layer were digested with collagenase Ⅳ and scraped repeatedly .HGBECs were isolated and seeded in cell cul-ture plates containing medium supplemented with or without 10 ng/mL EGF or with 10 ng/mL HGF and 10 ng/mL EGF respec-tively .Then the morphologic changes of the cells were observed and taken photos with inverted phase contrast microscope ,and counted number of cells ,MTT assay detected vigor of cells in different groups .Results The number of the HGBECs of the HGF+EGF group was obviously more than the EGF group ,the duration of the HGBECs of the HGF+ EGF group was obviously longer than the EGF group(19 .3 ± 2 .5)d vs .(14 .2 ± 2 .4)d ,P< 0 .05 .And the HGBECs of the group with HGF+ EGF had better cell vigor .Conclusion HGF combines with EGF added to medium can obviously promote the proliferation of HGBECs and prolong the duration and stabilize morphology of HGBECs in vitro .

Objective:To investigate the relationship between serum carcinoembryonic antigen (CEA) and the predictive value of epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer patients, as well as to analyze further EGFR muta-tions and CEA levels affecting patient survival. Methods:From March 2009 to March 2011, a total of 387 cases were treated in the Lung Cancer Department in Tianjin Cancer Hospital. Preoperative CEA tumor marker and postoperative EGFR gene mutation were used for routine detection. The influence of CEA tumor marker on EGFR mutation and its relationship with the prognosis were ana-lyzed further. Results:A total of 168 cases involved EGFR mutations, the incidence of which is more frequent in women, non-smokers, adenocarcinoma patients, and patients below 60 years old (P<0.05). This study also determined that EGFR mutation was related with tu-mor markers and chemosensitivity indicators. Elevated Cyfra21-1, SCC, and ERCC1-positive are more common in wild-type patients (P<0.05). However, abnormal CEA was more common in EGFR mutation patients (P=0.015). The rate of EGFR gene mutations signifi-cantly increased as the serum CEA level increased. Serum CEA levels were divided into three groups (<5, 5-20, and>20). The positive rates of EGFR mutations were 40.1%, 47.5%and 66.6%(P=0.003). Logistic regression analysis determined that CEA levels are inde-pendent factors in predicting EGFR mutations and independent prognostic factors in patients with non-small cell lung cancer. Conclu-sion:Serum CEA levels can independently predict the prognosis of resected non-small cell lung cancer patients, which is has a close re-lationship with EGFR mutations.

Objective To investigate the effects of limited resuscitation on hepatic ischernia-reperfusion in-jury in rots with hemorrhagic shock. Method Uncontrolled hemorrhagic shock was induced in 54 rats by transect-ing the middle branch of the splenic artery to produce standardized massive splenic injury. Resuscitation started when the mean arterial pressure (MAP) reached 40 mmHg. The rats were randomly divided (n = 9per group) in-to sham-operated group (SS), or one of five treatment groups in which infusion of Ringer' s solution was continually administrated to maintain MAP at 40 (RS40), 50 (RS50), 60 (RS60), 80 (RS80) or 100 mmHg (RS100) for 45 minutes (T45 point). After the bleeding was controlled, resuscitation was continued with Ringer's solution and whole blood (2:1) to increase the MAP to 100 mmHg for 120 minutes (T165 point), which was followed by obser-vation for 240 minutes (T405 point). All animals were observed for 240 minutes or until death. Blood specimens were collected at TO, T45, T165 and T405 for determination of blood lactate levels. At the end of the experiment,a small amount of hepatic tissue was collected to measure tissue blood perfusion, total antioxidative capacity (TAOC), Na+K+ ATPase activity and malondialdehyde (MDA) levels. Results At T405, the blood lactate lev-els in the RS80 and RS100 groups [(3.60±0.68) and (3.84 ± 1.09) mmol/L, respectively] were significantly higher than those in the SS, RS40, RS50 and RS60 groups [(2.00±0.66),(2.74±1.45),(2.43 +0.94) and (2.07±0.95) mmol/L, respectively;all:P < 0.05]. The MDA levels were significantly higher in the RS80 and RS100[(7.32±0.31) and (7.71±0.23) nmol/mg,respectively] than those in the SS, RS40, RS50 and RS60 groups[(4.95±0.80),(6.14±0.94),(6.42±0.48) and (6.84±0.36) nmol/mg, respeetively;all: P <0.05]. The Na+ K+ ATPase and TAOC levels were significantly lower in all of the RS groups than those in the SS group (all: P < 0.05), and those in the RS80 and RS100 groups was significantly lower than those in the RS40,RS50 and RS60 groups (all: P <0.05). Blood perfusion in the RS80 and RS100 groups was significantly lower than that in the other groups (all: P < 0.05). Conclusions If hemorrhage is uncontrolled, limited resuscitation appears to balance the needs for organ perfusion, improve the microcircttlation and decrease lactate levels. Fur-thermore, limited resuscitation could decrease ischemia-reperfusion injury in liver tissue.

Objective To evaluate the incidence of net adverse clinical and cerebral events (NACCE) 1 year after implantation of biodegradable polymer stents (BP-SES) in elderly patients with coronary artery disease.Methods The clinical data of patients inserted with BP-SES in I-LOVE-IT 2 Trial were retrospectively analyzed, including 1829 elderly patients admitted in the General Hospital of Shenyang Military Command from Oct. 2012 to Jun. 2013, of which 62 cases aged equal to and more than 65 years (elderly group) and 1202 cases less than 65 years (non-elderly group). The primary end-point of this research was target lesion failure (TLF) rate on 12 months and the secondary end-point was the incidence of NACCE, including all-cause death, all myocardial infarction, stroke and severe hemorrhage (BARC type ≥3), and then the multiple regression analysis was performed.Results The Baseline conditions of the two groups were significantly different (P<0.05) including BMI, diabetes, hypertension, hyperlipidemia, family history of coronary heart disease, smoking history, past stroke history, history of peripheral vascular disease and stable angina pectoris. When comparing elderly group with non-elderly group, marked differences existed on the incidence of NACCE (10.0%vs. 5.2%,P<0.01), all-cause mortality (2.7%vs. 0.7%,P<0.01), myocardial infarction (5.6%vs. 3.5%,P=0.03), stent thrombosis (1.9%vs. 0.5%,P<0.01) and stroke (2.2%vs. 0.8%,P=0.01). Multiple regression analysis revealed that elderly (age ≥65) was the independent predictive factor for NACCE (OR=1.904, 95%CI 1.304-2.781,P=0.001).ConclusionThe incidence of NACCE is increased significantly in elderly patients (age ≥65), and elderly is an independent predictive factors for 12-month NACCE in patients implanted with BP-SES

This study inquired into the mechanisms of co-immobilized cytokines and free cytokines-induced apoptosis on HeLa cells. With the use of photochemical fixed method, TNF-alpha/IFN-gamma were co-immobilized on a 24-well polystyrene culture plate. HeLa cells were stained with fluorescent probe JC-1 to detect the changes of mitochondrial membrane potential (deltapsim), and then were examined by flow cytometry. The results showed that co-immobilized cytokines could induce the apoptosis of HeLa cells in a dose-independent manner. When treated with low-dose of co-immobilized cytokines (20ng/ml), the mitochondrial membrane potential (deltapsim) of HeLa cells continually decreased in 6 days. These indicate that low dose co-immobilized cytokines have a long-term of apoptosis-inducing effect on HeLa cells. We assume that there is close relationship between the mitochondrial membrane potential decrease and the apoptosis of HeLa cells.
Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Immobilized Proteins ; pharmacology ; Interferon-gamma ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondrial Membranes ; drug effects ; physiology ; Tumor Necrosis Factor-alpha ; pharmacology

Objective To evaluate the long-term effect of surgical procedures for congenital choledochal cyst (CCC).Methods From 1986 to 2000, 120 cases of CCC were admitted and 73 of them underwent the primary operations in General Hospital of PLA. Three types procedures were performed,type I: external drainage of CCC in 7 cases; type II:cystojejunal Roux-en-Y anastomosis in 5 cases; type III: cyst excision with cystojejunal Roux-en-Y anastomosis or cystoduodenostomy in 57cases,and other procedures in 4 cases.Results 68 cases were followed-up for 6 months to 5 years (median 2.7 years). Three cases undergoing type I operations accepted reoperations;two cases undergoing type II operations accepted reoperations due to severe complications as cholongitis and hepatolithiasis; 57 cases treated by type III operation with the good results 88.7% and none reoperation.Conclusions External drainage is only a first-aid management on emergency basis. Internal drainage should never be done,because the effect is temporary,and severe complications result in reoperations. Cyst excision with biliary tract reconstruction is recommended as the optimal treatment of CCC.

OBJECTIVETo develop a new method to rapidly determine and identify Guizhi Fuling capsule by portable acousto-optic tunable filter-near infrared spectroscopy.

METHODThe qualitative model was set up using principal component analysis. The correlation models between the NIR spectra and the reference values of five major constituents were obtained with partial least squares method.

RESULTThe identifying model accurately identified Guizhi Fuling capsule, and quantitative analytical models could precisely predicted the content of ellagic acid, baicalin, benzoylpaenoniflorin, cinnamaldehyde, and paeonol. The correlation coefficients of the calibration models were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, the validation set coefficients of the calibration were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, and the RMSEP were 1.138%, 3.014%, 0.751%, 0.625%, 3.455%, 1.363%, respectively. The results of external validation showed no significant difference between the predictive and the determining values by t-test.

CONCLUSIONThe method is accurate, rapid and non-destructive, and can be used for determining and identifying Guizhi Fuling capsule.

Acetophenones ; analysis ; Acrolein ; analogs & derivatives ; analysis ; Calibration ; Capsules ; chemistry ; Drug Evaluation ; methods ; Drugs, Chinese Herbal ; analysis ; Ellagic Acid ; analysis ; Flavonoids ; analysis ; Least-Squares Analysis ; Models, Chemical ; Principal Component Analysis ; methods ; Spectroscopy, Near-Infrared ; methods

Lung cancer is the most common malignant tumor in the world. In order to improve the survival rate of patients with advanced lung cancer, more effective treatment methods are needed,in which immunotherapy has a broad therapeutic prospect. In recent years, immune-checkpoint inhibitors have received extensive attention in the treatment of lung cancer. Significant progress has been made in the development of a variety of first-line and second-line treatments, and significant advances have been made in the treatment of advanced lung cancer. With the successful application of immune-checkpoint inhibitors, neoadjuvant therapy has attracted extensive attention. In addition, the successful application of combined therapies such as immune combined immunization, immune combined tyrosine kinase inhibitor (TKI) and immune combined chemotherapy improved the survival rate of patients to some extent. However, pseudo progression and drug resistance has become a non-negligible problem in the immunotherapy of non-small cell lung cancer, which is worthy of further study. Although immune-checkpoint inhibitors have once again brought attention to tumor immunotherapy, their side effects are also worthy of attention. The recent advances in the application of immune-checkpoint inhibitors in lung cancer were summarized in order to provide a theoretical basis for its clinical application.

OBJECTIVEThe aim of this study was to investigate the effects of combination of icotinib and cetuximab on the acquired drug resistance caused by T790M mutation of EGFR in NSCLC, and provide experimental evidence for rational treatment of NSCLC.

METHODSThe effects of these two agents on cell proliferation, apoptosis, and EGFR-dependent signaling were evaluated using 3-(4, 5-dimethylthiazol-2-yl)- 5-diphenyltetrazolium bromide (MTT) assay, annexin V staining, and Western blotting. The expression of molecular markers of tumor proliferation PCNA and Ki-67 protein was further examined by immunohistochemistry, and the expression of EGFR-signaling-related proteins in tissue sections taken from H1975 tumor xenografts was assessed by Western blot assay. Sensitivity to EGFR inhibitors was detected in human H1975 tumor xenograft in nude mice.

RESULTSThe in vitro experiment showed that the proliferative ability of H1975 cells was inhibited in a dose-dependent manner, along with the increasing doses of cetuximab and icotinib, and the combination of cetuximab with icotinib resulted in a more pronounced growth inhibition of the H1975 cells. The apoptosis rate of H1975 cells after treatment with 0.5 µmol/L icotinib and 1 µg/ml cetuximab was (22.03 ± 2.41)% and that after treatment with 5 µmol/L icotinib and 10 µg/ml cetuximab was (42.75 ± 2.49)%, both were significantly higher than that after treatment with the same dose of icotinib or cetuximab alone (P < 0.05). The nude mouse experiment showed that the transplanted tumor was growing to (614.5 ± 10.8) mm(3) in the blank control group and to (611.2 ± 8.7) mm(3) at 28 days after icotinib treatment, but (30.8 ± 2.0) mm(3) in the cetuximab treatment group and 0 mm(3) in the cetuximab combined with icotinib group. There was a significantly decreased expression of Ki-67 and PCNA proteins and down-regulation of phosphorylation of EGFR signaling-related proteins in the cetuximab combined with icotinib group.

CONCLUSIONSThe combination of icotinib with cetuximab can exert synergistic inhibitory effect on the acquired drug resistance caused by T790M mutation of EGFR in NSCLC H1975 cells, interrupts the EGFR-downstream signaling pathway, and enhances the anticancer activity of chemotherapeutic drugs. Our results provide further experimental evidence for the clinical studies of combination of icotinib with cetuximab in the treatment of NSCLC patients associated with secondary drug resistance caused by T790M mutation of EGFR.

Animals ; Antibodies, Monoclonal, Humanized ; administration & dosage ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Apoptosis ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Cetuximab ; Crown Ethers ; administration & dosage ; therapeutic use ; Down-Regulation ; Drug Resistance, Neoplasm ; genetics ; Genes, erbB-1 ; genetics ; Humans ; Lung Neoplasms ; drug therapy ; Mice ; Mice, Nude ; Mutation ; Quinazolines ; administration & dosage ; therapeutic use ; Receptor, Epidermal Growth Factor ; Signal Transduction

BACKGROUND: The HELIOS stent is a sirolimus-eluting stent with a biodegradable polymer and titanium oxide film as the tie-layer. The study aimed to evaluate the safety and efficacy of HELIOS stent in a real-world setting. METHODS: The HELIOS registry is a prospective, multicenter, cohort study conducted at 38 centers across China between November 2018 and December 2019. A total of 3060 consecutive patients were enrolled after application of minimal inclusion and exclusion criteria. The primary endpoint was target lesion failure (TLF), defined as a composite of cardiac death, non-fatal target vessel myocardial infarction (MI), and clinically indicated target lesion revascularization (TLR) at 1-year follow-up. Kaplan-Meier methods were used to estimate the cumulative incidence of clinical events and construct survival curves. RESULTS: A total of 2998 (98.0%) patients completed the 1-year follow-up. The 1-year incidence of TLF was 3.10% (94/2998, 95% closed interval: 2.54-3.78%). The rates of cardiac death, non-fatal target vessel MI and clinically indicated TLR were 2.33% (70/2998), 0.20% (6/2998), and 0.70% (21/2998), respectively. The rate of stent thrombosis was 0.33% (10/2998). Age ≥60 years, diabetes mellitus, family history of coronary artery disease, acute myocardial infarction at admission, and device success were independent predictors of TLF at 1 year. CONCLUSION: The 1-year incidence rates of TLF and stent thrombosis were 3.10% and 0.33%, respectively, in patients treated with HELIOS stents. Our results provide clinical evidence for interventional cardiologists and policymakers to evaluate HELIOS stent. CLINICAL TRIAL REGISTRATION ClinicalTrials.gov, NCT03916432.
Humans ; Middle Aged ; Sirolimus/therapeutic use* ; Drug-Eluting Stents/adverse effects* ; Prospective Studies ; Cohort Studies ; Treatment Outcome ; Risk Factors ; Time Factors ; Percutaneous Coronary Intervention/adverse effects* ; Cardiovascular Agents/therapeutic use* ; Coronary Artery Disease/therapy* ; Myocardial Infarction/etiology* ; Thrombosis/complications* ; Polymers ; Registries