Objective To investigate the effects of co-transfection of the plasmid containing the wildtype (wt) INK4a/ARF gene on the chemosensitivity of human lung adenocarcinoma cell line A549. Methods Recombinant eukaryotic expression vectors pcDNA3-p16 INK4a and pcDNA3-p14 ARF were co-transfected into A549 cells by cationic liposome, in which the site of both genes was lost. By RT-PCR, immunocytochemistry and Western blotting analysis, G418 positive clone was obtained. The parental cell and negative control cell with plasmid pcDNA3-LacZ were used as controls. The 50% inhibition concentration (IC 50 ) of five commonly-used chemotherapy drugs and the apoptosis index (AI) of IC 50 were analysed respectively. Results Compared with the parental cells and negative control cells, the influence of the five drugs on IC50 was different. The values of IC 50 of doxorubicin and cisplatin decreased significantly but that of taxol, topotecan and vinorelbine remained unchanged. The apoptosis index of doxorubicin and cisplatin was much higher than that of other three drugs. Conclusion The chemosensitivity of a part of anti-cancer drugs could be changed by transfection of exogenous INK4a/ARF gene into lung adenocarcinoma cell A549.

Objective To evaluate the therapeutic efficacy and side effects of cyclopnosphamide, adriamycin, cisplatin (CAP) or CAP combined with verapamil(VPL) in chemotherapy of patients with advanced lung adenocarcinoma. Methods A total of 56 patients(male: 27, female: 29, average age: 48 years old, age range: 32-78 years old) with stage Ⅲb/Ⅳ lung adenocarcinoma confirmed pathologically from April 1998 to December 2001 were divided into two groups: group A(30 patients), treated with cyclophosphamide (CTX, 600 mg?m -2 ?d -1 ) on days 1 and 8, adriamycin (ADM, 40 mg/m 2) on day 1, and cisplatin (CDDP, 30 mg?m -2 ?d -1 ) on days 1, 2 and 3 and group B(26 patients), treated with the same dose of CTX, ADM and CDDP and an additional oral VPL treatment (60 mg t.i.d . on days 1-7). Each of the 3 cycles was repeated every 21 days. Results The patients in group A had a lower response rate (26.7% vs 38.5%, P

Objective To investigate the influence of age on liver histopathological feature of patients infected with chronic hepatitis B. Methods Liver biopsies were performed on 114 patients with chronic hepatitis B. The biochemical tests were measured by routine automated techniques. Serum hepatitis markers including HBsAg, anti-HBs, HBeAg, anti-HBe, and anti-HBc were assayed by using a microparticle enzyme linked immunosorbent assay. HBV-DNA was measured with quantitative polymerase chain reaction assay. The grades of liver necrosis/inflammation and fibrosis were compared between four groups with different age: younger than 20years, 20 ～ 30, 31 ～40, and older than 40 years. Results All 114 patients had liver histological changes with different degree. 75.4 percent (89/114) of cases had 2/over 2 grades of liver necrosis/inflammation, and 47.4 percent (54/114) of cases had 2/over 2 stages of liver fibrosis.There was no significant relation between the level of ALT and age or between the level of ALT and the grades of liver necrosis/inflammation and fibrosis( P ＞0. 05 ). The significant difference in degrees of liver inflammation and fibrosis was found among three age groups ( x2 = 30. 86, P ＜ 0. 01; x2 = 21.17; P ＜0.05 ). The grades of 1 iver inflammation and fibrosis increased with the increased age of the patients. Conclusion These results suggested that age was an independent factor for the disease progression. It was very important to undertake liver biopsy for patients with CHB more than 30 years to reveal the liver histopathological characteristics and guide the treatment.

OBJECTIVETo compare the clinical effect of the double eyelid blepharoplasty with central minimal incision and with three minimal incisions. Methods: From Jul. 2010 to May 2012, 268 cases (Group A) received double eyelid blepharoplasty with central minimal incision, while 102 cases (Group B) underwent double eyelid blepharoplasty with three minimal incisions. Photos were taken immediately, and 1,2, 4, 8,12 weeks after operation. Operation time, recovery time and postoperative complications were evaluated and recorded. The operation time and recovery time were analyzed by Wilcoxon rank sum test. The postoperative complications were analyzed by chi square test. The satisfactory degree was analyzed by t test.

RESULTSThe operation time in Group A was (25.63 ± 3.74) min, compared with that (29.90 ± 4.13) min in Group B (Z = -8.011, P <0.01). Meanwhile, the recovery time in Group A was shorter than that in Group B (Z = -15.887, P <0.01). The occurrence rate of postoperative complications,including hematoma,recurrence and scar hyperplasia in Group A was also lower than that in Group B. At the same time, the satisfactory degree in Group A was(97.302 ± 1.764), which was higher than that(88.628 10.880) in Group B (t = 12.650, P <0.05).

CONCLUSIONSThe double eyelid blepharoplasty with central minimal incision, which is suitable for all cases except those who has serious blepharochalsais, has more advantages than double eyelid blepharoplasty with three minimal incisions.

Blepharoplasty ; adverse effects ; methods ; Cicatrix ; pathology ; Eyelids ; surgery ; Hematoma ; Humans ; Hyperplasia ; Photography ; Postoperative Complications ; Prospective Studies ; Recurrence

BACKGROUNDp16INK4a and p14ARF, encoded by gene INK4a/ARF located at chromosome 9p21, are cyclin dependent kinase (CDK) inhibitors. Both p16INK4a and p14ARF are cell cycle regulatory proteins and play an important role in Rb and p53 passways respectively. In this study, wild-type INK4a/ARF gene was transfected into human lung adenocarcinoma cell line A549, in which this gene site was lost, and the effects on the cell's biological behavior were investigated.

METHODSThe recombinant eukaryotic expression plasmids pcDNA3-p16INK4a and pcDNA3-p14ARF were transfected into A549 by cationic liposome method. By RT-PCR, immunocytochemistry and Western blot after G418 selection, A549 cells that could stably express p16INK4a and p14ARF were obtained. As a control, the parental cell and negative control cell with plasmid pcDNA3-LacZ were used. Inhibition of proliferation was measured by MTT assay. The cell growth curve was drawn according to cell counts. Cell cycle distribution was measured by flow cytometry (FCM), the apoptosis indexes were observed at the same time. The colony formation rate was counted by staining the cells with Coomassie brilliant blue.

RESULTSThe introduction of exogenous INK4a and ARF caused significantly growth inhibition of A549. By FCM, more percentage of A549-p16INK4a-p14ARF cells couldn't pass through the checkpoint G1. The percentage of A549-p16INK4a-p14ARF cells inhibited at G0/G1 was 59.9%, 50.3% for A549-vector and 51.2% for A549. The statistical differences were significant between A549-p16INK4a-p14ARF cell and A549-vector cell (P=0.025) and between A549-p16INK4a-p14ARF cell and A549 cell (P=0.043). The apoptosis index of A549-p16INK4a-p14ARF cell was 8.0% and 2.7% for both A549-vector and A549 cell (P < 0.01). The colony formation ability of A549-p16INK4a-p14ARF was weaker than that of A549-vector and A549, they were 63%, 87% and 85% respectively.

CONCLUSIONSThe wild-type INK4a/ARF gene can be co-introduced effectively into A549 cell by cationic liposome method. The reexpression of p16INK4a and p14ARF in A549 can inhibit the growth and enhance the apoptosis. This trial will be helpful in using gene therapy of lung cancer in the future.

Objective: To explore the gene lifestyle interaction of ATP2B1-eNOS pathway gene polymorphisms on blood pressure. Methods: Using the convenient cluster sampling method, a total of 872 junior middle school students from 3 school in July to August 2019, were included in the final analysis. The survey included questionnaire investigation, anthropometry measurement and blood sample collection. After DNA was extracted from peripheral blood samples, the gene polymorphisms ( ATP2B1 gene rs 17249754 and rs 2070759, eNOS gene rs 1799983 and rs 2070744) were genotyped. Logistic regression model was used to analyze the association between gene polymorphism and blood pressure phenotypes. Results: The prevalence of high blood pressure was 9.52 % in adolescents(9.15% in boys and 9.87% in girls),with no significant sex difference ( χ 2=0.13, P =0.72). There were statistically significant differences between boys and girls in age, systolic blood pressure (SBP), diastolic blood pressure (DBP), body mass index (BMI) classification, birth weight, daily school physical exercise time and daily playing video games time ( P <0.05). Logistic regression analysis showed that, eNOS gene rs 2070744 polymorphism was associated with high blood pressure (HBP) under the recessive model, and the risk of HBP in CC genotype carriers were higher than that TT/TC genotype carriers ( OR=3.88, 95%CI =1.00-15.02, P < 0.05 ). The results of gene lifestyle interaction showed that ATP2B1 gene rs 2070759 polymorphism gene had an interaction with the time of physical exercise in school ( P interaction =0.05). In the subgroup with daily physical exercise time at sch ool ＜1 hour , the TT/TG genotype carriers were associated with increased risk of HBP compared with GG genotype carriers( OR= 2.65 , 95%CI =1.11-6.30, P <0.05). But in the subgroup with daily physical exercise time in school ≥1 hour, rs 2070759 was not significantly associated with HBP. Conclusion eNOS/rs 2070744 polymorphisms are associated with risk of HBP among adolescents. There is significant interaction between ATP2B1 gene rs 2070759 polymorphism and physical exercise time in school on HBP. Adolescents should spend more time on physical activity in school, which will help to maintain normal blood pressure level.

Objective: To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the detection of serum oleic acid (OA), and preliminarily evaluate the role of OA in insulin resistance (IR) of type 2 diabetes (T2DM). Methods: OA-［ 13 C 5 ］ was used as isotope-labeled internal standard, and the ion pairs of OA and OA-［ 13 C 5 ］ were 281.3/281.3 and 286.3/286.3, respectively. The ultrapure water was used as mobile phase A and methanol: acetonitrile (1∶1, v/v) as mobile phase B in a ZORBAX SB-Aq C18 reversed phase column. Meanwhile, the gradient elution system with a flow rate of 0.3 mL/min was used. According to the CLSI guidelines (EP15-A3), the reliability of the established method was evaluated by detecting the performance indicators such as precision, trueness, linear range, stability and carrying contamination rate. Serum OA levels were detected by the established HPLC-MS/MS method in 109 patients with clinically diagnosed T2DM and 100 healthy controls. The insulin resistance index (HOMA-IR) was calculated to evaluate IR, and the relationship between OA and IR was further analyzed. Results: The established HPLC-MS/MS method for the detection of serum OA had good specificity and linearity in the range of 10-1 000 μmol/L (y=0.007 55x+0.004 83,r=0.997 7), and the low limit of quantification (LLOQ) was 10 μmol/L. It also had good precision, and the within-run coefficient of variation (CV) and total CV were not more than 1.62% and 1.73%, respectively, indicating that the method was suitable for the detection of serum OA. The serum OA levels in T2DM patients ［(425.58 ± 220.17) μmol/L］ were significantly higher than that in the healthy controls ［(113.20±58.00) μmol/L］, and serum OA levels were significantly correlated with HOMA-IR in T2DM patients and healthy controls. The area under the receiver operating characteristic (ROC) curve (AUC) of OA for the diagnosis of IR was 0.689. When the cut-off value identified by Youden index was 235.8 μmol/L, the sensitivity and specificity were 70.4% and 63%, respectively. When OA combined with fasting blood glucose (FBG) to diagnose IR, the AUC increased to 0.806, which was significantly higher than that of OA (P＜0.05). Conclusion A scientific and efficient HPLC-MS/MS method for the quantitative detection of serum OA is established successfully, which provides a reliable method for the dynamic monitoring of the changes of OA levels in the patients with metabolic diseases.