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 discuss the clinical treatment methods of Kaposiform hemangioen-dothelioma (KHE).Methods:Eleven children with KHE admitted to the Union Hospital of Fujian Medical University from November 2009 to November 2015 were retrospectively analysed. Children aged from 5 days to 2 years old, 8 males and 3 females. In the treatment of cases 1-5, glucocorticoids and propranolol were routinely used for treatment regardless of the platelet count. The glucocorticoids was taken orally 4.0-4.5 mg/kg every other day, and the maximum daily dose of propranolol was 2 mg/kg given by three times a day. In the treatment of cases 6-11, the critical value of platelets number was 20×10 9/L. For those higher than the critical value, glucocorticoids and propranolol were routinely given. For those lower than the critical value, under the premise of taking glucocorticoids and propranolol, sirolimus was givenorally by the dose of 0.8 mg/m 2 twice a day. Two of the children had been treated with vincristine at a dose of 0.05 mg/m 2. The treatment effect was observed and followed up. Results:Followed up for three to eight years. Three of eleven cases did not get thrombocytopenia, and the effect of glucocorticoids combined with propranolol treatment was good. One case’s platelets number was 20×10 9/L. The tumor dissappered after treated with glucocorticoids combined with propranolo. Seven cases with platelets was lower than the critical value. Five in seven cases were treated with glucocorticoids, propranolol and sirolimus. The effect was good. One case, treated with glucocorticoids, propranolol and vincristine, died of cerebral hemorrhage. Another one case was lost to follow-up after treatment with glucocorticoids and propranolol. Conclusions:With the critical value of platelet criticality, patients above this value are routinely treated with glucocorticoid and propranolol first while those below this value are treated with sirolimus while taking glucocorticoids and propranolol. Through this method, better results can be achieved.

Objective:To discuss the clinical treatment methods of Kaposiform hemangioen-dothelioma (KHE).Methods:Eleven children with KHE admitted to the Union Hospital of Fujian Medical University from November 2009 to November 2015 were retrospectively analysed. Children aged from 5 days to 2 years old, 8 males and 3 females. In the treatment of cases 1-5, glucocorticoids and propranolol were routinely used for treatment regardless of the platelet count. The glucocorticoids was taken orally 4.0-4.5 mg/kg every other day, and the maximum daily dose of propranolol was 2 mg/kg given by three times a day. In the treatment of cases 6-11, the critical value of platelets number was 20×10 9/L. For those higher than the critical value, glucocorticoids and propranolol were routinely given. For those lower than the critical value, under the premise of taking glucocorticoids and propranolol, sirolimus was givenorally by the dose of 0.8 mg/m 2 twice a day. Two of the children had been treated with vincristine at a dose of 0.05 mg/m 2. The treatment effect was observed and followed up. Results:Followed up for three to eight years. Three of eleven cases did not get thrombocytopenia, and the effect of glucocorticoids combined with propranolol treatment was good. One case’s platelets number was 20×10 9/L. The tumor dissappered after treated with glucocorticoids combined with propranolo. Seven cases with platelets was lower than the critical value. Five in seven cases were treated with glucocorticoids, propranolol and sirolimus. The effect was good. One case, treated with glucocorticoids, propranolol and vincristine, died of cerebral hemorrhage. Another one case was lost to follow-up after treatment with glucocorticoids and propranolol. Conclusions:With the critical value of platelet criticality, patients above this value are routinely treated with glucocorticoid and propranolol first while those below this value are treated with sirolimus while taking glucocorticoids and propranolol. Through this method, better results can be achieved.

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 investigate a simple,minimally invasive and effective operative method for the facial rejuvenation in the upper eyelid and midface region.Methods Blepharoplasty was combined with suspending orbicularis oculi muscle flap and fixing it on the periosteum underneath the eyebrow through eyebrow incision.Meanwhile,for midface rejuvenation,inferior palpebral margin incision was performed and prezygomatic interspace was separated completely under the orbicularis oculi muscle.The under-eye puffiness and tear trough deformity were corrected through releasing orbital fat,reposition and fastening orbital septum,and transposition of orbicularis oculi muscle flap.And the deep sulci nasolabialis and cheek anetoderma were relieved by dual lifting of malar fat pad and orbicularis oculi muscle flap.Follow-up was taken at the 1 week,3 months,6 months,1 year,2years and 3 years after operation.Each case was evaluated with postoperative effect,reprocessing time and postoperative complications and underwent photography.Results From Feb.2010 to Oct.2014,190 patients (9 male,181 female,an average age of 49.03 ± 5.67 years) underwent this operation.Obvious improvement on the upper eyelid and midface region was achieved in all the patients after operation without serious or irreversible complication.Conclusions Combining eyebrow and inferior palpebral margin incision,through suspending the malar fat pad and orbicularis oculi muscle flap at the same time,as a simple,minimally invasive and reliable method,can strengthen the effect of the facial rejuvenation in the upper eyelid and midface region markedly.

Objective:To investigate the effect of orbital septal fat replacement combined with orbicularis oculi muscle flap transposition in correction of tear trough deformity.Methods:The clinical data of patients with tear trough deformity underwent surgical correction admitted to Fujian Medical University Union Hospital from January 2009 to January 2020 were retrospectively analyzed. According to the surgical methods, they were divided into the combined group (orbital septal fat replacement combined with orbicularis oculi muscle flap transpose) and the simple group (orbital septal fat replacement). Surgical methods: using percutaneous approach, the orbital septum fat was separated and replaced under the orbicularis oculi muscle to fill the tear trough deformity, and the orbicularis oculi muscle flap with the transfer pedicle on the medial side was added to fill the medial tear trough deformity in the combined group. Before and 6 months after surgery, Hirmand scale was used to rate the tear trough deformity(grade Ⅰ, Ⅱ, Ⅲ, severity is increasing from medial to lateral), and the lower eyelid without obvious tear trough deformity was rated as grade 0. The improvement rate and the complete improvement rate were calculated in the two groups. The related complications were followed up within 1 month after operation. Chi-square test or Fisher’s exact test was used to compare Hirmand improvement rate at all levels and complete improvement rate between groups. P< 0.05 was considered statistically significant. Results:A total of 175 patients (350 lower eyelids) were enrolled. There were 104 patients (208 lower eyelids) in the combined group [16 males and 88 females, aged (34.3±8.3) years old], including 47 cases of Hirmand grade Ⅰ, 54 cases of Hirmand grade Ⅱ and 107 cases of Hirmand grade Ⅲ before operation. There were 71 patients (142 lower eyelids) in simple group [7 males and 64 females, aged (33.2±8.6) years old], including 24 cases of Hirmand grade Ⅰ, 32 cases of Hirmand grade Ⅱ and 86 cases of Hirmand grade Ⅲ before operation. At 6 months after operation, the improvement rate of grade Ⅰ cases in the combined group was 91.5% (43/47), which was higher than that in the simple group (54.2%, 13/24) ( P<0.01), while there was no significant difference in the improvement rate of grade Ⅱ and Ⅲ cases between the two groups ( P>0.05). The complete improvement rates of grade Ⅰ, Ⅱ and Ⅲ cases in the combined group were 91.5% (43/47), 77.8% (42/54) and 72.0% (77/107), respectively, which were higher than those in the simple group [54.2% (13/24), 25.0% (8/32) and 24.4% (21/86)], and the differences were statistically significant (all P<0.01). Postoperative complications included ectropion, hematoma, ecchymosis, and local edema for more than 2 weeks. There was no significant difference in the incidence of complications between the two groups ( P>0.05). Conclusions:Orbital septal fat replacement combined with orbicularis oculi muscle flap transposition can improve all levels of tear trough deformity, and the improvement effect of medial lacrimal groove depression is significantly better than that of orbital septal fat replacement alone.