AIM:To explore the effects of small interfering RNA(siRNA) targeting PCNA gene on nasopharyngeal carcinoma CNE2 cells growth and cycle.METHODS:Three synthesized siRNA targeting PCNA gene was transfected into CNE2 cells by using LipofectamineTM reagent.The PCNA mRNA and PCNA protein were detected by real-time polymerase chain reaction(RT-PCR) and immunohistochemical method.Inverted phase contrast microscope was used to determine the CEN2 cells growth before and after PCNA-siRNA transfected.Flow cytometry was used to observe the cell cycle.RESULTS:In CNE2 cells after PCNA-siRNA transfection,the expressions of PCNA mRNA and protein were down-regulated at different degree.Inhibition ratio of PCNA mRNA was 98.5%.Meanwhile,the cell cycle was suffocated at G0/G1 stage.CONCLUSION:The synthesized PCNA-siRNA effectively interferes nasopharyngeal carcinoma cells by down-regulating the expressions of the PCNA mRNA and its protein,therefore inhibits the growth of CNE2 cells.Future application of PCNA-siRNA in the gene therapy of nasopharyngeal carcinoma might be expected.

Adolescent ; Diagnosis, Differential ; Disorders of Sex Development ; Female ; Humans ; Male ; Neoplasms, Germ Cell and Embryonal ; pathology ; radiotherapy ; surgery ; Seminoma ; pathology ; Testicular Neoplasms ; pathology ; radiotherapy ; surgery

This paper introduces a new method which controls discharge energy accurately. It is achieved by calculating target voltage based on transthoracic impedance and accurately controlling charging voltage and discharge pulse width. A new defibrillator is designed and programmed using this method. The test results show that this method is valid and applicable to all kinds of external defibrillators.
Defibrillators ; standards ; Equipment Design

With the development of medical education,the traditional cell biology teaching modes and methods need constant adjustment to adapt to the current teaching.In view of the present high-speed development of cell biology,we seriously picked some representative themes to carry out thematic teaching.Students were encouraged to read some references about the corresponding content and thought primarily before the class.After the lesson,the teacher guided students to discuss and find the answers to the questions they asked before.Participation in class discussion and homework completion accounted for 10％ of the final assessment results.The thematic teaching helped to optimize classical teaching contents and frontier progress.This teaching mode not only stimulated learning interest but also fully exercised learning ability.

ObjectiveTo investigate the protein expression of ERK1/2,p-ERK1/2,HIF-1α and α-enolase in hypertrophic cardiomyocytes induced by ET-1 and explore the regulation mechanism of overexpression of α-enolase in hypertrophic cardiomyocytes.MethodsET-1-induced abnormal cardiomyocytes were used as model of cardiac hypertrophy.Cellsurface area, [<'3>H]-leucine incorporation and the actin staining were measured to determine the extent of hypertrophy. Cultured cardiomyocytes were divided into 4 groups at random, control group, PD98059 treated group, ET-1 treated group and PD980594- ET-1 treated group. The protein expressions of ERK1/2, p-ERK1/2,HIF-1α and α-enolase were detected by immunoblotting analysis.ResultsCompared with the control group , cell surface area and [<'3>H] leucine incorporation were increased in ET-1 treated group ((1350.7±107.5)μtm<'2> vs. (896.1±70. 2)μtm<'2> , P<0.05; (1387.9±14.8) dpm vs. (787.7±10.2)dpm,P<0.013. Actin staining showed that ET-1-treated cardiomyoeytes had more intense actin staining and clear cross-striations than did control group, which suggested that myocardial cell hypertrophy could be induced by ET-1 in WKY neonatal cardiomyocytes. After MEK 1/2 inhibitor PD98059 was used, the cell surface area and [<'3>H] leucine incorporation were decreased in PD980594-ET-1 treated group compared with ET-1 treated group[(907.0±92.5)μm<'2> vs. (1350.7±107.5)μm<'2>;(841.5±10. 5)dpm vs. (1387.9±14.8)dpm, both P<0.05], which suggested that myocardial cell hypertrophy could be regulated by ERK1/2 signal pathway. Immunoblotting analysis showed that the protein expressions of p-ERK1/2, HIF-1α and α-enolase increased after ET-1 treatment,while PD98059 as an inhibitor of the upstream kinase of ERK1/2 was used, the protein expressions of HIF-1α and α-enolase were partially inhibited.ConclusionsET-1 induces hypertrophic cardiomyocytes through ERK1/2 phosphorylation in cultured neonatal rat cardiac myocytes.ERK1/2 and HIF-1α signal pathway may play an important role in the overexpression of α-enolase in the hypertrophic cardiomyocytes.

ObjectiveTo investigate changes of the cellular immune function in the elderly patients with nonsmall-cell lung cancer (NSCLC) after chemotherapy.Methods T-lymphocyte subsets and natural killer cell were detected in 29 elderly patients with NSCLC,20 adults with NSCLC and 22 healthy elderly,and their levels were compared between pre-chemotherapy and at the end of 2 cycles of chemotherapy in the elderly patients with NSCLC.ResultsThe levels of CD3,CD4,CD8,CD4/CD8andNK cell were (58.9±15.8),(32.3±12.7),(22.0±9.8),(1.3±0.7),(21.6± 7.7),respectively in the elderly patients with NSCLC,(65.9 ± 7.2),(38.5 ± 7.6),(23.1 ± 9.2),(1.5±0.7),(16.8±6.2),respectively in adults with NSCLC and (67.3±9.0),(39.0±7.8),(23.9±9.3),(2.0±1.6),(22.5±5.8),respectively in healthy elderly.The levels of CD3 and CD4 were decreased (t=2.109,2.159,P＜0.05) and NK cell was increased (t=2.273,P＜0.05) while CD8 and CD4/CD8 had no difference(t = 0.406,0.736,P＞ 0.05 ) in the elderly patients with NSCLC as compared with adults with NSCLC.The levels of CD3,CD4,and CD4/CD8 were lower (t = 2.234,2.200,2.016,all P＜ 0.05) in elderly patients with NSCLC than in healthy elderly,with no significant change in the levels of CD8 and NK cell(t= 0.700,0.474,P＞0.05) between the two groups.The levels of CD3 (51.6 ±10.3)was reduced(t=2.067,P＜0.05) and CD4 (31.7 ± 11.7),CD8(21.6 ± 6.5),CD4/CD8 (1.3 ± 0.7),NK cell (26.0 ±12.7)had no remarkable difference (t =0.186,0.180,0.289,1.570,all P＞ 0.05)after chemotherapy in elderly patients with NSCLC.ConclusionsThe cellular immune function in the elderly patients with NSCLC is lower than in adults with NSCLC and healthy elderly,and further decreases after chemotherapy.

Objective The aim of this study is to establish a practical VX2 tumour model less than or equal to 10 mm from large blood vessel(as standard) for HIFU ablation .Methods 15 New Zealand rabbits were involved ,VX2 tumour blocks were inoculated near postcava through spine path ,and tumour rate was observed two weeks later by anatomy and MRI .Results Three experimental rabbits did not survive ,all the rest of the 12 into the tumour ,assembly tumour rate was 100% (12/12);The tumour rate reaching the standard accounted for 75% (9/12) ,the average distance between the tumour and the inferior vena cava was (5 .6 ± 3 .4)mm . Conclusion It is feasible to establish the VX2 tumor model less than or equal to 10 mm from large blood vessel through spine path .

Objective To explore the relationship between sex hormone-binding globulin (SHBG) of gestational diabetes mellitus ( GDM ) pregnant women with well-controlled glucose and pregnancy outcomes. Methods Two hundred and fifty-one GDM pregnant women of 24 - 28 weeks in Shengjing Hospital of China Medical University were recruited from Mar. 2005 to Mar. 2010. Two hundred and sixteen cases of GDM with well-controlled glucose were defined as glycemic satisfied group, and they were treated by diet therapy ( 169 cases) or insulin therapy (47 cases) . Thirty-five cases with unsatisfied glucose were defined as glycemic unsatisfied group. One hundred and ninety-two healthy pregnant women of 24 - 28 weeks were defined as healthy control group. Serum SHBG and homeostasis model analysis of insulin resistance ( HOMA-IR) at 24 - 28 weeks and above 36 weeks were measured. GDM was diagnosed bytwo-step method according to the National Diabetes Data Group ( NDDG) criteria. The pregnancy outcomes and complications of the three groups were recorded. Results ( 1 ) Comparison of pregnancy outcomes and complications: glycemic satisfied group was less likely to develop hypertensive disorders in pregnancy ( 10. 6% ) , premature birth(8. 3% ) ,large for gestational age ( LGA) (8. 8% ) , neonatal asphyxia(3. 7% ) and neonatal hypoglycemia ( 2. 3% ) compared to glycemic unsatisfied group ( 42. 9% , 34. 3% , 31. 4% , 22. 9% and 11. 4% ,respectively). And the difference was statistically significant (P ＜0. 05 or P ＜0. 01). There was no significant difference for incidence of polyhydramnios, pueperal infection, postpartum hemorrhage, neonatal hyperbilirubinemia between the two groups ( P＞ 0. 05 ) . When compared to healthy control group(7. 3% ,2. 1% ,4. 2% ,2. 1% and 1. 6% ) ,no significant difference was found for incidence of premature birth( 8. 3% ) , pueperal infection ( 3. 2% ) , postpartum hemorrhage (5. 1% ) , neonatal asphyxia (3. 7% )and neonatal hypoglycemia(2. 3% ,P ＞0. 05). (2) Comparison of results of 24 - 28 weeks and above 36 weeks: serum SHBG of glycemic satisfied group [( 384 ± 88 ) , (457 ± 48 ) nmol/L]was significantly higher than that of glycemic unsatisfied group[(313 ±45) ,(401 ±73) nmol/L];HOMA-IR of glycemic satisfied group (5. 3 ±1.1,5.5 ±1.1) was significantly lower than that of glycemic unsatisfied group (7. 0 ± 1. 3 ,7. 6 ± 1. 7 ; P ＜ 0. 01). Serum SHBG of glycemic satisfied group was significantly lower than that of healthy control group [( 492 ± 95 ) , (565 ± 40 ) nmol/L]; and HOMA-IR of glycemic satisfied group(5. 3 ± 1. 1,5. 5 ± 1. 1) was significantly higher than that of healthy control group (3. 6 ±0. 6,3. 9 ± 0. 5 ;P ＜ 0. 01 ) . FPG of glycemic satisfied group [( 5. 84 ± 0. 28 ) , ( 5. 16 ± 0. 13 ) mmol/L]was significantly lower than that of glycemic unsatisfied group [(6. 13 ± 0. 16 ) , ( 5. 68 ± 1. 14) mmol/L; P ＜ 0. 01]. FINS of glycemic satisfied group [( 20. 4 ± 2. 1 ) , ( 24. 1 ± 4. 2 ) mmol/L]was significantly lower than that of glycemic unsatisfied group [(24. 7 ± 4. 5 ) , ( 29. 9 ± 2. 7 ) mmol/L; P ＜ 0. 01]. ( 3 ) Correlation analysis. Between 24 - 28 weeks, SHBG was negatively correlated with HOMA-IR in the three groups ( r = -0. 952, P ＜0. 01) ; and SHBG was negatively correlated with HOMA-IR in glycemic satisfied group ( r = -0. 903, P ＜0. 01). Conclusions Well-controlled glucose can not completely improve maternal and fetal outcomes of GDM pregnant women. High insulin resistance and low serum SHBG can influence pregnancy outcomes.

Objective To investigate the effects of sex hormone-binding globulin (SHBG)gene in the apoptosis of human trophoblastic cells.Methods The siRNA specific-targeting SHBG gene was transfected into human trophoblastic cells and they were divided into six groups:trophoblasts without transfection in normal control groups(group Ⅰ);transfect liposome in blank control groups(group Ⅱ);transfect nonspecific siRNA in negative control groups(group Ⅲ);transfect SHBG siRNA-Ⅰ,SHBG siRNA-Ⅱ,SHBG siRNA-Ⅲ respectively in trans-fection group(group Ⅳ,Ⅴ,Ⅵ).Hoechst 33258 dying method was used to detect cell apoptosis.SHBG and Caspase-3 mRNA profiling and the level of SHBG and caspase-3 protein were detected by real-time PCR and Western blot.Results There was no statistical significant difference in the gene expression and protein level of SHBG and caspase-3 in group Ⅰ,Ⅱ and Ⅲ (P >0.05).In Ⅳ,Ⅴ and Ⅵ group,there was no statistical significant difference in the expression level of SHBG and caspase 3 (P >0.05).Compared with group Ⅰ,Ⅱ and Ⅲ,the a-mount of SHBG gene expression decreased obviously,the caspase-3 mRNA and protein level increased obviously and the trophoblast cell ap-optosis increased markedly (P <0.05).Conclusion Through siRNA interference technology can reduce SHBG gene expression in human trophoblastic cells,and it can lead to excessive apoptosis of human trophoblasts cells.

Objective To investigate the cellular immune function changes and the effect of thymosin alpha-1 on the changes in elderly patients with severe pneumonia. Methods T cell subset and natural killer (NK) cell were detected in 66 elderly patients with severe pneumonia and 34 elderly patients with common pneumonia. The severe pneumonia patients were randomly divided into 2 groups: the treatment groups (34 cases) and the control group (32 cases). All patients received conventional therapy of pneumonia. The treatment group received 1.6 mg of thymosin alpha-1 through subcutaneous injection once a day for a week and twice a week later. Results The levels of CD3, CD4, CD8 and NK cell were lower in elderly patients with severe pneumonia than in patients with common pneumonia [(43.54%±18.97%) vs. (45.46%±10.43%), (25.43%±12.72%) vs. (38.47%±8.20%), (16.68%±9.30%) vs. (22.36%±8.06%), (13.52%±4.66%) vs. (17.87%±7.11%), t=-6.779、-5.85、-3.161、-3.285 respectively all P＜0.05]. The levels of CD3, CD4, CD4/CD8 and NK cell increased significantly after treatment in treatment group [(64.22%±5.53%) vs. (61.53%±13.41%), (31.70%±4.38%) vs. (26.07%±4.31%), (1.27%±0.91%) vs. (0.97%±0.22%), (17.67%±4.56%) vs. (15.44%±3.82%), F=5.591,11.526,8.934,4.564 respectively, all P＜0.05]. The duration of antibiotic injection and length of stay were lower in treatment group than in control group [(14.17±2.51) d vs. (14.42±2.79) d, (12.69±2.80) d vs. (15.04±3.58) d, t=-3.152、-2.690 respectively, all P＜0.05]. Conclusions The immune function of the elderly patients with severe pneumonia is lower. Thymosin alpha-1 can improve the immune function of the elder patients with severe pneumonia and is helpful for controlling an infection.