MicroRNAs (miRNAs) are a class of small non-coding RNAs which involve in the regulation after gene transplantation.Researches show that miRNA is closely related to the development and progression of non-small-cell lung cancer (NSCLC).They caused a significantly change in NSCLC cells through downregulate tumor suppressor gene,or act as oncogene or work on some targets which is important in cell signal pathway,in the end it lead to oncogenesis,cell proliferation,cell apoptosis and resistance to chemotherapy or radiotherapy.Therefore,miRNA will be hopefully used in the diagnose and therapy of cancer.

BACKGROUND: Osteosarcoma cell OS-9607 is a cell line cultured from human osteosarcoma tissue. To construct cDNA library of this cell line so as to clone target gene from it.OBJECTIVE: To construct cDNA library of osteosarcoma cell OS-9607 by switching mechanism at 5' end of RNA transcript (SMART) technique and analyze its quality.DESIGN: Verified experimental study SETTING: Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA MATERIALS: This experiment was carried out at the Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA in May 2005. Osteosarcoma cell OS-9607 was cultured in CO2 incubator. Cells were pre-treated with DM for 30 minutes, recovered for 24 hours in standard culture medium. The whole RNA of osteosarcoma cell OS-9607 was isolated by RNAease reagent kit. The quantity and integrity of total RNA was detected by ultraviolet spectrometer and electrophoresis on a denaturing formaldehyde agarose gel stained by ethidium bromide (EtBr). METHODS: Total RNA was extracted from human osteosarcoma cell OS9607 and mRNA was isolated, cDNA of OS-9607 cells was acquired by in situ polymerase chain reaction (PCR) and long-distance PCR (LD-PCR),then the cDNA was ligated with dephosphorylated λ phage vector. The recombinants were constructed with SMART cDNA library construction kit.The size of cDNA inserts and the diversity of library were detected by PCR.MAIN OUTCOME MEASURES: The quality and integrity of total RNA, the titer of un-amplified and amplified library and recombination fraction; analysis of cDNA length of library RESULTS: ①The concentration of the isolated total RNA was determined and the measuring absorbance was between 260 nm and 280 nm. Its absorptance was about 1.9 A. 0.8％ denaturing formaldehyde agarose gel electrophorosis showed clear 28S rRNA and 18S rRNA,and the brightness rate of 28S rRNA and 18S rRNA (28S/18S) was 2:1. ②After linked to λ phage in vitro, packed and transfected host bacteria, the titer of unamplified library was 2.2×107 pfu/mL, and the titer of amplified 1 ibrary arrived to 4.5×1010 pfu/mL. The recombination fraction analysis showed that there were 8 blue plaques and 1 360 colorless plaques in one plate, so the recombination fraction was 99.5％. ③ The obtained cDNA library consisted of 1.6×106 recombinant bacteriophages and the average exogenous inserts of the recombinants was 1.5 kb.CONCLUSION: These results suggest that the obtained osteosarcoma cell OS-9607 cDNA library has high quality, which sets up a firm foundation for the further research on osteosarcoma cell OS-9607 and its elated genes screen.

Objective To study the significance of thyroid stimulating antibody (TSAb) and thyroid stimulating-blocking antibody (TSBAb) in children with Graves' disease (GD) or Hashimoto's thyroiditis (HT).Methods Five hundred and twenty-seven cases of serum from 180 children with autoimmune thyroid disease (AITD) children were divided into 282 cases of GD and 245 cases of HT.According to the status of thyroid function,they were divided into 157 cases of hyperthyroidism,91 cases of hypothyroidism and 279 cases of normal thyroid.GD group was subdivided into 127 GD hyperthyroidism and 155 GD remission;HT group was subdivided 30 HT hyperthyroidism,124 HT remission and 91 HT hypothyroidism.Seventy-nine healthy children were taken as the healthy control group.Free triiodothyronine(FT3),free thyroxine(FT4) and sensitive thyroid stimulating hormone(TSH) were detected by chemoluminescence.Serum TSAb and TSBAb were detected by serum TSAb or TSBAb enzyme linked immunosorbent assay (ELISA),respectively.The differences in TSAb and TSBAb among each group were compared and analyzed of find out the relationship between TSAb and TSBAb was performed.Beside,the correlation between TSAb and TSBAb with FT3,FT4,and TSH were analyzed.Results (1) TSAb levels were significant (F =11.995,all P =0.000):the GD group (0.727 ± 0.157) ＞ HT group (0.605 ± 0.148) ＞ healthy control group (0.350 ± 0.105);the difference was significant(F =109.165,P =0.000) among hyperthyroidism group (0.745 ± 0.169) ＞ normal thyroid group (0.647 ± 0.153) ＞hypothyroidism group(0.612 ±0.144) ＞healthy control group (0.350 ±0.105);the difference was significant(F=156.712,P =0.000) in the GD hyperthyroidism group(0.747 ±0.17) ＞ GD remission group (0.640 ± 0.16) ＞ healthy control group (0.350 ± 0.105);the difference was significant (F =109.165,P =0.000) in the HT hyperthyroidism group(0.739 ±0.140) ＞HT remission group(0.655 ±0.135) ＞ HT hypothyroidism group(0.612 ± 0.140) ＞healthy control group (0.350 ±0.105).(2) TSBAb levels were significantly different(F =15.610,P =0.000):the HT group(0.704 ±0.633) ＞ GD group(0.567 ±0.178) ＞ healthy control group (0.334 ±0.104);the difference was significant(F =13.311,P =0.000) in the hypothyroidism group (0.693 ± 0.125) ＞ remission group (0.648 ±0.446) ＞hyperthyroidism group(0.562 ±0.181) ＞healthy control group(0.334 ±0.104);the difference was significant(F =19.269,P =0.000) in the GD remission group (0.672 ±0.572) ＞ GD hyperthyroidism group (0.550 ± 0.187) ＞ healthy control group (0.334 ± 0.104);HT hypothyroidism group (0.693 ± 0.725) was higher than HT hyperthyroidism group(0.618 ±0.142) and HT remission group (0.619 ±0.199),the difference was not significant between HT hyperthyroidism group and HT remission group(F =12.208,P =0.000).(3) TSAb level was positively correlated with TSBAb,FT3 and FT4(r =0.162,0.091,0.194,all P ＜ 0.05) and was negatively correlated with TSH (r =-0.224,P ＜ 0.05).TSBAb levels were negatively correlated with FT3 (r =-0.155,P ＜ 0.05) and was positively correlated with TSH (r =0.131,P ＜ 0.05).Conclusions Thyroid function was related to the serum levels of TSAb and TSBAb.TSAb and TSBAb could be regarded as an important predictive index for children with AITD during the treatment period.

Objective To evaluate the diagnostic value of the diameter ratio of left subclavian artery to aortic isthmus(LSA/AoIS)in fetuses with aortic coarctation(COA).Methods A retrospective study of 79 fetal echocardiographic data was undergone on 49 COA fetuses.The COA cases were divided into four groups according to different gestational week (cases in each group including follow-up review of different gestational weeks):group 1 ,24-27+6 W;group 2,28 -31+6 W;group 3,32 -35+6 W;group 4,36 -39+6 W.The normal fetuses with gestational age matched were also divided into four control groups.The diameter of left subclavian artery(LSA) and aortic isthmus (AoIS) were measured,the ratio of LSA/AoIS of each group were calculated,and the data between the COA and control groups were compared.ROC curve analysis was used for LSA/AoIS to predict the demarcation point of postnatal COA.Results ①There was no statistic difference in the LSA diameter between COA groups and control groups (P >0.05):group 1 (1 .87±0.42)mm vs (1 .75 ±0.25)mm;group 2 (2.25 ±0.36)mm vs (2.21 ±0.22)mm;group 3 (2.74±0.32)mm vs (2.90 ±0.29)mm;group 4 (2.83 ±0.28)mm vs (3.06 ±0.30)mm;②The ratio of LSA/AoIS was significantly increased in COA groups than those in control groups:group 1 (0.88± 0.15) vs (0.66±0.06),group 2 (0.85±0.13)vs(0.64±0.05),group 3 (0.94±0.17)vs(0.73±0.07), group 4 (0.94±0.18)vs(0.70±0.07),the difference was statistically significant(P <0.05);③The cut-off value of LSA/AoIS ratio were 0.78,0.73,0.83,0.78 in each group,respectively.Conclusions The ratio of LSA/AoIS can be used as a useful echocardiographic parameter which suggest the presence of aortic coarctation.This ratio≥0.8 may have a certain diagnostic significance.

Objective To explore the effect of glucocorticoid on glucose transport activity and the expression of insulin signaling peptides in the primary cultured rat adipocytes .Methods Isolated rat adipocytes were cultured for 2h,8h,16h and 24h respectively at 5 mmol glucose with dexamethasone (Dex) 0 3?mol. Then the glucose uptake, cellular contents of insulin receptor substrate (IRS) 1/2, phosphatidylinositol 3-kinase 85 subunit (p85) and protein kinase B(PKB)were measured by Western blotting.Results These adipocytes treated with Dex have shown to impair the basal and insulin-induced glucose uptake with a maximum inhibition at 2h and 24h respectively; Dex down-regulated IRS1 protein expression in a time-dependent manner and up-regulated IRS2 content. The cellular p85 and PKB contents were also decreased by Dex. Conclusion Chronic exposure to glucocorticoid can inhibit glucose uptake and induce insulin resistance. The mechanism may be involved in affecting the expression of insulin signaling peptides.

Objective To investigate the relationship between the levels of total serum IgE and IL-4 in patients with seasonal contact dermatitis. Methods Total serum IgE was tested by immunoradiometric assay(IRMA) in 29 cases of seasonal contact dermatitis, 35 cases of allergic contact dermatitis and 24 cases of healthy controls. Production of IL-4 was detected by ELISA from peripheral blood mononuclear cells (PBMC) cultured with PHA for 48 hours. Results The levels of IL-4 secreted by PBMC and total serum IgE were significantly higher in patient with seasonal contact dermatitis than those in control group (P

Objective To explore the molecular mechanism of glucocorticoid and high glucose-induced insulin resistance(IR).Methods Isolated rat adipocytes were cultured for 24h at 5 or 25 mM glucose with or without dexamethasone (Dex) 0 3?M. Then the glucose uptake , the phosphorylation of tyrosine of insulin receptor substrate (IRS)1/2 and expression of IRS 1/2 and protein kinase B(PKB) protein were measured by western blot analysis with molecular dynamics. Results These adipocytes treated with 25 mM glucose have shown to impair glucose uptake, IRS1 phosphorylation and the protein expression; Combined treatment with Dex enhanced high glucose-induced the suppression, and inhibited IRS1 tyrosine phosphorylation; High glucose increased IRS2 protein expression, Dex abolished partly its the effect. Conclusions High glucose can induce IR, Dex can aggravate the effect induced with high glucose. The mechanism may be involved in affecting the phosphorylation and expression of insulin signaling proteins.

Objective To discuss blood collection tubes with different additives and their effects on the testing results of alcohol concentration in blood sam ples. Methods Blood sam ples from 10 volunteers were collected 2 hours after drinking with seven different types of disposable vacuum blood collection tubes, including ordinary tube without anticoagulant, coagulant tube, separating gel-coagulant tube, sodi-um citrate (1∶4) tube, sodium citrate (1∶9) tube, sodium citrate (9∶1) tube and EDTA-K2 tube. The al-cohol concentrations in these blood sam ples were analyzed by headspace gas chrom atography. Results The concentration testing results of the sam e blood sam ples in different types of tubes were different from one to another. The sequence was as follows:separating gel-coagulant tube>coagulant tube>ordi-nary tube without anticoagulant>EDTA-K2 tube>sodium citrate (1∶9) tube>sodium citrate (1∶4) tube, whereas the results of the sam e blood sam ple in sodium citrate (1∶9) tube and sodium citrate (9∶1) tube showed no obvious difference. Conclusion It is better to collect a suspicious drunk driver’s blood sam-ple using a disposable vacuum blood collection tube, with the EDTA-K2 tube being preferred.

Objective] To sum up professor He Ruoping’s clinical experience and academic views in the treatment of bone marrow suppression after chemotherapy in patients with malignant tumor. [Method] Through the way of clinical practice with teacher, arranging medical records and clinical observation, systematically arranging He Ruoping’s clinical experience and academic perspective in the aspects of etiology and pathogenesis, treatment ideas and drug characteristics. And further explain the characteristics of the treatment of He through case analysis.[Result] He Ruoping believes that the treatment of bone marrow suppression after chemotherapy related closely with spleen and kidney, and is also related with interweaving of toxicity, deficiency and blood stasis. The treatment should be based on kidney, spleen, centralizer, make Qi and blood sufficient. Besides, supplemented by the Quxie treatment, make Qi and blood flow.[Conclusion] Using Yishen Jianpi Fuzheng, supplemented by eliminating method in treating post chemotherapy bone marrow suppression, has significant clinical effect, has the value of popularization and application.

Objective To investigate the protective effects of cardiomyopeptidin (CMP) pretreatment against hippocampal neuronal injury caused by anoxia-reoxygenation and the underlying mechanism. Methods Hippocampal neurons were isolated from neonatal SD rats and cultured for 9-10 days. The cultured primary hippocampal neurons were randomly divided into 4 groups: ( I ) control group; ( II ) anoxia-reoxygenation group (A-R); (I) CMP 10?g?ml-1 group (CMP1) and (IV) CMP 100 ?g?ml-1 group (CMP2). In A-R group hippocampal neurons were subjected to 4 h anoxia (cultured in 95 % N2 + 5 % CO2 ) and 48 h reoxygenation (cultured in aerobic environment). In group III and IV CMP was added to culture media with end-concentration of CMP 10 ?g?ml-1 or 100?g?ml-1 before oxygen-deprivation. Neuronal viability was assessed by MTT method and Bcl-2 protein expression was determined by immune-cytochemistry.Results The optical density (OD) value of hippocampal neurons and the Bcl-2 expression were significantly higher in group IV (CMP2) than those in group Q (A-R) and III (CMP1) (P