To study effects of 9 cis retinoic acid (9 cisRA) on biological characteristics of lung squamous cancer cells, cell growth, cell differentiation and apoptotic indexes were observed in 9 cisRA treated L 78 and A 2 lung squamous cancer cells with flow cytometry. The results showed that 9 cisRA exerted marked inhibitory effect on the growth of two lung squamous cancer cell lines, 9 cisRA also had significant inducing effect on the differentiation of L 78 and A 2 cell lines, whereas the percentages of apoptosis of two cell lines in the treatment group were significantly higher than the control group. We conclude that 9 cisRA could inhibit growth inhibition, induce cell differentiation and apoptosis of L 78 and A 2 lung squamous cancer cells

Objective To evaluate the prognostic value of MTV measured by pretreatment 18F-FDG PET/CT imaging in patients with pancreatic cancer (PC).Methods The clinical data of 52 patients (31 males,21 females,median age 58.5 years) with pathologically or clinically proved PC from January 2011 to October 2014 were retrospectively analyzed.Serum CA19-9 was measured within 1 week before PET/CT examination.SUVmax and MTV were measured respectively,and PET/CT staging was obtained simultaneously.All patients were followed up until January 2015.The CA19-9,MTV,SUVmax,PET/CT staging and clinical factors(age,gender,treatment methods) were assessed by ROC curve analysis,Kaplan-Meier analysis and multivariate Cox model.Results The median survival time of 52 patients was (11.20±7.25) months.ROC curve analysis showed that the AUC of MTV,SUVmax and CA19-9 were 0.735,0.614 and 0.527 respectively.Kaplan-Meier survival analysis manifested that the survival times were significantly different between patients with different MTV (＜ 12.14 cm3 vs ≥ 12.14 cm3),different SUVmax (＜ 8.95 vs ≥ 8.95),different PET/CT staging and different treatment methods (x2 =4.272-11.693,all P＜0.05).The survival time of patients with MTV ＜ 12.14 cm3 and that of patients with MTV ≥ 12.14 cm3 were (13.44±8.40) and (7.00± 4.82) months,respectively.Cox single-factor analyses indicated that MTV,PET/CT staging and SUVmax were risk factors of survival,the hazard ratios (HR) were 0.393,0.503,0.547 respectively (P=0.002,0.020,0.027).Cox multi-factor analyses indicated that MTV and PET/CT staging were independent risk predictors of survival.Conclusion MTV and PET/CT staging are significant factors in prognosis prediction of patients with PC,which would be helpful to make individual treatment for patients with high risks.

OBJECTIVE:To develop an HPLC method for the determination of Tanshinone ⅡA in Lianchuang pills. METHODS:The chromatographic separation was performed on Agilent Zorbax SB-C18 (250mm?4.6mm,5?m) column with column temperature at 25℃. The mobile phased consisted of methanol-water (75∶25) at a flow rate of 1.0mL?min-1.The detection wavelength was set at 270nm.RESULTS:The linear range of Tanshinone ⅡA was 0.020 2～0.202 0?g(r=0.999 9).The average recovery was 99.79%(RSD=1.01%,n=6). CONCLUSION:The method is simple, rapid and accurate, and it could be used for the quality control of Lianchuang pills.

Objective To investigate dynamic changes of extracellular signal regulated protein kinase (ERK) 1/2 in lung fibroblast of newborn rats with chronic lung disease (CLD) caused by hyperoxia.Methods Full-term newborn rats were randomly divided into two groups:air-exposed group and hyperoxia - exposed with 90％ oxygen group.Rats were sacrificed separately 3 d,7 d and 14 days after exposure to air or 90％ oxygen. Then lung fibroblasts of rats were isolated and primarily cultured. By using Immunocystochemistry,Western-blot and RT-PCR methods,the levels of ERK1/2 protein and expressions of ERK1/2 mRNA were measured. Results The levels of p-ERK1/2 protein in lung fibroblast in the hyperoxia group were significant higher on the 7th day and 14th day after exposure to 90％ oxygen compared with those in the air-exposed group (P ＜0.01 ).And the levels of total ERK1/2 protein and expressions of ERK1/2 mRNA did not change noticeably and were not significantly different between two groups (P ＞0.05 ).Conclusions The activation of phosphorated ERK1/2 may lead to lung fibrosis caused by hyperoxia in newborn rats.

ObjectiveTo study the expression and the role of α-smooth muscle aetin (α-SMA) and collagen Ⅰ ( Col Ⅰ ) in lung fibroblasts of newborn rats with hyperoxia-induced lung injuries.Methods Thirty full-term newborn Wistar rats were randomly assigned to hyperoxia group (90％ oxygen exposure,n =15 ) and air group (room air exposure,n =15) within 12 h after birth.Then lung fibroblasts were isolated and primary cultured from rat lungs on postnatal 3 d,7 d,and 14 d.The distribution of α-SMA protein were measured by immunohistochemistry.The levels of Col Ⅰ were detected by ELISA.ResultsThere were no significant differences in the levels of α-SMA and Col Ⅰ expressions between the two groups at 3 d ( P＞0.05 ).While the expression of α-SMA ( 112.60 ± 4.61 vs 94.69 ± 2.38,200.30 ± 3.97 vs 103.04 ± 1.91,P＜0.01 ) and Col Ⅰ protein [ ( 28.66 ± 1.15 ) μ.g/L vs ( 24.62 ± 3.15 ) μg/L,( 30.60 ± 0.65 ) μg/L vs (27.46 ± 1.68 ) μg/L,P ＜ 0.05 ] in lung fibroblasts caused by hyperoxia were significantly higher than those in air-exposed group on postnatal 7 d and 14 d.There was positive correlation between α-SMA and Col Ⅰ protein ( r =0.72,P＜0.01 ).ConclusionHyperoxia promotes differentiation of lung fibroblasts into myofibroblasts,and synthesis of type Ⅰ collegen in neonatal rats,which leads to lung fibrosis finally.

Objective To analyze the clinical characteristics of two patients with 3β-hydroxysteroid dehydrogenase deficiency and to explore their molecular genetic defects.Methods The clinical features and laboratory data of two patients were collected.The exons of HSD3B2 gene were amplified by PCR and sequenced by Sanger sequencing.Results Patient 1, aged 5 yrs old, was raised as a girl with 46, XY karyotype, presented with hyperpigmentation, female infant vulva, clitoral hypertrophy, and bilateral cryptorchidism;Patient 2, aged 11 yrs old, was raised as a girl at birth but as a boy after 1 yr old for known 46, XY karyotype, presented with hyperpigmentation, micropenis and severe hypospadias.Both patients had markedly elevated adrenocorticotropin and decreased cortisol.Two homozygous missense mutations in HSD3B2 gene were identified:conversions of codon Pro155 toLeu(p.P155L)inpatient1,andcodonAla82toThr(p.A82T)inpatient2,bothofwhichwerereportedforthe first time in China.Conclusion The patients with 3β-hydroxysteroid dehydrogenase deficiency in 46,XY karyotype mainly present with male pseudohermaphroditism and adrenocortical deficiency, and the diagnosis should rely on the steroids detection and HSD3B2 gene screening.

This paper is to report the exploration of the activation of Rho/ROCK signal pathway in 5-HT-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and the inhibitory effect of m-Nis on this pathway. PASMCs were cultured with the explant technique. MTT assay was used to explore the proliferation of PASMCs after 5-HT treated for different time and the intervening effect of m-Nis. RT-PCR and Western blot were used respectively to explore the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 in 5-HT-treated PASMCs and intervening effect of m-Nis. The results of MTT assay suggested that 5-HT (1 µmol · L(-1)) treatment for 12-72 h significantly induced the proliferation of rat PASMCs (P<0.05 or P < 0.01), which were inhibited by m-Nis (1 x 10(-5), 1 x 10(-6), l x 10(-7), 1 x10(-8) mol · L(-1)) in dose-dependent manners (P < 0.05 or P < 0.01). Similarly, the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 were also inhibited by m-Nis in different degrees (P < 0.05 or P < 0.01). Thus, the results of this study suggested that Rho/ROCK pathway played an important role in 5-HT-induced proliferation of rat PASMCs, m-Nis inhibited 5-HT-induced proliferation obviously, which may be related to the blockage of Rho/ROCK signal pathway.

Aim To explore the changes of Apelin/APJ system in LPS-induced injury of rat pulmonary mi-crovascular endothelial cells( PMVECs) , and the effect and mechanism of Apelin. Methods PMVECs were cultured with the explant technique, and the identifica-tion of rat PMVECs was carried out by immunocyto-chemical staining of factorⅧrelated antigen. MTT as-say was used to evaluate the viability of PMVECs. The mRNA expression of Apelin and APJ was detected by RT-PCR. The protein expression of PCNA and the phosphorylation of Akt was analyzed by Western blot. Results The mRNA expression of Apelin and APJ showed a compensatory increase after LPS treatment for a short period of time ( P<0. 01 ) , but with the exten-sion of time, which was significantly inhibited, even lower than the control group ( P<0. 05 or P<0. 01 ) , suggesting that Apelin/ APJ system might be involved in LPS-induced PMVECs injury. MTT results showed that 10 -6 ~10 -9 mol · L-1 Apelin obviously promoted the proliferation of rat PMVECs ( P <0. 05 or P <0. 01 ) , and with certain concentration and time de-pendence. Moreover, Apelin also improved the LPS-induced PMVECs injury in different degrees ( P<0. 05 or P < 0. 01 ) . In addition, Western blot analysis showed that Apelin significantly reversed the decrease of the protein expression of PCNA and the Akt phos-phorylation level induced by LPS ( P <0. 05 or P <0. 01 ) . Conclusions The Apelin/APJ system is in-volved in LPS-induced PMVECs injury. Apelin plays an important role in protecting the pulmonary microvas-cular endothelial function and reversing the LPS-in-duced PMVECs injury, which might be related to the activation of Akt phosphorylation pathway.

Objective To clarify the relationship between CT120,a novel human plasma membrane-associated gene,and the proliferation of lung adenocarcinoma cell line A549.Methods Expression vector(pcDNA3.1)containing antisense oligonucleotides of CT120 was constructed and transfected into the adenocarcinoma cell line A549.RT-PCR and Western blot detected the expression of CT120.Meantime,flow cytometry and soft agarose colony formation were applied for cell proliferation,and P53,CyclinD1 and CDK4 were detected by RT-PCR.ResultspcDNA3.1 containing antisense oligonucleotides of CT120 was successfully constructed and inhibited the expression of CT120 effectively by RT-PCR and Western blot.The expression of P53 was up-regulated and the expression of CyclinD1 and CDK4 were down-regulated.Conclusion The down-regulation of CT120 expression by antisense oligonucleotides technique may be a potential drug target for treatment of lung cancer.