Objective The purpose of this study was to investigate the expression of human mitochondrial transcription termi-nation factor-3 ( hMTERF3) in non-small cell lung cancer ( NSCLS) and to analyze its clinicopathological significance. Methods The paraffin block samples used in this study included 65 cases of NSCLC and 32 cases of normal alveolar epithelial tissues. We determined the expressions of hMTERF3 in NSCLC and normal alveolar epithelial tis-sues by immunohistochemistry, calculate the survival rate using the Kaplan-Meier method, and analyzed the risk factors affecting the prognosis of NSCLC using the Cox Proportional Hazard Model. Results In the 65 cases of NSCLC, 31 ( 47. 69%) showed positive expression of hMTERF3. The total survival time was significantly shor-ter in the patients with a high than in those with a low hMTERF3 ex-pression ([30.39±3.35] vs [57.61±7.12] mo, P<0.05). The riskfactors affecting the prognosis of NSCLC included positive expression of hMTERF3 (HR=3.302, 95% CI:1.598-6.905) and lymph node metastasis (HR=4.052, 95% CI: 1.212-12.398). Conclusion hMTERF3 is overexpressed in NSCLC. Highly expressed hMTERF3 and lymph node metastasis reduce the survival time of NSCLC patients, suggesting that hMTERF3 may be a potential bio-marker for the prognosis of NSCLC.

Objective To investigate the meaning of expression of apoptosis related molecules NFKBp65 and p53 and GPX1-mRNA in patients with Keshan disease(KSD).Methods Sixteen chronic Keshan Disease patients were enrolled in KSD group according to electrocardiogram,chest X ray film and clinical examinations on 15,September in 2009,and 23 healthy people were included in control group from physical examination taken in The Second Affiliated Hospital of Xi'an Jiaotong University.Fresh blood(5 ml)was collected from antecubital vein of all subjects in the fasting state.Total mRNA and protein of blood sample were isolated using Trizol.GPX Assay Kit was used to detect GPX enzyme activity,and GPX1-mRNA expression was determined by SYBR Real-Time PCR.Meanwhile,expression of apoptosis related molecules NFKBp65 and p53 were determined by Western blot.Results GPX enzyme activity decreased significantly in KSD group[(108.61±14.10)U]compared with control group[(122.78±11.89)U,t=2.874,P<0.05],GPX1-mRNA level of KSD group(0.553±0.299)notably KSD group(0.802±0.057)compared with control group[(1.065±0.355),t=6.829,P<0.01].p53 increased in KSD group(1.604±0.191)compared with control group[(1.137±0.186),t=3.033,P<0.05].Conclusiom Decreased GPX1-mRNA expression may result in lower GPX enzyme activity of patients with KSD.Thus oxidative damage increases and cadioeyte apoptosis is activated by activating apoptosis signal pathway.

Object: To investigate the effects of Huaiqihuang Granule, a compound Chinese herbal medicine, on expressions of nephrin and podocin of slit diaphragm of glomerular podocytes in rats with adriamycin-induced nephrosis and to explore the mechanism in reducing the proteinuria. Methods: Twenty SD rats were randomly divided into five groups: control group, model group, glucocorticoid group, Huaiqihuang Granule group and Huaiqihuang Granule plus glucocorticoid group. The 24-hour urine was collected 7, 14, 21 and 28 days after adriamycin injection respectively to measure 24-hour urinary protein, and all rats were sacrificed after 28-day treatment. Pathological changes in renal tissues were observed under a light microscope and an electron microscope. Expressions of nephrin and podocin mRNAs in renal cortex were determined by real-time polymerase chain reaction, and protein levels of nephrin and podocin were detected by Western blotting. Results: (1) In the model group and the treatment groups, the level of urinary protein increased significantly from the 14th day. (2) Under the light microscope, inflammatory cells and slight fibroplasia were found in renal interstitium of the model group, but there were less inflammatory cells in renal interstitium in the intervention groups than in the model group. Under the electron microscope, 29 days after adriamycin injection, extensive fusion of foot processes was observed. (3) The expressions of nephrin and podocin were higher in treatment groups than in the model group. (4) Proteinuria level was negatively correlated with the expressions of nephrin mRNA and nephrin and podocin proteins. Conclusion: The above results indicate that Huaiqihuang Granule can maintain the integrity of the slid diaphragram in podocyte, alleviate the lesion of glomerular filtration membrane, and decrease the proteinuria by up-regulating the expressions of nephrin and podocin. Huaiqihuang Granule plus glucocorticoid maybe has better effects than glucocorticoid alone.

Objective To explore the miRNA regulating the potential cancer-promoting gene CCL18 in cutaneous malignant melanoma.Methods Bioinformatics analysis was conducted by using online software miRanda and TargetScan,so as to predict the miRNA targeting CCL18 gene.Three kinds of C CL18 3'UTR dual-luciferase reporter vectors,including mutant 3'UTR vector (mutant 3'UTR group),wildtype 3'UTR vector (wild-type 3'UTR group) and empty vector (blank control group),as well as miRNA vectors carring selected miRNAs were constructed according to human gene sequence analysis,and then were used to co-transfect 293T cells.After 48-hour treatment,the cells were lysed for detection of luciferase activity.Real-time fluorescence-based quantitative PCR was performed to measure the expression of CCL 18 and selected miRNA in 14 fresh malignant melanoma tissue specimens and 14 paracancerous normal skin tissue specimens (control tissues),and their correlations were analyzed.Results Online software analysis showed that some miRNAs were identified to target the 3'UTR of CCL18 gene,including miR-183,miR-128 and miR-33a.Luciferase reporter vectors and miRNA vectors were constructed successfully.As luciferase activity assay showed,when miR-183 and miR-128 were bound to the CCL18 3'UTR,the luciferase activities were significantly higher in their mutant 3'UTR groups (11.63 ± 0.42;8.80 ± 0.49) than in their wild-type 3'UTR groups (4.86 ± 0.39;5.01 ± 0.54;both P ＜ 0.05) and blank control groups (2.41 ± 0.13;2.39 ± 0.05;both P ＜ 0.01),while there were no significant differences between miR-33a-hinding mutant 3'UTR group (6.41 ± 0.47) and miR-33a-binding wild-type 3'UTR group (6.16 ± 0.22,P ＞ 0.05).Real-time fluorescence-based quantitative PCR revealed higher mRNA expression of the CCL18 gene (3.52 ± 1.68),but lower expression of miR-183 (0.49 ± 0.32),miR-128 (0.30 ± 0.20) and miR-33a (0.46 ± 0.40) in the malignant melanoma tissues compared with the control tissues.The mRNA expression of the CCL18 gene was negatively correlated with the expression of miR-128 (rs =-0548,P ＜ 0.05),but showed no significant correlations with the expression of miR-183 and miR-33a (both P ＞ 0.05).Conclusion miR-128 may play a role in regulating the potential malignant melanoma-promoting gene CCL18.

Objective To study the radiosensitizing effect of 2'-hydroxyflavanone (2'-HF) on prostate cancer cells,and to preliminarily investigate its mechanism.Methods Colony formation assay,tert-butylhydroperoxide (TBHP) oxidative stress assay,Hoechst staining,and apoptosis flow cytometry using Annexin V-FITC and propidium iodide (PI) were performed to measure the impact of 2'-HF on the radiosensitivity of VCaP prostate cancer cells.Western blot was used to determine the effects of 2'-HF on expression of AKT,phosphorylated AKT (p-AKT),and aldo-keto reductase 1 C3 (AKR1 C3) in VCaP cells and preliminarily investigate the mechanism.Data were analyzed by t test and factorial analysis of variance.Results The results of colony formation assay indicated that after exposure to radiation,VCaP cells treated with 2'-HF had a significantly lower proliferation level than cells in the control group (P=0.010),yielding a sensitization enhancement ratio of 1.19.The resuhs of TBHP oxidative stress assay suggested that VCaP cells treated with 2'-HF had significantly weaker anti-oxidative capacity than cells in the control group (P=0.015).Hoechst staining and apoptosis flow cytometry with Annexin V-FITC and PI indicated that 2'-HF treatment plus irradiation significantly enhanced apoptosis in VCaP cells (P=0.001.The results of Western blot suggested that 2'-HF treatment significantly inhibited the protein expression of p-AKT and AKR1C3 in VCaP cells (P=0.013 and P=0.016).Conclusions 2'-HF can enhance the radiosensitivity of prostate cancer cells,which is probably associated with its inhibitory effects on AKT pathway and AKR1C3 expression in prostate cancer cells.

BACKGROUND:Increasingly studies report that the normal balance of bone metabolism may be destroyed in the case of postmenopausal osteoporosis or osteoarthritis. The concrete metabolic process of bone turnover could be revealed sensitively by measuring the bone turnover markers in the serum or urine.
OBJECTIVE:To study the bone density and bone metabolic index of knee osteoarthritis (KOA) and postmenopausal osteoporosis (PMO), and to discuss the characteristics of bone density and bone metabolic index in KOA and PMO.
METHODS:A total of 248 postmenopausal women were detected for bone mineral density and knee X-ray. Final y 180 patients were included in this study and were divided into three groups:KOA group, PMO group, and control group. The levels of bone turnover markers (bone alkaline phosphatase, bone gla protein, col agen type I cross-linked C-telopeptide, and tartrate-resistant acid phosphatase 5b) in serum from the participants were measured. The correlation between bone turnover markers and the disease progression was analyzed by Logistic regression analysis.
RESULTS AND CONCLUSION:The bone mineral density in the KOA group was higher than the control group but col agen type I cross-linked C-telopeptide was lower. The levels of bone gla protein, col agen type I cross-linked C-telopeptide, and tartrate-resistant acid phosphatase 5b in serum from PMO group were higher than the control group. The decrease of col agen type I cross-linked C-telopeptide was associated with the incidence of KOA, and the increases of bone gla protein, col agen type I cross-linked C-telopeptide, and tartrate-resistant acid phosphatase 5b were associated with the incidence of PMO. The lower bone absorption can be seen in postmenopausal women with KOA. PMO patients showed a higher bone turnover rate. The difference of bone metabolism between patients with KOA and PMO led to negative relationship of bone mineral density. The serum levels of bone gla protein, col agen type I cross-linked C-telopeptide, and tartrate-resistant acid phosphatase 5b can assist clinical diagnose and therapeutic effect detection of both KOA and PMO.

A UPLC-MS/MS method based on metabonomic skills was developed to study the serum metabolic changes of rats after acute liver injury induced by CCl4 and to evaluate the action mechanism of Si-Ni-San. The integrated data were exported for principal components analysis (PCA) by using SIMCA-P software, in order to find the potential biomarkers. It showed that clear separation of healthy control group, model group, silymarin group, Si-Ni-San group was achieved by using the PCA method. Nine significantly changed metabolites were identified as potential biomarkers of acute liver injury. Compared with the health control group, the model group rats showed higher levels of phenylalanine, tryptophan and GCDCA together with lower levels of LPC 16 : 0, LPC 18 : 0, LPC 18 : 1, LPC 16 : 1, LPC 20 : 4 and LPC 22 : 6. These changes of serum metabolites suggested that the disorders of amino acid metabolism, lipid metabolism, bile acid biosynthesis and anti-oxidative damage were related to acute liver injury induced by CCl4. Si-Ni-San might have the anti-liver injury effect on all these four metabolic pathways.

Poly ?-glutamate is a biopolymer material that has a good application prospect.The Vitreoscilla hemoglobin(VHb) gene was integrated into the chromosome of Bacillus licheniformis WX-02 by integrative vector pDG1730-vgb.The expression of VHb was confirmed by CO-difference spectra analysis.It was shown by the results of batch cultures in a 3 L bioreactor that biomass and ?-PGA obtained in the recombinant M2 were 25.5 % and 20% higher than those of the control respectively.

Objective To study the action of apolipoprotein E genetic polymorphism on the efficacy of Tanshinol.Methods Through detecting the expression of low density lipoprotein receptor mRNA and 3-hydroxy-3 -methylglutaryl coenzyme A reductase mRNA in L -02 cells by reverse transcription-polymerase chain reaction , to compare the effect of different genetic subtypes of ApoE on Tanshinol.Results Apolipopro-tein E can weaken the induction of low density lipoprotein receptor ex-pression caused by Tanshinol.The weakening extent of apolipoprotein E 2 (Arg112Cys)was stronger than apolipoprotein E3 and apolipoprotein E4 ( Cys 158 Arg) ( P<0.01 ).Apolipoprotein E also can enhance the inhi-bition of 3-hydroxy -3-methylglutaryl coenzyme A reductase expres-sion, especially apolipoprotein E4 (P<0.01).But apolipoprotein E2 or apolipoprotein E3 can not have statistical significance ( P >0.05 ).Conclusion Both 112 and 158 site might be the key site that have effect on the efficacy of Tanshinol.

Objective To observe the steps of vascular smooth muscle cells (VSMCs) calcification induced by high phosphate enviroment in vitro. Methods VSMCs were incubated with high phosphate (2.5 mmol/L or 3.5 mmool/L) medium for different times. Expression of core binding factor α1(Cbfα1), osteopontin(OP), collagen type Ⅰ(Col Ⅰ), osteocalcin(OC) and α-smooth muscle actin (α-SMA) was investigated by Western blot, immunofluorcscencc staining and real time PCR. Mineral deposition was assessed by von Kossa aad Alizarin red staining. Ultrastructure of VSMCs calcification was observed by electron microscopy (EM). Results Up-regulated expression of osteoblast-specific transcription factor Cbfα1 in the nuclei oceured at as early as 12 hours. The protein of Col Ⅰ and OP was up-regalated when VSMCs were incubated in high phosphate medium for 3 days, and content of OC increased at the time of 6 days. When cultured in 2.5 mmol/L phosphate medium for 15 days, VSMCs lost their lineage marker α-SMA, developed granular calcium deposits. Moreover, the results of real time PCR indicated mRNA level of OP and Col Ⅰ increased at day 1, OC increased at day 5 and α-SMA level decreased at day 10, respectively. Ultrastructural analysis also confirmed the presence of collagen and matrix vesicles in the cells. Conclusion VSMCs phenotype transformation induced by high phosphate enviroment is an orchestrated, highly regulated process.