Background and purpose:Overexpression of human telomerase reverse transcriptase(hTERT)plays a critical role in the process of immortalization of tumor cells.This study aims to investigate siRNA's effects on endogenous hTERT mRNA level and growth of breast cancer cells by targeting hTERT with its specific siRNA.The ultimate goal was to further elaborate the possible mechanism of immortalization in breast cancer cells.Methods:Real-Time RT-PCR was used to determine the hTERT transcripts in various breast cancer cells.hTERT siRNA was transfected into MCF7 cells with lipofectamine 2000.Cell growth was illustrated by growth curve,and cells apoptosis percentage was measured with Flow Cytometry(FACS).The relative expression levels of immortalization-related genes were evaluated with Real-Time RT-PCR.Results:Overexpressions of hTERT were demonstrated in all tested breast cancer cell lines.The inhibitory effects of hTERT siRNA were shown on both hTERT mRNA levels and cell growth from day 4 post transfection.Apoptosis was induced by hTERT siRNA as well.Certain immortalization-related genes were reduced by more than 50%,such as RAC1,PCYT2,FDFT1 and ATP5G2.Conclusions:hTERT siRNA specifically inhibited hTERT at mRNA level as well as cell growth.The apoptosis and downregulation of immortalization-related genes due to hTERT siRNA was demonstrated.

Background and purpose:The effects of triiodothyronine(T3)on breast cancer remain unclear.The aim of this study was to investigate the expression and correlation of estrogen receptor-?(ER-?)and thyroid hormone receptor-?(TR-?)in breast cancer cell lines,and to determine the changes of relevant receptors expression and its possible mechanism due to T3 treatment.Methods:Real-Time PCR was used to analyze the expressions of ER-? and TR-? at mRNA level in seven breast cancer cell lines.The expression level of retinoid X receptor-?(RXR-?)and these two receptors after T3 treatment were determined as well.The gene profile was further determined in SK-BR-3 cells treated with T3.Results:The expressions of ER-? and TR-? were quite low in SK-BR-3,MDA-MB-231 and MDA-MB-435 cells,but high in MCF7,BT20,BT474 and T47D cells.ER-?,TR-? and RXR-? were upregulated due to T3 treatment in ER-?(-)cells,but not in ER-?(+)cells.In SK-BR-3 cells,T3 treatment led to up-regulation of E2F1,TGF-?1R,TGF-?2R and hTERT,but down-regulation of TGF-?1 and p21.Conclusions:The expressions of ER-? and TR-? are correlated in breast cancer cells;the effect of T3 treatment on hormone receptors is associated with endogenous ER-? level.

Objective To evaluate the potential effects of RS504393, CC chemokine receptor (CCR) 2b and CCR1 antagonist, on LPS-induced acute lung injury (ALI) and to investigate the underlying mechanisms. Method A549 cell line was stimulated with LPS (10 μg/mL) and then treated with RS504393 (10 μg/mL) for 6 hours. ALI model was established with intranasal administration of LPS (5 mg/kg) in C57BL/6J mice. RS504393 (5 mg/kg) was administered 30 min before LPS dripped nasally. IL-8, IL-1β, plasminogen activator inhibitor (PAI)-l,monocyte chemoattractant protein (MCP)-2,and the expressions of CCR1 and CCR2b were studied by using Realtime-RT-PCR, ELISA and cyto-flowmetry. Results In A549 cell line treated with RS504393,the expressions of CCR1, CCR2b and IL-8 were significantly inhibited after LPS stimulation. In rats with LPS-induced ALI, treatment with RS504393 significantly protected mice against lung injury by attenuating influx of leukocytes and protein into bronchoalveolar space and by lessening pathological changes of lung. Treatment with RS504393 down-regulated IL-1β and PAI-1 expressions in bronchoal veolar lavage fluid (BALF) and lungs at mRNA and protein levels along with up-regulation MCP-2 expression compared to rats of vehicle-treated groups. Conclusions CCR2b and CCR1 play pivotal roles in the development of ALl,and RS504393 as a antagonist can halt the development of ALI.

Objective To evaluate the effect of intravenous Gd-DTPA on DWI of breast. Methods DWI was performed before and after Gd-DTPA injection on 25 patients with breast cancer confirmed by core or fine-needle aspiration biopsy. The signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR) and ADC values were measured on DWI when b = 0 and 1000 s/mm2 were selected. A paired t-test and non-parametric test were used to compare the differences between SNR, CNR and ADC values before and after enhancement in breast cancer and normal breast. Results There were no significant differences between the SNR(34. 56 ± 11.34 and 33.60 ± 14. 34 for cancer, 9. 88 ± 3.16 and 10. 42 ± 4. 18 for normal breast)and CNR( 24. 16 ± 9. 05 and 22. 26 ± 10. 05 ) before and after administration of contrast medium in breast cancer and normal breast, and no significant differences between the ADC values before and after administration of contrast medium in breast cancer [(0.96±0. 13) × 10-3 vs (0.95±0. 14) × 10-3 mm2/s] and normal breast[(1.90±0.47) × 10-3 vs (1.91±0.61) × 10-3 mm2/s], either (P =0.20 and 0.97). Conclusion It is possible to repeat DWI acquisition after administration of contrast medium in breast MRI.

Objective To explore the relationship between lesion sites of stroke and swallowing function. Methods Ischemic stroke patients consecutively admitted into the stroke unit were screened in this study. The new and previous lesion sites of stroke, dysphagia and aspiration under videofluoroscopy (VF) were recorded and their relation was analyzed. Results 211 patients were included and 169 patients had completed the MRI and VF examination. 159 patients had dysphagia and 94 had aspiration under VF. 72 patients (9 missing) were found disorder in oral phase and 150 in pharyngeal phase. There was no significantly difference in dysphagia under VF and in aspiration among different lesion sites (P>0.05). Patients with stroke in medulla tended to happen aspiration (P=0.056). Stroke sites above the tentorium of cerebellum (P=0.028) or above medulla (P=0.005) may cause disorder in oral phase. Conclusion The stroke injury in brain cortex, white matter, brainstem and cerebellum or hemisphere could lead to dysphagia and aspiration. The injury of medulla may cause aspiration. Lesion of brain hemisphere or above the medulla could result disorder in oral phase of swallowing.

Objective To investigate the advantage of detaining peripherally inserted central catheter through the veins of lower extremity in neonatal intensive care unit.Methods 165 cases of infants were randomized into veins of lower extremity group (first saphenous veins,then popliteal vein) and veins of upper extremity group (first basilic vein,then median cubital vein,cephalic veins) from January to December 2012.We observed the success rate of detaining,catheter tip location,and complications such as phlebitis in the two groups.Results The success rate of detaining of lower extremity group was higher than that of upper extremity group,and ectopic rate of lower extremity group was lower than that of upper extremity group.There was no significant difference in complications such as phlebitis between two groups.Conclusions Peripherally inserted central catheter through the veins of lower extremity has more advantage over through the veins of upper extremity group.

Objective To evaluate the feasibility of active breathing control (ABC) in conformal radiotherapy (CRT) for patients with non-small cell lung cancer (NSCLC). Methods From Feb 2005 to Mar 2008, 29 patients with inoperable NSCLC (stage Ⅱ-Ⅳ) were evaluated. For each patient, two series of CT scans were obtained with free breathing (FB) and ABC system during simulation, respectively. Then two confonnal radiotherapy (CRT) plans were finished based on the two sets of reconstructed images. The pattern of post-inspiratory breath-hold was triggered at 80% of the peak of inspiration curve. The margin of clinical target volume (CTV) to planning target volume (PTV) was 0. 6 cm for lesions of the superior lobe, and 1.0 cm for the lesions of middle and inferior lobes. Three to five coplanar fields were performed in conformal radiotherapy. The gross tumor volume (GTV), CTV, PTV, volume of the bilateral lungs (Volume_(lung)), V_(20) and mean lung dose (MLD) of two plans were evaluated by dose-volume histogram (DVH). The World Health Organization criteria and National Cancer Institute Common Toxicity Criteria 3.0 (NCI-CTC3.0) scale were used to assess the immediate response and acute side-effect, respectively. Results Significant differences of GTV, CTV, FIN, Volum_(lung), V_(20) and MDL were observed between the two plans (36. 35 cm~3 vs. 31.40 cm~3, t = 9. 70, P <0. 001 ;82. 33 cm~3 vs. 70. 83 cm~3, t = 8. 19, P < 0. 001 ; 230. 73 cm~3 vs. 197.59 cm~3 ,t=5.72,P <0. 001 ;21.66% vs. 18. 76% ,t = 11.16,P <0. 001 ;1329. 07 Gy vs. 1143. 14 Gy, t = 13. 24, P < 0. 001). With ABC, all patients completed their treatment successfully except one patient for financial problems. The median radiation dose to the GTV was 64 Gy (60 -64 Gy). The overall immediate response rate was 64% (18/28). According to the NCI-CTC 3.0, grade 1 and 2 acute radiation-related toxicities occurred in 68% (19/28) and 18% (5/28) of patients for esophagitis, 82% (23/28) and 7% (2/28) for pneumonitis, respectively. Grade 1, 2 and 3 bone marrow suppression occurred in 57% (16/28), 25% (7/28) and 14% (4/28) of patients, respectively. Grade 1 and 2 acute cardiac injuries occurred in 86% (24/28) and 14% (4/28) of patients. Conclusions During CRT for patients with NSCLC, the use of ABC can decrease the radiation dose and acute complications of normal tissues.

Aim To investigate the expression and im-plication of HIF-1α, ROCK-2 , FoxM1 in PC12 cell in-jury induced by lead acetate. Methods PC12 cells were treated with lead acetate at the doses of 100 , 200 and 400 μmol·L-1 . The cell viability was determined by MTT reduction assay and LDH assay, the intracellu-lar production of oxygen species was measured by as-sessing SOD and MDA levels, cell apoptosis was deter-mined by Hoechst 33342 staining, the expressions of HIF-1α, ROCK-2 , FoxM1 , Bcl-2 and Bax were deter-mined by immunoblotting analysis. Results Lead ac-etate induced cell injury in PC12 cells in a dose-de-pendent manner, and it potentiated oxygen radical pro-duction and cell apoptosis. In addition, lead acetate enhanced HIF-1α and ROCK-2 expressions, increased Bax/Bcl-2 ratio and decreased FoxM1 expression. Conclusion Lead acetate can induce PC12 cell apop-tosis, which may be related with the expressions of HIF-1α, ROCK-2 and FoxM1 . Cellular oxidative stress may contribute to the injury as well.

Objective To evaluate the efficacy of Lianhua Dingchuan Tablets in bronchial asthma.Methods Fifty BALB/C mice were randomly and equally divided into control (Con) group,ovalbumin (OVA) group,dexamethasone (DEX) group,high-dose Lianhua group,low-dose Lianhua group.The mice were sensitized and challenged with OVA plus aluminium hydroxide to establish asthmatic model and were pre-treated 30 minutes before challenge.Specific airway resistance (sRaw) was used to evaluate airway hyperresponsiveness,and airway inflammatory changes were measured.ELISA and Magnetic Luminex(R) were used to quantified the levels of IL-4,IL-13 and INF-γ.Results Airway resistance significantly decreased in DEX group and High-dose Lianhua group (P＜0.05).Levels of inflammatory cells and IL-13 in BALF evidently reduced in DEX group,high-dose Lianhua group and low-dose Lianhua group (P ＜ 0.05),while IL-13 level in serum only decreased in DEX group.There was no significant changes in the levels of IL 4 and INF γ among those groups.Conclusions Lianhua Dingchuan Tablets might relieve the symptoms of asthma by reducing IL-13 level and inhibiting the airway inflammation.

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein hormone that is essential for the development of ovarian follicles and testicular seminiferous tubules. The relatively short half-life of FSH in vivo requires daily injections for more than 10 days that is inconvenient and possibly contribute to the stress perceived by the patients. The goal of the present study was to increase FSH glycosylation, in order to develop a long-acting recombinant FSH. The cDNA of native alpha and beta subunit of human FSH was linked by a sequence with two N-linked glycosylation sites, and the resulted DNA was inserted into pcDNA3.1 vector to generate a recombinant vector of pcDNA3.1-FSH. The pcDNA3.1-FSH was linearized and transfected into CHO-K1, positive transformants were selected by G418 and confirmed by PCR and Western blotting. A single chain recombinant FSH was expressed, with molecular weight of about 49 kDa. The recombinant FSH expression level in CHO-K1 cell strain in serum-free culture was 3 mg/L. Single injection of this recombinant FSH could induce folliculogenesis and ovulation in rats, the efficacy was similar with the commercially available FSH preparation (Folltropin-V) administrated 8 times consecutively. The results suggested a long-acting FSH was produced successfully.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Female ; Follicle Stimulating Hormone, Human ; biosynthesis ; Genetic Vectors ; Half-Life ; Humans ; Ovarian Follicle ; drug effects ; Ovulation ; drug effects ; Rats ; Recombinant Proteins ; biosynthesis ; Transfection