Objective To detect CK-19 mRNA expression by quantitative real-time RT-PCR in axillary drainage fluid of rectal cancer and investigate its clinical significance.Methods Axillary drainage fluids were collected from 59 patients with rectal cancer and 15 patients with benign abdominal lesion from Sep.2010 to Dec.2010.Level of CK-19 mRNA in axillary drainage fluid was detected using specific primers by real-time RTPCR.The data were statistically analyzed to investigate the relationships between CK-19 mRNA level and tumor invasion,lymph node status,tumor stage and tumor differentiation level.Results The positive rate of CK-19 mRNA expression in patients with rectal cancer was 67.8％,which was significantly higher than that in patients with benign abdominal lesion.The expression of CK-19 mRNA was significantly correlated with the depth of tumor invasion,lymphnode status,tumor stage and histopathological differentiation( P ＜ 0.05 or P ＜ 0.01 ).Ck129 mRNA expression was associated with the pathological level,the higher of the lymph node translation level,the higher expression in the axillary drainage fluid after rectal cancer surgery (r =0.674,P =0.021 ).The lower of the lymph node differentiation level,the higher expression in the axillary drainage fluid after rectal cancer surgery (r =-0.741,P =0.014).Conclusion Quantitative detection of CK-19 mRNA in axillary drainage fluid of rectal cancer by RT-PCR could enhance the diagnostic sensibility of colorectal cancer micrometastases.RT-PCR assay is suitable for predicting peritoneal micrometastasis of rectal cancer,which is a reference for postoperative treatment and prognosis prediction.

Background and purpose:This study aimed to investigate the effect of glucagon-like peptide-1 receptor agonists on proliferation, secretion of calcitonin and energy metabolism of medullary thyroid cancer (MTC) cell.Methods:The MTC cell line (TT) was culturedin vitro. After treatment with exenatide and liraglutide (0, 1, 10 and 100 nmol/L) for 24, 48 and 72 h, the proliferation of TT was analyzed by CCK-8 kit, the calcitonin was measured by calcitonin assay kits, and the energy metabolism of TT was measured by Seahorse XF instrument.Results:When compared with control group, neither exenatide nor liraglutide had effects on proliferation of TT (P>0.05); the calcitonin levels did not change signiifcantly after treatment with GLP-1 receptor agonists (P>0.05). Exenatide and liraglutide did not alter glycolysis and mitochondrial respiration in TT cells in a dose- and time-dependent manner.Conclusion:GLP-1 receptor agonists have no effect on the development of TT. Further collection of the safety data of exenatide and liraglutide on thyroid is still needed.

The pathway of 2-methyl-D-erythritol-4-phosphate (MEP) is the exclusive isoprenoid precursor biosynthetic pathway in Escherichia coli, with a higher theoretical yield than mevalonate (MVA) pathway. However, due to lack of information about the regulation of MEP pathway, only engineering MEP pathway in E. coli achieved limited improvement of heterologous isoprenoid production. We used exogenous MEP pathway genes to improve MEP pathway in E. coli and optimized the glucose feeding to release the potential of MEP pathway. The results demonstrate that co-expression of dxs2 from Streptomyces avermitilis and idi from Bacillus subtilis can increase amorphadiene production with 12.2-fold compared with the wild-type strain in shake flask fermentation. Then we established a high-cell density fermentation process for the engineered strain, and found that the phase from 24 to 72 h is important for product biosynthesis. The optimization of glucose feeding rate during 24 to 72 h significantly improved product accumulation, which was improved from 2.5 to 4.85 g/L, within the same process time. Considering the attenuation of strain metabolism after 72 h, this study further modulated the glucose feeding rate during exponential phase to control strain growth and the amorphadiene yield eventually reached to 6.1 g/L. These results provided useful information to develop engineered E. coli for isoprenoid production through MEP pathway engineering.
Bacillus subtilis ; Biosynthetic Pathways ; Culture Media ; chemistry ; Erythritol ; analogs & derivatives ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Engineering ; Glucose ; chemistry ; Industrial Microbiology ; Sesquiterpenes ; metabolism ; Sugar Phosphates ; metabolism ; Terpenes ; metabolism

Objective To analyze the efficacy and safety of erlotinib associated conformal intensity modulated radiotherapy in treatment (IMRT) of locally advanced pancreatic carcinoma. Methods The clinical data of 23 patients with locally advanced pancreatic carcinoma were retrospectively analyzed. The patients′ therapeutic methods: erlotinib was taken continuously and orally at 100 mg/time, 1 time/d until disease progressed or serious adverse reactions happened; intensity modulated radiotherapy (IMRT) was used combined with erlotinib at 50.4 Gy, 1 time/d, 1.8 Gy/time, 5 times/week, total 28 times. Tumor response was evaluated at the end of radiotherapy after 4 weeks. Results In 23 patients, there was partial response in 10 cases, stable disease in 9 cases and progress disease in 4 cases. The objective response rate was 43.5%(10/23), and the median survival time was 11.3 months. Adverse reactions included fatigue, rash, bone marrow suppression, nausea and diarrhea. The adverse reactions were mostly tolerable with grade 1-2. Conclusions Erlotinib combined with IMRT is safe and effective in patients with locally advanced pancreatic carcinoma, which is worthy of further study.

Objective To assess the efficacy and toxicity of topoteean hydllochloride plus carbo-platin(TC)versus etoposide plus carboplatin(CE)in patients for previously untreated small cell lung cancer (SCLC).Methods Sixty-nine patients with previously untreated SCLC,TC group(34 eases)were treated with topotecan 1 mg/m2 from day 1 to day 5 and carboplatin 300 mg/m2on day 1.CE group(35 cases)were treated with etoposide 100 mg/d from day 1 to day 5 and carboplatin 300 mg/m2 on day 1.Treatment was repeated every 3 weeks.The efficacy and toxicity were ev Mumed in patients who received two cvcles of chemotherapy.ResMN The total effective rate Was 76.5%in TC group and 71.4%in CE group(P＞0.05). The progression-free survival interval was 4.1 months in TC group and 2.6 months in CE group(P＜0.05). There was no significant difference in the most common toxieities between two groups(P＞0.05).Conclusion Compared with etoposide plus carboplatin,topotecan plus carboplatin has similar total effective rate,and in- different toxieities,and the longer progression-free survival interval,so it is a safe and effective first-line treatment for SCLC.

Objective To investigate the feasibility and efficacy of the intensity modulated radiotherapy (IMRT) with oral S-1 for locally advanced pancreatic cancer.Methods Forty-two patients with locally advanced pancreatic cancer were selected,and the patients were divided into 2 groups by random digits table method with 21 cases each.The patients in treatment group were treated by IMRT combined with oral S-1 40 mg/m2 twice daily from day 1 to day 14 of a 21-d cycle;the patients in control group were treated by IMRT combined with 30 min intravenous infusions of gemcitabine 1 000 mg/m2 on day 1,8,21 and 29.Radiation was concurrently delivered at a dose of 50.4 Gy (1.8 Gy/d,5 times per week,28 fractions).The dose of intensity modulated radiotherapy was 1.8 Gy/time (5 times/week,50.4 Gy/28 times).The recent curative effect and untoward reaction were assessed at the end of radiotherapy after 4 weeks.Results There were no statistical differences in efficient rate and disease control rate between treatment group and control group:57.1％ (12/21) vs.47.6％ (10/21) and 85.7％ (18/21) vs.76.2％ (16/21),P ＞ 0.05.The neutropenia rate and thrombocytopenia rate in treatment were significantly lower than those in control group:23.8％ (5/21) vs.57.1％ (12/21) and 28.6％ (6/21) vs.66.7％ (14/21),and there were statistical differences (P ＜ 0.05).There were no statistical differences in the incidences of anemia,nausea,vomiting,elevated aminotransferase and fatigue (P＞ 0.05).Conclusions The IMRT with oral S-1 for locally advanced pancreatic cancer is safe and effective.It is worthy of clinical promotion.

Objective:To explore the influence of blood transfusion on cellular immunofunction in patient with laryngeal carcinoma.Method:EPICS-XL flow cytometry was used to measure T cell subgroup,NK cell and CD28 in 36 patients with laryngeal carcinoma pre-operation and 2 weeks post-operation.Patients were divided into allotransfusion group and non-transfusion group.Comparison was conducted between the 2 groups.Result:①Comparsion with normal population,decreasing of CD3,CD4,NK cell and CD28 in the 36 patients pre-operation was statistically significant (P＜0.01).②Decreasing of CD3,CD4,NK cell and CD28 was statistically signficant post-operation(P＜0.05).③In the non-transfusion group change of CD3,CD4,CD8,NK cell and CD28 post-operation was statistically insignificant (P＞0.05).Conclusion:While cellular immunofunction is generally low in patients with laryngeal carcinoma,allotransfusion will reduce further.It makes contribution to spreading and metastasis of carcinoma easier.

The aim of this study is to develop a synthetic medium suitable for 13C metabolic flux analysis (13C-MFA) of Streptomyces rimosus. The cell growth rate and oxytetracycline production by S. rimosus M4018 were compared when M4018 cells were growth on the optimized chemically defined media with organic nitrogen sources or inorganic nitrogen sources. First, a synthetic medium contained KNO3 as the main nitrogen source was screened, then optimized by a response surface method. Using this new medium, the oxytetracycline yield was increased from 75.2 to 145.6 mg/L. Furthermore, based on the 13C-MFA, we identified that Entner-Doudoroff pathway does not exist in S. rimosus cells cultured in a chemically defined medium with feed of 100% 1-13C labeled glucose. This study is helpful for subsequent 13C-MFA application of S. rimosus.
Carbon Isotopes ; analysis ; Culture Media ; chemistry ; Metabolic Flux Analysis ; Nitrogen ; chemistry ; Oxytetracycline ; biosynthesis ; Streptomyces rimosus ; metabolism

Aim To screen the potential inhibitors of post-transcriptional activity of pro-inflammatory media-tor TNF-α from the lipophilic constituents in Chinese Medicine Salvia miltiorrhiza Bunge ( Danshen) , we es-tablished dual luciferase reporter gene system pGL3-TNF-α3′UTR ( 3′untranslated region ) co-transfected with Renilla control gene. Methods Complementary DNA ( cDNA) template was obtained from human um-bilical vein endothelial cells ( HUVECs ) . The full length DNA of TNF-α 3′-UTR was amplified through PCR, and then connected the luciferase reporter vector pGL3-control after enzyme digestion. pGL3-TNF-α 3′UTR constructs were co-transfected with pSVRenilla into the mononuclear macrophages RAW264. 7 cells. The relative activity of reporter genes was measured by dual luciferase reporter ( DLR ) assay system after the stimulus of lipopolysaccharide ( LPS ) in presence or absence of tanshinones compounds. Results The pGL3-TNF-α3′UTR luciferase reporter gene was suc-cessfully constructed. The cloning DNA fragment and sequence were both consistent with the GENBANK da-tabase. LPS significantly induced the relative reporter activityof RAW264 . 7 cells transfected with pGL3-TNF-α 3′UTR. Among four tanshinones compounds, we found only cryptotanshinone could significantly de-crease LPS-induced relative reporter activity. Conclu-sion The pGL3-TNF-α 3′UTR construct combined with DLR assay system was successfully established, which can be used to discover the agents such as cryp-totanshinone that regulate the post-transcription of TNF-α in treatment of inflammatory and malignant dis-eases.

Sertoli cell (SC) is intrinsic to the testis and provides an appropriate growth environment for the germ cells. It was separated from rat's testis and identified by hematoxylin and eosin staining(HE) and immunocytochemical reaction, then cultivated in vitro. Culture conditions such as pH, osmotic pressure and metabolic parameters that include consumption rates of glucose, glutamine, amino acids and formation rates of lactic acid, ammonium ion were investigated. It was showed that adhesion process of SCs was accomplished within 2-4 hours after inoculation. It was also observed that the SCs entered into the decline phase when the concentration of ammonium ion and lactic acid were above 2.3 mmol/L and 14 mmol/L, respectively, which caused osmotic pressure above 326 mosm/kg and pH below 6.8 in the medium. As the changes of amino acids during culture were concerned, Glu and Ala accumulated rapidly, while Val, Leu, Ile reduced slightly and at the same time Ser, Arg, and Gly were stable. The restrict factors for SCs grown in static culture might be high osmotic pressure and low pH, which were generated when glutamine and glucose were metabolized into lactic acid. The findings could be fundamental in the process optimization of large scale Sertoli cells in vitro culture.
Amino Acids ; metabolism ; Animals ; Animals, Newborn ; Cell Culture Techniques ; Cells, Cultured ; Culture Media ; Male ; Rats ; Sertoli Cells ; cytology ; metabolism ; Testis ; cytology