Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) comprise an effective therapy for advanced non-small cell lung cancer patients with EGFR-activating mutations. Unfortunately, most patients eventually develop resistance to EG-FR-TKIs, probably due to a secondary point mutation of EGFR T790M. Thus, a sensitive method for accurate detection of T790M mu-tation is essential. Peripheral blood detection has gained our attention because it is convenient, making dynamic noninvasive quantita-tive detection of T790M mutation an optimal means of monitoring the efficacy of EGFR-TKIs. To date, the clinical significance of T790M mutation and EGFR-TKI resistance remains controversial. Several EGFR-TKIs targeting EGFR mutation, which have been in-troduced in recent years, showed better response in patients with T790M mutation, indicating that T790M may be a biomarker for con-quering resistance. This review introduces the methodology of T790M detection and its role in clinical practice.

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) have shown great efficacy in the treatment of non-small cell lung cancer (NSCLC) patients with EGFR-mutation positive tumors.However,the response to EGFR-TKI is quite different even in EGFR-mutation positive patients.Besides that,different lesions in same patient can also show different response to EGFR-TKI.These phenomena might be associated with the heterogeneity of EGFR mutations,which involves intratumoral heterogeneity,intertumoral heterogeneity,and the heterogeneity before and after treatment.The article introduces the advance in heterogeneity of EGFR mutations from these three aspects.

Objective To establish a method of large-scale cryopreservation of porcine hepatocytes to meet the need of the biological artificial liver. Methods Porcine hepatocytes were isolated from Chinese miniature swines by collagenase disgestion. The cells were suspended in self-made nutrient solution supplemented with 10% DMSO. The hepatocytes (1?10 10~2?10 10) were stored in liquid nitrogen (-196℃) after being treated with two kinds of freezing methods. The cryopreserved hepatocytes were thawed in 39℃ water after one month and cultured. The morphology, viability and the function of thawed hepatocytes were observed. Results The cell viability of cryobreserved hepatocytes after both freezing methods were high. However, synthesis of urea nitrogen and glucose was higher in stepwise temperature lowering group compared with programmed temperature lowering group. Conclusion The results indicate that the cryopreservation method is feasible and simple, large amount porcine hepatocytes can be cryopreserved.

AIM: To investigate the changes of apoptosis in isolated pancreatic islet cells, insulin secretion, expression of Bcl-xL and Bax induced by combination of IL-1?, TNF-? and IFN-?, and effects of taurine on them. METHODS: Isolated pancreatic islet cells from Wistar rat were incubated in monolayer in vitro. NO-2/ NO-3 production, NOS activity, insulin secretion, the protein expression of Bcl-xL and Bax, percentage of islet cell apoptosis and DNA fragmentation in pancreatic islet cells incubated with combination of IL-1?, TNF-? and IFN-? were measured, and the effects of taurine on the changes of them were further investigated. RESULTS: Combination of IL-1?, TNF-? and IFN-? induced a significant increase in percentage of pancreatic islet cell apoptosis, NO-2/ NO-3 production and NOS activity, DNA ladder appearance, a decrease in insulin content, up-regulation in the protein expression of Bax and down-regulation in the protein expression of Bcl-xL (P

This study was aimed to establish the quality standard of Pyrethri Tatsienenis Flos. The medical material was identified by the microscopy and the thin layer chromatography ( TLC ) methods . The moisture , total ash , acid-insoluble ash and alcohol-soluble extract were determined according to procedures recorded in the Chi-nese Pharmacopoeia (2010 edition). The content of luteolin was determined by the HPLC method. The results showed a strong characteristic microscopic of Pyrethri Tatsienenis Flos , and its TLC identification had a good resolution with clear spots . The mass fractions of luteolin was 0 . 036%~0 . 104% ( average of 0 . 078%) , moisture was 9 . 32%~15 . 82% ( average of 13 . 11%) , total ash was 6 . 65%~8 . 29% ( average of 7 . 45%) , acid-insoluble ash was 0 . 23%~0 . 59% ( average of 0 . 42%) , and the extraction was 21 . 42%~30 . 15% ( average of 24 . 86%) . It was concluded that this established standard was simple to operate with good stability and reproducibility , which can be used for quality evaluation of Pyrethri Tatsienenis Flos .

BACKGROUND: Heme oxygenase-1 (HO-1) promotor region has a pair of dinucleotide(guanosine thymidine, GT) repeats with a lengthy polymorphism, also named microsatellite polymorphism. Experiments in vitro have shown that we can indirectly learn about the level of gene transcription by measuring the number of GT repeats.OBJECTIVE: To investigate if an association exists between restenosis after percutaneous coronary intervention(PCI) and microsatellite polymorphism in HO-1 gene promoter.DESIGN: A case-control study based on the observation of the patients with coronary heart disease after undergoing coronary stenting.SETTING: Wards of the department of cardiology of a university hospital.PARTICIPANTS: A total of 118 patients were admitted from April 1996 to May 2002 at the Department of Cardiology of the First Hospital of Peking University who underwent successful coronary stenting. Inclusion criteria: The patients with coronary heart disease who underwent coronary stent implantation for more than 3 months now came to perform coronary angiography in follow-up. Exclusion criteria: Angiography showed that the stenosis of lumen in diameter in the patients with coronary heart disease was less than 50%and the follow-up in angiography was less than three months. There were 92males and 26 females aged(62±10) years old and the informed consents were obtained. The patients were divided into two groups according to the criteria stipulated by American Heart,Lung and Blood Association: in-stent restenosis(68 cases) and non-restenosis (50 cases).METHODS: DNA of the peripheral blood was isolated from the whole blood. The length of GT repeat was confirmed by PCR amplification and Spreadex Gel electrophoresis. Selected samples were sequenced with Sanger's method.MAIN OUTCOME MEASURES: Microsatellite gene frequency of HO-1promoter and its relationship with restenosis RESULTS: Patients with GT repeats <25 GT in the HO-1 gene promoter on either allele had significantly less often restenosis than patients without (47.5% vs. 68.4% ,P<0.05). After controlling some possible confound ing factorsfor coronary heart diseases, multivariate analysis indicated that still there was a significant difference between the two groups in restenosis rate(odd ratio 0. 418,95% CI: 0. 197 to 0. 887,P<0.05).CONCLUSION: The present study indicated that short(GT) n repeats of HO-1 gene promoter is associated with reduced post-PCI restenosis, which suggests the genetic contribution to in-stent restenosis after stent implantation. It may have important meanings to prevent the occurrence of restenosis.

Objective To verify the trueness and assess the traceability of results from a routine chemistry system procedure for measurement of urea and ereatinine in serun.Methods Series of fresh frozen patieot sera,whose values of urea or creatinine were assigned by isotope dilution gas chromatography mass spectrometry (ID-GC/MS) or isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS),were chosen to be analyzed by a routine chemistry system.The measurement results of urea and creatinine by the routine chemistry system were used for linear regression analysis against the assigned values bv the ID-MS method to calculate the percentage deviation and assess the expected bias.Results For urea and creatinine,the linear regression equations between the routine chemistry system and ID-MS methods were Y =0.9890X + 0.0192 (R2 =0.9990) and Y =0.9815X-6.4794 (R2 =0.9989),and the average percentage bias were-0.41％ (P ＞0.05) and-4.20％ (P ＜ 0.05),respectively.The expected percentage bias at three medical decision levels were-0.46％,-0.83％ and-0.96％ for urea and -15.90％,-5.87％ and-2.95％ for creatinine.Conclusions The results of urea analyzed by the routine chemistry system were consistent with the ID-MS method,which suggested that the results of the routine system procedure could be traced to ID-GC/MS method.For creatinine,the bias between the results of routine procedures and the assigned values met the minimum acceptance criteria' derived from biologic deviations,which would be better if its specificity improved.

Objective To investigate radioactivity levels in the main agriculture products around a uranium mine in Northern Guangxi.Methods The agriculture products and soil samples were collected and analyzed by using HPGe gamma ray spectrometer.Results The specific activity of 226Ra in radish (including leaf),radish leaves and radish,collected in one place,were 45.0,66.7 and 32.3 Bq/kg,respectively.Those of 226Ra and 23SU in the radish soil collected in the same place were 19 672 and 85 917 Bq/kg,respectively.The transfer coefficients of soil-to-radish and soil-to-leaves were 1.61 × 10-3 and 3.40 × 10-3,consistent with those reported in relevant literature.Radioactivity levels in agricultural products in another survey was in consistence with those in the national survey for food products.Radioactivity levels in soil elsewhere near the radish site was consistent with the results of the national soil radioactivive background survey.Conclusions The soil in this place has been contaminated by the nearby uranium mine.It is important to investigate this place further and take the necessary measures.

Objective To apply iTRAQ technology to observe changes in protein expression group in the process of inducing C2C12 cells differentiation towards osteoblast by BMP-2.Methods The myoblast C2C12 cells were seeded in BMP-2 induced differentiation system for differentiation induction.In the 7th day,differentiation protein was extracted and labeled with iTRAQ reagent.Then,mass spectrometric detection,data analysis of differentially expressed proteins,and analysis of biological information were carried out.Results 23 significantly differentially expressed protein spots were screened by BMP-2-induced myoblast C2C12 differentiated cell protein expression profile analysis,where the protein was labeled with iTRAQ reagent.8 protein points were up-regulated,and 15 protein points were down-regulated.Trend classification found that the above differential protein had differential expression in each period of C2C12 cell osteogenic differentiation (1-7 days).Part of up-regulated protein in the early differentiation period showed high expression level;part of up-regulated protein in the late differentiation period showed high expression level;similarly,part of down-regulated protein in the early differentiation period presented low expression level;part of down-regulated protein in the late differentiation period showed low expression level.Preliminary identification showed SERCA3,Cytochrome bS,S100A4,ATPase inhibitor and ATPIF1 presented dynamic changes,which suggests that these proteins may be related to inducing osteogenic differentiation mechanism.Conclusion The results of differential protein expression trend show the necessity of full monitoring of C2C 12 cells osteogenic differentiation and indicate that iTRAQ technology is an effective method of studying protein changes of cellular molecule.Five proteins including SERCA3,Cytochrome b5,S100A4,ATPase inhibitor and ATPIF1 can be used as candidate targets for osteogenic differentiation mechanism research.

Attracting overseas medical consumption is the driving force behind the development of the medical industry in China's current"dual circulation"new development pattern.The progress of domestic medical technology and the improvement of service quality,the development of the economy and society,the enhancement of residents'health awareness,the increase of government investment in the health field,the advantages of the medical market and price competition,the guidance and support of government policies,international medical exchanges and cooperation,and the improvement of the medical insurance system are the main driving factors for attracting overseas medical consumption.Improving the level of medical technology,stimulating the vitality of attracting medical consumption,increasing the supply of medical consumption,unleashing the potential of attracting medical consumption,improving the quality of medical services,building support for attracting medical consumption to return,optimizing the medical consumption environment,creating an atmosphere for attracting medical consumption,and regulating overseas medical consumption,injecting the momentum of attracting medical consumption to return are the main implementation ways to attract overseas medical consumption to return.