Objective: To investigate the effects of rhTNF alone and in combination with antitumor chemicals on Walker-256 cell line in vitro. Methods: the cytotoxic activity in vitro were be examined by MTT method and the flow cytometry(FCM) analysis and alteration of every phase of cell cycle for rhTNF or (and) Adriamycin(ADM) against Walker-256 cell line. Results: rhTNF had a strong activity of antitumor in vitro and good dose-effect relationship(r=0.9811). Compared with negative control group, the effect of rhTNF was significant (P

This article summarized recent correlative literatures focusing on international standards on glycated hemoglobin.The basic concept,determination of glycated hemoglobin,the present review in laboratory measurement and metrological traceability was introduced.The international community has established reference system and metrological traceability to the International System of Units on HbA1c.Determination in glycated hemoglobin is still in incipient stage in our country.Both clinical laboratorians awareness and clinical determination need to be strengthened.

Objective:To establish the RPB5-mediating protein (RMP)-silenced stable cell lines and study the inhibitory effects of small interfering RNA (siRNA) targeting RMP gene on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods:Three RMPi siRNAs were designed and synthesized in vitro and transfected into SMMC-7721 cells. The inhibitory effect of siRNA on RMP gene expression was measured by RT-PCR to select the best siRNA. The expression vector pGPU6-Neo-RMP-484 was transfected into SMMC-7721 cells by the lipofectamine and the cells stably expressing the siRNA were selected by G418. RT-PCR was used to detect the interference efficacy against RMP gene. Cell proliferation and adhesion were measured by MTT assay. Wound healing test was used to observe the migration ability of cells. Results:The SMMC-7721 cell lines with down-regulated RMP expression were established by using RNA interference technology. Compared with the negative control cells, expression of RMP mRNA was down-regulated by(83.67±2.56)% .The proliferation of stable-transfected cells was inhibited by(74.33±0.58)% . The adhesion capability of stable-transfected cells was enhanced but the migration capacity was decreased compared with the negative control cells. Conclusion:The pGPU6-Neo-RMP-484 cell lines with stable transfection of RMP siRNA recombinant vector are successfully screened,which can be used as a cellular model for studying the molecular mechanism of RMP. Down-regulation of RMP gene expression can effectively inhibit the proliferation, enhance the adhesion, and decrease the migration of SMMC-7721 cells.

Objective To establish a method for measuring serum cotinine by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS) and provide an assay that can be applied to theevaluation of the level of smoke exposure and to the risk analysis of smoking related diseases.Methods Blood samples were collected from 94 apparently healthy subjects from October to December in 2010 and centrifuged,and the sera were separated.Serum samples were mixed with [ D3 ] -cotinine ( as the internal standard) and treated with acetonitriles to precipitate protein.After centrifugation,the supernatants were transferred and evaporated under a stream of nitrogen until dryness and reconstituted with mobile phase.Then the residuals were analyzed by LC/MS/MS system with multiple reaction monitor model; the concentration of cotinine were quantified by the isotope internal standard method and the stand curve was employed with a series of calibration.To estimate the precision of the method,five frozen serum pools were repeatedly analyzed in five runs,and every pool was analyzed in triplicate.In addition,the recovery rates were analyzed with the serum sample added with different levels of standard.The stability of cotinine in serum preserved at room temperature,4 ℃ and - 80 ℃,respectively.Finally,the levels of cotinine of 94 healthy subjects were measured to evaluate the distribution of cotinine with different smoke statuses.Results Serum cotinine measured by ID-LC/MS/MS was separated well with few interferences.The correlation coefficients between the peak area ratios and cotinine concentrations were higher than 0.9993.The values of within-run coefficients of variation (CV) of five frozen serum pools (0.68,48.42,94.34,250.95 and 287.04 μg/L) were 2.19％,0.78％,0.75％,0.65％ and 0.67％,respectively.The values of total CV were 4.71％,1.40％,1.98％,1.10％ and 1.03％,respectively.The limit of detection (LOD) and limit of quantitation ( LOQ ) were 0.013 and 0.050 μg/L,respectively.The analytical recoveries ranged from 99.22％ to 102.67％.The samples could maintain stability within 2 d at room temperature,7 d at 4 ℃ and 3 months at -80 ℃ resulting the accuracy of measurements from 99.28％ to 100.87％ and the CV＜5％.The levels of cotinine of 94 healthy subjects were measured and shown skewed and leptokurtic distribution.The concentrations of twenty smokers,fourteen former smokers and sixty non-smokers were 116.40 (63.17 -241.12),0.67 (0.15 - 0.95 ) and 0.22 (0.15 - 0.42 ) μg/L,respectively.Furthermore,the level of cotinine of former smokers (Z =-2.12,P ＜0.05) and smokers (Z =-6.67,P ＜0.001) were statistically higher than non-smokers.Conclusions An ID-LC/MS/MS method for serum cotinine detection has been established.It is hoped that the method will be applied to the assessment of smoke exposure and its association with the risks of smoking related diseases since it is simple,specific,precise,sensitive and accurate.

Objective To evaluate the analytical performance of 7 homogeneous HDL-cholesterol reagents. Methods An altracentrifugation-HPLC method was used as the comparison method. Fourty fresh patient samples were analyzed by homogeneous methods and the comparison method. The homogeneous methods were all performed on a Hitachi 7170A chemistry analyzer according to the manufacturer's instructions, Precision, accuracy and total errors were analyzed. Results The homogeneous assays typically demonstrated within run coefficient variance(CV) of < 1%, and total CV of < 3%. Methods A, B and D showed average bias, bias at the medical decision points and total errors all within the NCEP performance criteria and method C and F unacceptable biases (-19.74% and 11.46%, respectively) and total errors according to the NCEP criteria. However, all the homogenous methods (A-F) had total errors of < 30%, as required by the US Clinical Laboratory Improvement Amendment (CLIA). Conclusions Homogeneous HDL-C assays have been shown to be reasonably precise, but discrepant results have been observed with some of the assays. Clinical laboratories should pay more attention on selecting and validating homogeneous HDL-C reagents.

Objective To standardize total cholesterol (TC) and HDL cholesterol (HDLC) analytical systems with the US CDC TC reference method and HDLC designated comparison method (DCM). Methods CDC TC reference method and HDLC DCM were set up and the quality was controlled by participating in the CDC Cholesterol Reference Method laboratory Network (CRMLN) bimonthly survey. The performance of 21 TC or HDLC analytical systems from 3 manufacturers were tested with the methods according to the CRMLN certification protocols. Results The coefficient variation (CV) of TC analyses with the reference method in 18 surveys averaged 0.29% and the bias versus CDC target value 0.1%. The DCM HDLC CV in 17 surveys averaged 0.010 mmol/L(0.39 mg/dl) and the averaged biases versus CDC target and group mean were - 0.019 mmol/L (-0.72 mg/dl) and - 0.006 mmol/L (-0.25 mg/dl), respectively. Most of the TC and HDLC analysis events (> 90%) satisfied the CRMLN accuracy and precision criteria for the reference method and DCM. Eighteen of the 21 tested TC or HDLC systems met the performance criteria for analytical systems and were certified for traceability by CDC. Conclusions A reference method for cholesterol and a DCM for HDLC and performed within an international reference laboratory network have been established and used for certification of TC and HDLC analytical systems, Further application of the methods to the standardization of lipid analysis are expected.

Objective To develop an HPLC method for the measurement of fractional and molar esterification rate of serum HDL(FER HDL and MERHDL).Methods Blood samples were mixed with 5,5'-dithiobis-(2-nitrobenzoic acid)(DTNB)and the sera were separated.Serum HDL fractions were prepared by precipitation with Dextran sulfate and magnesium and the fractions were incubated at 37℃for 1 h in the presence and absence of 2-mercaptoethanol(ME).Free cholesterol levels of the HDL fractions were analyzed by HPLC and FERHDL and MERHDL were calculated.Results Under the selected conditions,serum free cholesterol could be stabilized by inhibition of LCAT with DTNB and the inhibition be reversed by ME. The total CVs for FERHDL and MERHDL were 1.59%-3.74% and 1.64%-2.88%,respectively.The averages of FERHDL and MERHDL in 70 apparently healthy subjects were 18.7%/h and 42.7 μmol·L-1·h-1 with standard deviations of 7.2%/h and 11.8 μmol·L-1·h-1·respectively,and the medians were 16.1%/h and 11.8 μmol·L-1·h-1.Close correlations of FERHDL.and MERHDL with other cardiovascular disease risk factors were observed.Conclusion A new method for the measurements of FERHDL and MERHDL by HPLC has been estabished. The method is safe, precise and simple and applications in the assessment of cardiovascular diseases risks are expected.

Objective To solve problems of ice bag in process of physical cooling for high fever patient, which is contacted area small and fixed difficulty and so on, and develop a new ice capsule for all kinds of special-site cooling. Methods Selecting glycol liquor, thin aluminum plate, high-density sponge, flexible plastic and other materials and according to clay characteristics, producing 12 different types of shaping ice capsule. Results "Wear-capsule of medical care" was developed in 2002 and assessed to national patents of utility model. The flexible shaping liquid ice capsule was declared national invention patents in 2007. Conclusion The liquid flexible ice capsule can not be coagulated, wear conveniently, applicable to each kind of different spot and had good cooling effect.

BACKGROUND:To date, it is stil unclear whether the intramuscular injection of heterogeneous umbilical cord mesenchymal stem cel s (UC-MSC) can cause cardiac ectopic pathological angiogenesis as wel as increase col agen synthesis to promote myocardial fibrosis. OBJECTIVE:To explore the effects of intramuscular injection of human UC-MSCs on myocardial micrangium and col agen expression in normal Wistar rats. METHODS:After 2 weeks of feeding, 60 male SPF Wistar rats were randomly assigned to receive intramuscular injection of PBS (normal group), DMEM (culture medium group), human UC-MSCs supernatant (supernatant group), 0.25×105, 1.0×105, 4.0×105 human UC-MSCs (low-, moderate-and high-dose groups), respectively (n=10 per group). Al the rats were subjected to second injection (same dose) at 4 weeks after first intramuscular injection. Then, the rats were kil ed under anesthesia at 4 weeks after second injection, to take heart tissues from the left ventricle for pathological observation, immunohistochemical examination and Masson staining. RESULTS AND CONCLUSION:No alteration of the response, activity, victualage, faeces, weight growth, and fur was found, and there was no death in rats during the experiment. Al the rats had no symptoms of molt, inflammation, skin ulcer, scleroma. Strong positive expression of CD34 for the micrangium in the myocardial tissue was observed, and positive expression of the col agen in the myocardial tissue observed by Masson staining. There were no significant differences in the microvessel density and col agen expression in the myocardium among the groups (F=0.110 and 0.585, P>0.05). To conclude, hUC-MSCs or its supernatant via intramuscular injection has no effect on the micrangium and col agen expression in normal rats.

Objective To develop an HPLC method for the measurement of n-3 fatty acid index of serum cholesteryl esters.Methods Serum triglycerides were hydrolyzed with ethanolic sodium hydroxide and cholesteryl esters (CEs) were extracted with hexane.The extracted CEs were analyzed by reversed phase HPLC with a UV detection at 205 nm.Cholesteryl eicosapentaenoate and docosahexaenoate ( major n-3 fatty acid cholesteryl esters) were identified by liquid chromatography-tandem mass spectrometry and cholesterol in each CE fraction was measured.Peak areas of CEs were corrected for cholesterol and CE n-3 index was calculated using the corrected peak area and expressed as the percentage of n-3 fatty acid CEs in total CEs.Results The HPLC analysis can be finished in 6 minutes.Triglycerides which interfere with the determination of n-3 fatty acid index, were hydrolyzed with ethanolic sodium hydroxide (4 mol/L) in 30 seconds.The within-run and total CVs for CE n-3 index averaged 0.66% and 0.90%, respectively.CE n-3 indexes of 70 volunteers and 36 coronary heart disease patients apparently healthy subjects and patients with coronary heart disease in Beijing Hospital appeared to be positively skewed and leptokurtic distribution ( skewness = 1.25, kurtosis = 1.70 ).The median of n-3 indices were 0.98% ( 0.37% - 2.40% ).The logarithm of n-3 index appeared to be normal distribution and the average is 0.003 7% with standard deviations of 0.15.The distribution of n-3 indices of gender groups was similar with the total.The medians of females and males were 1.08% (0.60% -2.40%) and 0.95% (0.37% -2.11%) respectively, and the former were significantly higher than the latter( t = - 3.021, P = 0.003 ).Conclusion A new method for the measurement of n-3 index of serum cholesteryl esters by HPLC has been established.It is simple and precise and can be used in predicting cardiovascular diseases risks and monitoring dietary intake of n-3 fatty acids.