Objective To analyze the plasma free fatty acid (FFA) composition in patients with T2DM. Methods All subjects were from Zhongnan hospital of Wuhan university, and they were divided into three groups: normal control ( n = 94 ), T2DM ( n = 101 ) and T2DM with hyperlipidemia ( n = 77 ). Fasting blood samples were taken from the participants, and plasma FFA were separated using a modified Doles method with the bromoacetophenone, pre-column-derivative. The quantitation of FFA was performed on were (355.63 ± 100. 35) μmol/L, (421.21 ± 200. 83 ) μ mol/L, ( 473.04 ± 213.40 ) μmol/L in healthy controls, T2DM group and T2DM with hyperlipidemia group, respectively. The significant differences were observed among the 3 groups(x2 = 13.08, P <0.01 ). However, there was no significant difference of UFA concentrations among the 3 groups [(206.29± 61.94) μ mol/L, (218.11 ± 110.28) μmol/L and ( 240.94 ± 116.79 ) μmol/L, x2 = 2.17, P > 0.05]. Compared to normal control [( 355.63 ± 100.35 )μmol/L], the FFA concentration[(421.21 ±200.83) μmol/L] in T2DM has significantly increased (x2 =FFA concentrations were higher in T2DM with hyperlipidemia [(473.04 ±213.40) μmol/L] (x2 =27.93,P <0.01 ). The RSD values for intra- and inter-day precision were less than 5%, and the minimal detection limits ranged from 0.05 μmol/L to 0.35 μmol/L The recoveries of high, intermediate and low-level materials were 96.4% -104.8%. Conclusions The total FFA concentration in T2DM has increased, most of which are saturated FFA. The unsaturated FFA has not significantly increased. They seem to be related to the development of T2DM, and might be a new biomarker for clinical monitoring of metabolic disorder of T2DM.

Objective To Evaluate the environment quality in underground construction site based on the measurement of atmosphere composition and microclimate. Method Concentrations and composition of gases were analyzed by combined detection with mass spectrometer and gas chromatograph. Microbial content was detected by striking method. A microclimate model was built based on the measurements of temperature, humidity and wind speed in two years. Result Concentrations of inorganic gas was higher than national standard. Total volatile organic compounds were in the range of 4～6 mg/m3, which highly exceeded the national standard. Concentration of O2 descended obviously at the time when personnel went into the construction site, and was lower than the national standard of 299 mg/m3 after continuous working for 3 h. When an aerator was started, the average noise level was in the range of 70～93 dB, which was higher than the national standard(55～70 dB). In the course of working, the average microbial content reached 4 000 cfu/m3 which was 3 times as high as the background level. The microclimate characteristic with high humidity, low temperature and inadequate sunshine was unsuitable for dwelling. Conclusion Environment in the underground construction site is unfavorable as compared with that on the ground. The physiological function of working staff may be influenced obviously due to cumulative effect of the above mentioned factors, and may lead to aprosexia, hypomnesis, fatigue, etc. Six kinds of protection means are suggested in the paper.

Case-based learning model was applied in teaching the course of Clinical Biochem-istry and Diagnosis for postgraduates with professional degree. Dyslipidaemia was chose as teaching content and one distinctly characterized case along with four typical cases were selected. Information of cases and core problems were submitted to students to prepare before the class. In the class, stu-dents were grouped and discussed independently. Students put forward the pathogenesis of the disease and the clinical laboratory test items for diagnosis and differential diagnosis. Teachers should adhere to the following principles: making the class student-centered and highlighting consciousness of stu-dents; creating clinical environment to stimulate enthusiasm for learning; problem-oriented and taking capacity building as the priority. After the class, teaching evaluation was designed to promote the con-tinuous improvement of teaching cases.

125 strains of the symbiotic and epiphyte microorganisms were isolated from marine organisms (sea cucumber, sea urchin,anemone, sea actinia, Ulra, Sargassum, Undaria). Among them,21 strains of bacteria,8 strains of actinomycetes and 2 strains of fungi showed antagonistic activity on bacterial or fungal growth. In the 21 strains of bacteria, 7 strains belong to Bacillus sp., 11 strains Vibrio sp., and 3 strains Pseudomonas sp.. In the 8 strains of actinomycetes, 5 strains belong to Streptomyces sp., 3 strains Micromonospora sp., 2 strains fungi Penicillum sp..

Objective To investigate the necessity and feasibility of setting up elective course of children's paychological problems in medical colleges and universities.Methods The course was practiced by using varied teaching methods such as theory teaching,psychological chat show video,seminar,students speaking,writing paper and etc.Results The attendance was more than 95％ and more than 90％ students submitted the lesson summary and spoke positively in classroom.The scores of 90％ students were more than 80 points and the course evaluation results were good.Conclusion It's important to set up this elective course and this course is worth further developing.

Objective To investigate the effects of enriched environment and impoverished environment on brain function after hypoxic-ischemic brain damage(HIBD).Methods Three-day-old SD rats,which were divided into enriched environment group(EE),standard environment group(SE) and impoverished environment group(IE) by random digits table were used to establish HIBD model.The sham-operation rats served as control group.Different environment stimulations were administrated to the rats respectively since the 2nd day after HIBD.On the 32nd day,hanging test and incline plane test were carried out to evaluate the sensorimotor function.Morris water maze was used to evaluate the learning and memory ability.The expression of GFAP in the cortex and hippocampus was measured by the method of immunohistochemistry.Results The brain function of IE group was much worse than that of SE group(P0.05).Immunohistochemical analysis showed the expression of GFAP in the cortex and hippocampus of Sham group was significantly lower than that of other groups(P

Objective To study on the association of apolipoprotein E(apoE)genotype with coronary heart disease(CHD)and type 2 diabetes mellitus(T2DM).Methods PCR-RFL,multiplex amplification refractory mutation system(muli-ARMS)and PCR-SSCP methods were used to detect the genotype of apoE,and DNA sequencing technique were used for further confrm the genotype and gene variations in 2 446 Chinese individuals,including 238 cases of CHD,316 cases of T2DM and 1 892 healthy controls.Fasting blood glucose(FBG)and plasma lipids levels[TC,TG,HDL-C,LDL-C,apoA I,apoB and Lo(a)]were measured by usual methods.Results Compared with the controls,plasma HDL-C(t=2.66)and apoA I(t=2.30)levels in the CHD group were significantly lower(P<0.05),but not in T2DM group;plasma TC level(t=5.22)in the T2DM group were significantly higher(P<0.05),but not in CHD group;systolic pressure(t=8.48,5.74)diastolic pressure(t=5.66,3.35),plasma TG(t=3.38, 4.56),LDL-C(t=2.48,7.00),apoB(t=1.67,2.24),Lp(a)(t=4.16,4.15)and FBG(t=7.04, 16.93)levels were significantly higher in both CHD group and T2DM group(P<0.05).The distributions of apoE ε2/2,ε2/3,ε3/3,ε2/4,ε3/4 and ε4/4 respectively were 0.4%,13.4%,58.0%,1.3%, 26.5%,0.4% in the CHD group;0.6%,5.7%,72.8%,1.9%,14.9%,4.1% in the T2DM group; 0.5%,10.5%,69.6%,1.6%,16.8%,1.1% in the control group.Significant differences were found between the CHD group(χ2=14.90,P=0.00),T2DM group(χ2=7.08,P=0.03)and the control group for the frequencies of apoE genotype.The distribution of ε3/4 was higher(26.5% vs 16.8%)and ε3/3 Was lower(58.0% vs 69.6%)in the CHD group.In the T2DM group.the distribution of εε4/4 Was higher (4.1% vs 1.1%),and 2 cases of ε3/3 with Arg 150 His mutation in exon 4 of apoE gene were firstly reposed in China,which is none in the CHD and control groups.Conclusions The results suggested that apoE ε3/4 and ε4 genotypes might be associated with the susceptibility of CHD and T2DM.respectively. To some extent,apoE ε3/3 may not be a good genotype for T2DM because of the Arg 150 His mutation. Blood pressure and plasma lipids could be used for diagnosis of the two diseases.

Objective To detect eight kinds of clinical common pathogenic bacteria by DNA microarray.Methods Eight kinds of common pathogenic bacteria,including Staphylococcus aureus,Pseudomonas aeruginosa,Klebsiella pneumoniae,Escherichia coli,Proteus mirabilis,Enterobacter aerogenes,Pseudomonas fluorescens,Shigella sonnei were collected.Universal primers were designed to amplify 16S rRNA gene fragment from the genomic DNA of the eight bacteria,and probes were designed in the highly variable regions.DNA microarray detection system was established and used for detection of colleted bacteria.A total of 50 samples were collected from the Zhongnan Hospital of Wuhan University,including 6 blood samples,32 sputum samples,9 feces samples and 3 bronchoscope lavage samples.DNA were extracted and detected by the established DNA microarray system.Results The desired fragments were well amplified by the self-designed universal primers.The selected probes had good detection results according to repeated detection.Of the 50 samples detected,pathgenic bacteria were accurately detected in 47 samples.Other three samples were not detected as those bacteria were not included in the chip.By optimizing the detection process,the results could be reported within 8 hours.Observation of probe signal attenuation indicated that even attenuated after 60 days,but the attenuation did not affect the results.Conclusion A microarray system was established for detection of clinical common bacteria accurately and quickly,which provided foundation for its clinical application.

Objective To develop a gene microarray system for detection of clinical common pathogenic fungi.MethodsThere were 8 clinical common fungi chosen as the subjects including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilokis,Candida pseudotropicalis,Aspergillus terreus,Aspergillus flavus,Aspergillus oryzae,Aspergillus fumigatus.Universal primers,probes and specific probes for the PCR amplification and microarray preparation were designed in ITS region of the fungi genomic DNA.The PCR products amplified from those fungi's genome DNA were denatured and hybridized with the probes in gene microarray.The rapid detection of fungi was based on the investigation on the fluorescent signal intensity in the chip.The detection results of gene microarray system were verified by true positive and negative clinical samples.Results There were totally 25 positive samples identified by clinical routine microbiological methods.The 10 samples identified as bacteria positive were determined as negative without fluorescent signal by the fungi gene microarray,while the 12 samples identified as fungi positive were determined as positive with certain fungus by the fungi gene microarray.And 3 artificial Candida krusei samples were detected as fungi positive,while they were failure to be identified as certain fungus.There was no fluorescent signal in positions of the 8 fungi specific probes,but there was fluorescent signal in the position of fungi universal probe.It indicated that there were fungi in the samples but it couldn't identify the species of the fungi,because the Candida krusei wasn't included in the detection fungi list of the fungi gene microarray.ConclusionsThe fungi gene microarray established by the study could detect the common fungi in clinic rapidly and accurately.This study lays technology foundation for clinical application of gene chip.

Objective To investigate the antitumor effect of Haishengsu injection extracted from marine shellfish. Methods Transplant tumor models of sarcoma 180 (S 180), Ehrlich ascites carcinoma (EAC), and hepatoma (Heps) in mice were established. Different doses of Haishengsu injection were given to the mice and the tumor inhibition rates of Haishengsu injection, life span of the mice were calculated. Results The tumor inhibition rates of haishengsu injection (490～1000mg?kg -1?d -1,iv) were 41.10%～49.08% in mice with S 180 and 36.29%～49.19% in mice with hepatoma,respectively. The same doses of Haishengsu injection prolonged the life spans of EAC-bearing mice by 22.93%～69.98%. Conclusion The haishengsu injection has the antitumor effects on the tumor-bearing mice without evident side effects.