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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.
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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.
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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.
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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.
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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.
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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.
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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
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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..
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AIM: To explore the relationship between various mitochondrial (mt) DNA tRNA Leu (UUR) and ND1 gene mutations and type 2 diabetes mellitus (T2DM) among Chinese in Hubei Province. METHODS: PCR restriction fragment length polymorphism (PCR-RFLP) analysis was used to screen point mutations of mtDNA ( 3 243, 3 256, 3 290, 3 316, 3 394, 3 421, 3 426, 3 460, 3 593) in 174 T2DM and 207 healthy controls. Then, DNA sequencing, reverse dot blot hybridization and Genchip were used to compare and confirm mutations. All mutations were analyzed by DNASTAR and Antherprot softwares. RESULTS: In diabetic group, there were 5 carriers (2.9%) of 3 316 G→A (Ala→Thr) mutation, 4 (2.3%) of 3 394 T→C (Tyr→His) mutation, 1 (0.6%) of 3 593 T→C(Val→Ala) mutation, and 1 (0.6%) of 3 618 T→C(Phe→Phe) mutation. Among 3 316 (G→A) mutations , there were more than 1 point mutations in 2 cases, one accompanied with 3 256 C→T(Arg→Arg) and 3 688 G→C (Ala→Pro) mutations, another accompanied with 3 606 A→G(Leu→Leu) mutation. 3 606 (A→G), 3 618 (T→C) and 3 688 (G→C) were novel mutations, GenBank accession number is DQ092356. In controls, only 3 316 (G→A) mutation was found in 1 subject (0.5%). There was significant difference between two groups for 3 394 (T→C) mutation frequencies (P