Table of Content

21 April 2012, Volume 33 Issue 2

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体外诊断专题

Association between serum CXCL16 levels and carotid atherosclerosis in stroke

WANG Ke-di;LIU Zhi-zhong;WANG Rui-min;KANG Xi-xiong

2012, 33(2):  139-142.  doi:10.3969/j.issn.1006-7795.2012.02.001

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Objective To study association between serum CXCL16 levels and carotid atherosclerosis in patients with stroke. Methods A total of 135 cases with acute atherosclerotic stroke were examined using ultrasound to evaluate their carotid atherosclerotic lesions, the patients were then divided into normal(intima-median thickness,IMT) and increased IMT group, without plaque, stable plaque and vulnerable plaque group, 4 groups according to carotid plaque area(CPA) quartiles. Results Serum CXCL16 levels were closely associated with increased IMT, vulnerable plaque and increased CPA, logistic analysis showed that increased serum CXCL16 levels was independent risk factor of increased IMT, vulnerable plaque and increased CPA. Conclusion CXCL16 is an independent risk factor of carotid atherosclerosis in stroke, detection of it may be a new way to find early atherosclerosis.

Distribution and antibiotic resistance of bacteria isolated from cerebrospinal fluid of patients in neurology and neurosurgery departments

JING Nan;TANG Ming-zhong;LIU ZHi-zhong;KANG Xi-xiong

2012, 33(2):  143-147.  doi:10.3969/j.issn.1006-7795.2012.02.002

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Objective To determine the distribution and antibiotic resistance of bacteria isolated from cerebrospinal fluid so as to provide evidence for clinical selection of antibiotics. Methods The bacterial identification and antibiotic resistance tests were performed for cerebrospinal fluid culture-positive specimens.Results 1) Comparison of bacteria constituent ratio: the ratio of Gram-positives was higher in Neurosurgery Department(67.2%) than Neurology Department(56.1%). The ratio of Pseudomonas aeruginosa in Gram-negatives was higher in Neurology Department. The ratio of Acinetobacter baumanii was higher in Neurosurgery Department. 2) Bacterial susceptibility: The Gram-positives were sensitive to vancomycin and Linezolid, while Gram-negatives were sensitive to imipenem and amikacin. Pseudomonas aeruginosa, isolated from Neurosurgery Department, had higher antimicrobial susceptibility to ciprofloxacin than those from Neurology Department. Staphylococcus epidermidis in Neurosurgery Department were more sensitive to Rifampicin. Conclusion Central nervous system infection was basically nosocomial infection and the prevailing pathogens were Gram-positives. The observation of bacterial distribution and antibiotic resistance will provide valuable information for prophylactic and empirical antibiotic use in intracranial infection.

Evaluation of a rapid and efficient method for detecting norovirus infection

LV Hong;Gao Zhi-yong;ZHANG Guo-jun;YAN Hui-ping;KANG Xi-xiong

2012, 33(2):  148-152.  doi:10.3969/j.issn.1006-7795.2012.02.003

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Objective To find a quick and efficient method to detect norovirus(NV) by comparing different approaches using reverse transcription-polymerase chain reaction(RT-PCR) and transcription reverse transcription concerted(TRC). Methods From February to March 2007, the fecal specimens were collected from 364 sporadic cases of acute viral gastroenteritis in Beijing area. RT-PCR and TRC were used to detect human NVs in stool specimens; RT-PCR was performed to evaluate the results by TRC and analyzed the sensitivity and specificity of results by TRC. The consistency of TCR was evaluated with RT-PCR by kappa test. Results A total of 160 positive cases were detected by TRC out of 364 stool specimens collected from infants and young children or adults with acute viral diarrhea, and NV positive rate was 43.96%, among which 3.85%(14/364) were identified as infection by NV GGI and 40.11%(146/364) as infection with NVs GGⅡ. In positive cases, 78.58% NVs GGI and 89.73% NVs GGⅡ were confirmed within 20 minutes. Men and women had no significant difference in the percentage of HuCV infection. The highest positive rate(63.63%) was found in group aged from 50 to 59. The lowest positive rate(18.93%) was found in the group aged from 10 to 19. No significant difference was seen among age groups except for the group aged from 10 to 19. Conclusion By comparing with RT-PCR, TRC presented a good sensitiveness and specificity which seem to be more convenient and time saving, indicating a good broader application.

Diagnosis of bacterial vaginosis by 16SrRNA genes PCR

LV Zhi;PENG Guo-li;SU Jian-rong

2012, 33(2):  153-157.  doi:10.3969/j.issn.1006-7795.2012.02.004

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Objective To determine the organism's isolation rate in bacterial vaginosis (BV), to establish a molecular diagnostic method for BV. Methods Totally 173 vaginal-fluid samples(91 with BV and 82 normal) were screened by clinical Amsel criteria. Bacterial genome DNA was extracted from the vaginal secretion samples. Eleven major vaginal bacterial species were detected by using 16SrRNA gene PCR assays to confirm its association with BV, and the utility of PCR for the microbiological diagnosis of BV. Statistical analysis was used to establish the best bacterial combination for diagnosis of BV. Results Leptotrichia/Sneathia, Atopobium vaginae, Megasphaera species, Mobiluncus mulieris and Prevotella among the bacterial species were significantly associated with BV, the organism's isolation rate between two groups had significant difference(P<0.01). As a diagnostic criteria of BV, getting a sensitivity of 93.41% and a specificity of 78.05% by co-detecting the three species, Atopobium vaginae, Leptotrichia and Megasphaera. Conclusion BV is a polymicrobial disease. PCR detection of one or more fastidious bacterial species is an effective reliable molecular diagnostic method.

Analysis of false positive results of Sysmex UF-1000i urine sediment analyzer

LIU Chang-jun;LIU Ting;DONG Jin;JIANG Hui;TIAN Ya-ping

2012, 33(2):  158-160.  doi:10.3969/j.issn.1006-7795.2012.02.005

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Objective To evaluate positive results of UF-1000i Urinary sediment analyzer and analyze false positive results. Methods A total of 500 urine samples were collected randomly and determined by UF-1000i full automatic urinary sediment analyzer(UF-1000i), urine strips and microscopic analysis. positive results of samples were deemed as false when results were abnormal with UF-1000i analyzing but normal with urine strips and microscopic analysis. Two results of urine formed elements were contrasted between UF-1000i and microscopic analysis in samples with false positive results. Results Totally 178 of 500 samples showed positive results with UF-1000i analysis, in which 20 samples were determined as false positive results in main examined items with microscopic analysis. In 178 positive results, the false positive rate of erythrocyte(RBC), leukocyte(WBC), cast and epithelial cells(EC) were 10.8%, 8.2%, 23.1% and 14.3% separately. Conclusion The analyses on urinary sediments by UF-1000i full automatic urinary sediment analyzer was effective. But it is important to observe sediments by microscope to avoid false positive results when results were abnormal with UF-1000i analysis.

Laboratory study on GyrA gene mutation in enterococci and multi-drug resistance efflux pumps result in enterococcal resistance to fluoroquinolones

YAN Dong-hui;XU Shu-zhen;SU Jian-rong

2012, 33(2):  161-164.  doi:10.3969/j.issn.1006-7795.2012.02.006

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Objective To study the relations between enterococcal GyrA gene mutation, enterococcal multi-drug resistance efflux pumps and fluoroquinolones resistant in enterococci. Methods All strains were identified using VITEK-2 compact and GPI cards. Mutation of GyrA gene in strains of 30 E.faecalis and 10 E.faecium of levofloxacin-resistant and ciprofloxacin-resistant were detected using PCR and single strand configuration polymorphism(SSCP), testing results and mutant types were confirmed by sequencing analysis. At the same time, the minimal inhibitory concentration(MIC) of 30 strains of enterococci against ciprofloxacin were tested using M-H agar including reserpine that final concentration was 20 μg/mL. Results All the 40 strains of enterococci resistant to Levo and Cip, were successful to amplify GyrA gene; 24 strains of E. faecalis and 4 E. faecium single-strand electrophoresis band appeared abnormal compared with ATCC 29212. The sequencing of 2 strains of E. faecalis had codon alterations at 87:GAA→GGA; 2 strains of E. faecium had the codon alteration at 83: AGT →TAT. Alterations of codons lead to the change of amino acids. Parts of E. faecalis' MICs were decreased adding reserpine in the M-H agar. Conclusion All 40 strains enterococci GyrA gene were detected by PCR-SSCP and confirmed by sequencing analysis, 70%(28 strains) had gene mutation. This result indicated GyrA gene mutations are main causes, which resulted in fluoroquinolones-resistance in enterococci. In addition, Multi-drug resistance efflux pumps were also existent in enterococci.

Preparation and stability of the mixed human candidate reference standard serum for gamma-glutamyltransferase

XU Jing;ZHANG Jian-ping;WANG Qing-tao

2012, 33(2):  165-168.  doi:10.3969/j.issn.1006-7795.2012.02.007

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Objective To investigate the feasibility of freezing admixing human serum which was the selected standard materials of gamma-glutamyltransferase(GGT), and to observe and analyze their stability at -80 ℃. Methods The GGT serum which had been collected was mixed, filtered and subpackaged according to five levels. Then the selected standard materials were measured at 0, 7, 30, 60, 120, 180, 330, 360, 390 and 750 days. In order to evaluate the stability, the results were statistic analyzed by linear regression. Results In the 750d, the P of the linear regression about C1~C5 was more than 0.05(the P of C1~C5 was 0.886, 0.753, 0.585, 0.571 and 0.945). Conclusion The preparation method of the freezing admixing human serum which was the selected standard materials of GGT was simple and convenient, otherwise the stability of the selected standard materials was well in 750 days.

Analysis of resistance features of anti-candida drugs

ZENG Zhao-ying;SU Jian-rong

2012, 33(2):  169-176.  doi:10.3969/j.issn.1006-7795.2012.02.008

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The prevalence of invasive fungal infection is increasing day by day. A significant number of candida isolates resistant to antifungal agents have been identified. Here we provide a brief presentation on the present status of drug resistance features of anti-candida drugs.

Advances in studies on the relationship between Toll like receptors and autoimmune diseases

MA Rui-min;ZHANG Guo-jun;KANG Xi-xiong

2012, 33(2):  177-181.  doi:10.3969/j.issn.1006-7795.2012.02.009

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Toll like receptors(TLRs) are a class of transmembrane receptors. They are expressed mainly in antigen presenting cells as well as epithelial cells, and can activate immune reaction by recognizing specific ligands. Besides exogenous ligands, TLRs can also recognize some of the endogenous ligands. Overactivation of TLRs' endogenous signal will induce autoimmune diseases. In this review, we summarize the types of TLRs and their ligands, and discuss the TLRs pathways and functional regulation. We focus on the recent researches about the association of TLRs with some autoimmune diseases, and prospectively discuss the roles of TLRs in the therapy of these diseases.

基础研究

Effect of echinomycin on the cell viability of PC12 cells after it inhibits hypoxia inducible factor-1α

LI Ran;WANG Yong;GONG Xiao-li;FAN Ming;WANG Xiao-min;ZHU Ling-ling;WANG Xuan

2012, 33(2):  182-186.  doi:10.3969/j.issn.1006-7795.2012.02.010

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Objective To clarify the role of hypoxia inducible factor(HIF)-1α in the process of hypoxia-induced cell death through inhibiting its transcription activity. Methods 1 PC12 cells were seeded in the 12-well plate and treated for 24 h in normoxia(20% O₂) or hypoxia(3% O₂). Then photomicrographs under light microscope were taken and analyzed for situation of cell growth by cell-counting method. 2 PC12 cells were seeded in 96-well plate and treated for 24 h in 20% O₂ or 3% O₂. A test for cell survival called MTS was further used to detect the cell viability of PC12 cells. 3 PC12 cells were treated with HIF-1α inhibitor-echinomycin for 24 h or 48 h at 5 nmol/L, 50 nmol/L and 500 nmol/L, respectively. Then MTS was used to detect the cell viability. Results 1 3% O₂ induced the death of PC12 cells significantly contrary to 20% O₂. 2 Echinomycin administration eliminated the cell survival difference between 20% O₂ and 3% O₂ at 24 h. 3 After a 48 h-treatment with echinomycin, it even reversed the hypoxia-enhanced cell death relative to normoxia. Conclusion Hypoxia could promote the death of PC 12 cells. After we used HIF-1α inhibitor echinomycin for 24 h, the death-promotion role of hypoxia was weakened. It even reversed the hypoxia-enhanced cell death after a 48 h-treatment with echinomycin. All these data suggested that the excessive transcription activation during hypoxia was detrimental to PC 12 cells.

Establishment of stably transfected cell line expressing BβArg448 or BβLys448 fibrinogen

LIN Yuan;LI Ji-feng;LIU Jie;NIU Meng-lin;SUN Ran;WANG Jun;LIU Yan;WANG Chen;

2012, 33(2):  187-192.  doi:10.3969/j.issn.1006-7795.2012.02.011

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Objective To investigate the effect of BβArg448Lys polymorphism on the function of fibrinogen, and to establish the stably transfected cell line expressing fibrinogen with BβArg448 and BβLys448 polymorphism. Methods Mutant fibrinogen Bβ chain cDNA(BβLys448) pMLP-FGB(448A) was generated from wild type Bβ chain cDNA(BβArg448) pMLP-FGB(448G), which contains the entire fibrinogen Bβ chain cDNA using site-directed mutagenesis method. The expression vectors encoding the entire fibrinogen Aα chain, wild type/ mutant type Bβ chain and γ chain cDNA were introduced into CHO-K1 cells respectively using Lipofectamine. The expression of all three chains in the transfected cell line was tested by RT-PCR. Results The stably transfected cell line expressing fibrinogen with BβArg448 and BβLys448 polymorphism was successfully established, which provide a useful cell model to the further functional study.Conclusion The stably transfected cell line expressing fibrinogen with BβArg448Lys and BβLys448 polymorphism was successfully established, which provide a useful cell model to the further functional study.

Co-expression of β₂ adrenoceptor and mucin2 in rat colonic mucosa

LI Xiao-feng;ZHANG Yue;LI Yun;ZHU Jin-xia

2012, 33(2):  193-197.  doi:10.3969/j.issn.1006-7795.2012.02.012

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Objective To investigate the effect of noradrenalin on the colonic mucus secretion and the distribution of β adrenoceptors in the colonic mucous cells. Methods Enzyme-linked immunosorbent assay(ELISA) was used to investigate the effect of norepinephrine(NE) on the colonic mucus secretion; immunofluorescence and laser scanning confocal microscope were used to observe the distribution of mucin2(MUC2) and β adrenoceptors in rat colonic mucosa. The mRNA expression of the β adrenoceptors in the rat colonic mucosa was analyzed by real-time PCR. Alcian blue/periodic acid schiffs(AB/PAS) staining was used to detect the distribution of mucous cells. Results NE stimulated the secretion of colonic MUC2 from(0.19±0.03) ng/mg to (0.47±0.09)ng/mg, increased by 247%. The mRNA expression of β₁ adrenoceptor and β₃ adrenoceptor was weak and β₂ adrenoceptor was the highest among the adrenoceptors expression in the colonic mucosa. Immunofluorescence staining results indicated the co-expression of MUC2 and β₂ in the colonic mucous cells. Conclusion NE is able to stimulate muc2 release from rat distal colonic mucosa.

Effects of overexpression of apolipoprotein A-Ⅰ on lipid accmulation in BEL-7402 cells

QIN Ling;WANG Yu-tong

2012, 33(2):  198-204.  doi:10.3969/j.issn.1006-7795.2012.02.013

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Objective To explore a new therapy to change the lipid metabolism and prevent non-alcoholic steatohepatitis(NASH), to establish a cell model that overexpresses apolipoprotein A-Ⅰ(apoA-Ⅰ), which promotes cellular cholesterol efflux by interacting with ATP-binding cassette transporter A1(ABCA1). The effect and its molecular mechanism of apoA-Ⅰ overexpression on cellular lipid levels were evaluated in lipid-loaded cells to investigate the therapeutic effect of apoA-Ⅰ on NASH. Methods 1 Human liver cancer cell line BEL-7402 were treated with different concentrations and types of fatty acids for 6, 12 and 24 h, cellular lipid accumulation was determined by Oil Red O staining, and optimal concentration and time of fatty acid treatment were then established; 2 BEL-7402 cells were treated with different concentrations and types of fatty acids for 6, 12 and 24 h, and cell activity was determined by MTT; 3 Control plasmid and pcDNA3.0/apoA-I plasmid were transfected into BEL-7402 hepatocytes, and intracellular and extracellular apoA-Ⅰ protein levels were determined by Western blotting; 4 Cellular cholesterol, free fatty acid and triglyceride content of control cells and apoA-Ⅰ-overexpressive cells were determined respectively. Results 1 The lipid accumulation in BEL-7402 cells could be achieved by treatment with 200 μmol/L or 400 μmol/L fatty acids for 6, 12 and 24 h , and was both dose- and time-dependent. 2 Saturated fatty acid palmitic acid decreased the cell activity, and the inhibitory effect of cell growth is coming with increasing time and concentration of palmitic acid. 3 ApoA-Ⅰ can be overexpressed intracellularly and excreted extracellularly to facilitate ABCA1 to mediate the transport of cellular cholesterol and phospholipids. 4 The cholesterol, free-fatty acid and triglyceride content in apoA-Ⅰ-overexpressive cells were less than that in control BEL-7402 cells. Conclusion Compared with the control cells, the lipid accumulation including cholesterol, free-fatty acid and triglyceride in human hepatocytes was markedly down-regulated by apoA-Ⅰ overexpression. These findings suggest that apoA-Ⅰ overexpression may increase ABCA1-mediated lipid transport, and may have a therapeutic effect on NASH.

Whole brain electroencephalogram difference research: effects of unilaterally closed and open eye

WANG Kun;CUI Wei-qi;LI Guo-zhen;CHEN Andrew CN

2012, 33(2):  205-208.  doi:10.3969/j.issn.1006-7795.2012.02.014

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Objective Electroencephalogram(EEG) is a modern examination method, which amplifies tiny bioelectrical signals of brain itself. This study focused on the differences of seven EEG spectral regional field powers: Delta(0.5~3.5 Hz), Theta(4~7 Hz), Alpha-1~(7.5~9.5 Hz), Alpha-2(10~12 Hz), Beta-1(13~23 Hz), Beta-2(24~34 Hz) and Gamma(35~45 Hz)) when the unilateral eye(right and left eye) is open and closed. Methods Fifteen healthy subjects(all were men; age: 19~27 years), in the resting state, in a quiet and comfortable room, who were asked to close and open unilateral eye(left-closed/open, right-closed/open) in counter-balanced order stimulation, while respective EEG was recorded and analyzed. Results In the resting state, regional EEG field powers of mid-frequency band Alpha-1 and Alpha-2 field powers were obviously higher in eyes-closed than in eyes-open state(Alpha-1, P=0.038; Alpha-2, P=0.02). Compared unilateral eye-closed and eye-open, there were no significant differences in alpha bands or other bands. Conclusion Alpha band power was significantly higher when subjects closed eyes than opened their eyes, however, when we compared unilateral eye-closed and eye-open differences of seven bands, significant differences was uncovered. We conclude that bilateral EEG powers are balanced via corpus callosum in persistent 2 min unilateral stimulation.

Pharmacokinetics of recombinant human endostatin in rats

CAI Yong-ming;ZENG Yong;JIANG Ling;ZHANG Jun;LIU Chang-xiao;ZHANG Zong-peng;

2012, 33(2):  209-213.  doi:10.3969/j.issn.1006-7795.2012.02.015

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Objective To characterize the pharmacokinetics, tissue distribution, and excretion profiles of recombinant human endostatin(rhEndostatin) following a single intravenous administration to rats. Methods The rhEndostatin was labeled with ¹²⁵I and the pharmacokinetics of ¹²⁵I-rhEndostatin was investigated by two assays: the total radioactivity assay(RA method) and the radioactivity assay after precipitation with trichloroacetic acid(TCA-RA method). Results The rats were randomly assigned into three groups in which the rats were given intravenously rhEndostatin at doses of 5, 10 and 20 mg/kg, respectively. The serum rhEndostatin concentrations and the area under the concentration-time curve(AUC) increased with the dose increment. The serum elimination half-life(t_1/2β) and clearance(CL) did not display markedly dose-dependence and were relatively consistent in the range of 243~265 min and 2.4~2.6 mL·kg^-1·min^-1 using TCA-RA method, respectively. In the rats with 10 mg/kg rhEndostatin(iv), the levels were the highest in the kidneys, and the lowest in the brain, muscle and adipose tissue. The accumulative excretion percentages of ¹²⁵I-rhEndostatin from urine and feces reached 86.9% and 5.8% of the administered radioactivity within 96 h, and 1.3% from bile within 24 h, respectively. Conclusion RhEndostatin has linear pharmacokinetic properties within the therapeutic dose range. The rhEndostatin has characteristics of targeting special tissue distribution(the highest level of the drug was found in the kidneys), which may reflect that urinary excretion is the dominant route of elimination via the kidneys in rats following iv administration of rhEndostatin.

Double negative T cells and lymphocyte subtype in chronic hepatitis C

LIANG Qi;JIAO Yan-mei;JI Yun-xia;LI Tian;WU Hao;ZHANG Guo-yuan;TANG Zhong

2012, 33(2):  214-217.  doi:10.3969/j.issn.1006-7795.2012.02.016

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Objective To investigate the expression and significance of CD3⁺CD4^-CD8^-(double negative T, DNT) cells and T lymphocyte subsets in chronic hepatitis C cases. Methods DNT cells and lymphocyte subsets of peripheral blood were detected by flow cytometry in 92 chronic hepatitis C patients and 93 healthy people, the differences between them was comparatively analyzed. Results The percent of DNT cells(8.57%±6.16%) were higher in patients with chronic hepatitis C than those in normal control group(6.78%±4.50%)(P<0.05), However, there was no significant difference in the absolute value of DNT cells between them(P>0.05). The CD4 cells, the ratio of CD4⁺/CD8⁺ were decreased but CD8 cells increased in comparison with normal control group(P<0.001). Conclusion The significant increase of DNT cells and the change of T lymphocyte subsets's percentage in chronic hepatitis C patients might be one of the important reason for chronicity and long-term liver damage of hepatitis C.

Eight-year change in dietary pattern and metabolic status among the elderly people in Beijing

WANG Wei;TANG Zhe;LI Xia;GAO Qi;SUN Fei;DIAO Li-jun;GUO Xiu-hua

2012, 33(2):  218-222.  doi:10.3969/j.issn.1006-7795.2012.02.017

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Objective To understand dietary pattern and to explore the development of dietary pattern and the related metabolic indices for elderly people in Beijing. Methods The baseline information in 1992 and information in the follow-up study in 2000 for Beijing Longitudinal Study of Aging, which was collected by Xuanwu Hospital, Capital Medical University was used. There were 698 elderly subjects in this analysis who fulfilled the blood test and with full information on dietary habits both in the year of 1992 and 2000. Factor analysis and cluster analysis were used to reveal dietary pattern. Latent class analysis(LCA) was a technology that discriminate people to different latent class with different answering pattern for items(manifest variables or observed variables). LCA was first used in psychiatry, education or psychology. Recent, the use of LCA in dietary pattern analysis was reported. In this research, a self-made questionnaire for dietary habits was used to collect the information in recent 3 month. It covered the frequency for eating breakfast, the amount of different types of foods. The entire variable used were turned to be binary to get a clearer result. LCA was used to extract diet pattern: ANOVA, paired t test and chi-square test were used to analyze the difference of metabolic indicator among different patterns. Results Three types of diet were found: group 1(n=314, 45.0%) had sufficient nutrients; group 2(n=259, 37.1%) and group 3(n=125, 17.9%). As compared to 1992, had excessive intake of meat. In 2000, there was a rise in animal oil consumption, fresh fruit intake, fish or shrimp intake in group 2 and group 3. There was a decrease in egg intake in group 1 and group 3. Generally, the intake of milk fruit bean product was on an increase. There was statistically significant increase in BMI, SBP, FPG, TC and TG in 2000 compared with the value in 1992. Conclusion LCA might be a solution for dietary pattern extraction. There were mainly three type of dietary pattern that developed with age and impacted the related metabolic indices. Multivariable analysis was especially needed to exclude the impact of potential confounding factors.

c-Met expression in esophageal squamous cell carcinoma tissues and its correlation with clinicopathological characteristics

LIU Chun-tao;ZHU Sheng-tao;TIAN Yue;ZHANG Shu-tian

2012, 33(2):  223-226.  doi:10.3969/j.issn.1006-7795.2012.02.018

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Objective To detect the expression of c-Met in esophageal squamous cell carcinoma tissues(ESCC) and paired non-cancerous tissues, and to investigate the association between c-Met and clinicopathological parameters of ESCC patients. Methods Eighty seven pairs of paraffin-embedded ESCC tissues and paired non-cancerous esophageal tissues were collected from the Department of Pathology, Beijing Friendship Hospital Affiliated To Capital Medical University. Sections from each specimen were examined by a pathologist and graded histologically. Patients' clinical parameters were collected meanwhile. Immunohistochemistry(IHC) was applied to detected the the expression of c-Met in the tissues. Results c-Met positive signals showed brown-yellow granules in the membrane. The expression of c-Met in non-cancerous esophageal tissues was always negative, whereas strong positive staining of c-Met was observed in 29/87(33.3%) of ESCCs. The expression of c-Met was correlated with tumor invasion depth, lymph node metastasis, and TNM(tumor-node-metastasis) stage(P=0.017, P=0.000, P=0.000, respectively). No significant association between c-Met expression and other clinicopathological parameters(patients' gender, age, cell grading, and tumor size) was found(P>0.05). Conclusion The overexpression of c-Met was correlated with tumor invasion depth, lymph node metastasis, and TNM stage of ESCC patients. The overexpression of c-Met may be a marker for highly invasive ESCC.

Expression, purification and characterization of ribose 5-phosphate isomerase A from Streptococcus mutans

WU Wen-qi;CONG Xu-zhen;YIN Ai-hong;HU Jia;ZHAI Fang-li;LI Shen-tao

2012, 33(2):  227-232.  doi:10.3969/j.issn.1006-7795.2012.02.019

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Objective To express, purify and characterize the ribose 5-phosphate isomerase A(rpiA) from Streptococcus mutans. Methods A DNA fragment encoding S. mutans ribose 5-phosphate isomerase A was amplified by PCR using the genomic DNA of Streptococcus mutans UA159 as a template. The PCR product was cloned into vector pGEX-6p-1. The construct carrying the coding DNA sequence of rpiA fused with GST was transformed into E.coli BL21(DE3), and the fusion protein was expressed by induction with IPTG. The recombinant protein was purified by affinity chromatography and ion exchange chromatography. The purified target protein was identified by SDS-PAGE and MALDI-TOF MS. Results S. mutans ribose 5-phosphate isomerase A was successfully expressed in E.coli in soluble form. After a series of purifications, we got the recombinant protein with the purity higher than 95%. The product was identified to be S. mutans ribose 5-phosphate isomerase A by SDS-PAGE and MALDI-TOF-TOF MS. Conclusion S. mutans ribose 5-phosphate isomerase A was successfully expressed in E.coli. An effective purification protocol was established. Recombinant rpiA with a purity higher than 95% was obtained, which provided a basis for the further studies of the biological activities and functions of ribose 5-phosphate isomerase A.

Protective effect of insulin on delayed neuropsychologic sequelae after carbon monoxide poisoning in the rat and its mechanisms

LIAO Qiu-ju;WANG Jing;QIN Jian

2012, 33(2):  233-237.  doi:10.3969/j.issn.1006-7795.2012.02.020

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Objective To determine the effects of insulin on cognitive function and neuronal apoptosis in delayed encephalopathy rat model of carbon monoxide(CO) poisoning. Methods The rat carbon monoxide poisoning model was established with introducing CO into abdominal cavity. Rats were randomly divided into two groups: CO group and RI-treated group , simultaneously administered 2 g/kg of glucose and 2 U/kg of regular insulin(RI) by intraperitoneal injection for 7 days. The learning and memory in delayed encephalopathy rat model of carbon monoxide poisoning were studied by using a Morris water maze from 1~5 weeks. At 5 weeks after CO exposure brains were perfusion-fixed. Neuronal changes were detected by thionine stain, TUNEL stain was used to assess injury. The data were expressed as(x±s ) and analyzed by analysis of variance. A P value less than 0.05 indicated significance. Results The cognitive function and neuronal apoptosis of RI-treated-group were improved as compared with CO group. Conclusion Insulin could inhibit the cell apoptosis and improve the cognitive function. Insulin could protect the nerve functions and alleviate on delayed neuropsychologic sequelae(DNS) after carbon monoxide poisoning.

The effect of vitamin C given in late gestation on the lung of fetal rat with congenital diaphragmatic hernia

CAI Jing-jing;CHEN Yong-wei;ZHANG Jian-min

2012, 33(2):  238-241.  doi:10.3969/j.issn.1006-7795.2012.02.021

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Objective To induce a congenital diaphragmatic hernia model of fetal rat and to test the effect of vitamin C given in late gestation on the incidence of congenital diaphragmatic hernia(CDH) and lung development of fetal rat. Methods Sprague-Dawley female rats were randomly divided into four groups: control group(C), vitamin C group(W), CDH group(N) and treatment group(T). Nitrofen(100 mg) was given intragastrically to the rats of CDH and treatment groups on day 9.5 of gestation to induce a congenital diaphragmatic hernia model of fetal rat. Vitamin C(1.5 mg) was given intraperitoneally to vitamin C and treatment group from day 16.5 to 20.5. The offsprings were recovered on day 21(term=22nd). Fetal rats and their lungs were weighed. The incidence of CDH was examined. Immunohistochemical staining(Ki-67) was performed. The percentage of proliferative cells were counted by software. Results The incidences of CDH in CDH group and treatment group were the same. The mean body weight of treatment group was higher than those of the other three groups. The mean fetal lung weight was the highest in vitamin C group and treatment group and the lowest in CDH group. Percentage of proliferation cells in CDH group was lower than those of the other three groups. Conclusion By the use of nitrofen we induced congenital diaphragmatic hernia model in fetal rat. In the CDH group, the mean lung weight and the percentage of proliferation cells decreased. There may be lung hypoplasia in this group. There was no obvious evidence that vitamin C could change the incidence of CDH in rat model induced by nitrofen. Vitamin C increased mean lung weight and percentage of proliferation cells of CDH fetal rats, so it may improve lung development of fetal rats.

Tacrolimus protects rats from renal ischemia-reperfusion injury

WU Fei;LIU Zhi-zhong

2012, 33(2):  242-245.  doi:10.3969/j.issn.1006-7795.2012.02.022

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Objective To study the protective effects of tacrolimus on the rat renal ischemia-reperfusion injury. Methods Sixty male healthy Wistar rats were assigned randomly into three groups: sham operation group, control group, and experimental group. In each group there were 20 rats. Blood and kidney specimens were taken in different reperfusion time, and serum creatinine(Cr), urea nitrogen(BUN), tumor necrosis factor(TNF-α) were determined; kidney tissues B cell lymphoma protein(Bcl-2) and pathological changes were detected. Results After 45 minutes of ischemia, reperfusion was performed at 2 h, 6 h and 24 h in the experimental group serum creatinine(Scr), blood urea nitrogen(BUN) levels were significantly lower than those of the control group(P<0.05), however, Bcl-2 immune response intensity was significantly higher than those of the control group(P<0.05). As to reperfusion at the various time points, in the experimental group of TNF-α content was significantly lower than that of the control group(P<0.05). Under light microscope, the control group showed more severe pathological changes in renal tubules and tubular epithelial cells; the experimental group showed significantly less severe renal disease and renal epithelial cells pathological changes than the control group. Sham operation group did not show any significant tubular and tubular epithelial cell changes. Conclusion Tacrolimus on showed significant protective effects against renal ischemia-reperfusion protective effect that may reduce serum levels of TNF-α expression and increase the Bcl-2 protein expression to improve renal ischemia-reperfusion injury.

Survey of general practioner job-transfer training program in Beijing-focusing on trainees' background and training needs

WU Tao;HUANG Ya-fang;CHEN Rui;XIE Ru-xu;SANG Sheng-juan;LIU Shao-jie;YE Chun;GUO Ai-min

2012, 33(2):  246-250.  doi:10.3969/j.issn.1006-7795.2012.02.023

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Objective To understand the status of general practioner job-transfer training, the training needs of trainees and the main effect on trainings in Beijing. Methods A cross-sectional survey was made to general practice trainees in the first round of job-transfer training program in Beijing, including information of trainees' background and training needs. Quantitative data were analyzed with SPSS 13.0 software. Results Totally 86 trainees in the first round of job-transfer training program were investigated and 80 trainees responded. The education backgrounds and professional titles of trainees in outer suburban district were extremely higher than those in the urban and peri-urban areas. The people registering general practice account for 1.3%. Diagnosis and differential diagnosis skills of common diseases were most needed, so did the treatments of common and frequently-occurring diseases to the training knowledge. Conclusion The syllabus of the training should be further improved. Training time arrangement should be regulated reasonably according to training needs. Training methods should be adjusted appropriately according to general practice' principle.

Damage of hyperhomocysteinemia on generation oxidation in smooth muscle cells

LI Yan;ZHAGNG Chun-lai;LU Feng;WANG Li-zhong;JIANG Yu-fen

2012, 33(2):  251-254.  doi:10.3969/j.issn.1006-7795.2012.02.024

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Objective Many recent studies have indicated that homocysteine(Hcy) is an important and independent risk factor for atherosclerosis(AS). This study aimed to test the effects of Hcy on the vascular smooth muscle cells in rabbits' carotid artery by feeding New Zealand white rabbits with high concentration of Hcy. Methods The carotid artery was separated after feeding with high methionine foods for three months. The vascular smooth muscle cells(VSMCs) in rabbits' carotid artery were separated, cultured and passaged. The intracellular superoxide dismutase(SOD) activity was tested using the xanthine oxidation.The intracellular activity of catalase(CAT) was measured by visible colorimetric method. Results The pathomorphological change of the separated rabbit carotid artery with methionine diet group was compared with control group under optical microscope. I: Intima; M: Medial. The CAT and SOD activity of rabbit carotid artery smooth muscle cells increase under Hcy diet group. Conclusion High Hcy accelerated VMSC proliferation, and may increase peroxidizine damages of VMSCs within the scope of certain dose, eventually accelerate the process of AS.

临床研究

Clinical and prognostic value of serum amyloid A in patients with acute ischemic stroke

WU Xiu-xin;LI Li-xin;LIAO Pin-jun;YUAN Yuan

2012, 33(2):  255-258.  doi:10.3969/j.issn.1006-7795.2012.02.025

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Objective To investigate the clinical and prognostic value of serum amyloid A(SAA) in patients with acute ischemic stroke. Methods Blood samples were collected from the patients within the 24 h after admission. Enzyme-linked immunosorbent assay(ELISA) was used to determinate SAA levels among 100 cases of acute stroke patients before and after 21-day treatment and CRP was detected by immunoturbidimetry. Fifty healthy volunteers served as the control group. All patients were given comprehensive treatment. On admission and after 21-day treatment, the National Institutes of Health Stroke Scale(NIHSS) was employed to provide a quantitative measure of stroke-related neurologic deficit. Furthermore, correlation between SAA levels and change in NIHSS score before and after treatment was analyzed using the Pearson correlation coefficient in order to explore the association of SAA levels with clinical picture and prognostic value in patients with acute cerebral infarction. Results SAA levels were significantly higher in acute cerebral infarction patients before treatment(18.05±6.77) mg/L than in the patients after treatment(7.81±2.66) mg/L and in the control group(7.49±4.49) mg/L, respectively(P<0.01). CRP levels were significantly higher in acute cerebral infarction patients before treatment (15.05±4.32) mg/L than in the patients after treatment(6.05±2.20) and in the control group(5.5±3.34) mg/L, respectively(P<0.01). After treatment, NIHSS score decreased significantly at 21 d (11.07±4.2 vs 7.78±2.89, P<0.05). The SAA levels at the acute stage showed a positive correlation with the net change in NIHSS score(r=0.612, P<0.05). Conclusion Patients with acute cerebral infarction have significantly increased SAA and CRP levels, which both indicate active disease. The SAA levels, but not CRP levels, are positively correlated with the net change in NIHSS score, suggesting that SAA may predict the prognosis of patients with acute cerebral infarction and has better prognostic value than CRP.

Gangliogliomas arising from the thalamus: Report of 6 cases and literature review

GENG Su-min;ZHANG Li-wei;HAO Shu-yu;WU Zhen;JIA Gui-jun;ZHANG Jun-ting

2012, 33(2):  259-262.  doi:10.3969/j.issn.1006-7795.2012.02.026

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Objective Ganglioglioma(GG) is a less common primary neoplastic disease of the central nervous system that is typically located supratentorially. The Objective of this study was to discuss clinical characteristics, microsurgical treatment and prognosis of gangliogliomas(GG) arising from the thalamus. Methods The hospital data of 6 patients who had undergone microsurgery and were pathologically diagnosed as thalamic GGs were reviewed. Results The main clinical presentations were intracranial hypertension, hemiparesis and impairment of sensation on the contralateral side of the lesion. The mean age at presentation was 12.5 years. The operations consisted of total resection in two, subtotal resection in 3, and partial resection in 1. There were increased hemiparesis and impairment of sensation for three cases. All patients were followed up for a mean of 29 months, of which 2 lived a normal life, three lived independently, one patient with anaplastic GG died of tumor recurrence. Conclusion Because of lacking typical clinical manifestations and neuroimaging characteristics, preoperative diagnosis of thalamic GGs is difficult. Its precise diagnosis mainly depends on histopathology. Surgical resection is the principal treatment, the prognosis of anaplastic GGs is poor, long term and close follow-up is necessary.

The postnatal follow-up in women with gestational diabetes mellitus

ZHOU Li;WU Lian-fang;FAN Ling;LI Xiao-yu

2012, 33(2):  263-267.  doi:10.3969/j.issn.1006-7795.2012.02.027

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Objective To investigate the prognosis and related factors of glucose metabolism in postnatal women with gestational diabetes mellitus(GDM). Methods Data of 624 women with GDM during pregnancy delivered in our hospital from January 2008 to December 2010 were collected. According to the 6~8 weeks postnatal glucose metabolism the 624 women were divided into two groups, 444 glucose metabolism; 180 abnormal postnatal glucose metabolism. Results 1) Among the 624 subjects, 444(71.2%) women glucose metabolism return to normal 6~8 weeks after delivery; 180(28.8%) still had abnormal glucose metabolism 6~8 weeks after delivery, including 18 with diabetes mellitus(DM), 32 with impaired fasting glucose(IFG), 130 with impaired glucose tolerance(IGT). 2) The time point of diagnosing GDM was earlier in abnormal glucose metabolism group than in normal glucose metabolism group, the level of fasting blood glucose, body mass index(BMI) in prenatal and postnatal and the lipid levels were higher in abnormal glucose metabolism group than in normal glucose metabolism group. Significant difference was found in using insulin during pregnancy, accompanying related complications and with DM family history between the two groups(P<0.05), which was higher in abnormal glucose metabolism group. No significant difference was found in the ratio of breast feeding(P>0.05). 3) No significant difference were found in age, the times of pregnancy and delivery and body mass gaining during pregnancy(P>0.05). Conclusion 1) Most GDM women could restore normal glucose metabolism 6~8 weeks' after delivery, only 28.8% GDM women continued to have abnormal glucose metabolism. 2) The earlier diagnosis of GDM, fasting blood glucose, DM family history, using insulin in pregnancy, accompanying related complications, BMI in prenatal and postnatal and higher lipid were high risks in postnatal glucose metabolism.

综述

Research progress of psoriasis-like models

DI Ting-ting;ZHAO Jing-xia;LI Ping

2012, 33(2):  275-279.  doi:10.3969/j.issn.1006-7795.2012.02.030

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Lack of the naturally occurred psoriasis animal models once made the researches on human psoriasis phenotype and pathogenesis stagnated in last several years. But with the increasing understanding of psoriasis mechanism, a series of models has been explored at organic, cellular and molecular levels. We evaluated the current psoriasis models and analyze theirs advantages and limitations, to provide reference for psoriasis studying, drug screening and treatment.

Advances in studies on cancer stem cell in diagnosis and treatment of endometrial carcinoma

CUI Kai-yu;KONG Xian-chao

2012, 33(2):  280-283.  doi:10.3969/j.issn.1006-7795.2012.02.031

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With the characteristics of self-renewal, well-differentiated potency and oncogenicity, cancer stem cell(CSC) have drew the interests of researchers in the study of neoplastic hematologic disorder, breast cancer and prostatic carcinoma,and certain advances have occurred. However, the evaluation and isolation of CSC are still in the process of exploration. This article made a general review of endometrial carcinoma, CSC and endometrial stem cell. It gives a comprehensive description of the application CSC in diagnosis and treatment of endometrial carcinoma, and the future research prospect are discussed.