Objective To investigate the regulation effect of promoter methylation of deathassociated protein kinase (DAPK) on mRNA and protein expression of DAPK in tissue of primary gastric cancer (GC). Methods The cancerous and noncancerous samples from 62 patients with GC were determined by RT-PCR for mRNA expression of DAPK. The DAPK promoter methylation was detected by methylation-specific PCR. The protein expression of DAPK in 34 patients with methylation was determined by Western blot. Results mRNA and protein expre.ssions of DAPK in cancerous tissues were reduced significantly compared to noncancerous tissues (0. 2863d±0. 2027 vs 0. 57364±0. 1968,0. 2616±0. 0913 vs 0. 65294±0. 1808, P＜0.01). Methylation frequency of DAPK in cancerous tissues was higher than that in noncancerous tissues (54.8% vs 17.7%, P＜0.01). Furthermore, DAPK mRNA expression was decreased in methylation group compared to unmethylation group (0.1399±0. 0835 vs 0. 46404±0. 1569, P＜0. 01). Moreover, a significant correlation was demonstrated between the TNM stage and DAPK promoter methylation (P = 0. 04). Conclusion Expression of DAPK is down-regulated in cancerous tissues at mRNA and protein levels. Low expression of DAPK is associated with hypermethylation of the promoter of DAPK gene.

AIM: To investigate the early changes of retinal function in diabetic patients detected by multifocal electroretinogram (mfERG).METHODS: The first-order kernel responses of mfERG were recorded from eyes of 33 normal control subjects,63 diabetic patients without retinopathy and 43 diabetic patients with background retinopathy. The response densities and implicit times of N1 and P1 were compared among the control, diabetic patients without retinopathy and diabetic patients with retinopathy.RESULTS: The response densities of N1 and P1 in central 3 rings were reduced significantly in diabetic eyes with and without retinopathy. And the implicit times of N1 and P1 were delayed significantly only in diabetic eyes with retinopathy.CONCLUSION: mfERG can detect the early changes of retinal function quantitatively in diabetic patients. Analysis of response densities and implicit times of N1 and P1 can reflect the progress of local retinal dysfunction in diabetes.

AIMTo observe the differences of hemodynamics and nitric oxide synthase(NOS) activity of ventricular cardiac muscle in two septic shock models and explore the possible mechanism.

METHODSTwo rat models of septic shock[lipopolysaccharide(LPS)-induced and cecal ligation and puncture (CLP)-induced septic shock] were used. The hemodynamic parameters and nitric oxide synthase activity of ventricular cardiac muscle were measured.

RESULTSThe hemodynamic parameters in CLP-induced model were increased in the early stage and decreased in the late stage while in LPS-induced model the parameters showed the same change of the CLP late stage. Both LPS model and CLP model (late stage) showed significant increase in NOS activity, but there was no difference between the two models. After treatment of the NOS inhibitor N-nitro-L-arginine methyl ester (L-NAME), the parameters of CLP-late stage and LPS model increased significantly. The NOS activity reached the highest level in the CLP-middle stage. The production of nitrite/nitrate decreased significantly in LPS model and CLP model(late stage) after treatment of L-NAME, but the nitrite/nitrate produced by constitutive NOS in LPS model was higher than CLP model(late stage).

CONCLUSIONThe increase of the NOS activity may be the main reason to lead to the depression of the hemodynamic parameters. Inducible NOS may play the leading role in the LPS model while cNOS and iNOS have the same effect in the CLP model.

Animals ; Hemodynamics ; Lipopolysaccharides ; Male ; Myocytes, Cardiac ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Shock, Septic ; classification ; metabolism

To determine the optimum conditions of supercritical CO2 extraction of Xiaoyaosan, and establish its fingerprint by gas chromatography-mass spectrometry (GC-MS), the yield of extract were investigated, an orthogonal test was used to quantify the effects of extraction temperature, pressure, CO2 flow rate and time, and fingerprint analysis of different batches of extracts were by GC-MS. The optimal extraction conditions were determined as follows: extraction pressure 20 MPa, extraction temperature 50 degrees C, CO2 flow rate 25 kg x h(-1), extraction time 3 h, and average yield 2.2%. The GC-MS fingerprint was established and 27 common peaks were found, whose contents add up to 81.89% of the total peak area. Among them, 21 compounds were identified, accounting for 53.20% of the total extract. The extraction process is reasonable and favorable for industrial production. The GC-MS method is accurate, reliable, reproducible, and can be used for quality control of supercritical CO2 extract from Xiaoyaosan.
Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods

Objective To investigate the relationship between alteration of gene in cyclic adenosine monophosphate(cAMP) signal transduction system in rats after myocardial damage and changes of cardiac function and ventricular remodeling. Methods Twenty eight male Wistar rats weighing 220 g to 250 g were randomly divided into three groups: acute myocardial damage group(AMD, n = 10), chronic myocardial damage group (CMD,n = 9 ) and sham-operation group (control, n = 9). Animal model of acute myocardial damage was established by ligation of rats left coronary artery in the AMD and the CMD groups. Rats in control group were treated similarly, except that the coronary suture was not tied. Hemodynamics and echocardiography were measured before rats were sacrificed 24 hours after operation in control and AMD groups but those in CMD groups were sacrificed 8 weeks later. Cadiocyte apoptosis were estimated by TUNEL method, cAMP levels in heart were tested by radioimmunity and the mRNA expressions for inducible cAMP early repressor (ICER), cAMP response element binding protein (CREB), phosphodiesterase 3A (PDE3A) and bcl-2 were assayed by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Results The difference of left ventricular end diastolic diameter (LVEDD), left ventricular end diastolic pressure(LVEDP), maximal rising and falling rate of ventricular pressure,left ventricular systolic pressure (LVSP), eject fraction (EF) and fraction shortening (FS) were statistically significant among the three groups(F = 285.9, 196.8, 83.2, 80.4, 54.9, 196.6, 95.2, all P ＜ 0.01). LVEDD[(7.03 ±0.28), (8.20 ± 0.27)mm] and LVEDP[(11.19 ± 2.89), (19.76 ± 3.34)mmHg] in AMD and CMD groups were significantly increased, compared with those in control group[ (5.05 ± 0.30)mm, (- 5.62 ± 3.01 )mmHg, all P ＜0.01 ]. While maximal rising rate[ (2964 ± 449), (2214 ± 434)mmHg/s] and falling rate[(- 2617 ± 441),(- 1891± 424)mmHg/s] of left ventricular pressure, LVSP[ (94.19 ± 4.03), (85.85 ± 6.39)mmHg], EF[ (41.6 ±5.9)%, (35.9 ± 4.1 )%] and FS[ (36.9 ± 4.6)%, (23.1 ± 4.9)%] of left ventricular in the two groups were lower than those in control[(4759 ± 406)mmHg/s, (- 4327 ± 388)mmHg/s, (116.29 ± 8.25)mmHg, (80.9 ± 5.6)%,(53.1 ± 4.3)%, all P ＜ 0.01 ]. These changes in CMD group were more significant than those in AMD groups(P ＜0.05 or P ＜ 0.01 ). The difference of apoptotic index, cAMP and expression of ICER, CREB, PDE3A mRNA and bcl-2 mRNA were statistically significant among the three groups(F= 172.5, 141.0, 540.8, 246.8, 165.1, 563.9,all P＜ 0.01 ). Apoptotic index[ (32.8 ± 4.2)‰, (18.4 ± 3.9)‰] and cAMP in heart[ (9.95 ± 0.30), (5.60 ± 0.25)nmol/kg] in AMD and CMD groups were increased compared to control group[ (3.9 ± 1.7)‰, (2.48 ± 0.29)nmol/kg,all P ＜ 0.01 ], and those in CMD group were lower than in AMD group(all P ＜ 0.01 ). Expression of ICER mRNA (1.434 ± 0.093, 0.942 ± 0.076) and CREB mRNA(5.70 ± 0.50, 2.64 ± 0.51) in AMD and CMD groups were higher, and expression of PDE3A mRNA(48.98 ± 8.14, 16.68 ± 8.46) were lower than those in control group (0.154 ± 0.063, 1.08 ± 0.35, 105.94 ± 12.61, all P ＜ 0.01 ). The three genes in CMD group were fewer than those in AMD group(all P ＜ 0.01 ). bcl-2 mRNA was up regulated in AMD group(4.55 ± 0.27) and was down regulated in CMD group(0.35 ± 0.15) compared to control(2.18 ± 0.30, all P＜ 0.01). Conclusions There is PDE3A-ICER positive-feedback loop leading to myocyte apoptosis and heart failure after myocardial damage. The downregulation of PDE3A mRNA observed in chronic myocardial damage may play a causative role in the progression of ventricular remodeling and heart failure, in part, by inducing ICER mRNA and promoting cardiac myocyte dysfunction.

Objective To explore the early diagnosis of Alport′s syndrome(AS).Methods Renal and skin biopsy was carried out in 5 patients who manifested with isolated hematuria and nephritic syndrome(NS).By using indirect immunofluorescence method,the expression of type Ⅳ collagen ? chains was detected on epidermal basement membrane(EBM) and glomerular basement membrane(GBM).Results ?_1 chains on EBM and GBM were expression in all patients,but ?_5 chains on EBM and ?_3,?_5 chains on GBM form 2 female patients were segmental expression.Thus the goal for early diagnosis was achieved.Conclusions ? chains for EBM type Ⅳ collagen and GBM type Ⅳ collegan should be investigated if condition permits for those patients with isolated hematuria,NS(steroid-resistant) and thinned GBM in electron microscopy.It can be useful for diagnosis and differenial diagnosis of AS.

Objective To exlore clinical characteristics and changes of electroencephalography(EEG) in children with migraine.Methods Clinical data of 98 children with migraine provided by our hospitals clinic service were analysed.The cases′ history were inquired,physical examination made(EEG),Video-EEG(V-EEG) and transcranial doppler sonography(TCD) were examined.After(diagnosis) was made,the patients were treated and followed up.Results In 98 cases of migraine,27 cases were classical migraine and 71(cases) were ordinary migraine.EEG and V-EEG indicated most of them were in normal range and increase in diffused inactive echoes,and only 1 case of 98 EEG showed scattered epileptic waves;TCD indicated that the velocities of blood flow of intracranial arteries of 87 children with migraine were faster.Conclusions Clinical characteristics of children with migraine is similar with adault.Diagnosis of children with migraine and epilepsy,apply both EEG and TCD have great value in diagnosis of children with migraine.

OBJECTIVE: To study the characters of oscillatory potentials (OPs) of electroretinogram (ERG) after methanol intoxication in rats. METHOD: The SD rat models of methanol intoxication were established and divided into control group, 3-day intoxication group, 7-day intoxication group. The changes of OPs of ERG were recorded in a dark room. RESULTS: The total amplitudes of 3-day and 7-day intoxication groups decreased approximately 50% compared with that of the control group, while the schedule delayed approximately 16% and 61%, respectively. CONCLUSION The characters of methanol intoxication in rats included delay in schedule and decline in the total amplitude of OPs.
Animals ; Electroretinography ; Methanol/poisoning* ; Rats ; Retina/physiopathology*

alphaB-crystallin is the structural protein of vertebrate lens, which is widely expressed in non-lens tissue. As one of the heat shock protein family members, alphaB-crystallin possesses biological properties of molecular chaperones and anti-apoptotic effects. Multi-factor injuries, such as retinopathy, inflammation and nervous system diseases, have a closely relationship with alphaB-crystallin. This paper reviews the research progress of the expression and mechanism of alphaB-crystallin in retina and extraocular tissues and organs.
Crystallins ; Gene Expression Regulation, Developmental ; Heat-Shock Proteins/metabolism* ; Humans ; Lens, Crystalline ; Retina ; alpha-Crystallin B Chain/metabolism*

OBJECTIVE: To compare the correlation between contrast vision (LV) and sweep visual evoked potential acuity (SVEP-A) among people with emmetropia, mild myopia, and moderate myopia. METHODS: The CV and SVEP-A were tested individually in 96 eyes from healthy young volunteers, including 37 eyes of emmetropia, 27 eyes of mild myopia, and 32 eyes of moderate myopia. The statistic analysis was done by ANOVA analysis and rank sum test. RESULTS: (1) With the decrease of contrast, CV and SVEP-A decreased in every group. (2) At 100% contrast, the difference of CV between emmetropia and mild myopia had statistical significance (P<0.05). At 100%, 25% and 10% contrast, the difference of CV between emmetropia and moderate myopia had statistical significance (P<0.05). (3) In the same group, the difference of 100% and 25% contrast had statistical significance (P < 0.05). So was between 100% and 10% contrast. (4) At 100% and 10% contrast, the difference of CV and SVEP-A had statistical significance (P < 0.05). CONCLUSION The CV of myopia relates to many factors including ametropia and fundus lesions. The correction of ametropia is important to the values of CV and SVEP-A.
Evoked Potentials, Visual/physiology* ; Eye ; Fundus Oculi ; Humans ; Myopia/physiopathology* ; Neurologic Examination ; Severity of Illness Index ; Vision Tests/instrumentation* ; Visual Acuity/physiology*