Objective To develop new homogeneous enzyme immunosorbent assay for serum IgM antibody of EV71 infection.Methods After protein EV71 VP1 1-297 was purified and biotinylated,the optimal quantities of the antigen was obtained by ELISA.The homogeneous enzyme immunosorbent assay was valuated with the serum samples of both 20 healthy subjects and 30 samples from the patients of HFMD detected by ELISA.Results The reaction system of homogeneous enzyme immunosorbent assay for EV71 IgM was established in this study.Then the homogeneous enzyme immunosorbent assay was valuated with ELISA for EV71 IgM.The results revealed serum IgM were negative for the 20 healthy subjects by both ELISA and homogeneous enzyme immunosorbent assay.Of the 30 HFMD patients,positive serum EV71 IgM detected by both ELISA and homogeneous enzyme immunosorbent assay,and yet,6 IgM positive samples were identified with homogeneous enzyme immunosorbent assay in the 14 negative samples detected by ELISA.Conclusion A homogeneous enzyme immunosorbent assay for detection of serum EV71 IgM has been established.

Objective To investigate the genotype distribution of thalassemia intermedia , major and compound thalassemia in Peking Union Medical College Hospital from 2012 to 2015. Methods Retrospectively 1 084 suspected thalassemia cases were analyzed in recent four years .Three common deletions of αglobin chain were detected by GAP-PCR.Three common point mutations of αglobin chain and seventeen common mutations of βglobin chain were identified by PCR reverse dot blot hybridization . Hemoglobin electrophoresis was carried out by Capillary Electrophoresis System .RBC associated parameters and morphology were analyzed by hematology analyzer and blood smear .Results 702 cases were confirmed to be thalassemia, and the positive rate was 64.76% (702 /1084).19 types of gene defects were detected. There were 4 types of gene defects in 23 case with α-thalassemia intermeida, including -α3.7 /--SEA , -α4.2 /--SEA , αCSα/--SEA and αQSα/--SEA , -α3.7 /--SEA to be the most common genotype (18 cases) .3 cases with β-thalassemia intermeida were confirmed and the genotypes were βCD 17(A→T) /β-29(A→G) , β-28(A→G) /β-28(A→G) andβIVS-Ⅱ-654(C→T) /βCD17(A→T) , respectively.There were also 1 βCD 41 -42(-TTCT) /βCD17(A→T) thalassemia major case. The genotypes of 2 HbE/β-thalassemia cases were βCD41 -42(-TTCT) /βE and βCD17(A→T) /βE.5 αβ-thalassemia including 2 βCD 41 -42(-TTCT) /βA compounded with αα/-α3.7 , 1βIVS-Ⅱ-654(C→T) /βA compounded with --SEA /αCSα, 1βCD17(A→T) /βA compounded with -α4.2 /ααand 1βCD 41 -42(-TTCT) /βA compounded with αCS α/αα.Rare and untypical haematological results were found , such as normal level HbA 2 and undetectable HbH, in compound heterozygosity with --SEA /αCS α and βIVS-Ⅱ-654(C→T) /βA. Conclusions The genotypes of thalassemia intermedia, major and compound thalassemia in Peking Union Medical College were highly variable .

To investigate changes of 14-3-3beta from apoptosis induced by PrP106-126 polypeptide, HeLa cell was incubated with PrP106-126 for 4h or 8h. Nucleus changes and the expression of PARP were detected differently by Hoechst staining and Western blotting. Expressing of protein and mRNA from 14-3-3beta was determined by Western blotting and Real-time PCR. The results show that typical nucleus pyknosis and chip of apoptosis and degradation of PARP were induced by PrP106-126 peptide in HeLa cells. Degradation of 14-3-3beta appeared in apoptosis groups induced by PrP106-126 peptide. However, 14-3-3beta mRNA did not display any changes in apoptosis groups. This study indicated that degradation of antiapoptosis protein 143-3beta induced by PrP106-126 peptide may be one of pathogenesis mechanism of prion disease.
14-3-3 Proteins ; metabolism ; Apoptosis ; drug effects ; HeLa Cells ; Humans ; Peptide Fragments ; pharmacology ; Prions ; pharmacology ; Proteolysis ; drug effects

AIMTo investigate the neural network and cellular mechanisms of hippocampal epileptogenesis contralateral or ipsilateral to the side of acute tetanization (60 Hz, 2 s, 0.4 - 0.6 mA) of the posterior dorsal hippocampus (ATPDH).

METHODS10 trains of the ATPDH were administered into the CA1 basal dendritic region of the right hemisphere at an interval of 10 minutes.

RESULTS(1) The firing rate of CA1 single neuron in the right or the left hippocampus was inhibited respectively after the ATPDH, and the effects weakened gradually while the trains of the ATPDH increased. The inhibited firing rate and the transformed firing pattern from tonic one to clonic one were more obvious at the side contralateral to the stimulation (62.94% +/- 3.68%, 36.61% +/- 3.14%, P < 0.01). (2) Synchronous primary afterdischarges of depth EEG and single unit discharges were more commonly observed at the side ipsilateral to the ATPDH (P < 0.01). (3) Primary or secondary hippocampal network afterdischarges at high frequency were only found in CA1 region ipsilateral to the ATPDH. (4) Secondary afterdischarges of CA3 basal dendritic neural network were completely synchronized with those of subicular single neuron, which reoccurred and persisted several hours.

CONCLUSIONIt is possible that post-inhibition bursting of single neuron and recurrent network seizures in the hippocampus contralateral to the artificial focus be the important manifestation of the formation of "epileptic networks" across from one hemisphere to another.

Animals ; Electric Stimulation ; Hippocampus ; physiology ; Male ; Neural Pathways ; physiology ; Rats ; Rats, Sprague-Dawley ; Seizures ; etiology

AIMThe electrographic and behavioral kindling effects were induced by chronic tetanization of the right caudate-putamen (CPu) to study the target-behavior expression involved in the CPu or hippocampus (HPC) network abnormalities.

METHODSExperiments were performed on 58 SD rats. Tetanization (60Hz,0.4 - 0.6mA, 2s) was delivered into the CPu or the HPC, once a day, for 7-12 days. Animal behaviors were observed every day and depth electrographs were recorded at the beginning or at the end of the experiments.

RESULTSChronic tetanization of the CPu or of the HPC induced: (1) Rhythmic sharp waves in the CPu and paroxysmal epileptiform events in the HPC electrographs. (2) Primary behavioral seizures, secondary behavioral seizures, and kindling effects, including wet dog shakes (WEDS), rearing, face washing, immobility, chewing and head nodding. (3) Lower rate of primary WEDS (P < 0.01), and higher rate of secondary WEDS (P < 0.01) in the CPu-tetanized rats. (4) Longer silent period of behavioral seizures before kindling appeared in the CPu-tetanized rats.

CONCLUSIONKindling effects in the CPu-tetanized rats resembles those in the HPC-tetanized rats. The CPu might participate in the origin of epileptic focus and be involved in reestablishment of limbic epileptic networks, which may be responsible for the target-behavioral seizures.

Animals ; Behavior, Animal ; Caudate Nucleus ; Electric Stimulation ; Epilepsy ; physiopathology ; Kindling, Neurologic ; Male ; Rats ; Rats, Sprague-Dawley ; Seizures ; physiopathology

AIMTo study the characteristics of neural information encoding of the epileptic networks involved caudate-putamen(CPu) and the hippocampi induced by tetanization of the right CPu in rats.

METHODSExperiments were performed on 59 SD rats. Acute or chronic tetanization of the right CPu (ATRC or CTRC) (60Hz,0.4-0.6 mA, 2 s) was used to induce rat epilepsy.

RESULTS(1) The bursting or primary unit afterdischarges of single neurons were asymmetric in dual hippocampi induced by the ATRC. (2) Continuous sharp waves were observed in ipsilateral or contralateral CPu induced by the CTRC. The oscillatory network seizures with phase shift appeared between two sharp waves in ipsilateral CPu. The frequency of oscillatory waves was negatively correlated with the time and fluctuated from 70 Hz to 110 Hz, then to 35 Hz, and finally to 30 Hz. (3) In the contralateral side primary network after discharges in the CPu were induced by the CTRC. Therefore, the characteristic primary network afterdischarges could be shifted from the CPu or to the HPC, but amplified. On the other hand, HPC sharp waves could be depressed when the CPu network seizures occurred.

CONCLUSIONThe reestablishment of CPu-hippocampal epileptic networks could be transhemispherically promoted by over-activation of the right CPu network, in which bilateral hippocampal neuronal network and CPu neural network were involved in some particular pathophysiological information encoding.

Animals ; Caudate Nucleus ; metabolism ; Epilepsy ; physiopathology ; Hippocampus ; metabolism ; Male ; Neurons ; metabolism ; Putamen ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To explore the relationship between the course of disease and glycolipid metabolic parameters in drug-naive schizophrenia patients. Methods All 186 drug-naive schizophrenia patients,admired to our hospital from March 2010 to October 2011,were chosen in our study; relative glycolipid metabolic parameters at baseline were tested and Positive and Negative Syndrome Scale (PANSS) was performed on these patients; and the relationships between relative glycolipid metabolic indexes and both the course of disease and manipulated variable (age,gender,education level and severity of the disease) were assessed.Results Gender might play a significant role to some glycolipid metabolic parameters (waist-hip ratio [WHR]:β=0.364; high-density lipoprotein [HDL]:β=-0.248; triacyiglycerol [TG]:β=0.167 and lysophosphatidic acid (LPA):β=-0.198,P＜0.05); age might play an important role to some glycolipid metabolic parameters (body mass index [BMI]: β=0.213; WHR: β=0.286 and apolipoprotein B 100 [apoB100]:β=0.221,P＜0.05).Simultaneously,the severity of disease appeared to affect some glycolipid metabolic parameters (BMI:β-0.167; WHR:β=-0.150 and fasting blood-glucose [FBG]:β=0.172, P＜0.05). The course of disease hardly affected the majorities of relative glycolipid metabolic indices of drug-naive schizophrenia but LPa (β=0.173, P＜0.05). Conclusion The high metabolic abnormality incidence in schizophrenia patients maybe result from multi-factor interactions.

Objective To investigate the regulatory effects of fenofibrate on TNF-α-induced CD40 expression and matrix metalloproteinase (MMP) activity in human vascular endothelial cells (HUVECs). Methods Quantitative RT-PCR and flow cytometry were employed to evaluate the effect of fenofibrate on TNF-α-induced CD40 mRNA and cell surface CD40 expression in HUVECs, and gelatin zymography was used to determine the effect of fenofibrate on the gelatinolytic activities of MMP-2 and MMP-9 in TNF-α-stimulated HUVECs. Results Fenofibrate at the concentrations of5×10-5, 1×10-4 and 2×10-4mol/L significantly reduced TNF-α-induced increment of CD40 mRNA and cell surface CD40 expressions (P＜0.01), with the maximal inhibition achieved at the concentration of 1 × 10-4 mol/L. Fenofibrate at 2× 10-4 mol/L did not further decrease CD40expression induced by TNF-α. Fenofibrate significantly inhibited the stimulatory effect of TNF-α on MMP-2 and MMP-9 activities in HUVECs. Conclusion Fenofibrate reduces TNF-α-induced increment of CD40 expression and MMP-2 and MMP-9 activities in HUVECs.

Objective To investigate human enterovirus 71 (EV71) resistance to type Ⅰ interferon induced antiviral effect.Methods After type Ⅰ interferons (α,β ) were incubated with HeLa cells,recombinant type Ⅰ herpes simple virus ( HSV-1 ) with green fluorescent protein (GFP) was inoculated onto the HeLa cells.HSV-1 proliferation was observed by GFP expression and PCR.After EV71 was inoculated onto HeLa cells incubated with the same quantity of interferon,proliferation of EV71 were detected by RTPCR of 2A gene. Results Recombinant HSV-1 GFP expression and viral DNA replication obviously decreased in HeLa cells incubated with type Ⅰ interferon (α,β).However,EV71 effectively proliferated in the interferon irritated HeLa cell by RT-PCR.Conclusion HeLa cell irritated by type Ⅰ interferon (α,β) produced antiviral substance that inhibits HSV-1 proliferation.EV71 resisted the antiviral substance induced by type Ⅰ interferon and could significantly replicate in the HeLa cells.

Objective To develop an AlphaLISA method for detection of antibody of Hepatitis A virus.Methods After hepatitis A virus antigen was concentrated and biotinylated,optimal biotinylated HAV antigen,donor bead,acceptor bead concentration have been explored and determined by both dotblotting and AlphaLISA methods.97 samples including 23 serums from patients HAV infected and 70 serums from blood donors have been detected by new AlphaLISA method.Results The sensitivity and specificity for anti-HAV IgM were 78％ and 98.5％,the positive and negative predictive value were 95％ and 93％.Conclusion A homogeneous and fast AlphaLISA assay for detection of HAV IgM has been established by preliminary verification on samples.