Acupuncture Analgesia ; Acupuncture, Ear ; Adult ; Chemoembolization, Therapeutic ; Humans ; Liver Neoplasms ; complications ; surgery ; Male ; Pain, Postoperative ; etiology ; therapy

Objective To compare the significance of DNA-associated autoantibodies to cell membrane(cmDNA)in systemic lupus erythematosus(SLE)detected with indirect immunofluorescence on human B lymphoma cell line Raji and pmmyelocytic line HL60.Methods Indirect immunofluorescence assay both on cell line Raji and HL60 was used to measure anti-cmDNA antibodies in sera of 306 SLE patients.192 patients with other rheumatic diseases and 50 healthy controls.Results Indirect immunofluorescence assay on cell line Raji was used to measure anti-cmDNA antibodies.72.5% SLE and 10.4% other rheumatic diseases were positive for anti-cmDNA,but negative in 50 blood donors(P<0.01).Indirect immunofluorescence assay on cell line HL60 was used to measure anti-cmDNA antibodies,76.1% SLE and 16.7% other rheumatic diseases were positive for anti-cmDNA,but negative in 50 blood donors(P<0.01).The sensitivity of anti-cmDNA were 72.5%and 76.1%,respectively.The specificity of anti-cmDNA was 91.7% and 86.8%,respectively.There was no significant difference in sensitivity and spocificity(P>0.05).The methods of culture,freeze and resuscitation on the two cells were similar.but cell line Raji was easier to resuscitate than cell line HL60.Observing with fluorescence microscope.we find that cmDNA was expressed on the both cells and the staining was stronger on cellline Raji than HL60.Conclusion Anti-cmDNA antibody has high positivity which is one of the most valuable marker in the diagnosis of SLE.We recommend to measure anti-cmDNA antibodies with indirect immunofluorescence assay on cell line Raji rather than HL60.

ObjectiveTo compare the significance of anti-cmDNA antibody in systemic lupus erythematosus (SLE) patients detected with IIF on human's B lymphoma cell line Raji and promyelocytic line HL60.The diagnostic value of anti-cmDNA antibody in SLE was also explored.MethodsThree hundred and six patients with SLE were included in this study.As control groups,we included 192 patients with other rheumatic diseases and 50 healthy controls.The testing method for anti-cmDNA antibody was set up.The assessment of the significance of anti-cmDNA antibody in SLE detected with IIF on cell line Raji and HL60 was carried out andthe diagnostic value of anti-cmDNA antibody in SLE was investigated.ANA and antidsDNA antibody were measured by IIF at the same time.Anti-Sm was measured by immuno-diffusion andWestern blotting.AnuA was tested by enzyme linked immunosorbent assay.The statistical methods used in this study including McNemar X2 test,Spearman related test and Logistic regression analysis.Results The fluorescence brightness of Raji cell line was stronger than HL60 cell line.There was no statistically significant difference in the sensitivity and specificity of anti-cmDNA antibody in SLE detected with IIF with Raji or HL60 cell lines (P＞0.05).The sensitivity of anti-cmDNA antibody detected with IIF on Raji cell line was higher than anti-dsDNA antibody and anti-Sm antibody(P＜0.01),while the specificity of anti-cmDNA antibody was similar to anti-dsDNA antibody (P＞0.05) and was lower than anti-Sin antibody (P＜0.01).The sensitivity of anti-cmDNA antibody was similar to AnuA(P＞0.05) and the specificity was lower than AnuA (P＜0.01).The sensitivity of ANA was higher than anti-cmDNA antibody (P＜0.01) and the specificity was much lower than anti-cmDNA antibody(P＜0.01).The sensitivities of anti-dsDNA antibody,anti-Sm antibody and AnuA were much higher when combined with anti-dsDNA antibody than any one antibody only (P＜0.05).Anti-cmDNA antibody was correlated with mucosa ulcer in SLE patients(OR=2.343,P=0.029).The ESR of SLE patients was also correlated with anti-cmDNA antibody(OR=l.031,P=0.012).Anti-cmDNA antibody was not correlated with SLEDAI (r=0.070,P=0.600).ConclusionRaji cell line is better than HL60 cell line in detecting anti-cmDNA antibody with IIF.Anti-cmDNA antibody has higher sensitivity and specificity in SLE.Combined detection of anti-cmDNA antibody and other autoantibodies can further improve the diagnostic accuracy of SLE.

BACKGROUNDProteins or peptides can be directly transferred into cells when covalently linked to protein transduction domains (PTDs). TAT is one of the most widely studied PTDs. The effect of fusion protein TAT and heme oxygenase-1 (HO-1) on liver sinusoidal endothelial cells (SECs) apoptosis during cold storage is unknown. The present study aimed to determine whether fusion protein TAT-HO-1 would transduce efficiently into liver during cold storage, and, if so, to determine whether TAT-HO-1 would attenuate SECs apoptosis during preservation injury in rat.

METHODSLivers of Sprague-Dawley rats were harvested and randomly assigned to group 1 (HTK solution) and group 2 (HTK solution containing TAT-HO-1 fusion protein) according to the type of the preservation solution. The transduction efficiency of TAT-HO-1 was examined and the impairment of SECs was assessed during the period of cold storage followed by 1 hour of reperfusion.

RESULTSTAT-HO-1 can transduce efficiently into liver during cold storage. A significantly lower apoptotic index of SECs was observed in group 2, at 6, 12 and 18 hours of cold storage after 1 hour reperfusion, when compared with group 1. TAT-HO-1 reduced HA and ET levels in liver at each time point. Both Bcl-2 and Bax protein were expressed in hepatocytes and SECs at the periphery of the sinusoidal space. Moreover, higher Bcl-2 expression and lower Bax expression were observed in group 2.

CONCLUSIONSTAT-HO-1 can transduce efficiently into rat livers and shows a protective effect on SECs by attenuating apoptosis during cold ischemia/reperfusion injury. Protein transduction will be a novel therapeutic strategy to reduce the risk of preservation injury in liver transplantation.

Animals ; Apoptosis ; drug effects ; Endothelial Cells ; cytology ; drug effects ; Heme Oxygenase-1 ; genetics ; Immunohistochemistry ; In Situ Nick-End Labeling ; In Vitro Techniques ; Liver ; cytology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; bcl-2-Associated X Protein ; metabolism ; tat Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo summarize the phenotypes and identify SCN1A mutations in families with generalized epilepsy with febrile seizures plus (GEFS(+)), and analyze the genotype- phenotype correlations in GEFS(+) families.

METHODGenomic DNA was extracted from peripheral blood lymphocytes of the proband and other available members in the GEFS(+) families. The phenotypes of the affected members were analyzed. The coding regions and flanking intronic regions of the SCN1A gene were screened for mutations using PCR and direct DNA sequencing.

RESULTIn 39 GEFS(+) families, there were 196 affected members, ranging from 2 to 22 affected members in each family. Their phenotypes included febrile seizures (FS) in 92(46.9%), febrile seizures plus (FS(+)) in 62(31.6%), FS or FS(+) with partial seizures in 12(6.1%), afebrile generalized tonic-clonic seizures (AGTCS) in 11(5.6%), myoclonic atonic epilepsy in 8(4.1%), Dravet syndrome in 2(1.0%), childhood absence epilepsy in 1 (0.5%), FS(+) with myoclonic seizures in 1(0.5%), AGTCS and myoclonic seizures in 1 (0.5%), partial seizures in 1 (0.5%), unclassified seizures in 5 (2.6%). Four families were found with SCN1A mutations, including three families with missense mutation (N935H, R101Q, G1382R) and one family with truncation mutation (C373fsx378). In three families with missense mutations, the phenotypes include FS, FS(+), FS(+) with partial seizures, and AGTCS. In one family with truncation mutation, the phenotypes included FS, FS(+), and Dravet syndrome. The mother of proband in the family with missense mutation (R101Q) and the father of proband in the family with truncation mutation (C373fsx378) were both somatic mosaicism. Both of their phenotypes were FS(+).

CONCLUSIONThe most common phenotypes of GEFS(+) were FS and FS(+), followed by the FS/FS(+) with partial seizures and AGTCS. The most severe phenotype was Dravet syndrome. SCN1A mutation rate in GEFS(+) was about 10%. Missense mutation was common in GEFS(+) families, few with truncation mutation. Few members of GEFS(+) families had somatic mosaicism of SCN1A mutations and their phenotypes were relatively mild.

Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Epilepsies, Myoclonic ; genetics ; Epilepsy, Generalized ; genetics ; Female ; Genotype ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; genetics ; Mutation, Missense ; NAV1.1 Voltage-Gated Sodium Channel ; genetics ; Pedigree ; Phenotype ; Seizures, Febrile ; genetics

OBJECTIVETo summarize and analyze the clinical characteristics and diagnostic and therapeutic measures for the first human case of H5N1 avian influenza pneumonia in mainland of China.

METHODSThe clinical data of the first case of H5N1 avian influenza virus infection in China were analyzed and summarized.

RESULTSThe case is a 9-year old boy, who developed acute symptoms of a light common respiratory infection, including fever and dry cough without obvious catarrh. On the 7th day after onset, his temperature reached 40 degrees C, tachypnea occurred, distinct rales could be heard and large areas of consolidation were seen in the lungs on chest X-ray. The patient's peripheral blood leukocyte count was 2.81 x 10(9)/L and neutrophils dominated. After comprehensive therapeutic approaches, including antiviral therapy (amantadine) and use of low-dosage glucocorticoid, the patient's temperature returned to normal on the 3rd hospitalization day, chest X-ray showed absorbed inflammatory change on the 5th day after admission, and leukocyte count became normal on the 6th day. No complication occurred during the whole course. The case was diagnosed by the 4 fold raised antibody to the H5N1 influenza virus in recovery stage serum because the H5N1 nucleic acid test in early stage was negative. The case was cured and discharged after 3 weeks comprehensive treatment.

CONCLUSIONSIt is very important for clinicians to pay enough attention to epidemiological history, especially history of exposure to avian influenza virus contaminated material, which will be very helpful for early detection, early diagnosis of the disease, and also very important for effective treatment and better prognosis.

Amantadine ; therapeutic use ; Animals ; Antibodies, Viral ; blood ; immunology ; Antiviral Agents ; therapeutic use ; Birds ; Child ; China ; Glucocorticoids ; therapeutic use ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; isolation & purification ; Influenza in Birds ; transmission ; Influenza, Human ; complications ; diagnosis ; Male ; Pneumonia ; diagnosis ; drug therapy ; physiopathology ; virology ; Treatment Outcome

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

Objective To investigate the microbleeding incidence of healthy eldery population and patients with stroke.Methods 30 cases of healthy eldery population,32 cases of cerebral hemorrhage,46 cases of patients with ischemic cerebral vascular diseases were performed of MRI and GRE-T_2 ~* WI examination.Results The microbleeding incidences was 37.5% in cerebral hemorrhage group,28.1% in multiple cerebral infarction group,25.0% in Binswanger's disease group.The most frequently seen microbleeding foci located in ganglia areas,then in thalamus areas,subcortical areas and brain stem,last in cerebellar.Conclusion GRE-T_2 ~* WI,helpful for finding microbleeding and indicating lesion degree of microblooding vessels,plays an important role in the diagnosis of stroke and decision making of treatment.

OBJECTIVETo analyze and review the characteristics of leukoencephalopathy with vanishing white matter (VWM).

METHODSThe clinical features including clinical manifestations, neurologic signs, cranial MRI and laboratory tests in 9 patients with the diagnosis of VWM were analyzed and the characteristics of the disease were reviewed.

RESULTS

CLINICAL MANIFESTATIONS8 cases had symptoms involving central nervous system, 1 case only showed abnormal cranial MRI findings. The onset of the disease occurred between 6 months to 3 years of age. Family history was positive in 5 cases. Almost all cases had normal psychomotor development before the onset of the disease. The initial symptom was usually movement disorder with predominant involvement of lower limbs. The onset or deterioration of the disease was followed by respiratory tract infection in 6 cases and minor head trauma preexisted in 3 cases. The course of the disease was progressive in 7 cases and there was episodic deterioration in 4 cases. Mental abilities were relatively better preserved. Head circumference was normal in 7 cases. Positive upper motor unit signs were found in 8 cases and ataxia in 4 cases. Bilateral optic nerve atrophy was found in 3 cases. Cranial MRI indicated diffuse and symmetrical involvement of deep white matter which showed long T(1) and T(2) signal. Subcortical white matter was also involved with predominance in frontal and parietal lobes. Flair image showed symmetrical high signal intensity in cerebral white matter with low signal intensity similar to that of CSF in partial area or low signal in most area of white matter with only meshwork of higher signal preserved. The results of all the laboratory tests including the enzyme and biochemical test specific for some well-known leukoencephalopathy were normal.

CONCLUSIONSThe clinical features of VWM include: 1. Initial symptom is usually movement disorder; 2. Movement disorder is more prominent compared to mental retardation; 3. Cranial MRI shows symmetrical abnormal T(1) and T(2) signal in deep white matter with signs of vanishing white matter. Exclusion of other hereditary and acquired leukoencephalopathy is necessary for diagnosis. Final diagnosis should be made on the basis of genetic evidence.

Adolescent ; Age of Onset ; Brain ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukoencephalopathies ; pathology ; Magnetic Resonance Imaging ; Male ; Movement Disorders ; physiopathology

To define the molecular basis of ethanol dependence, changes in the phosphorylation of cAMP response element binding protein (CREB) in the nucleus accumbens of rats after acute and chronic ethanol administration were detected using immunohistochemistry. The results demonstrate that the expression of phospho-CREB (p-CREB) protein in the rat nucleus accumbens significantly increased after 15 min of acute ethanol exposure, reaching a peak at 30 min after ethanol administration. The increment remained after 1 or 6 h of ethanol exposure compared to the control rats. In contrast, chronic intake of ethanol solution obviously decreased the expression of p-CREB protein compared to the control rats. The decrement remained 24 h or 72 h after ethanol withdrawal, and returned to the control levels after 7 d of ethanol withdrawal. The results suggest that an acute ethanol administration led to an increase in the phosphorylation of CREB in the nucleus accumbens, but chronic ethanol administration produced a decrement, which is possibly one of the molecular mechanisms of alcohol dependence.
Alcoholism ; metabolism ; physiopathology ; Animals ; Cyclic AMP Response Element-Binding Protein ; chemistry ; metabolism ; Ethanol ; pharmacology ; Male ; Nucleus Accumbens ; metabolism ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Substance Withdrawal Syndrome ; metabolism ; physiopathology ; Substance-Related Disorders ; metabolism ; physiopathology