OBJECTIVETo probe in the possible acting mechanism of Bizhongxiao Decoction (BZXD) for treatment of early active rheumatoid arthritis (RA) by way of observing the two-dimensional gel electrophoresis map of proteins in peripheral blood mononuclear cells (PBMCs) of healthy persons and RA patients (intervened or un-intervened with BZXD), analyzing the differential proteins and seeking out the RA associated proteins.

METHODSEighteen patients with early active RA were randomized into the BZXD group and the methotrexate (MTX) group, nine in each group, they were treated with BZXD (contained 15 Chinese herbs, as Herba Hedyotis diffusae, Herba Sarcandrae glabrae, Radix Salviae miltiorrhizae, Caulis Trachelosperi, Rhizoma Drynariae, Semen Coicis, etc.) and MTX combined with nimesulide Tablets respectively, three months as a treatment course, and their blood samples were collected for observation. Besides, blood samples from 9 healthy persons were taken as normal controls. PBMCs were isolated from blood using lymphozytes separation medium, and total protein in the cells was extracted through immobilized pH gradient two-dimensional gel electrophoresis. After Coomassie brilliant blue G250 staining, gel-image analysis was performed using PDQuest software. The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). Then partial proteins were validated by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe 2-DE protein profile of PBMCs from healthy persons and RA patients before and 3 months after treatment were obtained, and 23 differential protein spots were found, 14 from 18 differential protein spots were successfully identified, of which 8 proteins were up-regulated and 6 proteins were down-regulated in RA patients as compared with control. After 3-month treatment, 5 differentially expressed proteins showed more obvious in the BZXD group than in the MTX group. RT-PCR verified that the expression of ApoA-I in all the three groups was consistent with the outcomes of 2-DE.

CONCLUSIONSSome differentially expressed proteins exist in the PBMCs of RA patients, which may play a potential role in the pathogenesis of RA; BZXD may treat RA by way of regulating the expression of some differential proteins in patients.

Adult ; Aged ; Arthritis, Rheumatoid ; blood ; drug therapy ; Blood Proteins ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Methotrexate ; therapeutic use ; Middle Aged ; Phytotherapy ; Proteome ; analysis ; Proteomics ; methods

Determination of postmortem interval (PMI) is one of the most valuable subjects in forensic practice. It, however, is often very difficult to accurately determine the PMI in daily practice. Forensic DNA technology has recently been used to estimate the PMI. It has certain advantage to traditional methods. This article reviews this technology with respect to its invention, development, advantage, disadvantage, and potential future applications with emphasis on correlation of DNA degradation and PMI.
Animals ; Bone Marrow Cells/metabolism* ; Cell Nucleus/metabolism* ; DNA/metabolism* ; Flow Cytometry ; Forensic Medicine/methods* ; Hepatocytes/metabolism* ; Humans ; Image Processing, Computer-Assisted/methods* ; Myocardium/metabolism* ; Postmortem Changes ; Spleen/metabolism* ; Time Factors

Objective To investigate the effects of dominant-negative truncation mutant?NTCF4, lacking the N-terminal form of TCF4 gene,on biological characteristics of renal cancer cell line GRC-I and explore the molecular mechanisms.Methods GRC-I cell was transfected with pCDNA3-?NTCF4 eukary- otie expression plasmid,pCDNA3 empty vector to construct the stable cell line GRC-I/?NTCF4 and GRC-I/ Mock respectively.The morphological changes of stable cells were observed and the cells growth curve was detected through light microscope.The cellular proliferation activities were determined using the MTT assay. The protein expression of Wnt pathway downstream target gene C-Myc and Cox-2 was evaluated by immuno- cytoehemieal method and Western Blot analysis.Results After the dominant-negative?NTCF4 gene was permanently expressed,the GRC-I/?NTCF4 stable cells morphologically showed that appearance changed from circular to long-spindle shape,growth rate decreased with less karyosehisis found,malignant pheno- types reversed to normal renal tubular cells.MTT assay revealed that the proliferation activities of GRC-1/?NTCF4 cells were inhibited by 11.2%-35.5% compared with GRC-I cells (P＜0.05),while the GRC- I/Mock cells have no difference with the control cells.Immunocytochemical analysis and Western Blot showed that the C-Myc and Cox-2 protein expression level of GRC-I/?ANTCF4 cells were significantly sup- pressed in comparison with that of GRC-I/Mock and GRC-I cells.Conclusions The dominant-negative truncation mutant?NTCF4 could partially inhibit the growth of renal cancer cells and down-regulate the pro- tein expression of Wnt pathway target gene C-Myc and Cox-2.These findings provide a experimental founda- tion for applying cell signal therapy to renal cell cancer by blocking the Wnt signaling pathway.

OBJECTIVETo preliminarily study the essence of wind syndrome caused by Gan-yang hyperactivity (WSGH) in Chinese medicine at the protein expression level.

METHODSWSGH was strictly differentiated from wind stirring due to yin deficiency syndrome in patients with intracerebral hemorrhage (ICH) and those with cerebral infarction (CI); from Gan-yang hyperactivity syndrome in patients with cervical spondylosis (CS); from wind syndrome induced by blood deficiency in patients with Parkinson's disease (PD) according to Chinese medicine syndrome typing standard. Control studies were performed. Peripheral blood mononuclear cells (PBMCs) were isolated from all patients of the aforesaid syndromes and healthy subjects. The total proteins were extracted, two-dimensional gel electrophoresis (2-DE) conducted and analyzed by PDQuest software. The peptide mass fingerprint (PMF) was determined using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The SwissProt database was inquired using Mascot reference system. Proteins of different and same expressions in PBMCs of patients suffering from the same disease of different syndromes, different diseases of the same syndrome, and syndromes of the same kind were compared.

RESULTSThe 2-DE map of PBMCs' total proteins in the aforesaid syndrome groups and healthy subjects was established. Through comparison, analysis, and appraisement, there was 1 protein dot of the same expression and 22 protein dots of different expressions between ICH patients of WSGH and ICH patients of wind stirring due to yin deficiency syndrome. There were 6 protein dots of the same expression and 21 protein dots of different expressions between CI patients of WSGH and CI patients of wind stirring due to yin deficiency syndrome. There were 3 protein dots of the same expression and 12 protein dots of different expressions between CS patients of WSGH and CS patients of Gan-yang hyperactivity syndrome. There was no protein dot of the same expression and 12 protein dots of different expressions between PD patients of WSGH and PD patients of wind syndrome induced by blood deficiency. There were 13 protein dots of the same expression in different diseases of the same syndrome. There was 1 protein dot (Thioredoxin-dependent peroxide reductase, TPx) of the same expression in the four diseases of the same kind syndrome.

CONCLUSIONSDifferent connotations of the essence existed (having multiple different protein expressions) in patients with the same disease of different syndromes. Syndromes of the same kind share the same material bases (having the same protein expression). These suggested that Chinese medicine syndrome has its own material bases and essence findable.

Adult ; Aged ; Cerebral Hemorrhage ; diagnosis ; metabolism ; Cerebral Infarction ; diagnosis ; metabolism ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Proteins ; metabolism ; Proteomics ; methods ; Stroke ; diagnosis ; metabolism ; Yin-Yang ; Young Adult

OBJECTIVETo investigate the effects of ursolic acid (UA) on T-cell proliferation and activation, as well as to examine its effect on nuclear factor-κB (NF-κB) signaling pathway in T cells.

METHODST-cells isolated from BALB/c mice were incubated with UA at concentrations ranging from 5-30 μmol/L in the presence of phorbol 12-myristate 13-acetate (PMA) or PMA plus ionomycin. The proliferation of T cells was measured by the MTT assay. The expressions of CD69, CD25, and CD71 on T-cell surface were analyzed using flow cytometry. The level of interleukin-2 (IL-2) in the culture supernatant of activated T cells was quantified by enzyme-linked immunosorbent assay (ELISA). The level of phosphorylated IκB-α (p-IκB-α) in total protein and p65, a subunit of NF-κB, nuclear translocation were measured by Western blot analysis.

RESULTSUA in a dose-dependent manner significantly decreased the proliferation and inhibited the surface expressions of CD69, CD25, and CD71 in murine T lymphocytes upon in vitro activation (P<0.01). Significant reduction of IL-2 production was found in activated T cells treated with UA (P<0.01). The PMA-induced increase in p-IκB-α protein was inhibited, and nuclear translocation of p65 from the cytoplasm was blocked by UA.

CONCLUSIONUA is a potent inhibitor for T cell activation and proliferation; these effects are associated with the inhibition of NF-κB signaling pathway.

Animals ; Cell Nucleus ; drug effects ; metabolism ; Cell Proliferation ; drug effects ; I-kappa B Proteins ; metabolism ; Interleukin-2 ; secretion ; Ionomycin ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Mice, Inbred BALB C ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; Phosphorylation ; drug effects ; Protein Transport ; drug effects ; Signal Transduction ; drug effects ; T-Lymphocytes ; cytology ; drug effects ; immunology ; secretion ; Tetradecanoylphorbol Acetate ; pharmacology ; Triterpenes ; pharmacology

AIM To study the chemical constituents from the roots tuber of Aconitum ouvrardianum H..METHODS The ethyl acetate fraction of methy extract from A.ouvrardianum roots tuber was isolated and purified by silica,aluminium oxide column,Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Eighteen compounds were isolated and identified as kongboendine (1),anisoezochasmaconitine (2),lipo-14-O-anisoylbikhaconine (3),franchetine (4),talatisamine (5),chasmanine (6),crassicauline A (7),chasmaconitine (8),14-dehydrotalatisamine (9),lipoindaconitine (10),indaconitine (11),yunaconitine (12),lipoyunaconitine (13),liljestrandisine (14),transconitine B (15),ouvrardiantine (16),atropurpursine (17),8-deacetylyunaconitine (18).CONCLUSION Compounds 1-4,9-10,13,15,17 are isolated from this plant for the first time.

OBJECTIVETo evaluate the expression of epidermal growth factor receptor (EGFR) gene copy number and the expression of ERCC1 and BRCA1 proteins in patients with non-small-cell lung cancer (NSCLC) and the correlation between them.

METHODSThe status of EGFR gene copy number was determined by in situ hybridization (FISH), and the expression of ERCC1 and BRCC1 proteins was examined by immunohistochemistry (IHC). The relationship of EGFR gene copy number with the expression of ERCC1 and BRCA1 and the clinical pathologic features were analyzed.

RESULTSFISH-positive EGFR expression was identified in 40 of 166 samples (24.1%). More FISH-positive EGFR in the female than male patients (31.9% vs. 18.6%, P = 0.048), and non-smoker than smoker (32.8% vs. 16.7%, P = 0.045). FISH-positive EGFR was not associated with age, pathological type, clinical stage and metestasis status (P > 0.05). The expression of ERCC1 protein was identified in 60 of 132 samples (45.5%). The expression of ERCC1 protein varied significantly in tumors of different pathological types (P = 0.046), but not associated with age, gender, clinical stage, metestatic status and smoking status (P > 0.05). The expression of BRCA1 protein was identified in 46 of 131 samples (35.1%). The expression of BRCA1 was not associated with age gender, pathological type, clinical stage, metestatic ststus and smoking status (P > 0.05). There was a moderate correlation between the expressions of ERCC1 and BRCA1 (r = 0.449, P < 0.001), but EGFR gene copy number was not correlated with the expression of ERCC1 or BRCA1 protein.

CONCLUSIONSFISH-positive EGFR expression is associated with gender and smoking status, but not correlated with the expression of ERCC1 and BRCA1 proteins. There is a moderate correlation between the expressions of ERCC1 and BRCA1.

Adult ; Aged ; Aged, 80 and over ; BRCA1 Protein ; metabolism ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Endonucleases ; metabolism ; Female ; Gene Dosage ; Gene Expression Regulation, Neoplastic ; Genes, erbB-1 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Sex Factors ; Smoking ; Young Adult

BACKGROUNDTracheostomy should be considered to replace endotracheal intubation in patients requiring prolonged mechanical ventilation (MV). However, the optimal timing for tracheostomy is still a topic of debate. The present study aimed to investigate whether early percutaneous dilational tracheostomy (PDT) can reduce duration of MV, and to further verify whether early PDT can reduce sedative use, shorten intensive care unit (ICU) stay, decrease the incidence of ventilator associated pneumonia (VAP), and increase successful weaning and ICU discharge rate.

METHODSA prospective, randomized controlled trial was carried out in a surgical ICU from July 2008 to June 2011 in adult patients anticipated requiring prolonged MV via endotracheal intubation. Patients meeting the inclusion criteria were randomly assigned to the early PDT group or the late PDT group on day 3 of MV. The patients in the early PDT group were tracheostomized with PDT on day 3 of MV. The patients in the late PDT group were tracheostomized with PDT on day 15 of MV if they still needed MV. The primary endpoint was ventilator-free days at day 28 after randomization. The secondary endpoints were sedation-free days, ICU-free days, successful weaning and ICU discharge rate, and incidence of VAP at day 28 after randomization. The cumulative 60-day incidence of death after randomization was also analyzed.

RESULTSTotal 119 patients were randomized to either the early PDT group (n = 58) or the late PDT group (n = 61). The ventilator-free days was significantly increased in the early PDT group than in the late PDT group ((9.57 ± 5.64) vs. (7.38 ± 6.17) days, P < 0.05). The sedation-free days and ICU-free days were also significantly increased in the early PDT group than in the late PDT group (20.84 ± 2.35 vs. 17.05 ± 2.30 days, P < 0.05; and 8.0 (interquartile range (IQR): 5.0 - 12.0) vs. 3.0 (IQR: 0 - 12.0) days, P < 0.001 respectively). The successful weaning and ICU discharge rate was significantly higher in early PDT group than in late PDT group (74.1% vs. 55.7%, P < 0.05; and 67.2% vs. 47.5%, P < 0.05 respectively). VAP was observed in 17 patients (29.3%) in early PDT group and in 30 patients (49.2%) in late PDT group (P < 0.05). There was no significant difference between the two groups in the cumulative 60-day incidence of death after randomization (P = 0.949).

CONCLUSIONSThe early PDT resulted in more ventilator-free, sedation-free, and ICU-free days, higher successful weaning and ICU discharge rate, and lower incidence of VAP, but did not change the cumulative 60-day incidence of death in the patients' anticipated requiring prolonged mechanical ventilation.

Aged ; Aged, 80 and over ; Critical Illness ; Female ; Humans ; Male ; Middle Aged ; Respiration, Artificial ; Time Factors ; Tracheostomy ; methods

OBJECTIVETo investigate the gene mutations of CHRNB2 and CHRNA2 in Chinese population with autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE).

METHODSOne hundred and six Han nationality patients (74 sporadic and 32 familial) were recruited and studied. Mutational screening was performed by sequencing all the 6 coding exons of the CHRNB2 gene and exons 6 and 7 of the CHRNA2 gene including the donor and acceptor splice sites.

RESULTSThe results excluded the involvement of all known published mutations of the CHRNB2 and CHRNA2 genes. However, a novel synonymous mutation c.483C>T (H161H) and a single nucleotide polymorphism (c.1407C>G) of CHRNB2 gene were detected in two ADNFLE sporadic patients respectively. The nucleotide variation H161H was heterozygous and absent in 200 healthy control samples. The mutation was also found in the proband's unaffected mother.

CONCLUSIONOur study suggests that the mutations of CHRNB2 and CHRNA2 genes may be rare in Chinese ADNFLE population. The novel synonymous mutation of H161H has not been reported previously and its impact on the pathogenesis of ADNFLE needs to be further studied.

Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; DNA Mutational Analysis ; Epilepsy, Frontal Lobe ; genetics ; Female ; Genes, Dominant ; Humans ; Male ; Mutation ; Receptors, Nicotinic ; genetics

Objective To understand the forming cause of the Oncomelania hupensis snail-existent non-endemic areas of schistosomiasis(SENEAS),and to verify the conclusion of previous studies,so as to provide the evidence for schistosomiasis monitoring in such areas in Nantong City,Jiangsu Province. Methods The controlled field tests were carried out to observe the O. hupensis snails artificially infected by schistosome miracidia in SENEAS. The influence of the soil from SENEAS and the en-demic areas on O. hupensis snails artificially infected by miracidia were observed. Results All the experimental snails could be infected by schistosome miracidia except the smooth-shell snails from Tangyuan Village in the controlled field test environment of SENEAS or the endemic areas. The infection rates of the smooth-shell snails were lower than those of the ribbed-shell snails , but there were no statistically significant differences. The mortality rates of the smooth-shell snails were higher than those of the ribbed-shell snails,which were statistically significant (χ2Xindian = 135.118,χ2Shuangdian = 122.836,χ2Baipu =154.436,χ2Dingyan =138.288,χ2Control=151.923,all P<0.01). There were no significant differences in the infection rates of snails between each test group of the soil from SENEAS and the endemic areas(χ2Rugao=0.071,χ2Rudong=0.216,both P>0.05). Also there was no signifi-cant difference between each test group and the control group without soil(χ2=7.148,P>0.05). Conclusion It is likely to form the spread of schistosomiasis in SENEAS in Nantong City with sufficient amount of infection source of schistosomiasis im-ported. It is still necessary to implement the surveillance of schistosomiasis and O. hupensis snails in Nantong City.