Objective　To compare the Immunoreactivity of various HEV Antigen with Anti-HEV IgM. Methods Solid-phase enzyme immunoassay( EIA ) was developed for detecting anti-HEV IgM by using synthetic peptides E30, E42, E33, and recombinant antigen from HEV ORF-2. Results Of 60 anti-HEV positive sera by using E30, E42, E33 and recombinant antigen as coating antigen, Anti-HEV IgM positive rates were 76.7%, 26.6%, 18.3% and 66.7% respectively. In Acute-phase and convalescence-phase sera of the patients with Hepatitis E, Anti-HEV IgM positive rate was 90% and 3.3% respectively. Conclusions The HEV E30-based EIA will be very useful in the early diagnosis of Hepatitis E.

AlM: To investigate the relationship between the anterior ischemic optic neuropathy ( AlON ) and the carotid artery change using doppler ultrasound.METHODS:Fifty-four cases of AlON patients and 54 cases of healthy control were observed, atherosclerotic spots were detected by the application of color ultrasound.RESULTS:ln AlON group of 54 patients, 38 cases appeared carotid atherosclerosis, accounting for 70%. The number of cases with hard plaque, soft plaque and mixed plaques were 18, 13, and 7 respectively, accounting for 33%, 24% and 13%. ln the control group, 20 cases were detected atherosclerotic change, accounting for 37%. And the number of cases with hard plaque, soft plaque and mixed plaques were 12, 5 and 3 respectively, accounting for 22%, 9%, 6%. Significant stenosis and velocity change were showed in neither AlON group nor control group. Compared with the control group, AlON group had more cases of atherosclerotic plaque, the difference was statistically significant (χ2=12. 836, P=0. 005)CONCLUSlON: The incidence of AlON is correlated with carotid atherosclerosis, and carotid ultrasonography is significantly valuable for AlON etiology and diagnosis.

Background: Recent studies indicate that C-X-C motif chemokine receptor 4 (CXCR4) and its ligand, C-X-C motif chemokine ligand 12 (CXCL12), stimulate expression of the cell cycle regulatory protein Cyclin D1 in neurofibroma-tosis 1-associated malignant peripheral nerve sheath tumor (MPNST) cells and promote their proliferation. In this study, we measured the expression of CXCR4, CXCL12, and Cyclin D1 proteins in sporadic MPNST tissues from Chinese patients and investigated their prognostic values. Methods: CXCR4, CXCL12, and Cyclin D1 protein expression in samples from 58 Chinese patients with sporadic MPNST was assessed with immunohistochemical staining. Their prognostic values were evaluated with Kaplan–Meier analysis and a log-rank test. Multivariate Cox regression analysis was used to identify independent prognostic factors. Results: High expression of CXCR4, CXCL12, and Cyclin D1 was observed in 19 (32.8％), 32 (55.2％), and 16 (27.6％) samples, respectively. CXCR4 expression was positively correlated with CXCL12 expression (r= 0.334,P= 0.010) and Cyclin D1 expression (r= 0.309,P= 0.018). Patients with high CXCR4 expression showed longer overall survival than those with low CXCR4 expression (χ2= 4.642,P= 0.031). Conclusion: High CXCR4 expression may define a specific subtype of sporadic MPNST with favorable prognosis.

OBJECTIVETo investigate the effect of SIRT6/NF-κB signal axis in delaying hematopoietic stem/progenitor cell senescence with ginsenoside Rg1, in order to provide theatrical and experimental basis for looking for methods for delaying HSC senescence.

METHODSca-1 + HSC/HPC was isolated by magnetic cell sorting (MACS) and divided into five groups: the normal control group, the aging group, the positive control group, the Rg1 anti-senescence group, and the Rg1-treated group. Senescence-associated β-galactosidase (SA-β-Gal) staining, cell cycle analysis and hemopoietic progenitor cell mix (CFU-Mix) were adopted to determine the effect Rg1 in delaying or treating Sca-1 + HSC/HPC senescence biology. The mRNA and protein of senescence regulation molecules SIRT6 and NF-KB were examined by realtime fluorescence quantitative PCR (FQ-PCR) and western blotting.

RESULTCompared with the senescence group, the Rg1 anti-senescence group and the Rg1-treated group showed lower percentage in SA-β-Gal-stained positive cells, decreased cell proportion in G1 phase, increased number of CFU-Mix, up-regulated in SIRT6 mRNA and protein expression, down-regulation in NF-KB mRNA and protein expression. The Rg1 anti-senescence group showed more evident changes in indexes than the Rg1-treated group.

CONCLUSIONRg, may inhibit Sca-1 + HSC/HPC senescence induced by t-BHP by regulating SIRT6/NF-KB signal path.

Animals ; Antigens, Ly ; analysis ; Cellular Senescence ; drug effects ; Female ; Ginsenosides ; pharmacology ; Hematopoietic Stem Cells ; drug effects ; Male ; Membrane Proteins ; analysis ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; physiology ; Signal Transduction ; physiology ; Sirtuins ; physiology

This study is to establish an HPLC-DAD-ELSD method for simultaneous determination of 5 flavones and saponins in Rhizoma Anemarrhenae including neo-mangiferin, mangiferin, timosaponin B II, timosaponin B III and timosaponin A III. Samples were analyzed on a Merck Purospher STAR column(4.6 mm x 250 mm, 5 μm). The mobile phase consisted of acetonitrile( A) and 0. 1% formic acid (B) with gradient elution at a flow rate of 1.0 mL · min(-1). The column temperature was set at 40 °C. The DAD detector wavelength was set at 254 nm. The ELSD conditions were as follows: the nebulizing gas flow rate was 2.0 L · min(-1) and temperature of drift tube was 105 °C. The volume was 10 μL. The five compounds were well separated with good linear correlations. The mean recoveries were between 102.0%-104.0%. This method was quick and reliable which provides a foundation for quality control of R. Anemarrhenae.
Anemarrhena ; chemistry ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Rhizome ; chemistry ; Saponins ; analysis

OBJECTIVETo study the effects of Bushen Zhuyun Decoction (BZD) on the reproductive capacity of mice.

METHODSTotally 60 female Kunming mice were randomly divided into the experimental group and the control group. BZD was perfused by gastric tube to mice in the experimental group, twice daily. Meanwhile, mice in the control group were administered with normal saline by gastrogavage. Mice were sacrificed on the 10th day and the 20th day of medication respectively, 15 mice each time in each group. The serum levels of estradiol and progesterone were detected by radioimmunoassay. The morphological changes of uterus and ovary were observed using HE staining. The expressions of leukaemia inhibitory factor (LIF), epidermal growth factor (EGF), and calcitionin (CT) were determined using immunohistochemical assay.

RESULTSCompared with the control group at the same time point, the weights of uterus and ovary increased, the serum levels of estradiol and progestogen increased, the expressions of LIF, CT, and EGF increased in the experimental group on the 10th day and the 20th day of medication (P <0.05, P <0.01). Better results were shown on the 20th day of medication (P <0.05). Histological results showed increased ovarian follicle numbers, increased endometrial gland numbers, endometrial hyperplasia (stratiform arranged), and increased stromal cells in the experimental group, especially on the 20th day of medication.

CONCLUSIONBZD could improve the reproductive capacity by advancing the development of generative organs, promoting the secretion of estradiol and progestogen and the follicular growth, and increasing the receptivity of endometrium.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Endometrium ; metabolism ; Estradiol ; metabolism ; Female ; Mice ; Mice, Inbred Strains ; Organ Size ; Ovary ; drug effects ; Progesterone ; metabolism ; Uterus ; drug effects

Objective To investigate the serum levels of endothelin-1 (ET-1), tumor necrosis factor- α (TNF-α) and their dynamic changes in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) and their relation with the condition variation of the patients.Methods The serum ET-1 and TNF-α levels were determined with enzyme-linked immunno-sorbent assay (ELISA) in 31 patients with DEACMP, and the dynamic changes of patients' condition were monitored by use of the activity of daily living (ADL) scale, the information-memory-concentration test (IMCT) and the Hasegawa's dementia scale (HDS). The comparisons between patients with DEACMP and both 30 patients with acute carbon momoxide poisoning (ACMP) but without DEACMP and 30 normal controls were also conducted. Results At the acute stage of the DEACMP group, the serum levels of ET-1 and TNF-α were both significantly higher than those in the normal control group (P＜0.05); that of TNF-α was significantly lower than that in the ACMP group (P＜0.05), but that of ET-1 was not significantly different from that in the ACMP group (P＞0.05). The serum levels of ET-1 and TNF- α in the ACMP group were both significantly higher than those in the normal control group(P＜0.05). In the DEACMP group, the serum level of ET-1 at the convalescent stage was significantly lower than that at the acute stage (P＜0.05), but the serum level of TNF-α was not significantly different from that at the acute stage (P＞0.05). At the acute stage of the DEACMP group, ADL scores were significantly higher than those in norms, and IMCT scores and HDS scores were significantly lower than those in norms (P＜0.05). In the DEACMP group, the ADL scores at the convalescent stage were significantly lower than those at the acute stage (P＜0.05), IMCT scores and HDS scores were significantly higher than those at the acute stage (P＜0.05). Significant correlations between scores of any 2 of 3 scales in patients with DEACMP at both acute and convalescent stage were noted (P＜0.05). Conclusion The dynamic detection of serum ET-1 and TNF-α level variations could be used as an indicator for condition severity in patients with DEACMP.

Background: According to the 7th edition of the American Joint Committee on Cancer (AJCC) staging system, over 50% of patients with nasopharyngeal carcinoma (NPC) have N1 disease at initial diagnosis. However, patients with N1 NPC are relatively under-researched, and the metastasis risk of this group is not well-stratified. This study aimed to evaluate the prognostic values of gross tumor volume of metastatic regional lymph node (GTVnd) and pretreatment serum copy number of Epstein–Barr virus (EBV) DNA in predicting distant metastasis of patients with N1 NPC, and to develop an integrated prognostic model that incorporates GTVnd and EBV DNA copy number for this group of patients. Methods: The medical records of 787 newly diagnosed patients with nonmetastatic, histologically proven N1 NPC who were treated at Sun Yat-sen University Cancer Center between November 2009 and February 2012 were ana-lyzed. Computed tomography-derived GTVnd was measured using the summation-of-area technique. Blood sam-ples were collected before treatment to quantify plasma EBV DNA. The receiver operating characteristic (ROC) curve analysis was used to evaluate the cut-off point for GTVnd, and the area under the ROC curve was used to assess the predicted validity of GTVnd. The survival rates were assessed by Kaplan–Meier analysis, and the survival curves were compared using a log-rank test. Multivariate analysis was conducted using the Cox proportional hazard regression model. Results: The 5-year distant metastasis-free survival (DMFS) rates for patients with GTVnd > 18.9 vs. ≤ 18.9 mL were 82.2% vs. 93.2% (P < 0.001), and for patients with EBV DNA copy number > 4000 vs. ≤ 4000 copies/mL were 83.5% vs. 93.9% (P < 0.001). After adjusting for GTVnd, EBV DNA copy number, and T category in the Cox regression model, both GTVnd > 18.9 mL and EBV DNA copy number > 4000 copies/mL were significantly associated with poor prognosis(both P < 0.05). According to combination of GTVnd and EBV DNA copy number, all patients were divided into low-, moderate-, and high-risk groups, with the 5-year DMFS rates of 96.1, 87.4, and 73.8%, respectively (P < 0.001). Multi-variate analysis confirmed the prognostic value of this model for distant metastatic risk stratification (hazard ratio [HR], 4.17; 95% confidence interval [CI] 2.34–7.59; P < 0.001). Conclusions: GTVnd and serum EBV DNA copy number are independent prognostic factors for predicting distant metastasis in NPC patients with N1 disease. The prognostic model incorporating GTVnd and EBV DNA copy number may improve metastatic risk stratification for this group of patients.

OBJECTIVETo study the immunogenicity and immunoprotection of the recombinant expressing product (rSpaO) of S. paratyphi A spaO gene, and to demonstrate the frequencies of spaO gene carrying and expressing in S. paratyphi A isolates.

METHODSThe spaO gene of a clinical S. paratyphi A strain JH01 was amplified and then cloned. After sequencing of the cloned spaO gene, a prokaryotic expression system of the gene was constructed. SDS-PAGE were applied to examine the rSpaO expression. Ni-NTA affinity chromatography was performed to collect rSpaO. Immunogenicity of rSpaO was determined by Western blot assay. A PCR assay and an ELISA were established to respectively detect the carrying and expressing frequencies of the spaO genes in 98 S. paratyphi A isolates. The immunoprotective effects of rSpaO in S. paratyphi A strain 50001 infected mice were observed.

RESULTSIn comparison with the reported corresponding sequences, the nucleotide and putative amino acid sequence homologies of the cloned spaO gene were 99.45%-99.89% and 99.01%-100%, respectively. The expression output of rSpaO was approximately 75% of the total bacterial proteins. S. paratyphi A antiserum could recognize as well as combine with rSpaO. rSpaO could efficiently induce rabbits to produce specific antibody. 94.9% (93/98) of the S. paratyphi A isolates had spaO gene and 91.4% (85/93) of the spaO+ strains could express SpaO. 58.3% and 50.0% of the mice that oral-taken or subcutaneous injected with 500 microg of rSpaO for immunization were survival after challenged by lethal dose of S. paratyphi A strain 50001. When co-immunized with 5 microg rLTB, the survival rates of the mice increased to 88.3% and 75.0%, respectively.

CONCLUSIONThe S. paratyphi isolates had relatively high carrying and expressing frequencies of spaO gene. rSpaO showed a fine immunogenicity and a certain immunoprotective effect, which could be used as an antigen candidate for developing genetic engineering vaccine of S. paratyphi.

Animals ; Antibody Formation ; Bacterial Proteins ; genetics ; immunology ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation, Bacterial ; Genetic Engineering ; Membrane Proteins ; genetics ; immunology ; Mice ; Polymerase Chain Reaction ; Recombinant Proteins ; Salmonella Vaccines ; immunology ; Salmonella paratyphi A ; genetics

Objective To evaluate the causes of DR waste film rates in different characteristic cases,and provide basis for quality control.Methods DR waste film rates of 98 503 cases from PACS were analyzed.All cases were grouped according to gender,age,X ray projection direction and projection position.The differences of DR waste film rates between the groups were compared.Results DR waste film rate in 98 503 cases was 4.89％ and factors that caused waste film were incorrect position,foreign materials,motion artifacts,machine problems and inappropriate X ray dosage.DR waste film rate was affected by gender,age,X ray projection direction,but not affected by projection position.Conclusion Analysis of DR waste film rate in different characteristic cases could provide basis for the quality control to reduce the DR waste film rate.