Objective To purify Shiga toxin Ⅱ (STX Ⅱ) of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 by affinity chromatography, and characterize its biological function. Methods The immno-affinity chromatography column was prepared by STX Ⅱ A subunit-specific antibody S1D8 coupling to Sepharose 4B matrix. The purity and specificity of STX Ⅱ molecule secreted by EHEC O157:H7 were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, respectively. The purified toxin was serially diluted and the toxic activities to Vero cell line and mice were observed. The 50% cytotoxic dose (CD50) for Vero cell line and 100% lethal dose (LD100) for mice were calculated. The protection effect of anti-STX Ⅱ polysera to the mice against the purified toxin challenge was also observed. Results STX Ⅱ was successfully purified from culture supernatant of EHEC O157:H7 using affinity chromatography scheme. The relative molecular weights of STX Ⅱ A and B subunits were 32 000 and 7 500 confirmed by SDS-PAGE, respectively. The purified toxin could react with monoclonal antibodies against STX Ⅱ A and B subunits, respectively.The toxin was cytotoxic to Vero cell with CD50 of 20 ng/L and lethal to mice with LD100 of 5 ng.The toxin could be neutralized by anti-STX Ⅱ polysera in vivo. Conclusion STX Ⅱ is successfully purified and its toxic effects are confirmed in both cell line and mouse model.

To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.
Alleles ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Medicine, Mongolian Traditional ; Molecular Sequence Data ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods

Objective To evaluate the effect of pain management training to junior nurses' practice of pain assessment,to provide evidence for continuing education of pain management.Methods 60 junior nurses were randomly assigned to experimental group(30 cases) and the control group (30 cases).Nurses in control group received convention education,whereas nurses in experimental group received a pain management training for four weeks.The practice of pain assessment of two groups were compared after 4-week training.Results After training,the knowledge and attitude of pain management of nurses in experimental group scored an average of (29.20 ± 3.25),whereas the average score of nurses in control group is (16.83 ± 2.87).There was significance difference between two groups (t =-15.625,P ＜ 0.01) ; in terms of the contents of pain assessment,nurses in experimental group were more sufficient and more accurate (t =-14.726,P ＜ 0.01).Regarding the practice of pain assessment the mean score of nurses in experimental group and control group were (6.70±1.47) and (2.93 ±0.87).There was significant difference between two groups (t =12.11,P ＜0.01).For using pain assessment tool,the nurses in experimental group get 100.0％ correct rate,whereas nurses in control group get 30.0％ correct rate,which shows significant differences statistically (x2 =27.81,P ＜ 0.01).Conclusions The pain management training can improve nurses' pain knowledge,attitude,as well as promote the accuracy of nurses 'assessment practice consequently.

OBJECTIVETo investigate the induction of apoptosis on human oral epidermoid carcinoma KB cells and multidrug resistant KBv200 cells by Matrine.

METHODSMTT assay was used to investigate the inhibition ability of Matrine on the cells in vitro. Transmission electron microscope was used to observe the ultrastructure feature of cells. after treated by Matrine. Acridine orange (AO)/Ethidium bromide (EB) fluorescent staining and flow cytometry were used to observe apoptosis induced by Matrine. Flow cytometry was applied to study the effects of the drug on cell cycles of the cells.

RESULTSWhen 0.50, 1.00, 1.50, 2.00 mg/ml of Matrine was used, the vital rates of KB and KBv200 cells were decreased according to Matrine's concentration. The IC50 concentrations of Matrine on KB and KBv200 cells were 1.35 mg/ml and 1.43 mg/ml individually. The results of AO/EB fluorescent staining and flow cytometry showed that Matrine could induce apoptosis of two kinds of cells. While observed by transmission electron microscope, there were more contraction of cells, condensation of nuclei, bubble of cytoplasm in both kinds of cells after treated by Matrine. Matrine could stop the growth of KB and KBv200 cells at S period and restrain mitosis of cells.

CONCLUSIONMatrine can inhibit the growth of KB and KBv200 cells by inducing apoptosis. The apoptosis effect is dose-dependent and it has certain relation to the blocking of S period cells.

Alkaloids ; Apoptosis ; Carcinoma, Squamous Cell ; Humans ; KB Cells ; Quinolizines

Licorice, one of the most commonly used medicinal materials in China, grows mainly in arid and semi-arid regions and has important economic and ecological values. Basic leucine zipper (bZIP) transcription factors in plants play an important role in regulating biological or abiotic stress responses, growth, and secondary metabolite synthesis. bZIP transcription factors in the published whole genome database of Glycyrrhiza uralensis were identified using bZIP sequences found in Arabidopsis thaliana genome as reference, and ABA-dependent bZIP genes were identified by using Illumina high-throughput sequencing. The physical and chemical properties, structure of the encoded proteins, and the gene expression patterns with exogenous ABA stress were analyzed. A total of 69 bZIP transcription factor genes were identified in G. uralensis, named Gubzip1-69, and they were divided into 10 subfamilies (A-I and S) according to their similarity to bZIPs of A. thaliana. By calculating the relative expression levels of the 69 GubZIPs genes under different concentrations of exogenous ABA stress, genes that may be involved in the regulation of ABA signaling pathways were identified, namely GubZIP1, GubZIP5, GubZIP8, GubZIP30, GubZIP33 and GubZIP56. The results of expression pattern analysis of these GubZIPs genes under exogenous ABA stress showed that the expression pattern of GubZIPs genes changed significantly with 50 mg·L^-1 ABA. The relative expression levels of these genes decreased 3 h after treatment, and gradually increased 6 h after treatment. Except for GubZIP8, the relative expression levels of these genes were significantly increased after 12 h. Further research on the function of bZIP transcription factors of G. uralensis and elucidating their regulatory mechanisms should be of interest and will provide a scientific basis for cultivating high-quality cultivars of G. uralensis through molecular breeding methods.

Objective To observe apelin and its receptor (APJ) expressions in human osteoblasts and evaluate the effect of apelin on osteoblasts.Methods The expressions of apelin and APJ in human osteoblasts were tested by RT-PCR and Western blot.After human osteoblasts were treated with apelin,cell proliferation was measured by [~3H] thymidine incorporation and cell counting.Cell function was measured by alkaline phosphatase (ALP) activity,the secreted osteocalcin level and typeⅠcollagen production .The activation of signaling cascades was tested by Western blot.Small-interfering RNA (siRNA) to blockade APJ was applied to observe effects of apelin on cell proliferation and the activation of signaling cascades.Results Both apelin and APJ were expressed in human osteoblasts.Apelin increased the proliferation and did not show the influences on ALP activity, osteocalcin secretion and type I collagen production in human osteoblasts.Apelin induced activation of phosphatidylinositol-3 kinase (PI3K) downstream effector (Akt),but not mitogen-activated protein kinase (MAPK) such as c-jun N-terminal kinase (JNK),p38 and ERK1/2 in human osteoblasts.Suppression of APJ with siRNA or LY294002 (PI3K inhibitor) abolished the apelin-induced cell proliferation and the activation of Akt.Conclusion Human osteoblasts express apelin and APJ.Apelin stimulates the proliferation of human osteoblast via APJ/PI3K/Akt pathway,but has no effect on osteoblast differentiation.

Objective To investigate the effect of transcatheter arterial chemoembolization(TACE) using As_2O_3 and Lipiodol on the growth and metastasis of the implanted hepatic tumor in rabbits and the correlation of metastasis with angiogenesis of the residual tumor.Methods Forty-eight New Zealand white rabbits were randomly divided into four groups and VX_2 carcinoma was implanted in the left lobes of the livers.Two weeks later,a catheter was inserted into the hepatic artery and infusion was performed via the hepatic artery using physiological saline(group A),Lipiodol(group B),ADM-Lipiodol(group C),and As_2O_3-Lipiodol(group D),respectively.One week after the treatment,the value of microvessel density (MVD)of tumors(samples got by biopsy)was examined by immunohistochemistry.Three weeks after the treatment,the volume and necrotic area of the implanted tumor were measured.The metastases in the liver, lungs and other organs were recorded.Results One week after the treatment,the value of MVD of the tumorswas(21.8?5.3),(23.4?3.9),(22.4?4.5),and(14.3?3.4)/400 power LM(F= 11.246,P=0.000).Three weeks after the treatment,the mean volume of the implanted tumor was (35.5?7.1),(21.2?8.3),(20.7?9.1),and(11.8?3.7)cm~3(F=21.203,P=0.0000)in groups A,B,C and D,respectively.There was significant difference between group D and group B(q= 4.398,P

OBJECTIVETo analyze the outcome and prognosis of autologous hematopoietic stem cell transplantation (AHSCT) combined with high dose radiotherapy/chemotherapy in 22 patients with nasal NK/T cell lymphoma.

METHODSFrom July 1992 to December 2005, 22 patients with nasal NK/T cell lymphoma were diagnosed pathologically. Immunophenotyping was performed in 13 cases. The patients were classified by Ann Arbor staging system and international prognosis index (IPI). The patients received cycles of chemotherapy every other two weeks or combined with radiotherapy for remission induction, followed high dose radiotherapy/chemotherapy, combined with autologous peripheral blood stem cell transplantation (APBSCT), or autologous bone-marrow transplantation (ABMT). Patients were given complementary radiotherapy after transplantation if they did not have it before. Twelve patients of IPI 3 -4 received consolidation chemotherapy, and one of them received the second transplantation.

RESULTSThe median follow-up duration was 64 (12 - 168) months. The 5 and 8-year overall survivals (OS) were 79.3% and 64.1%, and disease free survivals (DFS) were 36.4% and 27.3%, respectively. The 5-year OS were as follows: for stage I - II and III - IV disease were 90.0% and 70.0% (P = 0. 041); for patients without and with B symptom were 100.0% and 70.7% (P = 0.045); and for IPI 1 - 2 and 3 - 4 were 100.0% and 60.0% (P = 0.035), respectively. Multivariate analysis by COX regression revealed that disease stage, B symptom and IPI were independent prognostic factors.

CONCLUSIONAHSCT combined with high dose radiotherapy/chemotherapy is an effective treatment for patients with poor prognosis nasal NK/T cell lymphoma.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphoma, Extranodal NK-T-Cell ; diagnosis ; radiotherapy ; therapy ; Male ; Middle Aged ; Nose Neoplasms ; diagnosis ; radiotherapy ; therapy ; Prognosis ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVEUnderstand the public's current attitudes and knowledge about suicide and, thus, provide essential information to the development of targeted public education programs-important components of the suicide prevention effort.

METHODSSeventeen mental health professionals who were extensively trained in the methods of conducting focus groups used a pre-tested focus group outline on attitudes and knowledge about suicide to conduct 101 focus groups and 18 individual in-depth interviews with a total of 842 community respondents from 6 regions in northern China. The focus groups and in-depth interviews were audio-taped, transcribed and analyzed using the QSR Nvivo text analysis software.

RESULTSMost respondents believed that suicide was a greater problem in rural areas and among women and identified physical illnesses, economic problems and interpersonal conflicts (particularly family conflicts) as the main causes of suicide. Rural residents and women were believed to exhibit impulsive suicidal behavior because of their personal limitations and over-sensitiveness. Most thought that suicide was understandable and a small proportion felt that it was acceptable behavior in certain circumstances. Almost all felt that suicide resulted in the stigmatization and a loss of 'face' for the family. Most believed that one should show concern for persons who have suicidal behavior and their family members and expressed a willingness to have superficial social relationships with them but were unwilling to establish close personal relations with them. The vast majority believed that suicide was either very difficult or impossible to prevent.

CONCLUSIONSIn China the community is tolerant, sympathetic and, in some cases, accepting of suicide but there remains a substantial underlying stigmatization of suicide. Community members have some misunderstandings about suicide; the most obvious misunderstanding is the underestimation of the importance of mental illness as a cause of suicide. The content of public health messages used in suicide prevention programs should be developed by combining findings from both qualitative and quantitative research.

China ; Health Knowledge, Attitudes, Practice ; Humans ; Social Support ; Suicide ; prevention & control ; psychology ; Surveys and Questionnaires

Type B Niemann-Pick disease is an autosomal recessive sphingolipidosis due to mutations in the sphingomyelin phosphodiesterase 1 gene (SMPD1). Here we present molecular findings for two sibling patients. One mutation V36A due to c.107T>C in exon 1 is a single nucleotide polymorphism and the other N522S due to c.1565 A>G in exon 6 is a novel missense mutation. This non-fatal missense mutation leads to –20% residual lysosomal acid sphingomyelinase activity in vitro and only results in hepatosplenomegaly without neurologic involvement.
Female ; Humans ; Middle Aged ; Mutation, Missense ; Niemann-Pick Disease, Type B ; genetics ; Polymorphism, Single Nucleotide ; Siblings ; Sphingomyelin Phosphodiesterase ; genetics