Objective:A study was conducted to investigate the effect of Prunella vulgaris sulfated polysaccharide (PVSP) on the expression of angiogenic growth factors (bFGF, VEGF, and IL-8) and angiogenesis in hepatocellular carcinoma. Methods:ELISA as-say was used to observe the effects of PVSP and the negative control drug Lentinan (LNT, non-sulfate radical drug) on secretions of the angiogenic growth factors, namely, bFGF, VEGF, and IL-8, in HepG2 cells in vitro. In an in vivo experiment, the microvessel density in hepatocellular carcinoma tissue sections treated with PVSP and LNT was calculated, analyzed, and compared with the microvessel den-sity in the phosphate-buffered saline (PBS) control. Results:Compared with the PBS control group, PVSP at 200μg/mL inhibited bF-GF secretion (P<0.01), whereas LNT failed to affect bFGF secretion. Neither PVSP nor LNT affected the secretions of VEGF and IL-8. In vivo results showed that PVSP at 200 mg/kg reduced the microvessel density in tumor tissue sections (P=0.03), whereas LNT at 10 mg/kg failed to affect microvessel density. Conclusion:Inhibition of bFGF secretion is a probable mechanism underlying the preven-tive effect of PVSP on hepatocellular carcinoma angiogenesis.

Objective To investigate the action and mechanism of NF-κB pathway in up-regulating B cell-activating factor receptor (BAFF-R) expression in multiple myeloma cells induced by IFN-γ.Methods Activated NF-κB were detected with Western blot, while the expression of BAFF-R were measured with RT-PCR and ELISA, and investigated the effect of BAY11-7082 on transcription of BAFF-R mRNA and translation of protein in multiple myeloma cells stimulated by IFN-γ. Results IFN-γ can induce the degradation of IκB-α in time-dependent and dosage-dependent manner, and up-regulated BAFF-R expression in multiple myeloma cells. BAY11-7082, an NF-κB inhibitor, inhibited not only the transcription of BAFF-R mRNA but also the protein of regulated by IFN-γin dosage-dependent manner. Conclusion NFκB may play an important role in high expression of BAFF-R in multiple myeloma cells induced by IFN-γ.

Objective To determine the antigen-specific CTL in PBMC induced by a fusional family-gene vaccine of the immunoglobulin heavy chain variable gene framework region combined with the sequence of cytokine CM-CSF in vitro with MHC pentamer. Methods Peripheral blood samples were collected from two healthy donors and two patients. One was follicular lymphoma and another was hair cell leukemia. PBMC were isolated by density gradient centrifugalization with Ficoll and then subsequently differentiated into immature DCs (imDCs) induced by recombinant human GM-CSF and recombinant human IL-4. Gene gun was used to deliver the plasmids of the gene vaccine or the control plasmids into the imDCs. RT-PCR and ELISA assay were used to detect IgHVl-GM-CSF mRNA and GM-CSF in order to validate the transfection of the vaccine. After adding the cytokine cocktail, the imDCs became mature DCs. Then the mature DCs were co-cultured with lymphocytes from the blood samples for the induction of the antigen-specific CTL. The cultured cells were classified into vaccine group and control group and harvested at different time points of 0 d,7d, 17 d and 24 d after transfection. The subset of CD3+CD8+ T cells was analyzed by FCM assay. Finally, the CTL levels were detected with fluorescently labeled MHC pentamer antibody targeting vaccine epitopes. Results With the induction of cytokines, the imDCs with typical morphology were generated in PBMC. After delivering, the efficient expressions of the vaccine in the imDCs were determined by RT-PCR. And ELISA results also confirmed that GM-CSF was produced at a level of (28 ±6) ng/106 cells of the imDCs loaded with the vaccine, which was significantly different from that of control group (10 ± 3) ng/106 cells (t = 5. 191, P <0.01). FCM assay result showed that the CD3+ CD8+ T cells increased in a stepwise pattern during the culture. For control group, the levels at 0 d,7d, 17d and 24 d were ( 34. 24 ± 2. 72 )% , (46.06 ± 3.08)%, ( 65. 34 ± 4. 26 )% and (73.86 ±4.85 )% , respectively. For vaccine group, the results were (32. 28 ± 2. 08 ) % , (45. 32 ± 3. 81)% , ( 63. 37 ± 4. 21)% and (75. 01 ±3. 20)%. The differences between each time point had statistical significance (F = 176. 966 ,P <0.01) ,but there was no statistical differences between vaccine group and control group ( F = 0.657,P>0.05). The MHC pentamer analysis showed that the DCs loaded with IgHV1-GM-CSF fusional vaccine could efficiently induce the antigen-specific CTL response and the CTL levels increased gradually with the culture time, with the highest level of 4. 36% in the lymphoma blood and 3. 89% in the hair cell leukemia blood. Conclusions MHC pentamer assay could efficiently determine the antigen-specific CTLs response induced by the gene vaccine of family IgHV frame region in vitro. It could be a useful method for monitoring of anti-tumor cell immunity and evaluating of diagnosis and prognosis of the tumors in clinical application.

Morphology and molecular identification technology were used to identify 3 original plants of Fructus Elaeagni which was commonly used in Uygur medicine. Leaves, flowers and fruits from different areas were selected randomly for morphology research. ITS2 sequence as DNA barcode was used to identify 17 samples of Fructus Elaeagni. The genetic distances were computed by kimura 2-parameter (K2P) model, and the Neighbor-Joining (NJ) and Maximum Likelihood phylogenetic trees were constructed using MEGA5.0. The results showed that Elaeagnus angustifolia, E. oxycarpa and E. angustifolia var. orientalis cannot be distinguished by morphological characteristics of leaves, flowers and fruits. The sequence length of ITS2 ranged from 220 to 223 bp, the average GC content was 61.9%. The haplotype numbers of E. angustifolia, E. oxycarpa and E. angustifolia var. orientals were 4, 3, 3, respectively. The results from the NJ tree and ML tree showed that the 3 original species of Fructus Elaeagni cannot be distinguished obviously. Therefore, 3 species maybe have the same origin, and can be used as the original plant of Uygur medicineal material Fructus Elaeagni. However, further evidence of chemical components and pharmacological effect were needed.
DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Elaeagnaceae ; anatomy & histology ; classification ; genetics ; Fruit ; anatomy & histology ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Quality Control

Objective To explore the relationship between autism spectrum disorder (ASD) and gastrointestinal problems. Methods 172 ASD children were investigated with self-made questionnaire from July, 2014 to Augest, 2015. The frequencies of motor behaviors, vocal behaviors and behavioral problems, and the intensity of the behavioral problems in the period with or without gastrointestinal symptoms were compared. Results The frequencies of motor behaviors (F=67.4, P<0.001), vocal behaviors (F=43.6, P<0.001) and behavioral prob-lems (F=79.4, P<0.001), and the intensity of the behavioral problems (F=65.7, P<0.001) were all more in the period with gastrointestinal symptoms than without gastrointestinal symptoms. Conclusion There are more behavioral problems in the ASD children with the presence of gastrointestinal symptoms;while the behavioral symptoms may be a indicator of gastrointestinal problems in children with ASD.

Objective To explore cognition level on nuclear and radiation in Beijing high school students,which may provide evidence for promoting science popularization on nuclear and radiation.Methods Questionnaire-based survey was conducted in Beijing high school students,randomized cluster sampling was used to recruit study participants.Demographic information was collected,and cognition level on nuclear and radiation was evaluated by questionnaire.Results A total of 1 029 pieces of eligible questionnaires were collected.The correct rate for answering common sense about nuclear and radiation was 58％,with score of boys significantly higher than that of girls (t =4.131,P ＜0.05).About subjective cognition of nuclear and radiation knowledge,87 (8.5％) indicated ' quite clear',779 (75.7％) indicated 'know a little',163 (15.8％) indicated 'know nothing'.There was significant difference in score of common sense about nuclear and radiation among people with various subjective cognition level of nuclear and radiation (J-T =8.279,P ＜0.05).There was a linear correlation between support degree for nuclear power and subjective cognition level of nuclear and radiation (r =0.161,P ＜0.05).There was significant difference in score of common sense about nuclear and radiation among people with various support degree for nuclear power (J-T =7.508,P ＜0.05),whereas those who had got high scores tended to support nuclear power to a higher degree.Conclutions Students knew little about knowledge on nuclear and radiation.It is necessary to strengthen propaganda and education on nuclear and radiation,which may help enhance the students' comprehensive quality,and sustainable expansion of nuclear power more support in the long run.

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.

Objective To investigate the association of chromosome 8p copy number alteration (CNA) with postoperative survival of patients with hepatocellular carcinoma (HCC),and to screen for possible target genes in the survival-related CNA (s) in 8p.Methods 187 HCC patients were enrolled into the study,which included 66 patients whose follow-up data were available and the follow-up was 2.6 ～ 73.3 months.High-resolution Agilent 244K comparative genomic hybridization (CGH) and Affymetrix U133 Plus2.0 expression arrays were used to screen for CNAs and gene expression differences in 8p.The associations between CNAs in 8p and survival were analyzed using the log-rank test,Kaplan-Meier survival analysis and Cox proportional hazards models.The gene expression levels between the groups were compared by the Mann-Whitney U test.Results Copy number loss on 8p12 (31/66,47％) was significantly associated with reduced survival rate,and HCC patients with 8p12 loss had a 4.1-fold (95％ CI =1.8 ～ 9.4,P ＜ 0.05) increased hazard ratio (HR) for death from HCC,as compared to those without the loss.The mRNA expression levels of the 3 genes in 8p12,including TMEM66,DCTN6,and MAK16,were significantly decreased in HCCs with gene loss than in HCCs without the loss (all P ＜ 0.05),and in non-tumorous liver tissues (all P ＜ 0.05).Conclusion Loss of 8p12 is an independent prognostic marker of unfavorable survival for patients with HCC,and underexpression of genes TMEM66,DCTN6,and MAK16,owing to 8p12 loss,contributed to unfavorable prognosis.

Objective To explore the differences of the joint function and the patient's satisfaction between two different methods to reconstruct limb length for unilateral Crowe Ⅳ developmental dysplasia of the hip (DDH).Methods The clinical controlled study were used.21 cases with unilateral DDH Crowe Ⅳ treated with total hip arthroplasty were divided into 2 groups randomly.The patients in the first group were reconstructed the limb with the compensatory length and those in the second group were did with the real length.The patients were followed up 10 years.Two independent sample t-test was used to compare these two operations.Harris scores and SF-36 scores were used in the test.Survival analysis with Kaplan-Meier method was involved in the survival of the prosthesis and then test with Log-rank test.X-ray films of different period were compared to confirm the prosthetic loosening.Results Seventeen patients were followed up 8-10 years.There were no significant differences both in Harris score and most of the SF-36 scores at the median follow-up of 10 years.The item mental healthy of SF-36 expressed significant difference.Five patients required revision.There were 10 cases suffered with polyethylene wear,6 cases with severe osteoporosis in greater trochanter,and 3 cases with osteolytic reaction.Conclusion There are no significant differences between the two surgical method in the survival rate of prosthesis and the joint function in the median follow up.The satisfaction of the control groups is lower than the trial groups both at the early stage after the operation and the median follow up.The revision rate between the two groups is similar but the reason is different.

This study is to evaluate the effects of the metformin (Met) on β cell function of diabetic KKAy mice. Female diabetic KKAy mice selected by insulin tolerance test (ITT) were divided randomly into two groups. Con group was orally administered by gavage with water, Met group with metformin hydrochloride at a dose of 0.2 g x kg(-1) for about 12 weeks. ITT and glucose tolerance tests (OGTT) were determined. Beta cell function was assessed by hyperglycemic clamp. Pancreatic biochemical indicators were tested. The changes of gene and protein expression in the pancreas and islets were also analyzed by Real-Time-PCR and immunostaining. Met significantly improved glucose intolerance and insulin resistance in KKAy mice. Fasting plasma glucose and insulin levels were also decreased. In addition, Met markedly increased glucose infusion rate (GIR) and elevated the Ist phase and maximum insulin secretion during clamp. It showed that Met decreased TG content and iNOS activities and increased Ca(2+) -Mg(2+)-ATPase activity in pancreas. Islets periphery was improved, and down-regulation of glucagon and up-regulated insulin protein expressions were found after Met treatment. Pancreatic mRNA expressions of inflammation factors including TLR4, NF-κB, JNK, IL-6 and TNF-α were down-regulated, p-NF-κB p65 protein levels also down-regulated by Met. And mRNA expressions of ion homeostasis involved in insulin secretion including SERCA2 and Kir6.2 were up-regulated by Met. Met increased SIRT5 expression level in pancreas of KKAy mice under the hyperglycemic clamp. These results indicated that chronic administration of Met regulated pancreatic inflammation generation, ion and hormone homeostasis and improved β cell function of diabetic KKAy mice.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; drug therapy ; Down-Regulation ; Female ; Glucose Tolerance Test ; Homeostasis ; Inflammation ; drug therapy ; Insulin ; secretion ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Interleukin-6 ; metabolism ; Metformin ; pharmacology ; Mice ; NF-kappa B ; metabolism ; Pancreas ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism