Objective To explore the effect of Chinese herbal medicine formula "tonifying kidney plus strengthening vital energy" on regulating CD40, CD40L protein and mRNA expression in spleen cell in mice with SHENXU. Methods The interaction of CD40L expressed by activated T cells with CD40 on monocytes, macrophages, or B cells is essential for the production of IL-12 and other cytokines. An experimental model of mice with SHENXU by corticosterone was established to observe the effect of Chinese formula on CD40, CD40L protein and mRNA expression in spleen cells or in ConA stimulated spleen cells by immunohistochemistry and RT-PCR. Results No significant differences were found among 5 groups of mice about the CD40 and CD40L protein expression in spleen cells. Both CD40 and CD40L expression detected by immunohistochemistry decreased seriously in ConA induced cells from SHENXU mice (18.9%?3.3% vs 15.8%?2.3%) as compared to that of normal controls(29.7%?4.2% vs 23.3%?1.3%), and so did the mRNA expression on ConA stimulated cells from SHENXU mice. They were restored in different degrees when the SHENXU mice were cured by different Chinese medicine. The effect of "tonifying kidney plus strengthening vital energy" was stronger than that of "tonifying kidney" and "strengthening vital energy" alone. Conclusions "Tonifying kidney plus strengthening vital energy" enhanced the expression of CD40, CD40L mRNA and protein, and this may be one of the mechanisms of improving the status of immune inhibition resulted by Corticosterone.

BACKGROUND:Inducing factor and chondrogenic microenvironment is a primary factor, which influences chondrogenic differentiation and chondrogenesis of bone marrow-derived mesenchymal stem cells (MSCs). OBJECTIVE:To explore the feasibility of in vivo chondrogenesis by co-culture of bone marrow-derived MSCs and chondrocytes. DESIGN, TIME AND SETTING:A randomized controlled animal experiment was performed at Department of Pathology, Stomatological Hospital, Fourth Military Medical University of Chinese PLA between September 2004 and March 2005. MATERIALS:Fifteen New Zealand rabbits of clean grade were used for cell-scaffold construct transplantation. The rabbits were randomly divided into co-culture, chondrocyte, and bone marrow-derived MSC groups, with 5 rabbits in each group. Five neonatal New Zealand rabbits, aged 1-3 days, were used for isolation and culture of bone marrow-derived MSCs and chondrocytes. Polyglycolic acid (PGA) scaffold material (Shanghai Yikuo Company, China) has a fiber diameter of 15 μm, with an average interval of 150-200 μm, an interval porosity of 97% and 2-mm thickness. METHODS:In the co-culture group, bone marrow-derived MSCs and chondrocytes were mixed at a ratio of 3:1. The mixed cells were seeded onto a pre-wetted PGA scaffold (5 mm×5 mm )at the ultimate concentration of 6.0×1010 L-1. Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum was dropwise added to peripheral compound for 1 week of culture. In the chondrocyte, and bone marrow-derived MSC groups, chondrocytes and bone marrow-derived MSCs of the same ultimate concentration were seeded respectively onto the PGA scaffold. Then, the cell-scaffold constructs were transplanted into subcutaneous tissue of adult rabbits. MAIN OUTCOME MEASURES:Gross observation and hematoxylin-eosin & Masson staining of neo-cartilage were performed after in vivo culture for 8 weeks. RESULTS:Cell in all groups had a fine adhesion to the scaffold. In both co-culture and chondrocyte groups, the cell-scaffold constructs could maintain the original size and shape during in vivo culture and formed homogenous mature cartilage after 8 weeks of in vivo culture. Furthermore, the neo-cartilages in both groups were similar to each other in gross appearance and histological features. In the bone marrow-derived MSCs group, connective tissue rather than cartilage was found during in vivo culture. CONCLUSION:Chondrocytes can provide a chondrogenic microenvironment to induce a chondrogenic differentiation of bone marrow-derived MSCs and thus promote the chondrogenesis of bone marrow-derived MSCs in vivo.

Objective To examine the expression of follistatin-like protein 1 (FSTL1) in the peripheral blood and intestinal mucosa tissues of ulcerative colitis (UC) patients and analyze the correlation between its expression and the activity of UC.Methods From October 2010 to June 2012,sixty patients with UC were collected.From April 2012 to October 2012,thirty individuals without any obvious mucosa lesion under colonoscope and confirmed by pathological examination were set as control group.The serum expression level of FSTL1 of both UC group and control group were determined by enzyme linked immunosorbent assay (ELISA).t-test was performed for comparison between groups.The expression of FSTL1 in the intestinal mucosa of UC group and control group was detected by immunohistochemistry.Chi-square test was used for comparison between groups.The patients with UC were scored with ulcerative colitis disease activity index (UCDAI).Its correlation with plasma FSTL1 was analyzed by Pearson correlation coefficient.Results The serum expression level of FSTL1 of UC group ((14.37-±-1.80) μg/L) was higher than that of control group ((5.80±0.72) μg/L)and the difference was statistically significant (t=25.01,P＜ 0.05).The serum expression level of FSTL1 of UC group was positively correlated with UCDAI (r=0.814,P＜0.05).The positive expression rate of FSTL1 in the intestinal mucosa tissues of UC group (86.7％,52/60)was higher than that of control group (46.7％,14/30) and the difference was statistically significant (x2 =52.334,P＜0.05).Conclusions The expression of FSTL1 of UC patients increases and is positively correlated with disease activity.FSTL1 may play a role in the development of UC.

Objective To study the association of single nucleotide polymorphisms (SNPs) of solute carrier family 22 member 4 (SLC22A4) and SLC22A5 genes with Crhon's disease (CD) in Chinese Han nationality. Methods SNPs in the entire coding region of SLC22A4 and SLC22A5 genes were screened by direct DNA sequencing in 80 CD patients and 80 healthy subjects, and statistical in Han population. Five SNPs were found in entire coding region (2 in SLC22A4 gene and 3 in distribution of the alleles and genotypes of SLC22A4 and SLC22A5 polymorphisms between CD patients and healthy controls. Conclusion There is no correlation of SLC22A4 and SLC22A5 with CD in Chinese Han nationality.

OBJECTIVETo obtain the peptide that specifically binds to human osteosarcoma MG-63 cells from Ph. D. 7TM phage display peptide library.

METHODSHuman osteosarcoma MG-63 cells were used as the target cells with human embryonic kidney 293T cells as the control for screening the peptide from Ph. D. 7TM phage display peptide library. The enriched specially binding peptides were verified by cell enzyme-linked immunosorbent assay (ELISA). The location of the peptide in MG-63 cells was investigated using cell fluorescence staining, and targeting of the peptide was tested by organ immunohistochemistry with Osteosarcoma model.

RESULTSThe specifically binding peptides were enriched after 4 rounds of screening. The sequence SLTNLSK was confirmed as the most frequent peptide by DNA sequencing and showed strong specificity verified by cell ELISA, fluorescent staining and organ immunohistochemistry.

CONCLUSIONA peptide that specifically binds to MG-63 cells has been screened from Ph. D. 7TM phage display peptide library to serve as a potential candidate for osteosarcoma-targeting therapy.

Amino Acid Sequence ; Animals ; Bone Neoplasms ; drug therapy ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Osteosarcoma ; drug therapy ; Peptide Library ; Peptides ; analysis ; Protein Binding

In order to provide experimental data for the development and application of drug in clinic, we determined the effects of extracts from different parts of folium perillae (L. ) Britt. on hemorheological parameters, extracted from leaves (folium perillae), seeds (fructus perillae) and peduncles (caulis perillae). The results showed that all extracts from different parts of folium perillae (L. ) Britt. can significantly reduce the whole blood viscosity at low shear rate (10 s(-1)), erythrocyte aggregation index, erythrocyte electrophoresis index (P<0.05), and the whole blood reductive viscosity at low shear rate (10 s(-1)) (P<0.01). Extracts from folium perillae and caulis perillae can significantly decrease erythrocyte deformation index (P<0.05), whereas extracts from fructus perillae can not. Extracts from fructus perillae and caulis perillae can significantly decrease plasma viscosity at low shear rate(10 s(-1)), but extracts of folium perillae can not. Aspirin can only decrease the whole blood reductive viscosity at low shear rate and plasma viscosity (P<0.05). All extracts from different parts of folium perillae (L. ) Britt. had no significant effects on hematocrit, erythrocyte rigidity index, fibrinogen concentration , the whole blood viscosity and the whole blood reductive viscosity at middle and high shear rate (60 s(-1),120 s(-1)).
Animals ; Blood Viscosity ; drug effects ; Erythrocyte Aggregation ; drug effects ; Erythrocyte Deformability ; drug effects ; Male ; Perilla frutescens ; anatomy & histology ; chemistry ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Seeds ; chemistry

To investigate the postoperative serum triglyceride (TG) levels in predicting the risk of new-onset diabetes mellitus (NODM) in patients following allogeneic liver transplantation. One hundred and forty three patients undergoing allogeneic liver transplantation in Shanghai General Hospital from July 2007 to July 2014 were enrolled in this study. The NODM developed in 33 patients after liver transplantation. The curve of dynamic TG levels in the early period after liver transplantation was generated. Independent risk factors of NODM were determined by univariate and multivariant logistic regression analyses. The clinical value of TG in predicting NODM was analyzed by area under the ROC curve (AUC). Serum TG levels were gradually rising in the first week and then reached the plateau phase (stable TG, sTG) in patients after surgery. The sTG in NODM group were significantly higher than that in non-NODM group (=-2.31, <0.05). Glucocorticoid therapy (=4.054, <0.01), FK506 drug concentration in the first week after operation (=3.482, <0.05) and sTG (=3.156, <0.05) were independent risk factors of NODM. ROC curve analysis showed that the AUC of sTG in predicting NODM was 0.72. TG shows a gradual recovery process in the early period after liver transplantation, and the higher TG level in stable phase may significantly increase the risk of NODM in patients.
China/epidemiology* ; Diabetes Mellitus/etiology* ; Humans ; Liver Transplantation/adverse effects* ; Risk Factors ; Tacrolimus/adverse effects* ; Triglycerides

Objective To understand the prevalence of thyroid nodules of children lived in different water iodine areas in Cangzhou City. Methods From Oct. 2015 to Jan. 2017, 15 villages were selected as monitoring sites in Cangzhou,two drinking water samples were collected from each survey site(all had centralized water supply), and the water iodine content was determined. A total of 100 children aged 8 to 10 (half male and female) were examined for thyroid nodules, and at least 50 children (half male and half female) were selected to detect urinary iodine content. In the high iodine water counties, the monitoring sites of iodine salt was according to "National Iodine Deficiency Monitoring Program"; in the monitoring sites of iodine salt supplied counties, students in the monitored village were asked to detect urinary iodine and household salt samples were collected to monitor salt iodine. In the high iodine area, the salt iodine test was carried out by semi-quantitative method. In the non-high iodine area, the salt iodine content of the iodized salt monitoring sites was determined by direct titration, the salt iodine content of Chuan salt and other intensified edible salt was tested by arbitration(GB/T 13025.7-2012). Water iodine and urinary iodine were tested by arsenic and cerium catalytic spectrophotometry. Results Water iodine content was 28.2 - 1 128.0 μg/L in 15 villages; a total of 1 066 urine samples were examined, the median of uriary iodine in each village was 102.6-1 162.0 μg/L;a total of 1 575 children aged 8 to 10 years were examined,among them,125 cases of thyroid nodules were detected; thyroid nodules detection rate was 7.9%. The prevalence of male was 7.0% (61/871), and the prevalence of female was 9.1% (64/704), there was no significant difference in the detection rate of thyroid nodules between different sex (χ2=2.07,P>0.05); The detection rate of thyroid nodules were 4.5%(23/508),7.8%(4/51), 11.6%(59/507)in children with urinary iodine at the appropriate level (100 - <200 μg/L), the appropriate level (200 - < 300 μg/L) and iodine excess level (≥300 μg/L), the difference of thyroid nodules in children with different levels of urinary iodine detection rate was statistically significant (χ2=17.30, P < 0.01). The difference of prevalence of thyroid nodules in children aged 8 to 10 years with water iodine concentrations of 10 - < 100, 100 - < 300 and ≥300 μg/L was statistically significant[2.9%(13/448),7.9%(25/317), 10.7%(87/810),χ2=23.86,P<0.05].The patients with unilateral thyroid nodule accounted for 64.8% (81/125); the patients with multiple thyroid nodules counted for 58.4% (73/125), and 34.2%(13/38),69.0%(60/87)in areas with iodine content less than 300 μg/L and no less than 300 μg/L,the difference between the two was statistically significant (χ2= 13.14, P < 0.01). A total of 1 800 salt samples were collected from the high water iodine counties,of which 1 779 were iodine-free salt, the rate of iodine-free salt was 98.8%; a total of 190 salt samples were collected in student family, in the 4 iodized salt monitoring sites, the salt iodine median of resident's edible salt was 0.0 mg/kg. Conclusion The prevalence of thyroid nodules in children aged 8 - 10 years may be related to high water iodine in Cangzhou City; children with multiple thyroid nodules is also significantly higher in water iodine content greater than 300 μg/L areas.

Objective:To investigate the effect of myogenic differentiation protein 1(Myod1)on the proliferation inhibition and apoptosis of the SH-SY5Y cells induced by oxygen-glucose deprivation(OGD),and to elucidate its mechanism.Methods:Real-time quantitative fluorescence PCR(RT-qPCR)method was used to detect the mRNA levels of Myod1 and long non-coding RNA(lncRNA)small nucleolar RNA host gene 15(SNHG15)in peripheral blood of the subjects in normal group and the patients in ischemic cerebral infarction group as well as the normal cultured SH-SY5Y cells(control group)and the cells in OGD model(OGD group).After transfecting SH-SY5Y cells with si-Myod1,pcDNA3.0-Myod1,si-SNHG15,pcDNA3.0-SNHG15、si-NC,Vector,miR-NC,and miR-24-3p mimics,the cells were treated with OGD,and then the SH-SY5Y cells were divided into control group,OGD group,OGD+Vector group,OGD+Myod1 group,OGD+si-NC group,OGD+si-Myod1 group,OGD+si-SNHG15 group,OGD+si-SNHG15+Vector group,OGD+si-SNHG15+Myod1 group,OGD+miR-NC group,OGD+miR-mimics group,OGD+miR-mimics+Vector group,and OGD+miR-mimics+SNHG15 group.CCK-8 method was used to detect the cell activities in various groups;5-ethynyl-2'-deoxyuridine(EdU)staining was used to detect the rates of EDU positive cells in various groups;the rates of TdT-mediated dUTP nick end labeling(TUNEL)positive cells in various groups were detected by TUNEL staining;Western blotting method was used to detect the expression levels of cleaved caspase-3,cleaved caspase-9,B-cell lymphoma 2(Bcl-2)and Bcl-2 associated X protein(Bax)proteins in the cells in various groups;the association between Myod1 and SNHG15 was evaluated by chromatin immunoprecipitate(CHIP);dual luciferase reporter gene experiment was used to evaluate the targeting relationships between Myod1 and SNHG15 as well as SNHG15 and miR-24-3p.Results:Compared with normal control group,the expression levels of Myod1 and SNHG15 mRNA in peripheral blood of the patients in ischemic cerebral infarction group were significantly increased(P＜0.05).Compared with control group,the expression levels of Myod1 and SNHG15 mRNA in the SH-SY5Y cells in OGD group were significantly increased(P＜0.05).Compared with OGD group,the cell activities and rates of EdU positive cells in OGD+Myod1 group at 48 and 72 h were decreased(P＜0.01),and the rates of TUNEL positive cells were increased(P＜0.05);the cell activities and rates of EdU positive cells in OGD+si-Myod1 group were increased(P＜0.05),while the rates of TUNEL positive cells were decreased(P＜0.01).Myod1 binded to the promoter sequence of SNHG15.SNHG15 could absorb miR-24-3p,and there were target relatronships between Myod1 and SNHG15 as well as SNHG15 and miR-24-3p.After SNHG15 knockdown,compared with OGD group,the cell activities and rates of EdU positive cells in OGD+si-SNHG15 group at 48 and 72 h were increased(P＜0.01),and the rates of TUNEL positive cells were decreased(P＜0.01),the expression levels of Bax,cleaved caspase-3 and cleaved caspase-9 proteins were decreased(P＜0.01),and the expression levels of Bcl-2 protein were increased(P＜0.01).Compared with OGD+si-SNHG15 group,the cell activities and rates of EdU positive cells in OGD+si-SNHG15+Myod1 group at 48 and 72 h were decreased(P＜0.05),the rates of TUNEL positive cells were(P＜0.05),the expression levels of Bax,cleaved caspase-3,and cleaved caspase-9 proteins were increased(P＜0.05),and the expression levels of Bcl-2 were decreased(P＜0.05).After over-expression of miR-24-3p and SNHG15,compared with OGD group,the cell activities and rates of EdU positive cells in OGD+miR-mimics group at 48 and 72 h were increased(P＜0.01),the rates of TUNEL positive cells were significantly decreased(P＜0.01),the protein expression levels of Bax,cleaved caspase-3 and cleaved caspase-9 were decreased(P＜0.05),and the expression levels of Bcl-2 were increased(P＜0.01).Compared with OGD+miR-mimics group,the cell activities and rates of EdU positive cells in OGD+miR-mimics+SNHG15 group at 48 and 72 h were decreased(P＜0.05),and the rates of TUNEL positive cells were increased(P＜0.05),the expression levels of Bax,cleaved caspase-3 and cleaved caspase-9 proteins were increased(P＜0.05),and the expression levels of Bcl-2 protein were decreased(P＜0.05).Conclusion:Myod1 can promote the proliferation inhibition and apoptosis of OGD-induced SH-SY5Y cells by binding to the SNHG15 promoter region and then absorbing miRNA-24.

Objective To investigate the effects of acute sleep deprivation on the behavior and synaptic protein expression of rats.Methods Seventy healthy male Wistar rats were randomly divided into seven groups,a Control group and sleep deprivation groups(24,48,72,96,120 and 144 hours).The sleep deprivation rat model was established by the modified multiplatform water environment sleep deprivation method.Spatial learning and memory were assessed by the Morris water maze.Anxiety was assessed by the open field test.The morphology and quantity of hippocampal neurons were observed by Nissl staining.Western blot and Real-time PCR were used to determine the expression of synaptophysin(SYN),post-synaptic density protein-95(PSD-95),and brain-derived neurotrophic factor(BDNF)in rats.Results Compared with the Control group,the numbers of standing and modification were significantly increased by prolongation of the sleep deprivation time(P＜0.05).The escape latency and path length were significantly increased in 120 and 144 h groups(P＜0.05),whereas the number of platform crossings and the percentage of the target quadrant time were significantly decreased(P＜0.01)and negatively correlated to the sleep deprivation time.The expression levels of BDNF,SYN,and PSD-95 were significantly decreased with the prolongation of sleep deprivation time(P＜0.01).Conclusions With the increase in sleep deprivation time,cognitive dysfunction and anxiety gradually deteriorated,which may be related to decreases in the expression of synaptic biomarkers.