OBJECTIVETo provide molecular evidences for its breeding by studying the genetic relationship among varieties of Rehmannia glutinosa.

METHODNineteen varieties were detected by Randomly Amplified Polymorphic DNA(RAPD) markers.

RESULTThe 20 selected primers produced 163 bands, among which 114(69.9%) were polymorphic. A DNA molecular dendrogram was established based on Hierarchical cluster analysis of 163 DNA bands amplified by 20 primers, which divided the 19 varieties into four groups: Group Beijing, Group 85-5, Group Guolimao and the other Group.

CONCLUSION8 varieties of Group Beijing have a close genetic relationship, and so have varieties of Group 85-5, which provides information for Rehmannia glutinosa's breeding.

DNA, Plant ; genetics ; Plant Leaves ; genetics ; Plants, Medicinal ; genetics ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; Rehmannia ; genetics

OBJECTIVEAnalyzing the relationships between peripheral blood CD4+ CD25hi regulatory T (Treg) cells and peripheral blood immune status or plasma HIV-lviral load in HIV-infected individuals,so as to determine whether Treg were related to the progression of HIV-infected disease.

METHODS116 HIV-infected patients in different stages and 21 healthy control individuals were included in this study. The CD4+ and CD8+ T cell counts were determined by a standard 4-color flow cytometry technique. The Treg cells were examined with 3-color immune staining flow cytometry. The plasma HIV-1 viral load was detected by real time PCR.

RESULTSThe frequencies of Treg cells decreased in HIV-infected individuals with high CD4+ T cell counts( > 300/microl) compared with normal controls. With the progression of disease the frequencies of Treg cells were raised gradually, until were increased in HIV-infected individuals with low levels of CD4+ T cell counts ( < 100/microl). In addition, the frequencies of Treg cells were inversely related to CD4+ T cell counts and CD4+ /CD8+ ratio, data showed a statistically significant (respectively, r = -0.564, P < 0.001; r = -0.377, P < 0.001). Furthermore, the proportions of Treg cells were closely related to plasma HIV-1 RNA viral load (r = 0.514, P < 0.001).

CONCLUSIONCD4 CD25hi Treg cells should be a kind of important cells participating the immunopathogenesis of AIDS. It may play different roles in different stages of HIV-infected disease. The exact mechanism of Treg cells in the progression of the HIV-infected disease needs to be investigate further.

Adult ; Case-Control Studies ; Cells, Cultured ; Disease Progression ; Female ; HIV Infections ; immunology ; pathology ; virology ; HIV-1 ; genetics ; immunology ; isolation & purification ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; Viral Load

BACKGROUND:Functional graded biomaterials promote the development of human hard tissue replacement.OBJECTIVE:To review the research progress of functional graded biomaterials in human hard tissue replacement.METHODS:The first author retrieved the PubMed and CNKI databases for relevant articles published from January 2010 to April 2017 using the keywords of "functional graded biomaterial,hard tissue replacement implants,preparation methods,performance evaluation,dental implants,osseous tissue" in English and Chinese,respectively.RESULTS AND CONCLUSION:Functional graded biomaterials refer to a kind of heterogeneous composite materials with controllable and programmable gradient properties on account of continuous or quasi-continuous changes in the structure and chemical composition.Hydroxyapatite is the primary choice for the material surface.Serving as an emerging biomaterial,the functional graded biomaterial has unique structure mechanism and excellent properties.It gives full play to the performance advantages of each component and reduces internal stress interface between components.Therefore,the functional graded biomaterial will be an issue of concern in the future because of the optimization of its design,preparation and performance.

No abstract available.

OBJECTIVE: To compare the iterative decomposition of water and fat with echo asymmetry and the least-squares estimation (IDEAL) method with a fat-saturated T2-weighted (T2W) fast recovery fast spin-echo (FRFSE) imaging of the spine. MATERIALS AND METHODS: Images acquired at 3.0 Tesla (T) in 35 patients with different spine lesions using fat-saturated T2W FRFSE imaging were compared with T2W IDEAL FRFSE images. Signal-to-noise ratio (SNR)-efficiencies measurements were made in the vertebral bodies and spinal cord in the mid-sagittal plane or nearest to the mid-sagittal plane. Images were scored with the consensus of two experienced radiologists on a four-point grading scale for fat suppression and overall image quality. Statistical analysis of SNR-efficiency, fat suppression and image quality scores was performed with a paired Student's t test and Wilcoxon's signed rank test. RESULTS: Signal-to-noise ratio-efficiency for both vertebral body and spinal cord was higher with T2W IDEAL FRFSE imaging (p < 0.05) than with T2W FRFSE imaging. T2W IDEAL FRFSE demonstrated superior fat suppression (p < 0.01) and image quality (p < 0.01) compared to fat-saturated T2W FRFSE. CONCLUSION: As compared with fat-saturated T2W FRFSE, IDEAL can provide a higher image quality, higher SNR-efficiency, and consistent, robust and uniform fat suppression. T2W IDEAL FRFSE is a promising technique for MR imaging of the spine at 3.0T.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Magnetic Resonance Imaging/*methods ; Male ; Middle Aged ; Signal-To-Noise Ratio ; Spinal Diseases/*diagnosis/pathology ; Statistics, Nonparametric

The objective of this study was to observe the expression of hoxc4 and hoxc6 genes in the process of differentiation of hematopoietic stem cell (HSC) to colony forming unit-T Lymphocyte (CFU-TL) in vitro. and to explore the possible mechanism of HCMV-induced maldevelopment of human cord blood CFU-TL on genetic level through effecting the differentiation progress by human cytomegalovirus (HCMV) with and/or all-trans retinoic acid (ATRA), Normal CFU-TL culture was used as blank control. After detection with MTT, mRNA expression levels in the human cord blood CFU-TL hoxc4 and hoxc6 genes following HCMV infection and ATRA treatment were detected by fluorogenic quantitative reserve transcription polymerize chain reaction (FQ-RT-PCR) method. HCMV of 10(6) plaque formation unit (PFU)/ml was diluted to 0.1 ml 10(5) PFU/ml and added into the infected group. The results showed that the expression of hoxc4 and hoxc6 genes in the differentiation process increased slightly on day 3, and were up to the most on day 7 (p < 0.05), while became lower on day 12 respectively in normal group, HCMV group and ATRA group. Compared with the expression of hoxc6, the expression of hoxc4 was obviously higher in each group (p < 0.05). Compared with the expression of hoxc4 and hoxc6 genes in normal group, the expressions of hoxc4 and hoxc6 in ATRA group were up-regulated remarkably (p < 0.05), while the expressions of hoxc4 and hoxc6 in group HCMV were down-regulated (p < 0.05). It is concluded that the regular expression of hoxc4 and hoxc6 genes mRNA appeared in each group. A positive co-relationship exits between hoxc4/hoxc6 genes and lymphocytic progenitor hematopoiesis. Compared with the expression of hoxc6 gene, the expression of hoxc4 gene is obviously higher in each group. HCMV can down-regulate the expression of hoxc4 and hoxc6 genes and lead to suppression effect on cell morphology, which confirms that the normal hematopoietic lineage determination and maturation rely on the stable and consistent expression of homeobox gene. At the same condition, ATRA (6 x 10(-8) mol/L at 60 nmol/ml) can up-regulate hoxc4 and hoxc6 genes expression. ATRA can up-regulate the expression of hoxc4 and hoxc6 genes.
Cell Line ; Cell Proliferation ; Cytomegalovirus ; genetics ; Cytomegalovirus Infections ; genetics ; Homeodomain Proteins ; genetics ; Humans ; Lymphoid Progenitor Cells ; cytology ; Tretinoin ; pharmacology

Objectives To identify ABO blood type from a child having discrepant results in forward and reverse ABO blood grouping by serological identification and genetic testing.Methods After routine serological detection with ABO blood group,the ABO gene and ABO blood group-A subgroup genotype were tested by PCR-SSP method.Results The Serological results showed that the specimen was positive type A (anti-A:1 +w),the reverse type is O type (Ac:2 +,Bc:4 +);PCRSSP A-subtype typing showed that the genotype of the child was Ax14/O2.Conclusion Difficult blood type identification sometimes need to combine serology and molecular biology methods to confirm.In this case,the phenotype of the child was Ax,and the genotype was Ax14/O2.

OBJECTIVETo investigate the effect of hypoxia on the expression of matrix metalloproteinase (MMP) and tissue inhibitors of matrix metalloproteinase (TIMP) in human periodontal ligament fibroblasts (HPDLF).

METHODSHPDLF were cultured in α-minima essential medium (α-MEM) and subcultured at confluence. In the hypoxic groups, cells were incubated in a humidified atmosphere of 1%O(2), 5%CO(2), 94%N(2) at 37°C for 12, 24 and 48 h, respectively. In the normoxic control group, cells were incubated under normoxic conditions of 20%O(2), 5%CO(2), 75%N(2). The mRNA expression of MMP and TIMP was measured using reverse transcription-polymerase chain reaction (RT-PCR). The data was analyzed by Student's t test, one-way ANOVA and LSD test with SPSS 13.0 software package.

RESULTSThe expression of MMP-2, TIMP-1 and TIMP-2 mRNA in the hypoxia groups was higher than that in control. The expression of MMP-2 mRNA in hypoxic groups showed a significantly increasing trend. There was significant difference between the hypoxic group and the normoxic control group in the expression of MMP-2 mRNA in HPDLF (P < 0.01). The expression of TIMP-1, TIMP-2 mRNA in hypoxic groups of 12 h was momentarily increased. There was significant difference between the hypoxic 12 h group and the normoxic control group in the expression of TIMP-1, TIMP-2 mRNA in HPDLF (P < 0.05). However, with prolonged hypoxia time, the expression of TIMP-1, TIMP-2 mRNA in hypoxic groups showed a significantly declining trend, there were significant differences between the hypoxic 12, 24 and 48 h group and the normoxic control group in the expression of TIMP-2 mRNA in HPDLF (P < 0.05). The expression of MMP-1 mRNA in hypoxic groups of 12 h was momentarily decreased and then increased after 24 h of hypoxia. There were significant differences between the hypoxic 48 h group and the normoxic control group in the expression of MMP-1 mRNA in HPDLF (P < 0.05). There were significant differences between the hypoxic 12 h group and the normoxic control group in the ratio of MMP-1/TIMP-1 mRNA (P < 0.05). The ratio of MMP-2/TIMP-2 mRNA in the hypoxia group significantly increased compared with normoxic group. There were significant differences between the hypoxic group and the normoxic control group in the ratio of MMP-2/TIMP-2 mRNA (P < 0.05).

CONCLUSIONSHypoxia could change the expression of MMP and TIMP mRNA and other relevant growth factors and also lead to the imbalance of MMP-2/TIMP-2 mRNA expression. It is suggested that the imbalance of MMP-2/TIMP-2 expression may be closely correlated with the occurrence and development of periodontal disease and play an important role in the process of periodontal tissue destruction in periodontitis.

Adolescent ; Cell Hypoxia ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; Humans ; Matrix Metalloproteinase 1 ; genetics ; metabolism ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Periodontal Ligament ; cytology ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; metabolism

OBJECTIVETo clarify the association of EGFR expression with angiogenesis and chemoresistance in ovarian cancer.

METHODSImmunohistochemical PV-6000 staining was used to detect the expression of EGFR, LRP protein and MVD in 102 ovarian tumor specimens.

RESULTSEGFR, LRP positive rates and MVD in borderline and malignant ovarian specimens were significantly higher than those in the normal and benign ones (P < 0.01). EGFR positive expression rate in stage III-IV carcinoma tissues, poor differentiation and with ascites was higher than that in stage I-II carcinomas of well differentiation and without ascites (P < 0.05). MVD was related to histological grade, residual tumor and ascites, LRP positive expression had no correlation with the clinicopathologic parameters (P > 0.05). The effective rate of chemotherapy in patients with EGFR and LRP-positive expression were 57.1% and 53.7%, respectively, significantly lower than that in cases with EGFR and LRP-negative expression (85.0% and 90.9%, P < 0.05). In the 64 cases with complete data, the three-year survival rate was 53.0%. The survival time was shorter in the cases with EGFR and LRP-positive expression, poor differentiation, ascites and chemoresistance (P < 0.01), and only LRP-positive expression and chemotherapeutic effect were independently related to survival time (P < 0.05). There was a correlation between EGFR and MVD (r = 0.548, P < 0.01), EGFR and LRP positive expression (P = 0.020).

CONCLUSIONThe expression of EGFR in ovarian cancer is related to angiogenesis and chemoresistance. EGFR and LRP-positive expression are related to chemoresistance, and detection of the two proteins may be helpful in guiding chemotherapy choice for ovarian cancer. LRP-positive expression and chemotherapeutic effect may be independent prognostic factors.

Antigens, CD34 ; metabolism ; Ascites ; pathology ; Cystadenocarcinoma, Mucinous ; blood supply ; drug therapy ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; blood supply ; drug therapy ; metabolism ; pathology ; Cystadenoma, Mucinous ; blood supply ; drug therapy ; metabolism ; pathology ; Cystadenoma, Serous ; blood supply ; drug therapy ; metabolism ; pathology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; pathology ; Ovarian Neoplasms ; blood supply ; drug therapy ; metabolism ; pathology ; Receptor, Epidermal Growth Factor ; metabolism ; Survival Rate ; Vault Ribonucleoprotein Particles ; metabolism

OBJECTIVETo investigate mesenchymal stem cells (MSCs) immunosuppressive activity in the presence of interferon-gamma (IFN-γ) to reveal synergistic immunomodulatory effects of IFN-γ and MSCs.

METHODS① MSCs were cultured in the presence or absence of IFN-γ（100 ng/ml）, the supernatants were collected for measurements of PGE2、HGF and TGF-β1 by ELISA kits. ② MSCs were cultured in the presence or absence of IFN-γ （100 ng/ml）for 48 h. The cDNA was analysed for the expression of human indoleamine 2, 3-dioxygenase（IDO）mRNA by semiquantitative RT-PCR. ③ Mononuclear cells (MNCs) were extracted from peripheral blood of healthy donors. The T cell proliferation was tested in the co-culture system added with MSCs, recombinant human IFN-γ (100 ng/ml) and anti-IFN-γ mAb (5 μg/ml) by BrdU ELISA kit.

RESULTS①The immunosuppressive cytokines PGE2、HGF and TGF-β1 were detectable within 24-48 h in the supernatants. Their expressions were significantly up-regulated in the presence of IFN-γ. Concentrations of these cytokines were as of (1715.5±628.6) pg/ml vs (1344.5±709.4) pg/ml (P=0.001);(4031.8±1496.8) pg/ml vs (2452.4±1375.3) pg/ml（P=0.011）;(1753.5±413.8) pg/ml vs (1026.6±450.5) pg/ml（P<0.001）,respectively. ②The expression of IDO mRNA was undetectable when MSCs were cultured alone. In contrast, The IDO mRNA expression was remarkably enhanced in the presence of IFN-γ. ③Bone marrow-derived MSCs remarkably suppressed allogeneic T cell proliferation in vitro. Addition of exogenous IFN-γ had no significant effect on the inhibitory capacity of MSCs, the inhibitory ratios of T cell proliferation were （40.4±10.9）% vs（36.7±7.4）% (P=0.272). By contrast, the inhibitory ratio of T cell proliferation was significantly decreased in the presence of anti-IFN-γ mAb[(40.4±10.9)% vs (23.9±7.6)%,P=0.002].

CONCLUSION①Human MSCs constitutively expressed immunosuppressive concentrations of PGE2, HGF and TGF-β1, and their expressions were significantly up-regulated by IFN-γ. ②IFN-γ-induced expression of IDO on MSCs involved in tryptophan catabolism. ③MSCs notably suppressed allogeneic T cell proliferation in vitro. IFN-γ promoted the immunosuppressive capacity of human MSCs, indicating the synergistic immunomodulatory effect of IFN-γ and MSCs.

Bone Marrow Cells ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytokines ; immunology ; Humans ; Immune Tolerance ; Interferon-gamma ; pharmacology ; Mesenchymal Stromal Cells ; immunology ; T-Lymphocytes ; cytology