Ultra performance liquid chromatography (UPLC) was employed for simultaneous determination of three components and fingerprint analysis of Periplocae Cortex with gradient elution of mehtanol and water containing 0.1% phosphoric acid as mobile phase. Three components including chlorogenic acid, 4-methoxysalicylaldehyde and periplocoside were well separated under the analytical condition. Seventeen peaks were selected as the common peaks of 30 batches of Periplocae Cortex. The results showed that there is a significant difference in contents of periplocoside between the samples collected from Henan and Shanxi province. Based on the results of three components quantification and fingerprint analysis, hierarchical clustering analysis ( HCA) and principle component analysis (PCA) were used to further prove the differences between two group samples, and the results indicated that quality of Periplocae Cortex from Shanxi was more stable than that from Henan. The established UPLC fingerprint and quantitative analysis methods could be used efficiently in the quality control of Periplocae Cortex, and this study might contribute to the reasonable clinical application.
Benzaldehydes ; analysis ; China ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Ecosystem ; Periploca ; chemistry ; classification ; growth & development ; Plant Roots ; chemistry ; Quality Control

Objective To investigate the effect of peptide Tat-GluR6-9c on phosphorylations and protein expressions of mixed-lineage kinase 3 (MLK3),mitogen-activated protein kinase kinase 7 (MKK7) and c-Jun NH2-terminal kinase (JNK),and its effect on hippocampus CA1 region neuronal cell injury induced by cerebral ischemia followed by reperfusion.Methods Twenty four adult male SD rats were randomly divided into sham-operated group,ischemia-reperfusion group (I/R),Tat-GluR6-AA treatment group and Tat-GluR6-9c treatment group (n=6).Four-vessel occlusion method was employed to establish the cerebral ischemia models in the later 3 groups.The effects of peptide Tat-GluR6-9c on the phosphorylations and protein expressions of MLK3 (6 h after the reperfusion) and JNK (3 d after the reperfusion) were detected by Western blotting; the effects ofpeptide Tat-GluR6-9c on the phosphorylation and protein expression of MLK7 (1 d after the reperfusion) were detected by Western blotting and immunohistochemistry.Cresyl Violet (CV) staining was employed to examine the survival of CA1 pyramidal cells in the hippocampus.Results The phosphorylation of MLK3,MKK7 and JNK in Tat-GluR6-9c treatment group was significantly less than that in I/R group and Tat-GluR6-AA treatment group (P＜0.05).As compared with I/R group and Tat-GluR6-AA group,peptide Tat-GluR6-9c group could obviously increase the number of neuron cells (P＜0.05).Conclusion Peptide Tat-GluR6-9c has a protective effect on neuron in CA1 region of hippocampus following cerebral ischemia-reperfusion by significantly decreasing the phosphorylations ofMLK3,MKK7 and JNK.

In clinical cerebral perfusion CT examination, repeated scanning the region of interest in the cine mode increases the radiation dose of the patients, while decreasing the radiation dose by lowering the scanning current results in poor image quality and affects the clinical diagnosis. We propose a penalized weighted least-square (PWLS) method for recovering the projection data to improve the quality of low-dose cerebral perfusion CT imaged. This method incorporates the statistical distribution characteristics of brain perfusion CT projection data and uses the statistical properties of the projection data for modeling. The PWLS method was used to recover the data, and the Gauss-Seidel (GS) method was employed for iterative solving. Adaptive weighting is introduced between the original projection data and the projection data after PWLS restoration. The experimental results on the clinical data demonstrated that the PWLS-based sinogram restoration method improved noise reduction and artifact suppression as compared with the conventional noise reduction methods, and better retained the edges and details to generate better cerebral perfusion maps.
Algorithms ; Artifacts ; Cerebrum ; diagnostic imaging ; Humans ; Least-Squares Analysis ; Radiographic Image Interpretation, Computer-Assisted ; Tomography, X-Ray Computed

Objective To observe the influence of adrenomedullin (ADM) on neuron apoptosis, infarction volume of brain, and the expression of early growth response 1 (Egr-1) mRNA in ischemia-reperfusion rats. Methods The arteria cerebri media was tied for 2 h to construct the ischemia model. Infarction volume was detected by triphenltetrazolium chloride (TTC) staining, neuronal apoptosis and necrosis was detected with terminal deoxynucleotidyl transferase nick labeling (TUNEL) method, and the Egr-1 mRNA expression was examined by in situ hybridization (ISH). Results Infarction volume after ischemia-reperfusion is (269 +/- 20) mm(3). Infarction volume after injection of ADM through different ways are femoral vein (239 +/- 17) mm(3) (decreased by 11.2%), arteria carotis (214 +/- 14) mm(3) (by 20.4%) and lateral cerebral ventricle (209 +/- 13) mm(3) (by 22.3%), respectively. The results indicate that injecting ADM through arteria carotis and lateral cerebral ventricle is much more effective than it through femoral vein (P < 0.05). The TUNEL-positive cells in cerebral cortex or hippocampus are few in the sham operation group, but much more in the ischemia-reperfusion group. After being supplied with ADM, especially through arteria carotis interna or lateral cerebral ventricle way, the TUNEL-positive cells decreased obviously. Expression of Egr-1 mRNA was low in the cerebral cortex of the sham operation group rats, enhanced in the ischemia and reperfusion group rats, and enhanced markedly after treatment with ADM, especially through arteria carotis interna or lateral cerebral ventricle way (P < 0.01). Conclusion Injection of ADM through different ways could alleviate neural dysfunction, decrease neuron apoptosis and brain infarction volume, and increase the expression of Egr-1 mRNA.

OBJECTIVE: To explore the value of has-microRNA-155 (miR-155) in peripheral blood mononuclear cells (PBMNC) in prognostic evaluation of elderly patients with primary immune thrombocytopenia (PITP). METHODS: One hundred and thirty elderly PITP patients and 60 healthy volunteers in our hospital were selected. The relative expression level of miR-155 in PBMNC was detected by RT-PCR. Unconditional logistic multivariate regression analysis was used to analyze the relationship between miR-155 expression and prognosis of PITP patients, and Kaplan-Meier was further used to analyze the relationship between miR-155 and PITP recurrence. RESULTS: The relative expression level of miR-155 in PBMNC of elderly PITP patients was significantly higher than that in healthy volunteers, and increased significantly with the severity of the disease (P＜0.05). The overall effective rate of elderly PITP patients with miR-155 low-expression was significantly higher than that in the patients with miR-155 high-expression (96.92% vs 72.31%) by after treatment with glucocorticoid. Multivariate analysis showed that miR-155 was an independent risk factor for PITP patients. Elderly patients with high expression of miR-155 showed a higher risk of recurrence. CONCLUSION miR-155 in PBMNC has a high accuracy for PITP diagnosis, and the elderly patients with high level of miR-155 show a poor prognosis and a higher risk of recurrence.
Aged ; Biomarkers, Tumor ; Humans ; Kaplan-Meier Estimate ; Leukocytes, Mononuclear ; MicroRNAs ; Neoplasm Recurrence, Local ; Prognosis ; Purpura, Thrombocytopenic, Idiopathic

OBJECTIVE: To investigate the effect of ABO gene α-1,3-D galactosyl transferase mutation on B antigen expression and its molecular mechanism. METHODS: The proband and their family members were identified by routine serological methods, and ABO genotyping and sequence analysis were performed by polymerase chain reaction-sequence specificity (PCR-SSP) and direct sequencing of PCR products from exon 1-7 of ABO gene. The 3D structural simulation of mutant proteins was performed by bioinformatics software. The effect of gene mutation on protein structural stability was analyzed. RESULTS: The proband and his family members were subtype B. ABO genotyping indicated that the proband's genotype was Bw12/O. Gene sequencing results confirmed the presence of ABO*BW.12 characteristic variation c.278C>T in the 6th exon of allele B, leading to the replacement of polypeptide chain p.Pro93Leu. The 3D structure simulation analysis of the protein showed that the hydrogen bonds and water molecules connected to the protein changed after amino acid substitution. The family investigation found that the grandfather, father, uncle and brother of the proband all carried the same ABO*BW.12 allele. CONCLUSION The mutation of the 6th exon c.278C>T of ABO gene led to the substitution of polypeptide chain amino acids, which affected the stability of α-1,3-D galactosyl transferase protein, resulting in the change of enzyme activity, and the Bw.12 phenotype, which can be stably inherited.
ABO Blood-Group System/genetics* ; Alleles ; Amino Acids/genetics* ; Animals ; Base Sequence ; Exons ; Genotype ; Male ; Mutant Proteins/genetics* ; Mutation ; Phenotype ; Water

Adult ; Aged ; Aged, 80 and over ; Area Under Curve ; Esophagitis, Peptic ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; ROC Curve ; Surveys and Questionnaires

BACKGROUNDThe prevalence of Helicobacter pylori (H. pylori) infection varies by geographic locations. Studies indicate that the infection rate of H. pylori was previously high in China but that rates had been declining worldwide over recent decades.

THE AIMS OF OUR STUDY WERE(1) to determine the current prevalence of H. pylori infection among children and adults residing in areas with high (Muping County, Shandong) and low (Yanqing County, Beijing) incidences of gastric cancer in China, and (2) to compare the prevalence for 2006 with the prevalence for the early 1990s.

METHODSUsing Warthin-Starry silver staining of gastric mucosal biopsy specimens and H. pylori stool antigen tests (HpSA), we tested a total of 2065 asymptomatic children aged 8 - 15 years and adults aged 40 - 79 years in the above two regions from May to July 2006. We evaluated 520 children and 526 adults from Muping, and 516 children and 503 adults from Yanqing. Subjects were selected randomly and H. pylori status was determined by HpSA in children and either HpSA or histology of gastric biopsies in adults. Data obtained in the early 1990s in the same two areas of China were also collected and studied.

RESULTSFor children, the prevalence of H. pylori infection was significantly higher in Muping (37.69%) than it was in Yanqing (25.58%, P < 0.001). In both regions, the prevalence of H. pylori increased with age but was not related to gender. A significant difference was observed between 8 - 9-years old and 10 - 11-years old (P < 0.05), but not between other adjoining age groups (P > 0.05). From 1991 to 2006 H. pylori prevalence among 8 - 10-year-old children decreased in Muping (60.00% vs 32.07%, P < 0.001), but not Yanqing (24.06% vs 19.10%, P > 0.05). In the adult group, H. pylori prevalence was 50.95% in Muping, which was significantly higher than the 41.35% positive rate in Yanqing (P < 0.01). But there were no statistically significant differences between different age groups of 40 - 49, 50 - 59, and 60 - 79 years, or between males and females. A significant decrease in H. pylori prevalence in both regions was observed when the results of 2006 were compared with the data obtained in 1990 in Muping (50.95% vs 73.78%, P < 0.001) and in 1992 in Yanqing (41.35% vs 55.35%, P < 0.01).

CONCLUSIONSAfter fifteen years, the prevalence of H. pylori infection among both children and adults remained significantly higher in areas with a high incidence of gastric cancer in China compared with that in areas with a low incidence of gastric cancer. H. pylori infection rates have decreased in the general Chinese population during recent years.

Adolescent ; Adult ; Age Distribution ; Aged ; Antigens, Bacterial ; Child ; China ; epidemiology ; Female ; Helicobacter Infections ; epidemiology ; immunology ; Helicobacter pylori ; immunology ; physiology ; Humans ; Male ; Middle Aged ; Prevalence ; Stomach Neoplasms ; epidemiology ; immunology ; microbiology

OBJECTIVETo establish the stably lower expression of vascular cell adhesion molecule-1 (VCAM-1) in MSC cell line (C3H10T1/2) by siRNA technology, and explore the effect of knockdown of VCAM-1 on the immunologic regulation capacity of murine MSC.

METHODSThe mouse GV118-VCAM-1-RNAi retrovirus vector was constructed by gene recombination technology. The recombinant plasmid was identified by restriction analysis and sequencing, and then the recombinant plasmid GV118-VCAM-1-RNAi was transfected into 293 cells by Lipofectamine, and the supernatant was collected to transfect C3H10T1/2. Moreover, the VCAM-1 lower expression on MSC was evaluated by flow cytometry and fluorescent microscopy. The knockdown VCAM-1 MSC was sorted by flow cytometry. Furthermore, the inhibitory effect of the knockdown VCAM-1 MSC on lymphocyte proliferation was tested by lymphoblast transformation assay (LTT) and mixed lymphocyte reaction assay(MLR).

RESULTSThe recombinant retroviral vector of knockdown VCAM-1 (GV118-VCAM-1-RNAi) was successfully constructed and transfected into mouse MSC cell line C3H10T1/2. The knockdown VCAM-1/MSC was obtained by flow cytometric sorting. The LTT and MLR assay showed that the immunosuppressive effect of MSC lower-expressing VCAM-1 dramatically decreased (P<0.05).

CONCLUSIONKnockdown VCAM-1 in MSC can significantly down-regulate the inhibitory capability of MSC on the proliferation of T-cells. The data of this study laid an experimental foundation for studying effect of VCAM-1 transfecting into MSC on immune function.

Animals ; Cell Line ; Cell Movement ; Cell Proliferation ; Flow Cytometry ; Genetic Vectors ; Lymphocyte Activation ; Mesenchymal Stromal Cells ; Mice ; Plasmids ; RNA Interference ; RNA, Small Interfering ; T-Lymphocytes ; Transfection ; Vascular Cell Adhesion Molecule-1

OBJECTIVETo investigate the effect of vascular cell adhesion molecule-1 (VCAM-1) gene overexpression on adipogenic differentiation of mouse mesenchymal stem cells(MSC) and explore its molecular mechanism.

METHODSVCAM-1 overexpression MSC (MIGR1-VCAM-1/MSC) and the empty plasmid transfection MSC (MIGR1/MSC) were induced to adipogenic differentiation, oil-red-O staining and real-time PCR were used to detect the adipogenic differentiation ability and the mRNA expression level of key transcription factors C/EBP α and PPAR γ. The activation of P38, ERK and JNK pathways were analyzed by Western blot. Furthermore, the specific chemical inhibitors of MAPK pathway (SB203580, PD98059 and JNK inhibitor II) were added to the induced culture system and the alteration of the MSC adipogenic differentiation ability were evaluated.

RESULTSno matter in self or induced differentiation groups, the lipid droplets in MIGR1-VCAM-1/MSC became larger, the amount of adipocyte increased than that in MIGR1/MSC (P<0.01), the mRNA expression level of C/EBPα and PPARγ were upregulated, and JNK pathway were down-regulated while the P38 and ERK pathway were significantly up-regulated. The inhibition of JNK pathway of MIGR1-VCAM-1/MSC could lead to increased mRNA expression level of C/EBP α and PPAR γ, the amount of adipocytes increased (P<0.01), however, the inhibition of the P38 and ERK pathway of MIGR1-VCAM-1/MSC could lead to decreased mRNA expression level of C/EBP α and PPAR γ, and the lipid droplets and the number of adipocytes became smaller and less.

CONCLUSIONThe overexpression of VCAM-1 may promote MSC to differentiate into adipocytes through inhibiting JNK signaling pathway, activating P38 and ERK pathways.

Adipocytes ; Animals ; CCAAT-Enhancer-Binding Protein-alpha ; Cell Differentiation ; Down-Regulation ; MAP Kinase Signaling System ; Mesenchymal Stromal Cells ; Mice ; PPAR gamma ; Real-Time Polymerase Chain Reaction ; Transfection ; Up-Regulation ; Vascular Cell Adhesion Molecule-1