Objective: The fingerprint of Rehmannia glutinosa was established by HPLC to provide scientific basis for improving the quality of R. glutinosa. Methods: The characteristic silicagel C18 column-Atlantis T3 was used, and the mobile phase was 0.1% phosphate water and acetonitrile by gradient elution. Absorption wavelength was 203 nm, and flow rate was 1.0 mL/mL with column temperature of 35 ℃. Setting catalpol as the control peak, 20 batches of R. glutinosa fingerprint was established. Fingerprint similarity evaluation software was used for data evaluation and determinating the catalpol content of 20 batches of R. glutinosa. Results: The standard contrast chromatographic fingerprint similarity of 20 batches R. glutinosa reached above 0.917. The 20 batches catalpol content was above 0.2%. Conclusion: The established fingerprint chromatography was proved that it had good precision, stability, and repeatability. The catalpol content determination met requirement which can avoid interference of saccharide compared with China Pharmacopoeia method and provide scientific reference for improving the quality of R. glutinosa.

Twenty Newcastle disease virus(NDV)strains were isolated from diseased chicken and geese in field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi,and the antigenic analysis of the all NDV isolates had been done based on the reaction spectrum with a panel of monoclonal antibodies to the HN glycoprotein.The entire ORFs encoding HN protein of these NDV isolates were amplified by RT-PCR successfully,cloned and sequenced.The resultant sequences of HN genes of 13 isolates of chicken origin and 7 isolates of goose origin were gained and analyzed.The results of reaction spectrum showed that there were some distinct differences in the antigenic epitopes among the 20 NDV isolates.And the sequences revealed that the coding regions of the HN genes of these isolates all consisted of 1716 nt characteristic of virulent strains of NDV,coding for 571 amino acids.Neucleotides sequence homology were found to be from 94.8%to 100%among 18 NDV isolates of genotypeⅦ,and the neucleotides sequence homology between all the isolates and the other genotypeⅦstrains of recent years in China ranged from 92.1%to 99.6%.The deduced amino acid sequences and the receptor-binding regions of HN proteins between the NDV isolates of chicken origin and of goose origin were compared and analyzed.The results showed that some unique amino acid substitutions were found in the genome of the NDV isolates,and the close genetic similarity provided evidence for epidemiological linkage between the NDV isolates of chicken origin and of goose origin in the same period.

Objective To study the effects of the composite of propolis and magnolol on the growth of Streptococcus mutans (Sm).Methods The solution concentration of the propolis (250,125,62.5,31.25,15.625 g/L),magnolol (0.0625,0.0313,0.0156,0.0078,0.0039 g/L) and the composite of those two (propolis,magnolol) A1 (250 g/L,0.0625 g/L),A2(125 g/L,0.0313 g/L),A3 (62.5 g/L,0.0156 g/L),A4 (31.25 g/L,0.0078 g/L),A5 (15.625 g/L,0.0039 g/L) were chosen according to pretested minimum inhibitory concentration (MIC).The solutions were proportionally diluted to the presupposed concentration before operated on the Sm.Meanwhile,the solvent and the bacterial were also operated on the Sm as controls.Slip diffusion method was adopted to measure the inhibitory effect of those solutions on Sm.Results Bacteria inhibition zones were appeared at the concentration of 62.5 g/L prololis and 0.0156 g/L magnolol respectively.A inhibition zone appeared at the lower concentration of A5 (15.625 g/L,0.0039 g/L) of the propolis and magnolol composite.As general,the inhibition effects increased with concentration.The difference were statistically significant,P ＜ 0.05.Conclusions The inhibitory effect of the composite of propolis and magnolol would be much stronger than either of those two separately.

OBJECTIVEIn order to seek the functional evidence that there could be metastatsis suppressor gene for liver cancer on human chromosomes, the objective of this study is to establish a method of microcell mediated chromosome transfer (MMCT).

METHODSHuman chromosome 8 randomly marked with neo gene was introduced into highly metastatic rat liver cancer C5F cell line by treating the single human chromosome donor cells with sequential steps of micronucleation, enucleation and microcell fusion. Double selections of G418 and HAT were applied to screen positive microcell hybrids, which were cloned by single cell isolation. Microcell hybrid clones were confirmed by STS-PCR and WCP-FISH.

RESULTSMicrocell hybrids resistant to HAT and G418 were obtained, from which 15 clones were obtained by single-cell isolation cloning. STS-PCR and WCP-FISH proved that human chromosome 8 had been successfully introduced into rat liver cancer cell line C5F. The human chromosome 8 introduced into C5F was found to have random loss of chromosome fragments by STS-PCR and consistent recombination with rat chromosome by WCP-FISH.

CONCLUSIONThe successfulls introduction of human chromosome into highly metastatic rat liver cancer cell line has established the technical basis for functional localization of metastasis suppressor gene(s) for liver cancer on human chromosomes.

Animals ; Cell Line, Tumor ; Chromosome Mapping ; methods ; Chromosomes, Human, Pair 8 ; genetics ; Genes, Tumor Suppressor ; Genetic Techniques ; Humans ; In Situ Hybridization, Fluorescence ; Liver Neoplasms ; genetics ; pathology ; Polymerase Chain Reaction ; Rats

Objective To explore risk factors for surgical site infection(SSI) in colorectal surgery,and provide evidence for formulating measures for preventing SSI.Methods Patients who underwent colorectal surgery in the department of gastrointestinal surgery of a hospital from June 2013 to June 2016 were surveyed retrospectively,the related risk factors for SSI were analyzed by unconditional logistic regression analysis.Results Among 397 patients who underwent colorectal surgery,67 (16.88％) had SSI.Logistic regression analysis showed that smoking,low albumin,seniority of surgeons less than 5 years,irrational use of antimicrobial agents during perioperative period,and high National Nosocomial Infection Surveillance (NNIS) score were independent risk factors for SSI after colorectal surgery (all P＜0.05).Conclusion There are multiple risk factors for SSI after colorectal surgery,it is necessary to pay attention to it and formulate preventive measures,so as to reduce the occurrence of SSI effectively.

Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Humans ; Interleukin-1beta ; adverse effects ; Neoplasm Invasiveness

OBJECTIVETo identify the phenotype and immune function of dendritic cells derived from HBV-related HCC patients's peripheral blood monocytes pulsed with soluble tumor antigen, and their relation to immune escape.

METHODSPeripheral blood monocytes were isolated from 18 HBV-related hepatocellular carcinoma (HCC) patients, 11 HBV-related liver cirrhosis patients (LC) and 10 health blood donors; DCs were induced in the completed medium containing GM-CSF and IL-4. The morphology of DCs was studied using a confocal microscope and scanning electronic microscope, and the phenotype of DCs were detected by flow cytometric analysis. The mixed leucocyte reaction test was employed to determine the stimulatory capacity of DCs before and after being pulsed with soluble tumor antigen (prepared from HCCLM6 cell line). IL-12 ELISA kit was used to investigate IL-12 secretion of DCs in the supernate of MLR.

RESULTSThe amount of PBMC and DCs was significantly lower in LC and HCC compare to those in the healthy subjects; the expression levels of HLA-DR, CD1a, CD80 and CD86 on DC surfaces were lower in LC and HCC patients than those of the healthy group; the stimulating capacity of DC in MLR and levels of IL-12 in supernate of MLR were also lower in LC and HCC, but were enhanced after tumor antigen pulsed in all three groups, particularly in the LC group; the secretion of IL-12 in MLR supernate was still lower than that of the healthy group.

CONCLUSIONThe phenotype and function defects of DC derived from PBMC of LC and HCC patients might play a key role in immune escape in HBV infection and HCC. The function of DC of LC patients can be enhanced after the tumor was antigen-pulsed.

Antigens, Neoplasm ; immunology ; Carcinoma, Hepatocellular ; immunology ; virology ; Dendritic Cells ; immunology ; virology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Hepatitis B ; complications ; immunology ; Humans ; Interleukin-4 ; pharmacology ; Liver Neoplasms ; immunology ; virology ; Lymphocyte Culture Test, Mixed

OBJECTIVETo assess the feasibility and efficiency of eliciting leukemia-specific T cell responses in acute myeloid leukemia patients in complete remission (AML-CR) in vitro by dendritic cells (DC) pulsed with the leukemia cells total RNA.

METHODSThe immature DCs were generated from the adherent bone marrow mononuclear cell in vitro in the presence of combined cytokines (GM-CSF 100 ng/ml, IL-4 500 U/ml), and pulsed with total RNA isolated from autologous leukemic cells by cationic lipid 1,2-dioleoyloxy-3-trimethyl ammonium propane (DOTAP) at day 5 of culture. Then the cells were incubated for another 24 h in a medium containing 10 ng/ml of TNF-alpha for maturation of DC. After the total 7 days culture, the cells were harvested as the mRNA-DC and the expression of mature DC markers were determined by FACS. The proliferative capacity of T cell activated by mRNA-DC was determined by MTT assay. Meanwhile, the mRNA-DC was co-cultured with T lymphocytes at a ratio of 1:3 for 7 days. The activated T lymphocytes were harvested, the secretion of IFN-gamma was determined by ELISPOT assay, and the cytotoxicity was analyzed in vitro by LDH release assay.

RESULTSAfter culture, the BMMNC from 14 AML-CR patients developed morphologic and phenotypic characteristics of mature DC. At a stimulator/reactor ratio of 1:16, auto-T lymphocytes primed with mRNA-DC exhibited significant proliferative activity compared with T lymphocyte primed with non-pulsed DC [(36.84 +/- 5.68)% vs (12.20 +/- 3.16)%, (P < 0.05)]. An expansion of mRNA reacted T cell secreting IFN-gamma could be observed on ELISPOT assay. At an effector/target ratio of 20:1, the auto-T lymphocytes primed with mRNA-DC exhibited significant killing activity to auto-AML cells (45.46 +/- 6.34 )% as compared with that stimulated by IL-2 alone (13.26 +/- 2.28)% or primed by non-pulsed DC (12.32 +/- 1.32)% (P < 0.05).

CONCLUSIONImmunization with DC-leukemia cell RNA vaccines may be a simple, rapid and potent approach to elicitation of T cell-mediated anti-leukemia immunity.

Adolescent ; Adult ; Cell Communication ; Cells, Cultured ; Coculture Techniques ; Dendritic Cells ; drug effects ; immunology ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; immunology ; Male ; Middle Aged ; RNA ; pharmacology ; T-Lymphocytes ; immunology

OBJECTIVETo screen for mutations of fibrillin-1 (FBN1) gene in 4 patients with Marfan syndrome in order to provide prenatal diagnosis and genetic counseling.

METHODSPotential mutations of the FBN1 gene in the probands were detected with PCR and DNA sequencing. Subsequently, genomic DNA was extracted from amniotic fluid sampled between 18 to 20 weeks gestation. The mutations were confirmed with denaturing high-performance liquid chromatography - robust microsatellite instability (DHPLC-MSI) analysis with maternal DNA as reference. The products were further analyzed by direct sequencing and BLAST search of NCBI database.

RESULTSAn IVS46+1G>A substitution was identified in patient A at +1 position of intron 46 of the FBN1 gene. Two novel missense mutations were respectively discovered at positions +4453 of intron 35 in patient B (Cys1485Gly) and position +2585 of intron 21 in patient C (Cys862Tyr). In patient D, a novel deletion (c.3536 delA) was found at position +3536 of intron 28. In all of the 4 cases, the same mutations have been identified in the fetuses.

CONCLUSIONFBN1 gene analysis can provide accurate diagnosis of Marfan syndrome, which can facilitate both prenatal diagnosis and genetic counseling.

Adult ; Base Sequence ; DNA Mutational Analysis ; Female ; Fibrillin-1 ; Fibrillins ; Humans ; Introns ; Male ; Marfan Syndrome ; diagnosis ; embryology ; genetics ; Microfilament Proteins ; genetics ; Molecular Sequence Data ; Mutation, Missense ; Pregnancy ; Prenatal Diagnosis ; Sequence Deletion

The biological characterization, differentiation and regeneration of hepatic stem/progenitor cells are the one of very active and interested fields. In this report, intravenous injection of human umbilical cord blood (HUCB) cells into the BALB/c-nu and SCID mice, an animal model for transplantation and liver injury, was reported. Using of flow cytometry and tissue typing (HLA), it was found that the HUCB cells were survived in mouse liver for 9 weeks. After separation from perfused liver, HUCB cells were detected by hematopoietic colonies (CFU-GEM M) in hepatocyte culture. It was concluded that the transplanted HUCB hematopoietic stem/progenitor cells can be survived in the liver over a long period of time.
Animals ; Cell Division ; Fetal Blood ; cytology ; Flow Cytometry ; HLA-DR Antigens ; analysis ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; physiology ; Humans ; Infant, Newborn ; Liver ; cytology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, SCID