Acute Disease ; Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; Humans ; Infant ; Inpatients ; Insecticides ; poisoning ; Middle Aged ; Organophosphorus Compounds ; Pesticides ; poisoning ; Poisoning ; etiology ; mortality ; therapy ; Risk Factors ; Survival Rate

Objective To investigate the effects of cryopreservation on the growth and osteogenesis capa- bility of human bone marrow stromal cells(BMSCs)on demineralized bene matrix(DBM).Methods Bone marrow aspirates were obtained from the lilac crests of three donors.The BMSCs were isolated from the bone marrow by density gradient centrifugation.Cells of passage 3 were cryopreserved in liquid nitrogen for 24 hours,and then re- covered.The non-cryopreserved BMSCs were used as the control,The cryopreserved and control BMSCs were cul- tured in osteogenic media,collected and labeled with Dil to be seeded onto the DBM when cells were confluent.The percentage of BMSCs adhered to the DMB was detected.The cell morphology and matrices secreted by BMSCs on the DBM were observed by the inverted phase-contrasted microscope,fluorescence microscope and scanning electron microscope(SEM).The growth and viability of BMSCs on the DBM were determined using the modified MTT ashy. The osteogenesis ability of BMSCs on the DBM was determined by assessment of the alkaline phosphatase(ALP) activity and osteocalcin(OCN)content.Results The percentages of the cryopreserved and control cells adhered to DBM were(97.25?1.17)% and(97.00?1.09)% respectively.The cells adhered well to the DBM and grew rapidly.Large amounts of matrices on the DBM were observed by the light microscope and SEM.The cells embedded in the matrices could be observed by fluorescence microscope.There were no significant differences in the assay values of MTT,ALP and OCN between the cryopreserved and control BMSCs on the DBM.Conclusion Since cryopreservation does not affect the growth and osteogenesis capability of BMSCs on DBM,the cryopreserved BMSCs can be used as a cell source in bone tissue engineering.

0.05).In simple obesity group,there was positive correlation between BMI and urinary 24 h-Alb content(r=0.626,P

OBJECTIVETo investigate the Epstein-Barr virus (EBV) infection, the expression of EBV latent membrane protein 1 ( LMPl) and oncogene bcl-2 in lung cancer patients.

METHODSEBERI in 108 cases of lung cancer were detected with in situ hybridization. EBV positive and negative lung cancer tissues were analysed for the expression of LMP1 and Bcl-2 by immnohistochemistry. The average area (AA) and integral optical density (IA) of each sample was measured with the digital medical image analyzing system.

RESULTSIn 108 cases of lung cancer, 36 cases were EBER1 positive and 7 cases were LMP1 positive. The expression of Bcl-2 was higher in EBV positive lung cancer tissues than that in EBV negative. The AA value was 58014.23 +/- 6918.45 and 38156.22 +/- 4096.79, while the IA value was 11.00 +/- 1.48 and 8.03 +/- 0.78 respectively. No statistic difference was fund in the expression of Bcl-2 betwen LMP1 positive and negative lung cancer tisssues.

CONCLUSIONEBV infection in lung cancer increased the expression of bcl-2, which may play a role in the occurrence or development of lung cancer. The increased expression of Bcl-2 may not be induced by LMP1. The exact mechanism need further study.

Adult ; Aged ; Epstein-Barr Virus Infections ; genetics ; metabolism ; pathology ; virology ; Gene Expression Regulation, Viral ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; virology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism

OBJECTIVESTo examine the Epstein-Barr virus (EBV) in primary lung carcinoma tissue, and to investigate the relationship between EBV infection and tumorigenesis of lung cancer.

METHODSFormalin-fixed and paraffin-embedded lung tissue specimens from surgically resected lung carcinoma tissues of 108 cases treated in Tanshan area from 2001 to 2006, which were confirmed further by histopathological examination after hematoxylin-eosin (HE) staining, were used to observe the EBV encoded RNA-1 (EBER1) using in situ hybridization (ISH).

RESULTSEBER1 was detected in 36 of the 108 primary lung carcinoma cases, and in 1 of the 22 normal lung tissues. The positive rates of EBV infection in squamous cell carcinoma, adenocarcinoma, small cell carcinoma and large cell carcinoma were 35.9%, 31.6% 31.0%, 1/2, respectively. Gender, age and clinicohistopathological type were not found to have any correlation with EBER1 expression, but EBER1 expression in groups of cases with poorly and moderately differentiated carcinomas was significantly higher than those in the group of cases with well differentiated carcinoma, and the EBER1 expression in the right lung was higher than in the left lung.

CONCLUSIONSThe frequency of EBV infection in this series of patients from Tangshan area was 33.3%, the results suggest that there is a relationship between EBV infection and the occurrence of the primary lung carcinoma, EBV infection might be one of the potential causes to induce lung cancer.

Epstein-Barr Virus Infections ; diagnosis ; virology ; Herpesvirus 4, Human ; genetics ; Humans ; In Situ Hybridization ; methods ; Lung Neoplasms ; pathology ; virology ; RNA, Viral ; genetics

AIMTo clone and analyse the coding cDNA sequence of alpha1, beta2 and gamma2 subunit of GABAA receptor in American king Pigeon.

METHODSWithdrew total RNA from the American king pigeon brain, reverse transcribing general primers to acquire a gene set cDNA. Designing specific primers of three subunit mRNA of the GABAA receptor, by RT-PCR respectively expanded the conservative gene of al subunit, beta2 subunit and gamma2 subunit of GABAA receptor, and carried on clone, plastid identification and the sequence measurese of three genes.

RESULTSThe experiment on sequence measures has succeeded that sequence analysis indicated that lengths of the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor was respectively 899 bp, 597 bp and 563 bp, homology on reference sequence was respectively 94.99%, 94.64% and 96.28%.

CONCLUSIONHomology is high on the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor of brain tissue of pigeon and chicken but there is a discriminating characteristic in different kinds of animals.

Animals ; Brain ; metabolism ; Cloning, Molecular ; Columbidae ; DNA, Complementary ; genetics ; RNA, Messenger ; genetics ; Receptors, GABA-A ; classification ; genetics ; Sequence Analysis, DNA

OBJECTIVETo develop a new platform for genotyping human papillomavirus (HPV) and to investigate its effect in clinical application.

METHODSA pair of common primers of 18 HPV subtypes for PCR, was designed in HPV conservative L1 region. Genotyping probes for detecting 15 high-risk HPV subtypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68, together with 3 low-risk HPV 6, 11 and 42 were selected respectively from Genbank and fixed on membrane to make DNA chip. PCR amplification and DNA chip technology were optimized. 100 clinical samples were used to investigate the effect of HPV genotyping DNA chip. Veracity of the genotyping results was verified by sequencing.

RESULTSFrom the 100 clinical samples, 30 were found to be HPV positive, including high-risk HPV subtypes 16, 18, 33, 45, 51, 58, and 66, and low-risk HPV 6, 11 and 42. The sensitivity tested by standard samples was up to 10 copies of HPV DNA.

CONCLUSIONThe HPV genotyping system developed here with DNA chip showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types.

DNA Probes, HPV ; genetics ; DNA, Viral ; genetics ; Female ; Genotype ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; methods ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; virology ; Polymerase Chain Reaction ; Sensitivity and Specificity ; Uterine Cervical Neoplasms ; virology

The infection status of anisakid larvae was examined in 290 marine fish of 25 species and in 108 cephalopods of 3 species purchased in Bayuquan region, Yingko city nearby the coast of the Bohai Sea from may to August 1992. A total of 7,327 larvae were collected from 156 fish of 19 species and 8 squids of one species. The 3rd-stage larvae of Anisakis simplex were collected from 121 fish (63.4%) of 15 species (N = 191) and from 8 squids (14.8%) of one species (N = 54), and they were total, 5,992 (81.8%). Out of remaining 1,335 larvae, 154 (2.1%) were classified as Thynnascaris type B from 23 fish of 4 species, 1,013 (13.8%) as Thynnascaris type C from 79 fish of 13 species. 164 (2.2%) as Hysterothylacium China type V from 20 fish of 4 species, 3 (0.04%) as Raphidascaris from 3 fish of 2 species and one was Pseudoterranova decipiens larva.
Animal ; Anisakiasis/veterinary* ; Anisakiasis/parasitology ; Anisakiasis/epidemiology ; Anisakis/isolation & purification ; Anisakis/classification* ; China ; Fish Diseases/parasitology* ; Fish Diseases/epidemiology ; Fishes ; Larva ; Seawater ; Squid/parasitology*

OBJECTIVETo estimate the effect of the lymph duct ligation on systemic inflammatory factors and endotoxins during intestinal ischemia-reperfusion (I/R).

METHODSMale SD rats underwent occlusion of superior mesenteric artery for 60 min followed by reperfusion for 120 min plus lymph duct ligation or not. Forty rats were randomly divided into 4 groups: group A (blank); group B (sham); group C (intestinal I/R); group D (intestinal I/R plus lymph duct ligation). Mesenteric lymph nodes were harvested for standard bacteriologic cultures. The endotoxin, D-lactate, diamine oxidase (DAO), and cytokines in serum were detected.

RESULTSThe rates of bacterial translocation to mesenteric lymph nodes were 40% in group C and 20% in group D. No positive lymph node cultures were encountered in any of group A and B. The serum cytokines (except for sICAM-1) , D-lactate, DAO and endotoxin levels were lower in group D than those in group C (P<0.05), but both were higher than those in group A and B (P<0.05).

CONCLUSIONDuring intestinal I/R injury, blockage the lymph flow from gut into bloodstream decreases the levels of cytokines, and significantly attenuates the increase in intestinal permeability.

Animals ; Disease Models, Animal ; Inflammation ; Intestinal Diseases ; metabolism ; microbiology ; pathology ; Intestines ; blood supply ; pathology ; Ligation ; Lymph Nodes ; pathology ; Lymphatic System ; surgery ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; microbiology ; pathology

OBJECTIVETo clone and express the gene of Hgp44 in adhesin domains of gingipains from Porphyromonas gingivalis (Pg) and to purify the protein.

METHODSThe genomic DNA of Pg was isolated from PgATCC33277. The Hgp44 gene fragment was amplified by polymerase chain reaction (PCR) and then inserted into the cloning vector pMD18-T and sequenced. The correct fragment was linked with a prokaryotic expression vector pET-22b to construct the recombinant expression plasmid pET22b-Hgp44. The pET22b-Hgp44 confirmed by enzyme digestion was transformed into competent Escherchia coli (Ec) BL21 (DE3) cells. Expression of fusion protein was induced by isopropyl-β-D-thiogalactoside (IPTG), and purified by immobilized metal-chelating affinity chromatography (IMAC) using a Ni(2+) matrix column. SDS-polyaerylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis were used to examine the fusion protein.

RESULTSA 1 100 bp fragment was successfully amplified and verified by the agarose gel electrophoresis and sequencing. The generated recombinant expression vectors pET22b-Hgp44 were verified by enzyme digestion and agarose gel electrophoresis. The expression of fusion protein in Ec BL21 (DE3) cells was examined by SDS-PAGE and Western blotting analyses, and the data showed that the protein was 44 000 in size and expressed mostly in the form of inclusion body. The purification of fusion protein was achieved using Ni(2+) affinity chromatography. About 3.5 mg/L fusion protein was obtained.

CONCLUSIONSHgp44 was successfully expressed in the prokaryotic expression system and purified by IMAC using a Ni(2+) matrix column.

Bacterial Proteins ; biosynthesis ; genetics ; Base Sequence ; Blotting, Western ; Cells, Cultured ; Clone Cells ; Cloning, Molecular ; Genetic Vectors ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; genetics ; Recombinant Fusion Proteins ; Recombinant Proteins