Benzene ; toxicity ; Gene Expression Profiling ; Oligonucleotide Array Sequence Analysis

Objective To investigate the effect of brain-derived neurotrophic factor (BDNF) gene modified neural stem cells (NSCs)transplantation on cerebral ischemic injury in rats.Methods NSCs from newborn rat hippocampus were isolated,cultured in a medium containing fibroblast factor (FGF) in vitro. Their proliferation and differentiation were detected by immunohistochemistry.Virus vectors pLXSN-BDNF were built and BDNF were transfected into NSCs.Biological activity were detected to obtained engineering stem cells of BDNF protein with secretary activity.Middle cerebral artery occlusion (MCAO) models were made and transplanted with NSCs-BDNF by stereotaxic technique.The effect of transplantation on MCAO models was evaluated histologically and behaviorally.Results Statistical analysis showed that the behavioral performance of the animals improved after transplantation (Longa mark being 1.343?0.293 four weeks after stem cell transplantation).The number of hippocampal dentate gyrus neurons increased to 87.5%?6.6% , four weeks after stem cell transplantation on Nissle stained hippocampal sections,which was significantly different from that of controls.Positively BrdU stained neural stem cells revealed by immunohistochemistry in the cultured cells and in the transplanted brain sections,indicated that the engineering cells transplanted had survived in the host brain and began to proliferate.Conclusion Transplantation of BDNF genetically modified NSCs can effectively promote the recovery from cerebral ischemic injury.

OBJECTIVETo express P1B, P2A, P3AB and P3D cDNA gene fragments in prokaryotic system using thioredoxin fusion expression system; to investigate the antigenicity and application of recombinant protein.

METHODSBy using PCR technique, P1B, P2A, P3AB and P3D gene fragments were cloned. Choosing M47 as the expressive vector, the recombinant plasmid P1B, P2A, P3AB and P3D was constructed and expressed in Escherichia coli after inducing by IPTG. By anion exchange and affinity chromatography, purified recombinant protein was obtained. By using Western Blot analysis and indirect ELISA to detect its antigenic activity.

RESULTSFour recombinant plasmids was proved to be constructed successfully by sequencing and the correct molecular weight of their expression products. Recombinant proteins were obtained in BL21 (DE3) and purified after Ni2+ affinity chromatography. Western Blot analysis and indirect ELISA showed that P2a had specific antigenicity.

CONCLUSIONThe P2a protein expressed in prokaryotic system was proved to have specific antigenicity. The indirect ELISA distinguished 24 positive sera from 24 negative sera. It is very likely that P2a can be an antigen to diagnose acute patients of hepatitis A and differentiate inactivated vaccine-induced immunity from an infection.

Blotting, Western ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Gene Expression ; Hepatitis A virus ; genetics ; immunology ; metabolism ; Humans ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; metabolism ; Viral Proteins ; genetics ; immunology ; metabolism

OBJECTIVETo evaluate the intermediate and long-term follow-up effect of posterior dynamic lumbar stabilization in lumbar degenerative disease.

METHODSThe clinical outcomes of 96 patients (male 51, female 45, age from 21 to 68 years, mean 41.5 years) whose follow-up time were more than 2 years with lumbar degenerative disease treated by posterior decompression with Wallis posterior dynamic lumbar stabilization implant or combined with posterior lumbar fusion from August 2007 to January 2010 were retrospectively studied, and assessed with visual analogue scale (VAS) and spinal operative standard of Chinese Medical Association. The early and long-term follow-up effect and complications associated with Wallis posterior dynamic lumbar stabilization were recorded. The height of intervertebral space at the treated level in lateral plain film were measured at preoperatively, 3 month postoperatively and last follow-up, respectively. The finds of MRI obtained at over 6 month postoperative were recorded.

RESULTSThe operative procedure of Wallis posterior dynamic lumbar stabilization implant was easy and less invasive. The VAS scores were 78 ± 24, 28 ± 16 and 14 ± 12 preoperatively, 3 month postoperatively and last follow-up, respectively. The good or excellent result was 91.7% at the last follow-up. No complication related with Wallis posterior dynamic lumbar stabilization was found. The rate of patient's satisfaction with the Wallis implant operation was 95.8%. The disc height at the treated level in lateral plain film were (8.2 ± 3.7), (10.4 ± 2.6) and (10.1 ± 1.9) mm at preoperatively, 3 month postoperatively and last follow-up, respectively. There is no further degenerative change found in MRI obtained at over 6 month postoperative. MRI 1 year after Wallis procedure showed rehydration of the formerly black disc at the treated level.

CONCLUSIONSIt is easy and safe to use Wallis posterior dynamic lumbar stabilization in treatment of degenerative lumbar disease, and the effect of the intermediate and long-term follow-up more than 2 years is good. The Wallis system provides an alternative for treatment of lumbar degenerative disease.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Internal Fixators ; Intervertebral Disc Degeneration ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; instrumentation ; methods ; Treatment Outcome ; Young Adult

Objective To determine the main genotype of hepatitis B virus (HBV) in Xinjiang.Methods 200 HBsAg positive serum specimens were detected from more than 2000 serum of Xinjiang inhabitants, and HBV S gene was detected by using nPCR amplifying, and compared with the standard S region HBV nucleotide sequences of genotypes A-H retrieved from GenBank, then analyzed by MEGA3 and SAS3 software. Result Gene in 127 (63.5%) serum specimens was detected from 200 samples. Among 127 serum specimens, 10 (7.8%) was genotype B, 58 (45.7%) was genotype C, and 59 (46.5%) was genotype D. Han People were 51 (87.9%) in genotype D and 27(45.7%) in genotype D, Uygur were 24 in genotype D. Conclusion Genotype B(40.7%), C and D have been found in Xinjiang. Genotype C was more prone to causing severe liver disease.

BACKGROUNDTo study a new kind of adjuvant: transcutaneous immunization adjuvant.

METHODSThe full length gene of Heat-labile enterotoxin (LT) was amplified from E. coli H10407. The B subunit protein LTB and the nontoxic A subunit protein LTKA were expressed by genetic engineering manipulation. After purification, they were identified with SDS-PAGE, GM1-ELISA and so on.

RESULTSThe LTB protein still persisted its biologic activity that conjugated specifically with GM1 ganglioside, and the LTK63 protein lost its toxin activity.

CONCLUSIONThe results showed that LTB and LTK63 may be used as promising transcutaneous immunization adjuvant.

Adjuvants, Immunologic ; genetics ; isolation & purification ; metabolism ; Animals ; Bacterial Toxins ; genetics ; immunology ; metabolism ; Blotting, Western ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; immunology ; metabolism ; Gene Expression ; Genetic Engineering ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

OBJECTIVETo detect target genes for further study by way of analyzing the gene expression profiles of benzene poisoning by using cDNA microarray.

METHODSPeripheral mononuclear cells were isolated from seven patients with benzene poisoning of different degrees, and sevene age-and sex-matched normal subjects. Total RNA was extracted and purified, followed by revese transcription to cDNAs with concomitant incorporation of fluorescent dCTP (Cy3 or Cy5). The cDNAs were used as probes in microarray of 2780 cloned cDNA. Fluorescent signals were scanned to detect genes differentially expressed in patients and normal subjects.

RESULTSAmong 7 pieces of cDNA microarray of 2780 tumour related genes, the expression of 16 genes, such as GRO1, TGFBR3, LYN ctc was upregulated, whereas the expression of 28 genes, such as FOSB, DJ-1, MCT-1 etc was down-regulated.

CONCLUSIONAbnormal expression of tumour related genes of patients exposed to benzene suggests that they may be the key genes, which play important role in benzene-induced leucocythemia. cDNA microarray technique is useful to indicate the expression mode of benzene poisoning tumour related genes, and to find rapidly and effectively new research object and the way of gene therapy.

Benzene ; poisoning ; Case-Control Studies ; DNA, Complementary ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Oligonucleotide Array Sequence Analysis ; Oncogenes ; genetics ; RNA, Messenger ; genetics ; Transcriptome

OBJECTIVETo screen DNA replication, and damage repair genes associated with benzene poisoning by using gene expression profile analysis.

METHODSLeucocytes in peripheral blood of seven patients and seven normal controls were collected and total RNA was extracted. The cDNA probes were prepared by labeling cell RNA with Cy3-dUTP and Cy5-dUTP with reverse transcription. The arrays with 141 genes were hybridized against the cDNA probe mixture, and the fluorescent signals were scanned.

RESULTSTwenty five differentially expressed genes were screened out. Among these genes, 16 genes were up-regulated and 9 down-regulated. They were DNA replication genes such as PRIM2A, ORC1L etc; DNA synthesis, recombination and repair genes such as POLK etc; DNA damage signaling/repair proteins and DNA ligases such as RECQL, PRKDC, G22P1, ERCC3, ERCC1, CRY1, CHES1, BRCA2, APEX etc; proteins involved in recombination such as RECQL; and other DNA synthesis, recombination, and repair proteins such as SKIV2L, RBMS1, SON, SET.

CONCLUSIONSSome DNA replication, and damage repair genes associated with benzene poisoning show differential expression, which provides the basis for screening biomarkers of benzene poisoning.

Benzene ; poisoning ; Case-Control Studies ; DNA Damage ; genetics ; DNA Repair ; genetics ; Female ; Gene Expression Profiling ; Humans ; Oligonucleotide Array Sequence Analysis

To simultaneously compare and evaluate the examinations of bone marrow smear and trephine biopsy in differential diagnosis and therapeutical effect of pancytopenia patients, the differences between bone marrow smear and trephine biopsy in degree of cellularity, misdiagnosis rate and therapeutical effect for 71 patients with pancytopenia were analyzed. The results showed that the degree of cellularity in bone marrow biopsy for cytologic morphology was higher than that from the smear, but misdiagnosis rate in the biopsy was lower than that in the smear. In conclusion, bone marrow biopsy is necessary to the differential diagnosis and more valuable for evaluation of therapeutical effect and prognosis of pancytopenia patients.
Adolescent ; Adult ; Aged ; Biopsy ; Bone Marrow ; pathology ; Bone Marrow Examination ; Diagnosis, Differential ; Diagnostic Errors ; Female ; Humans ; Male ; Middle Aged ; Pancytopenia ; diagnosis

OBJECTIVEEplets mismatch based on HLAMatchmaker software evaluates the clinical application of kidney transplantation.

METHODSIn 239 cases of renal transplant,merits of methods of the traditional HLA six antigen matcheing criteria, cross reaction groups standard and Eplets mismatch based on HLAMatchmaker standard were compared respectively.

RESULTSThe number of mismatchs with three methods in 239 cases, were grouped according to low-high mismatchs. The results revealed that HLAMatchmaker algorithm could significantly increase the number of low mismatchs group 54 (22.6%), compared with the HIA group 19(7.9%) and CREGs group 32 (13.4%). The comparison was discovered statistical significance among the three groups (P<0.001), so the comparison between each group was.

CONCLUSIONHLAMachmaker of donor-recipients matching, is a more efficient, time-saving and high sensitivity matching solution to allograft renal transplantation.

Adolescent ; Adult ; Aged ; Female ; Histocompatibility Testing ; methods ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Software ; Transplantation, Homologous ; Young Adult