ObjectiveTo investigate the variation of peripheral blood CD34+ cells during the hematopoietic stem cell mobilization,and its influence on the collecting timing and results.Methods Twenty-seven cases of peripheral blood hematopoietic stem cell mobilization and collection from April 2010 to December 2011 were analyzed,including 13 autologous cases mobilized with chemotherapy combined with granulocyte colony-stimulating factor (G-CSF,10 μg· kg-1 · d-1) and 14 cases of healthy donors mobilized with only G-SCF (7.5 μg · kg- 1 · d- 1 ).The number of peripheral blood CD34+ cells was counted,and its correlation with the yield of mononuclear cells (MNCs) and CD34+cells was analyzed.ResultsMNCs (5.84 ± 1.48) × 108/kg and CD34+ cells (3.93 ± 2.16) × 106/kg were obtained in healthy donors,and (6.58 ± 3.72) × 108/kg MNCs and (3.98 ± 3.06) × 106/kg CD34+ cells were obtained in autologous cases,respectively.There was only 1 failure in autologous cases.The peak of peripheral blood CD34+ cells in autologous cases appeared at day 4 after the treatment of G-CSF,and in healthy donors the number of peripheral blood CD34+ cells at day 5 was still in ascendant phase.The CD34+ cells/kg in the collection products were positively correlated with the percentage and absolute value of peripheral blood CD34+ cells.The cases ratio of CD34+ cells≥2× 106/kg in the products of single collection was up to 76.2％ (16/21) in the cases with peripheral blood CD34+ cells absolute value greater than 20/μl.ConclusionThe number of peripheral blood CD34+ cells was an important monitoring indicator in hematopoietic stem cell mobilization and collection,CD34+ cell absolute value ≥20/μl could be used as collection threshold.

Objective To evaluate the feasibility of precise hepatic segmentectomy or subsegmentectomy using intraoperative image-guided interventional intravascular segmental vessel balloon catheter occlusion of the segmental hepatic artery and portal vein.Methods 6 patients with liver resection carried out from 2011.3-2011.8 were retrospectively analyzed.Results The mean operating time was (270.83±21.31) min,the median of blood loss was 800 ml,the median of intraoperative transfusion volume was 450 ml.The tumors were mainly located in segments Ⅴ,Ⅵ,Ⅶ,Ⅷ.The mean diameter of tumor was (5.67±1.03) cm.Postoperative liver function in the first postoperative day showed the mean alanine aminotranferase (ALT) was (570.00±157.76) U/L,the mean aspirate aminotrarsferase (AST) was (410.00 ±189.94) U/L,and the mean total bilirubin (TBIL) was (10.83± 1.60) mmol/L.Liver function recovered to normal within 7 days.There was intestinal leakage and wound dehiscence in one patient,pleural and effusion in two patients.Conclusion Imageguided interventional intravascular segmental vessel balloon catheter occlusion was a safe and efficacious maneuver.This technique allowed hepatic segmentectomy or subsegmentectomy to be carried out,decreased intraoperative bleeding,and protected the function of the liver remnant.

Total RNA was isolated from Siraitia grosvenorii fruit by the method of modified Trizol, according to S. grosvenorii fruit characteristics of rich phenols, polysaccharide, oil and proteins. The OD260/280, OD260/230, RNA integrity (RIN) and yield of the total RNA with this method were 2.01, 2.02, 9.50 and 260 mirog.g-1, respectively. The open reading frame (ORF) of dehydroascorbate reductase (DHAR), named as SgDHAR, was cloned by rapid amplification of cDNA ends (RACE) and RT-PCR method from S. grosvenorii. The GenBank accession number for this gene is KC907731. The SgDHAR gene contains a full-length cDNA of 1,252 bp including ORF of 819 bp and encodes a predicted protein of 272 amino acids. The molecular mass is 30.217 7 kD and the isoelectric point is 8.76. Homology comparison showed that it shared 87% nucleotide sequence homology with Cucumis sativus. Expression patterns using qRT-PCR analysis showed that SgDHAR was mainly expressed in fruit and stem, followed by flower, and was lowest in root, while the expression level was 6.83 times in triploid. T than that in diploid. Therefore, SgDHAR gene may be involved in abortion of triploid seedless S. grosvenorii.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Cucurbitaceae ; chemistry ; genetics ; DNA, Complementary ; genetics ; Flowers ; chemistry ; genetics ; Fruit ; chemistry ; genetics ; Molecular Conformation ; Open Reading Frames ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Roots ; chemistry ; genetics ; Plant Stems ; chemistry ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Protein Structure, Secondary ; RNA, Plant

OBJECTIVETo evaluate efficacy of adjuvant chemotherapy after D2 dissection on survival for patients with gastric cancer.

METHODSRandomized clinical trials (RCT) that compared adjuvant chemotherapy after D2 dissection with D2 dissection alone for gastric cancer were searched with Pubmed, Cochrane, Embase and CBM databases. Eligible trials published between 1990 and 2012 were included in the study. The quality of RCTs was assessed by the Jadad scale. Data synthesis and statistical analysis were performed by RevMan 5.1 software.

RESULTEight RCTs with 3633 patients were included in this study. Among them, 1824 patients received adjuvant chemotherapy and 1809 patients didn't. Adjuvant chemotherapy was associated with a significant benefit in terms of overall survival (RR = 0.76, 95% CI: 0.69-0.84), disease free survival (RR = 0.72, 95%CI: 0.66-0.80) and recurrence rate (RR = 0.69, 95% CI: 0.62-0.77).

CONCLUSIONAdjuvant chemotherapy was associated with survival benefit for gastric cancer after D2 dissection.

Chemotherapy, Adjuvant ; Disease-Free Survival ; Female ; Gastrectomy ; Humans ; Male ; Neoplasm Recurrence, Local ; Prognosis ; Randomized Controlled Trials as Topic ; Stomach Neoplasms ; drug therapy ; mortality ; surgery ; Survival Rate

Objective To investigate the variant distribution of voltage-gated sodium channel type Ⅰ (Nav1.1) on the different types of neurons in rat hippocampus. Methods In brain tissue sections from adult SD rats, the expression of Nav1.1 on the different types of neurons in rat hippocampus was detected and observed by immunohistochemistry (DAB staining) and laser confocal microscopy. Results Nav1.1 protein was expressed weakly on the soma of pyramidal cells and granular cells of the dentate gyrus, while strongly expressed on the scattered interneurons.Conclusions In the hippocampus of SD rats, the expression of Nav 1.1 protein on interneurons was strong, but weak on the projection neurons (pyramidal cells and granular cells of the dentate gyms). It can be inferred that on different types of neurons, different subtypes of sodium channels are predominately expressed.

Classical Swine Fever Virus (CSFV) E2 protein eukaryotic expression plasmid pVAXE2 was constructed. The plasmid pVAXE2 was transformed into Salmonella choleraesuis C500 (S. C500) attenuated vaccine strain by electroporation to generate Salmonella choleraesuis engineering strain S. C500/pVAXE2. The characterization of S. C500/pVAXE2 in morphology, growth, biochemistry and serology indicated that it retained the same properties as its original strain S. C500 with exception of kanamycin resistance originated from the plasmid pVAXE2. The plasmid stable in the bacteria after 15 passages. Kunming mice and rabbits were vaccinated three times at two weeks interval with S. C500/pVAXE2 in oral and intramuscular routes at the dosage of 1 x 10(8) CFU for mice and 2 x 10(9) CFU for rabbits each time. The specific antibody response against CSFV and Salmonella choleraesuis was detected by ELISA. Two weeks after the third boost the immunized rabbits were challenged with 20 ID50 of hog cholera lapinized virus (HCLV), followed by a virulent strain of Salmonella choleraesuis two week later than HCLV challenge. The results showed that all immunized mice and rabbits produced significant antibodies against CSFV and Salmonella choleraesuis, and the immunized rabbits demonstrated the effective protection against the challenge of HCLV and virulent Salmonella choleraesuis. These results indicated the potential of developing multiplex swine DNA vaccine by using this bacteria as the vector.
Animals ; Classical Swine Fever ; immunology ; prevention & control ; virology ; Classical swine fever virus ; genetics ; immunology ; Mice ; Rabbits ; Salmonella arizonae ; genetics ; Swine ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology ; Viral Vaccines ; immunology

OBJECTIVETo summarize and analyze the diagnosis and arthroscopic treatment of osteochondral lesion of talus (OLT).

METHODSFrom 2000 to 2005 the data of 34 patients of OLT of the talus were retrospectively studied, including the symptom, physical examination, image, arthroscopic treatment All patients took X-ray and MRI examination before the arthroscopic surgery. Arthroscopic debridement was performed for all patients, in addition to drilling in 5 cases, and microfracture in 18 cases. Before operation, ankle-hindfoot score of American Orthopaedic Foot and Ankle Society (AOFAS) was 71 +/- 8, and the score of pain (visual analogue scale, VAS) was 7.5 +/- 1.3.

RESULTSWeight-bearing pain of the ankle joint aggravated after exercise was the predominant complaint of OLT. X-ray examination was negative in 13 cases, and all lesions were detected by MRI, which was significantly better than X-ray (chi2 = 16.07, P < 0. 001). Thirty-one patients were followed up for an average of 28 months. The average post-operative AOFAS was 91 +/- 9 (t = 9.147, P < 0.001); And VAS was 2.4 +/- 2. 3, which was significantly lower than that in pre-operation (t = 10.853, P < 0.001). Of the 31 patients, 27 (87.1%) had good or excellent results.

CONCLUSIONSMRI could improve the accuracy of diagnosis. The results of arthroscopic treatment for OLT are satisfactory.

Adolescent ; Adult ; Ankle Injuries ; diagnosis ; surgery ; Arthroscopy ; methods ; Cartilage, Articular ; injuries ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Talus ; injuries ; Treatment Outcome

BACKGROUND/AIMS: The aim of this study was to investigate whether genetic variations at positions -1082, -819, and -592 in the interleukin (IL)-10 promoter affect IL-10 production in children with irritable bowel syndrome (IBS). METHODS: Ninety-four children with IBS and 102 children as healthy controls (HCs) were enrolled. Genomic DNA was extracted, and IL-10 -1082, -819, and -592 polymorphisms were detected by direct sequencing from all participants. Peripheral blood mononuclear cells (PBMCs) from 46 IBS children and 38 HCs were isolated and cultured with and without 5 ng/mL Escherichia coli lipopolysaccharide (LPS). IL-10 levels in the culture supernatants were measured by enzyme-linked immunosorbent assay. RESULTS: There were no significant differences in the distribution of IL-10 -1082, -819, and -592 polymorphisms or in the allele and haplotype frequencies between IBS children and HCs. PBMCs from children with IBS had significantly lower IL-10 levels after LPS stimulation than PBMCs from HCs (p=0.011); however, LPS-induced IL-10 levels in PBMCs with different genotypes of -819 and -592 polymorphisms were not significantly different between IBS patients and HCs. CONCLUSIONS: Although significantly lower LPS-induced IL-10 production by PBMCs was noted, it is unlikely that IL-10 production was fully genetically determined in our IBS children. ClinicalTrials.gov identifier: NCT01131442.
Alleles ; Child ; DNA ; Escherichia coli ; Genetic Variation ; Genotype ; Haplotypes ; Humans ; Interleukin-10 ; Interleukins ; Irritable Bowel Syndrome

BACKGROUNDLow-power helium-neon (He-Ne) lasers have been increasingly widely applied in the treatment of cardiovascular diseases, and its vasodilation effect has been proven. The aim of this study was to determine the effects of low-power He-Ne laser irradiation directed at the precardial region of Wistar rats on capillary permeability in the myocardium and the expression of myocardial vascular endothelial growth factor (VEGF).

METHODSSixteen rats were divided randomly into control and irradiated groups (n = 8, each). A He-Ne laser (632.8 nm) was applied to the irradiated group with a dose of 60.5 J/cm(2). Ferritin was perfused into the left femoral vein and capillary permeability was examined under an electron microscope. VEGF expression in the myocardium was investigated by immunohistochemical methods, RT-PCR, and image analysis.

RESULTSThe ultrastructures of the myocardial capillaries were examined. Compared to the control group, more high-density granules (ferritin), which were present within the capillary endothelium and the mitochondrions of myocardial cells in the internal layer of the myocardium, were observed in the irradiated group. VEGF staining of the myocardium was stronger in the irradiated group than that in the control group. The optic density of the irradiated group (0.246 +/- 0.015) was significantly higher than that of the control group (0.218 +/- 0.012, P < 0.05). Finally, the levels of RT-PCR products of VEGF165 mRNA were 2.79 times higher in irradiated rats than in the control rats.

CONCLUSIONSOur study demonstrates that He-Ne laser irradiation (in doses of 60.5 J/cm(2)) increases myocardial capillary permeability and the production of VEGF in myocardial microvessels and in myocardium. Our study provides experimental morphological evidence that myocardial microcirculation can be improved using He-Ne laser irradiation.

Animals ; Capillary Permeability ; radiation effects ; Female ; Heart ; radiation effects ; Immunohistochemistry ; Lasers ; Microscopy, Electron ; Myocardium ; metabolism ; ultrastructure ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; analysis ; genetics

BACKGROUNDRemodeling of the anterior cruciate ligament (ACL) graft usually takes longer than expected. Gene therapy offers a radical different approach to remodeling of the graft. In this study, the internal ribosome entry site (IRES) sequence was used to construct a new recombinant adenovirus which permits co-expression of transforming growth factor-beta1 (TGFbeta1) and vascular endothelial growth factor 165 (VEGF165) genes (named Ad-VEGF165-IRES-TGFbeta1). We investigated the effects of the new adenovirus on the migration of and matrix synthesis by ACL fibroblasts.

METHODSAdenoviral vector containing TGFbeta1 and VEGF165 genes was constructed. ACL fibroblasts were obtained from New Zealand white rabbits. After ACL fibroblasts were exposed to Ad-VEGF165-IRES-TGFbeta1, the expression of VEGF165 and TGFbeta1 proteins were assessed by enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis. Bioassay of VEGF165 and TGFbeta1 proteins were assessed by Western blotting analysis. Proliferation and migration of ACL fibroblasts were assessed by in vitro wound closure assay. Gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers were assessed by real-time PCR.

RESULTSThe results showed the successful construction of a recombinant co-expression adenovirus vector containing TGFbeta1 and VEGF165 genes. Co-expression of TGFbeta1 and VEGF165 can induce relatively rapid and continuous proliferation of ACL fibroblasts and high gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers.

CONCLUSIONCo-expression of TGFbeta1 and VEGF165 genes has more powerful and efficient effects on the migration of and matrix synthesis by ACL fibroblasts.

Adenoviridae ; genetics ; Animals ; Anterior Cruciate Ligament ; cytology ; metabolism ; Cell Movement ; Cells, Cultured ; Collagen ; genetics ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Fibroblasts ; physiology ; Fibronectins ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; Wound Healing