Animals ; Hyperbaric Oxygenation ; Inflammation ; Interferon-gamma ; metabolism ; Interleukin-6 ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; metabolism ; therapy ; Tumor Necrosis Factor-alpha ; metabolism

To investigate the feasibility to produce crocetin in tobacco plants.The coding region of zeaxanthin cleavage dioxygenase (CSzCD) gene from Crocus sativus L. was inserted into the downstream of the cauliflower mosaic virus (CaMV) 35S promoter of a binary vector pBI121, and integrated into the genome of Nicotiana tabacum L. Twenty-one transgenic lines were identified by genomic southern blotting analysis. Western blotting and HPLC analysis of the leaf extracts of transgenic tobacco showed that crocetin was produced in CSzCD gene-transformed plants, while no crocetin was found in the untransformed tobacco.

OBJECTIVETo explore the therapeutic methods of fracture and dislocation of coccyx and evaluate its curative effects.

METHODSFrom May 2002 to March 2010,56 patients with fracture and dislocation of coccyx were divided into surgical treatment group and non-surgical treatment group. There were 7 males and 20 females in surgical treatment group with an average age of (48.1 +/- 0.6) years (ranged, 29 to 62 years), treated with open reduction and mini-plate internal fixation. There were 8 males and 21 females in non-surgical treatment group with an average age of (47.5 +/- 0.9) years (ranged, 19 to 54 years),treated with manipulative reduction. All patients were underwent X-ray examination and were finally diagnosed before treatment. Clinical symptoms and Visual Analogue Scales (VAS) of all patients were statistically analyzed before and after treatment.

RESULTSThere was no significant difference between two groups in gender, age, BMI index and VAS evaluation. All patients were followed up from 12 to 25 months with an average of 17.2 months. In surgical treatment group,there were 26 cases with I/a incision and 1 case with II/a incision; the excellent rate of clinical symptom was respectively 92.6% and 100% at leaving hospital and final follow-up; the improvement rate of VAS was 97.6% and was excellent result;internal fixtures were removed at the 1 to 2 years after treatment and no unwell symptoms occurred; VAS of all patients in the group was 0 point. In non-surgical treatment group,the excellent rate of clinical symptom was respectively 72.4% and 82.8% at leaving hospital and final follow-up; the improvement rate of VAS was 72.1% and was good result. There was significant difference in clinical results between two groups (P<0.05).

CONCLUSIONThe results indicated that fracture and dislocation of coccyx should be treated in time. For the treatment of patients with manipulative reduction failures, instability reduction by X-ray examination and serious rectal irritation, open reduction and mini-plate internal fixation can obtain satisfactory results.

Adult ; Bone Plates ; Coccyx ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Joint Dislocations ; surgery ; Male ; Manipulation, Spinal ; Middle Aged ; Spinal Fractures ; surgery

This paper reviews the new progress in the research of fractional carbon dioxide laser in treating hypertrophic scar after burn injury, which remains a challenging problem for burn care surgeons. There have been many traditional therapeutic approaches, such as compression remedy, operation, and so on. However, a satisfactory method is lacking to date. In recent years, the newly developed fractional carbon dioxide laser has been employed to treat different kinds of scars, and it has been proved to be effective in terms of an improvement of scar color, texture, and rigidity. It seems to be a promising method for scar treatment in future.
Burns ; surgery ; Cicatrix ; surgery ; Cicatrix, Hypertrophic ; surgery ; Humans ; Lasers, Gas ; Treatment Outcome

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

BACKGROUNDO(6)-methylguanine-DNA-methyltransferase (MGMT) is a specific DNA revising enzyme transferring alkylated groups from DNA to its cysteine residue to avoid the abnormal twisting of DNA. Therefore, it is one of the drug resistant genes targeted in the treatment of cancer. This study explored the protective effect of MGMT gene transferred into mammalian cells.

METHODSMammalian expression vector containing the MGMT gene cloned from human hepatocytes by RT-PCR was constructed and transferred into K562 cells and human peripheral blood mononuclear cells (PBMCs) via liposome, then assayed for gene expression at RNA and protein levels. MTT assay was used to check the drug resistance of cells transfected with MGMT gene.

RESULTSMGMT gene was successfully cloned. Real-time PCR showed that the mRNA expression in gene transfected groups in K562 cell line and PBMC were 13.4 and 4.0 times that of the empty vector transfected groups respectively.

RESULTSof Western blotting showed distinct higher expression of MGMT in gene transfected group than in other two groups. The IC(50) values increased to 7 and 2 times that of the original values respectively in stable transfected K562 cells and transient transfected PBMC.

CONCLUSIONThe alkylating resistance of eukaryotic cells is enhanced after being transfected with MGMT gene which protein product performs the protective function, and may provide the reference for the protective model of peripheral blood cells in cancer chemotherapy.

Blotting, Western ; Cell Survival ; drug effects ; genetics ; physiology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Gene Expression Regulation, Enzymologic ; Green Fluorescent Proteins ; genetics ; metabolism ; Hepatocytes ; cytology ; drug effects ; metabolism ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; drug effects ; metabolism ; Microscopy, Fluorescence ; Nitrogen Mustard Compounds ; pharmacology ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; metabolism ; physiology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

It has need to separate red blood cells (RBC) from marrow graft in ABO group unmatched BMT and auto-BMT with purging tumor cells, the separating effect of methylcellulose was observed. The mixture of 0.5% methylcellulose and bone marrow was laid up in an open transfusion system, and then sedimentation of RBC was performed in the transfusion tube. The separating results of 18 marrow grafts showed that the recovery rates of mononuclear cells and CD34(+) cells were (83.8 +/- 55.2)% and (90.3 +/- 7.2)%, respectively. RBC residual rate was (4.3 +/- 1.5)%. The yield of CFU-GM was (60.8 +/- 22.4)/2 x 10(5) MNC, and there was no difference to [(69.8 +/- 23.4)/2 x 10(5) MNC] yielded from same marrow samples, separated by Ficoll-Hypaque separation. It is concluded that this method could be used for bone marrow transplantation.
Bone Marrow Transplantation ; methods ; Cell Separation ; methods ; Erythrocytes ; immunology ; Humans ; Methylcellulose ; pharmacology

OBJECTIVETo investigate the effects of transforming growth factor beta1 (TGF-beta1) on dendritic cells (DC).

METHODSMurine bone marrow cells were cultured with different cytokine combinations to develop immature DC (imDC, GM-CSF only) and TGFbeta-DC (GM-CSF + TGF-beta1), and their responses to lipopolysaccharide (LPS) stimulation were observed. The cell ultrastructure was observed by transmission electron microscopy and their phenotypes were assessed by flow cytometry (FCM). The allogeneic stimulating capacity of DC was assayed by mixed lymphocyte reaction (MLR) with BrdU incorporation. IL-12p70 protein was detected by ELISA and the expressions of Toll-like receptor 4 (TLR4) on DCs were analyzed with semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSCompared to imDC, the TGFbeta-DC had no significant alterations in ultrastructure after LPS stimulation. The expressions of CD80, CD86 were lower on TGFbeta-DC than on imDC [(4.14 +/- 0.95)% vs (13.90 +/- 7.22)%; (8.60 +/- 0.75)% vs (20.63 +/- 5.03)%, P < 0.05, both]. The TGFbeta-DC kept their immature morphology after LPS stimulation, but the expressions of I-Ab and CD80 were slightly increased. After 96 h MLR, TGFbeta-DC had weaker stimulating capacity than imDC did, especially when DC/T cells ratios were 1:4 and 1:1 (P < 0.05, both). TGFbeta-DC showed impaired IL-12p70 production and down-regulation of TLR4 expression.

CONCLUSIONSTGF-beta1 can inhibit the expression of co-stimulatory molecules on DC. The TGFbeta-DC is resistant to maturation stimulus (LPS) and might be linked with TLR4 down-regulation.

Animals ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dendritic Cells ; drug effects ; physiology ; Down-Regulation ; drug effects ; Lipopolysaccharides ; pharmacology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, Cell Surface ; immunology ; metabolism ; Toll-Like Receptor 4 ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1

OBJECTIVETo probe into the therapeutic effect of catgut implantation at acupoint on exercise fatigue and to study on the mechanism.

METHODSCatgut implantation was given at main points Guanyuan (CV 4), Shen shu (BL 23), Mingmen (GV 4), Zusanli (ST 36), Sanyinjiao (SP 6) in 41 players during training for 2 courses. Training-related condition and changes of blood testosterone (T) and hemoglobin (Hb) before and after training were observed.

RESULTSAfter treatment, sleeping quality, the mental state at the time of getting out of bed in the morning, appetite, training wish, training stamina, training quality, training attention, etc. in the player significantly improved, and blood T and Hb levels significantly increased.

CONCLUSIONCatgut implantation at acupoints can significantly improve exercise fatigue in the player and the mechanism is possibly related with increase of testosterone and hemoglobin levels.

Acupuncture Points ; Acupuncture Therapy ; methods ; Adolescent ; Adult ; Catgut ; Exercise ; Fatigue ; therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional

BACKGROUNDVertebrobasilar artery stenosis is an important cause of ischemic posterior circulation strokes. This study aimed at evaluating the safety and efficacy of treatment including conservative therapy alone and conservative plus endovascular therapy for elderly patients with symptomatic vertebrobasilar artery stenosis.

METHODSPatients older than 60 years with symptomatic vertebrobasilar artery stenosis (> or = 50%) confirmed by cerebral angiography were enrolled. All of them were treated with medical therapy and some with additional stent-assisted angioplasty (the stenting subgroup). Their clinical, imaging, intervention and follow-up data were analyzed.

RESULTSOne hundred and seventeen consecutive elderly patients (100 men, mean age (68.1 +/- 5.1) years) were enrolled and followed up for a mean time of 28.4 months; 81.7% of them were symptomatically resolved or improved; a stroke rate of 5.1% and a stroke-related death rate of 1.7% were found among them during the hospitalization and follow-up. In the stenting subgroup, 78 balloon expandable stents were employed in the 70 patients with a technical success rate of 98.7% and the mean degree of stenosis was significantly reduced from (81.7 +/- 14.3)% before stenting to (8.3 +/- 4.2)% after stenting (P < 0.001). Four (5.7%) periprocedural strokes occurred, of whom two led to death within 30 days after the procedure. During the follow-up (mean 27.7 months), sixty of the surviving 68 patients in the stenting subgroup were symptomatically resolved or improved. Only one (1.5%) posterior circulation stroke occurred, while duplex ultrasound scan of 34 patients demonstrated 10 (29.4%) in-stent restenosis.

CONCLUSIONSAppropriate utilization of conservative therapy alone and conservative plus endovascular therapy may improve short-term clinical outcomes for elderly patients with symptomatic vertebrobasilar artery stenosis. Furthermore, stent-assisted angioplasty is technically feasible and relatively safe in elderly patients.

Aged ; Aged, 80 and over ; Angioplasty, Balloon ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prospective Studies ; Stents ; Treatment Outcome ; Vertebrobasilar Insufficiency ; therapy