OBJECTIVETo investigate the effect of 5-aminolevulinic acid (ALA)-mediated photodynamic therapy (PDT) on human gastric cancer xenografts in vivo and to explore its potential tumoricidal mechanism.

METHODSCultured MGC-803 human gastric cancer cells were injected below the skins of the nude mice to develop the tumor model. The tumor-bearing nude mice were examined under the Leica LT-9 MACIMSYSPULS to detect the fluorescence. The tumor volume of day 1, 3, 7, 14, 21 after treatment were measured, and its histological changes were also studied. The tissues of the tumors in nude mice of the control group, light group, 5-ALA group and PDT group were examined with the electron microscope and apoptosis was detected by TUNEL assay.

RESULTSThe tumor model was successfully developed. The tumor in the nude mice emitted the red fluorescence under the Leica LT-9 MACIMSYSPULS. The tumor volumes were (0.189+/-0.010) cm(3), (0.183+/-0.011) cm(3), (0.185+/-0.019)cm(3), (0.182+/-0.015)cm(3) for the control group, light group, 5-ALA group, PDT group, respectively at day 1 after treatment, while at day 3, (0.294+/-0.010) cm(3), (0.280+/-0.013) cm(3), (0.278+/-0.016) cm(3), (0.183+/-0.014) cm(3); at day 7, (0.409+/-0.016) cm(3), (0.411+/-0.009) cm(3), (0.407+/-0.015) cm(3), (0.221+/-0.008) cm(3); at day 14, (0.970+/-0.055) cm(3), (0.976+/-0.054) cm(3), (0.981+/-0.032)cm(3), (0.318+/-0.005) cm(3); at day 21, (1.495+/-0.059) cm(3), (1.513+/-0.057) cm(3), (1.524+/-0.063) cm(3), (0.446+/-0.042) cm(3) (F=1003.086, P=0.000). The histology demonstrated that most tumor blood vessels were congested and necrosis developed after PDT while not in the control group, light group and 5-ALA group. Necrosis and apoptosis were observed in the cells of the tumors of the PDT group examined by TUNEL and electron microscope while not in the cells of the tumors of the other groups.

CONCLUSIONS5-aminolevulinic acid-mediated photodynamic therapy (PDT) can induce injury to human gastric cancer xenografts and inhibit the tumor growth while light only and 5-ALA only can not. 5-aminolevulinic acid-mediated photodynamic therapy (ALA- PDT) appears to be a promising therapy for human gastric cancer, whose mechanism involves in the destruction of the tumors partly by apoptosis other than necrosis.

Aminolevulinic Acid ; therapeutic use ; Animals ; Cell Line, Tumor ; Female ; Humans ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms, Experimental ; Photochemotherapy ; Stomach Neoplasms ; therapy ; Xenograft Model Antitumor Assays

OBJECTIVETo explore cell death and apoptosis in rat hippocampal neurons at different time points after ischemia, hypoxia and reperfusion injury and to elucidate time window characteristics in ischemia neuronal injury.

METHODSHippocampal neurons were obtained from rat embryo and were cultured in vitro. The ischemia and reperfusion of cultured rat hippocampal neurons were simulated by oxygen-glucose deprivation (OGD) and recovery. OGD at different time points (0.25 h to 3.0 h) and then the same recovery (24 h) were prepared. Annexin V-PI staining and flow cytometry examined neuron death and apoptosis at different time after injury.

RESULTSAfter OGD and recovery, both necrosis and apoptosis were observed. At different times after OGD, there were statistically significant differences in neuron necrosis rate (P < 0.05), but not in apoptosis rate (P > 0.05). At recovery, survival rate of hippocampal neurons further decreased while apoptosis rate increased. Furthermore, apoptosis rates of different time differed greatly (P < 0.05). Apoptosis rate gradually increased with significant difference among those of different time points (P < 0.05). However, 2 h after ischemia, apoptosis rate decreased markedly.

CONCLUSIONSApoptosis is an important pathway of delayed neuron death. The therapeutic time window should be within 2 h after cerebral ischemia and hypoxia.

Animals ; Apoptosis ; physiology ; Brain Ischemia ; pathology ; Cell Death ; physiology ; Cell Hypoxia ; Cells, Cultured ; Disease Models, Animal ; Female ; Fetus ; cytology ; Flow Cytometry ; Hippocampus ; pathology ; Neurons ; pathology ; Pregnancy ; Pregnancy, Animal ; Probability ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; Sensitivity and Specificity ; Time Factors

OBJECTIVETo construct an eukaryotic expression vector carrying rat brain-derived neurotrophic factor receptor trkB gene.

METHODSUsing the total RNA isolated from rat brain as template, the trkB gene was amplified by reverse-transcription-polymerase chain reaction (RT-PCR) with a pair of specific primers which contained the restrictive sites of EcoR I and BamH I. The amplified fragment of trkB gene was digested with EcoR I and BamH I, and then subcloned into cloning vector pMD18-T and expression vector pEGFP-C2 respectively. The recombinant plasmids were identified by restriction endonuclease enzyme analysis and PCR.

RESULTSThe amplified DNA fragment was about 1461 bp in length. Enzyme digestion and PCR analysis showed that the gene of trkB had been successfully cloned into vector pMD18-T and pEGFP-C2.

CONCLUSIONSThe trkB gene of rat has been amplified and cloned into the eukaryotic expression vector pEGFP-C2.

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; pharmacology ; Cloning, Molecular ; methods ; Eukaryotic Cells ; Female ; Gene Expression Regulation ; Genetic Therapy ; methods ; Genetic Vectors ; Male ; Models, Animal ; RNA ; analysis ; Rats ; Rats, Wistar ; Receptor, trkB ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Schwann Cells ; cytology ; Sensitivity and Specificity ; Templates, Genetic ; Transfection

Objective To estimate nursing effection of comfeel non adhesive foam dressing in the dressing tracheotomy.Methods 60 patients with tracheotomy were randomly divided into control and experimental group,each group had 30 cases respectively.The experimental group used the comfeel non-adhesive foam dressing for tracheotomy,The control group used ordinary sterile gauze dressing for tracheotomy.The skin condition after tracheotomy,incision infection,the average time between dressing changes and patient comfort were observed.Results 6 patients got skin lesion in experimental group and 12 in the control group,there was obviously different between the two groups(x2 =2.86,P ＜ 0.05).Dressing changing time were(3.63 ± 1.30)days and(1.28 ± 0.50)days in experimental and control group,respectively.there was statistically difference(t--9.24,P＜0.01); Extubation time in experimental group was less than the control group,it was(15.98 ±2.54)days and(21.05 ±3.15)days(t =6.86,P＜0.01).Conclusions Foam dressing is effective in reducing the incidence of skin reactions and wound infection.It can reduce the frequency of dressing change and increase the patient' s comfortable.

The aim of this study was to investigate the reconstitution of CD4(+)CD25(+) T cells after haplo-identical bone marrow transplantation (hiBMT) and its correlation with graft versus host disease (GVHD) and relapse. Peripheral blood samples from 27 patients after hiBMT were harvested and the percentage and absolute counts of CD4(+)CD25(+) T cells were detected by flow cytometry. The correlations of GVHD occurrence and disease relapse with the reconstitution of CD4(+)CD25(+) T cells were analyzed. The results showed that the percentage of CD4(+)CD25(+) T cells of peripheral blood samples increased significantly after G-CSF priming. At day 30 after hiBMT, CD4(+)CD25(+) T cells were recovered to the 20% of normal level, followed by a slowly process in 3 months, and up to one half of the normal level at 180 days. There was no evidence to prove relationship between CD4(+)CD25(+) T cells and acute GVHD, while CD4(+)CD25(+) T cells were increased significantly in the chronic GVHD group. The absolute count of CD4(+)CD25(+) T cells showed no relations with relapse of leukemia during the first year after hiBMT. In conclusions, chronic but not acute GVHD was in relation to the reconstitution of CD4(+)CD25(+) T cells based on the anti-CD25 antibody therapy model for the prevention of GVHD after hiBMT. Further investigation is needed to clarify whether the relapse of leukemia after hiBMT is related to the reconstitution of CD4(+)CD25(+) T cells.
Bone Marrow Transplantation ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; immunology ; Graft vs Host Disease ; etiology ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Leukemia ; immunology ; surgery

OBJECTIVETo investigate the chronic effects of intracoronary autologous bone marrow mononuclear cell (BM-MNCs) transplantation in patients with refractory heart failure (RIHF) after myocardial infarction.

METHODSThirty patients with RIHF (LVEF < 40%) were enrolled in this nonrandomized study, autologous BM-MNCs (5.0 +/- 0.7) x 10(7) were transplanted with via infarct-related coronary artery in 16 patients and 14 patients received standard medical therapy served as control. Baseline and follow up evaluations included complete clinical evaluations, plasma BNP, ANP, ET-1 measurements, echocardiography, PET, and Holter monitoring.

RESULTSBaseline characteristics were similar between the 2 groups. There were no major periprocedural complications. One patient developed ventricular premature contractions during cell infusion for several seconds and recovered spontaneously. Compared to pre-transplantation, plasma BNP and ET-1 significantly decreased and plasma ANP significantly increased at 7 days post transplantation; 6 minutes walking distance increased from (72.1 +/- 31.5) to (201.6 +/- 23.3) m (P < 0.01), LVEF increased 9.9% (P < 0.001) and FDG-PET revealed vital myocardium area increased (10.3 +/- 3.4)% (P < 0.01) at 3 months after BM-MNCs transplantation. At 6 months follow up, the NYHA class improved from (3.4 +/- 0.1 to 2.4 +/- 0.2, P < 0.001) and no patient died and 1 patient rehospitalized due to lower extremities edema. In control group, LVEF decreased 7.2% compared to baseline (P < 0.001) and was significantly lower than transplantation group at 3 months (P < 0.001). At 6 months follow up, the NYHA class increased from (3.5 +/- 0.1 to 3.9 +/- 0.1, P < 0.05), 2 patients died and 10 patients rehospitalized due to aggravated heart failure.

CONCLUSIONPresent study demonstrates that intracoronary transplantation of autologous BM-MNCs is safe and effective for treating patients with RIHF after myocardial infarction.

Bone Marrow Transplantation ; Coronary Vessels ; surgery ; Follow-Up Studies ; Heart Failure ; complications ; Humans ; Mesenchymal Stem Cell Transplantation ; Monocytes ; transplantation ; Myocardial Infarction ; surgery ; Myocardial Ischemia ; complications ; Transplantation, Autologous

Age Distribution ; Aged ; Aged, 80 and over ; Calcium ; blood ; Female ; Herpes Zoster ; blood ; epidemiology ; Humans ; Incidence ; Male ; Middle Aged ; Neuralgia, Postherpetic ; blood ; epidemiology

UNLABELLEDTo explore the occurrence patterns of neurological complications following haploidentical bone marrow transplantation, a series of clinical data as the onset time, etiology, choices of therapies and prognosis in 10 patients with neurological disorders were summarized. The results showed that complications occurred in 10 out of 74 patients after bone marrow transplantation, which include 3 with encephalorrhagia, 3 infection, 1 epilepsy, 1 Guillian-Barr's syndrome, 1 encephalopathy and 1 schizophrenia. 4 patients died of neurological complications.

IN CONCLUSIONneurological complications commonly occurred in haploidentical bone marrow transplantation, and encephalorrhagia might be the main indication that needs intensive care. Moreover, central nerve system infection and peripheral nerve diseases associated with slow immune reconstitution should have clinical interests.

Adolescent ; Adult ; Bone Marrow Transplantation ; adverse effects ; immunology ; Child ; Female ; Histocompatibility ; Humans ; Intracranial Hemorrhages ; etiology ; Leukemia ; surgery ; Male ; Nervous System Diseases ; etiology

OBJECTIVETo construct lentivector carrying Tie2-Small interfering RNA (SiRNA), so as to study its influence on malignant melanoma cells.

METHODSRecombinant plasmid pSilencer 1.0-U6-Tie2-siRNA and plasmid pNL-EGFP were digested with XbaI, ligated a target lentiviral transfer plasmid of pNL-EGFP-U6-Tie2-I or pNL-EGFP-U6-Tie2-II, and then the electrophoresis clones was sequenced. Plasmids of pNL-EGFP-U6-Tie2-I and pNL-EGFP-U6-Tie2-II were constructed and combined with pVSVG and pHelper, respectively, to constitute lentiviral vector system of three plasmids. The Lentiviral vector system was transfected into 293T cell to produce pNL-EGFP-U6-Tie2- I and pNL-EGFP-U6-Tie2-II lentivirus. Then the supernatant was collected to determine the titer. Malignant melanoma cells were infected by both lentiviruses and identified by Realtime RT-PCR to assess inhibitory efficiency.

RESULTSThe recombinant lentiviral vectors of Tie2-RNAi were constructed successfully which were analyzed with restriction enzyme digestion and identified by sequencing. And the titer of lentiviral vector was 8.8 x 10(3)/ml, which was determined by 293T cell. The results of Realtime RT-PCR demonstrated that the lentiviral vectors of Tie2-RNAi could infect malignant melanoma cells and inhibit the expression of Tie2 genes in malignant melanoma cells (P<0.01). There was no significant difference in the expression level (P>0.05) between the two lentiviral vectors of Tie2-RNAi.

CONCLUSIONSLentivector carrying Tie2-SiRNA can be constructed successfully and inhibit the expression of Tie2 gene in vitro significantly. The study will supply the theory basis for the further research on the inhibition of tumor growth in vivo.

Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Melanoma ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, TIE-2 ; genetics ; Transfection

To investigate the efficacy and feasibility of parent non-T cell depleted haploidentical bone marrow transplants (haploidentical BMT) for children with leukemia, the efficacy of haploidentical BMT was evaluated in 8 leukemia children (1.9-9 years) received hematopoietic stem cell transplantation, donors were their parents with HLA-mismatched for two or three loci. Five children were pre-conditioned with a myeloablative regimen consisting of high-dose cytarabine (Ara-C), cyclophosphamide (CY) and total body irradiation. Busulfan (BU), Ara-C and CY were used for preconditioning regimen in other three children. The donors were given G-CSF prior to marrow harvest and the non-T-cell depleted grafts were used. A combination of CsA, MTX, ATG, mycophenolate mofetil (MMF) and CD25 monoclonal antibody were used for GVHD prophylaxis. The results showed that rapid engraftment was observed in all cases after transplantation by cytogenetic evidence. The mean time of neutrophil count exceeded 0.5 x 10(9)/L and the mean time of platelet count exceeded 20 x 10(9)/L were 16 and 17 days after transplantation respectively. Incidence of lethal aGVHD was lower, II-III acute aGVHD was found only in one out of eight patients. Chronic GVHD was observed in five patients, 4 from which showed local cGVHD, one developed extensive cGVHD. During the follow-up of 33 months (range 7-56 months), two patients died from relapsed leukemia, including one relapsed as donor-origin leukemia. Disease-free survival was achieved in the remaining six patients. No death occurred during the follow-up of 6 months. It is concluded that above-mentioned preconditioning regimen and GVHD prophylaxis procedure in non-T-cell depleted bone marrow transplantation from HLA-mismatched parents are effective approaches and safe strategy for the treatment of children leukemia.
Bone Marrow Transplantation ; adverse effects ; methods ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; prevention & control ; Haploidy ; Humans ; Infant ; Leukemia ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; therapy ; T-Lymphocytes ; cytology