1.Effect of 5-aminolevulinic acid-mediated photodynamic therapy on human gastric cancer xenografts in nude mice in vivo.
Guang-jun ZHOU ; Zong-hai HUANG ; Jin-long YU ; Zhou LI ; Lian-shu DING
Chinese Journal of Gastrointestinal Surgery 2008;11(6):580-583
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
2.Time window characteristics of cultured rat hippocampal neurons subjected to ischemia and reperfusion.
Zhong XU ; Ru-xiang XU ; Bao-song LIU ; Xiao-dan JIANG ; Tao HUANG ; Lian-shu DING ; Jun YUAN
Chinese Journal of Traumatology 2005;8(3):179-182
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
3.Construction of eukaryotic expression vector with brain-derived neurotrophic factor receptor trkB gene.
Tao HUANG ; Xiao-dan JIANG ; Zhong XU ; Jun YUAN ; Lian-shu DING ; Yu-xi ZOU ; Ru-xiang XU
Chinese Journal of Traumatology 2005;8(3):142-146
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
4.A Study on the Connection between the Incidence of Postherpetic Neuralgia and Serum Ionized Calcium.
Xue-Ying ZHAI ; Rui-Yong CHENG ; Ling-Tao KONG ; Lei YANG ; Jin-Li LI ; Shu-Juan DING ; Lian-Ping LI
Chinese Medical Journal 2015;128(22):3106-3108
Age Distribution
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Aged
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Aged, 80 and over
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Calcium
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blood
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Female
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Herpes Zoster
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blood
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epidemiology
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Humans
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Incidence
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Male
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Middle Aged
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Neuralgia, Postherpetic
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blood
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epidemiology
5.Construction of recombinant lentiviral vector of Tie2-RNAi and its influence on malignant melanoma cells in vitro.
Xiu-ying SHAN ; Zhao-liang LIU ; Biao WANG ; Guo-xiang GUO ; Mei-shui WANG ; Fu-lian ZHUANG ; Chuan-shu CAI ; Ming-feng ZHANG ; Yan-ding ZHANG
Chinese Journal of Plastic Surgery 2011;27(4):277-283
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
6.Clinical research on reconstitution of CD4+CD25+ T cells after haploidentical bone marrow transplantation.
Lian-Nng DUAN ; Li DING ; Hong-Min YAN ; Mei XUE ; Jing LIU ; Ling ZHU ; Heng-Xiang WANG ; Shu-Quan JI
Journal of Experimental Hematology 2010;18(1):177-180
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
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CD4 Lymphocyte Count
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CD4-Positive T-Lymphocytes
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immunology
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Graft vs Host Disease
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etiology
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Humans
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Interleukin-2 Receptor alpha Subunit
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metabolism
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Leukemia
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immunology
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surgery
7.Clinical investigation on treatment of children leukemia with non-T-cell depleted haploidentical bone marrow transplantation.
Hong-Ming YAN ; Shu-Quan JI ; Hui-Ren CHEN ; Lian-Ning DUAN ; Ling ZHU ; Jing LIU ; Mei XUE ; Li DING ; Heng-Xiang WANG
Journal of Experimental Hematology 2008;16(1):101-105
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
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adverse effects
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methods
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Child
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Child, Preschool
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Female
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Graft vs Host Disease
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prevention & control
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Haploidy
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Humans
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Infant
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Leukemia
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therapy
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Leukemia, Myeloid, Acute
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therapy
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Male
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Precursor Cell Lymphoblastic Leukemia-Lymphoma
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therapy
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T-Lymphocytes
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cytology
8.Effect of photodynamic therapy with 5-aminolevulinic acid on human gastric cancer cells in vitro.
Zong-hai HUANG ; Guang-jun ZHOU ; Jin-long YU ; Zhou LI ; Lian-shu DING ; Ru-xiang XU ; Xiao-dan JIANG
Journal of Southern Medical University 2006;26(3):255-257
OBJECTIVETo investigate the effect of 5-aminolevulinic acid (5-ALA)-mediated photodynamic therapy (PDT) on MGC-803 human gastric cancer cells in vitro.
METHODSMGC-803 human gastric cancer cells were treated with 5-ALA at various concentrations followed by laser irradiation. The cells were also treated with 5-ALA at the same concentration before laser exposure at various doses. PDT-induced phototoxicity of the cells was determined by MTT assay.
RESULTSAfter laser exposure of the cells at the same dose (25.0 J/cm(2)), the cell survival rates decreased significantly with incubation of the cells with 5-ALA at 0.25, 0.5, 1.0, 2.0 and 4.0 mmol/L, respectively (F=266.39, P<0.001), but 2.0 and 4.0 mmol/L ALA showed no significant difference in lowering the cell survival rates (P>0.05). Following treatment with the same 5-ALA concentration (1 mmol/L), the cell survival rates decreased in response to increased laser doses (at 6.25, 12.5, 25.0, 50.0, and 100 J/cm(2), respectively, F=226.31, P<0.0001). Without laser exposure, the survival rate of the cells did not significantly change for different 5-ALA concentrations (F=0.79, P=0.5383), nor did it undergo obvious variation in response to different laser doses without 5-ALA incubation (F=0.61, P=0.6551).
CONCLUSIONSThe damage of MGC-803 cells by PDT increases with 5-ALA concentration within a relative lower range and is proportional to the laser doses delivered. Without 5-ALA treatment, the laser at the chosen dose cannot produce photodynamic effect and ALA itself is nontoxic. ALA-mediated PDT appears to be a promising therapy for gastric cancer.
Aminolevulinic Acid ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Dose-Response Relationship, Drug ; Dose-Response Relationship, Radiation ; Humans ; Lasers ; Photochemotherapy ; Photosensitizing Agents ; pharmacology ; Stomach Neoplasms ; drug therapy ; pathology
9.Application of fibrotic bronchoscopy in the diagnosis of pulmonary diffuse infiltration following bone marrow transplantation.
Heng-Xiang WANG ; Bo ZHANG ; Jing LIU ; Lian-Ning DUAN ; Li DING ; Mei XUE ; Ling ZHU ; Hong-Min YAN ; Hui-Ren CHEN ; Shu-Quan JI
Journal of Experimental Hematology 2008;16(4):946-949
In order to evaluate the diagnostic value of fibrotic bronchoscopy (FB) in the pulmonary infiltration following bone marrow transplantation (BMT), 18 patients with pulmonary complications after BMT from November 2003 to March 2006 were performed with FB. Bronchoalveolar lavage (BAL) and brushing were performed in patients who had received short-term empirical therapy without good response, and transbronchial lung biopsy (TBLB) was carried out in 3 cases. The results showed that 9 out of 10 cases with pulmonary infection, including bacterial pneumonia (n = 3), aspergillosis (n = 2), pneumocystis carinii pneumonia (n = 3) and viral infection (n = 1) were diagnosed by using FB. One case was diagnosed as tuberculosis after open lung biopsy following negative results from twice BAL. 2 out of 8 cases were diagnosed by TBLB as noninfectious pulmonary complications. In conclusion, FB, especially with BAL, is a safe and useful procedure for the evaluation of pulmonary complications, which is particularly suitable for diagnosis of pulmonary infection after BMT. Furthermore, TBLB should be recommended in order to avoid open lung biopsy, if the patients tolerate the operation.
Adolescent
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Adult
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Bone Marrow Transplantation
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adverse effects
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Bronchoalveolar Lavage Fluid
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microbiology
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parasitology
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Bronchoscopy
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Child
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Female
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Humans
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Lung Diseases
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diagnosis
;
etiology
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Male
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Middle Aged
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Pneumonia
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diagnosis
;
etiology
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microbiology
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Pneumonia, Pneumocystis
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diagnosis
;
etiology
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microbiology
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Young Adult
10.Neurological complications following haploidentical bone marrow transplantation.
Heng-Xiang WANG ; Shu-Quan JI ; Ling ZHU ; Jing LIU ; Mei XUE ; Li DING ; Lian-Ning DUAN ; Hong-Min YAN ; Hui-Ren CHEN
Journal of Experimental Hematology 2005;13(6):1128-1130
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