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1.
Chinese Journal of Hematology ; (12): 41-46, 2018.
Article in Chinese | WPRIM | ID: wpr-805982

ABSTRACT

Objective@#To explore effects of histone deacetylase inhibitor Belinostat on the immunologic function of dendritic cells (DC) and its possible mechanism.@*Methods@#Cultured mouse bone marrow-derived DC from C57BL/6 mouse in vitro. The experiments were divided into 0, 50, 100 nmol/L Belinostat + immature DC (imDC) group, and 0, 50, 100 nmol/L Belinostat mature DC (mDC). The changes of the ultrastructure of DC were observed by transmission electron microscope (TEM). Immunophenotype and CCR7 expression rate were detected by FCM, and the migration rate was observed by chemotaxis assay. The proliferation of lymphocytes stimulated by different DC was detected by mixed lymphocyte culture reaction. The cytokines in the culture supernatant, including TNF-α, IL-12 and IL-10, were examined by ELISA. RQ-PCR was used to examine the relative expression of mRNA in RelB.@*Results@#Successful cultured and identified the qualified imDC and mDC. Belinostat decreased the expression of CCR7 on imDC [(25.82±7.25)% vs (50.44±5.61)% and (18.71±2.00)% vs (50.44±5.61)%], meanwhile increased the rate on mDC [(71.14±1.96)% vs (64.90±1.47)%]. Chemotaxis assay showed that the migration rate of Belinostat+imDC and Belinostat+mDC group were both decreased, but the difference in imDC was not significant. T lymphocyte proliferation rate stimulated by 100 nmol/L Belinostat+imDC group was lower than imDC group in condition irritation cell∶reaction cell=1∶2 [(227.09±13.49)% vs (309.49±53.69)%]. Belinostat significantly suppressed the secretion of cytokines TNF-α, IL-12 and IL-10 (all P<0.01). The relative expression of mRNA in RelB was slightly decreased in Belinostat+imDC and Belinostat+mDC group (all P<0.05).@*Conclusion@#Belinostat could effectly suppress DC maturation and regulate immune tolerance of DC, which may be due to the down-regulation of mRNA level of RelB in DC.

2.
Article in Chinese | WPRIM | ID: wpr-606976

ABSTRACT

BACKGROUND: Bone marrow necrosis has unspecific clinical features, which is often misdiagnosed or missed due to a lack of the knowledge of the disease. OBJECTIVE: To improve the awareness and vigilance to bone marrow necrosis, and to further explore the clinical manifestations, hematological characteristics, pathogenesis and treatment of bone marrow necrosis. METHODS: The bone marrow necrosis, hematologic neoplasms, solid tumor, bone marrow puncture, bone marrow pathology in Chinese and English served as the search terms to search articles related to bone marrow necrosis in PubMed and Wanfang databases, published from 1941 to 2016. Totally 43 articles were selected for review. RESULTS AND CONCLUSION: Bone marrow necrosis is a rare complication caused by various diseases, clinically characterized by bone pain, fever, anemia, and nucleated red cells and immature neutrophilic leukocytes in the blood smear. Bone marrow aspiration and/or bone marrow biopsy show(s) necrotic features. Its pathogenesis is complex, and it is still poorly understood and needs further research. There is no good treatment for bone marrow necrosis, and the prognosis is poor. Early correct diagnosis and etiological treatment are crucial for the prognosis of bone marrow necrosis.With the improvement of disease awareness, bone marrow cytology, genetics, MRI and hematopoietic stem cell transplantation, bone marrow necrosis is expected to get a better prognosis.

3.
Article in Chinese | WPRIM | ID: wpr-638207

ABSTRACT

Background Laser peripheral iridoplasty (LPI) is widely used in the treatment of glaucoma by flattening the iris and widening angle of anterior chamber (AA).However,no evidence suggests the optimal site of LPI in iris.Objective This study was to compare the therapeutic effects of LPI at different sites of iris for glaucoma.Methods Glaucoma models were established in the right eyes of 40 healthy adult male pigment rabbits by intrachamber injection of 0.1 ml compound carbomer solution with 0.3% carbomer and 0.025% dexamethasone.The models were randomly divided into model control group,corneoscleral limbus group,one spot from corneoscleral limbus group and two spots from corneoscleral limbus group.LPI was performed at corresponding site of iris by 532 nm argon laser with the spot diameter 500 μm,energy 300 mW,exposure time 0.3 seconds and laser number 24 spots,and the rabbits in the model control group did not receive LPI.Intraocular pressure (IOP),coefficient of outflow facility (C value) were measured and calculated with Schi(o)tz tonometer before LPI and 2,4,7,14 and 30 days after LPI,and anterior chamber depth (ACD),AA,anterior chamber angle opening distance within 500 μm radius from scleral spur (AOD500) were measured with ultrasound biomicroscope (UBM).The eyeballs were extracted 30 days after LPI,and the chamber angle were observed under the optical microscope after hematoxylin and eosin staining.The use and care of the animals complied with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.Results UBM showed that compared with the model control group,the anterior chamber angle was evidently widened in all the LPI groups,with the best effectiveness in the one spot from corneoscleral limbus group and the worst one in the two spots from corneoscleral limbus group.Compared with the model control group,the IOP was evidently reduced,and C values,AA and AOD500 were significantly increased in the corneoscleral limbus group,one spot from corneoscleral limbus group and two spots from corneoscleral limbus group after LPI,showing significant differences among the four groups (IOP:Fgroup =16.848,P < 0.01;C value:Fgroup =9.629,P < 0.01;AA:Fgroup =62.336,P<0.01;AOD500:Fgroup =77.779,P < 0.01).IOP was reduced and C value,AA and AOD500 were increased in 2,4,7,14 and 30 days after LPI as compared with before LPI,with significant differences over time (IOP:Ftime =3.041,P =0.011;C value:Ftime =4.311,P<0.01;AA:Ftime =14.627,P<0.01;AOD500:Ftime =20.378,P<0.01).Compared with the model control group,the ACD was significantly increased in the corneoscleral limbus group and one spot from corneoscleral limbus group,and that in the two spots from corneoscleral limbus group was significantly reduced,and the ACD was insignificantly increased over time after LPI (Fgroup =18.017,P<0.01;Ftime =0.022,P =1.000).Hematoxylin and eosin staining showed that the trabecular meshwork and adhesion of tissure were reopened and the anterior chamber angle was widened after LPI.Conclusions LPI can widen anterior chamber angle and lower the IOP.The best therapeutic outcome for glaucoma is displayed when LPI is performed at the iris site corresponding to one spot from the corneoscleral limbus.

4.
Article in Chinese | WPRIM | ID: wpr-636752

ABSTRACT

Primary glaucoma is a group of blinding eye diseases,including of primary open angle glaucoma (POAG),primary angle-closure glaucoma (PACG) and primary congenital glaucoma (PCG).It is thought that the pathogenesis of primary glaucoma is a comprehensive action of genetic factor,environment factor and life style,and the genetic factor plays an important role.CYP1B1 gene was firstly identified as a causal gene for PCG in 1997.After that,thousands of reports on the pathogenesis of POAG focused on CYP1B1 gene mutation.With the developing of research,researches found that CYP1B1 gene to be one of the candidate genes of POAG.The structure and function of CYP1B1 gene,the relationship between CYP1B1 and POAG were reviewed.

5.
Article in Chinese | WPRIM | ID: wpr-403498

ABSTRACT

BACKGROUND: Tumor necrosis factor-α (TNF-α) is one of important cytokines to promote the maturation of dendritic cells. Blockage of TNF-α action by binding with soluble tumor necrosis factor receptor 1 (sTNFR1) may arrest dendritic cells in an immature state and induce stable, long-term tolerance. OBJECTIVE: To construct the lentiviral vectors carrying sTNFR1 gene and investigate sTNFR1 expression in immature dendritic cells. METHODS: Total RNA of human peripheral blood mononuclear cells was taken as a template. The sTNFR1 gene fragment was amplified by RT-PCR, subcloned to the lentiviral vectors pXZ208, and ligated to the enhanced green fluorescent protein (eGFP) reporter gene to establish lentiviral vector, called pXZ9-sTNFR1. DNA sequencing was performed for lentiviral vector identification. Lentivirus was prepared by transfection of 293 FT cells with pXZ9-sTNFR1. Viral titer was determined by eGFP expression. C57BL/6 mouse bone marrow-derived dendritic cells were in vitro cultured with low-dose granulocyte-macrophage colony stimulating factors and interleukin 4. On day 5 of culture, immature dendritic cells were transfected with pXZ9-sTNFR1 recombinant lentiviral supernatant, sTNFR1 transcription was detected by RT-PCR, sTNFR1 protein expression by Western blot analysis. Following sTNFR1 gene modification and lipopolysaccharide stimulation, the phenotype characteristics of dendritic cells were observed. RESULTS AND CONCLUSION: Recombinant plasmid pXZ9-sTNFR1 was successfully constructed. Twenty-four hours after 293 FT cell transfection, eGFP expression was observed and viral titer was over 10<'6> U/L. RT-PCR demonstrated that pXZ9-sTNFRl-transfected immature dendritic cells showed sTNFR1 positive expression. Western blot analysis revealed that sTNFR1 protein appeared in the immature dendritic cells and supernatant following 293 FT cell transfection. On day 5 of culture, dendritic cells expressed low level of class Ⅱ major histocompatibility complex (MHC Ⅱ), as well as CD40, CD86, CD80, molecules. However, following lipopolysaccharide stimulation, dendritic cells expressed high level of MHC Ⅱ, as well as CD40, CD80, and CD86, molecules, exhibiting the phenotype characteristics of mature dendritic cells. But after sTNFR modification, the expression level of MHC Ⅱ, as well as CD40, CD80, and CD86, molecules was not altered obviously. Lentiviral vectors carrying sTNFR1 gene and eGFP reporter gene were successfully constructed, and recombinant lentiviral plasmids with high titer were acquired. Following high efficacy of lentiviral gene transfection, immature dendritic cells stably express sTNFR1 mRNA and protein, which prevents immature dendritic cells from activation by exogenous lipopolysaccharide and maintains the immature state.

6.
Journal of Leukemia & Lymphoma ; (12): 681-683, 2009.
Article in Chinese | WPRIM | ID: wpr-472000

ABSTRACT

Objective To investigate the effect of recombinant human erythropoietin (rhEPO) in patients of hematologic malignancies with aneamia and its relationship of serum erythropoietin levels. Methods Serum EPO (sEPO) level in 80 patients with hematologic malignancies were detected by chemolumimiscence,and treated by recombinant human erythropoietin for patients with Hb<100 g/L. Results The effect on aneamia in tumor patients with remission were significantly higher than that with no-remission. The patients with lower level of sEPO had better respose to treatment by rhEPO than patients with higher level. Conclusion Higher level of sEPO in patients with no-remission hematopoietic tumor, with condition of marrow erythropoiesis aplasia, the effect of rhEPO was poor;, but sEPO level in patients with remission hematopoietic tumor were nearly normal with recovery of marrow erythropoiesis aplasia was effective by use of rhEPO.

7.
Article in Chinese | WPRIM | ID: wpr-407965

ABSTRACT

BACKGROUND: The principal deterrent to the success for hematopoietic stem cell transplantation (HSCT) is the complications after transplantation. The complications are associates with the conditioning regimens in the early stage. The highly-effective preparative regimens of proper dose and low-toxicity are the key to the successful HSCT.OBJECTIVE: To evaluate the curative effects and regimen related toxicity (RRT) of high-dose alkylating-agent-based chemotherapy as conditioning regimens for HSCT in the patients with hematological malignancies.DESIGN: Controlled study with observation.SETTING: Department of Hematology, Affiliated Hospital of Xuzhou Medical College.PARTICIPANTS: A total of 45 patients with leukemia and lymphoma hospitalized at Affiliated Hospital of Xuzhou Medical College from July 1997 to February 2006 were enrolled, including 31 males and 14 females. The median age was 31 years (from 7 to 52 years). The median course was 8 months (from 5 to 17 months) until transplantation.METHODS: Totally 45 patients with leukemia and lymphoma approached or got complete remission were treated by bone marrow transplantation and peripheral blood stem cell transplantation with preparative regimens of high-dose alkylating-agent-based chemotherapy. RRT was graded according to Bearman proposal, from grade 0 (no toxicity) to grade Ⅳ (fatal toxicity). The period of hematopoietic reconstitution, the rates of complete remission and relapse and disease-free survival were statistically observed in transplant recipients.MAIN OUTCOME MEASURES: Occurrence of RRT as conditioning regimens.RESULTS: ①Five patients did not show any toxicity. The greatest toxicity of grade Ⅲ was uncommon (13%, 6/45). Most of the cases with RRT were in grade Ⅰ - Ⅱ and severe oases in grade Ⅲ were rare. In grade Ⅰ - Ⅱ, stomatocace and gastrointestinal toxicity were common respectively of 73% (33/45) and 51% (23/45) which were recovered in short time after treatment; Heart toxicity was rare and only in grade Ⅰ, most of which were tachyoardia and changes of ST-T shape. The increase of transaminase was common in the clinical manifestations of liver RRT except two cases of HVOD.There were four oases of HC, in which one was delayed. RRT on kidney, lungs and CNS was uncommon. ②Totally 43 patients engrafted gained hematopoietic reconstitution, 2 patients died of implant failure (4%). Within the median follow-up period of 37 (8-102) months, 10 patients relapsed, 5 patients died of transplantation-related complications and 28 patients were alive in a disease-free situation (62.2%). The cause of death within 100 days after transplantation was ordinal as acute graft-versus-host disease (GVHD), cytomegalovirus (CMV) interstitial pneumonia, disseminated infections,multiple organ failure and early relapses.CONCLUSION: Alkylating-agent-based conditioning regimens may be well tolerated with low toxicities for HSCT in leukemia and lymphoma.

8.
Article in Chinese | WPRIM | ID: wpr-407901

ABSTRACT

BACKGROUND: The primary qualification of peripheral blood stem cell transplantation (PBSCT) is the effective mobilization and harvesting of hematopoietic stem cells. The mobilization efficacy is closely related to the selection of high-efficacy low-toxicity regimen, the timing of mobilization and harvesting as well.OBJECTIVE: To investigate the efficacy of mitoxantone (MIT) combined with high-dose arabinosylcytosin (Ara-C),followed by granulocyte colony-stimulating factor (G-CSF) alone or combination of G-CSF and granulocyte-macrophage colony-stimulating factor (GM-CSF) on mobilizing PBSCs in patients with hematological malignancies and solid tumors.DESIGN: Controlled study with observation.SETTTNG: Department of Hematology, the Affiliated Hospital of Xuzhou Medical College.PARTICIPANTS: Forty-two patients with hematological malignancies and solid tumors admitted to Department of Hematology, Xuzhou Medical College from September 1998 to December 2006 were involved in this study. They were diagnosed according to FAB classification criteria and new WHO proposals. The involved patients, 25 male and 17 female, averaged 29 years ranging from 7 to 54 years and weighted (52±18) kg. Among them, 12 were patients with acute myeloblastic leukemia, 6 were patients with acute lymphoblastic leukemia (ALL), 1 was patient with chronic granulocytic leukemia (CGL) at chronic phase, 15 were patients with non-Hodgkin lymphoma (NHL), 4 were patients with Hodgkin lymphoma (HL), 2 was patient with multiple myeloma (MM), 2 were patients with advanced breast cancer. All the patients apprcached to or got complete remission after conventional chemotherapy. No tumor cell infiltration was observed in bone marrow cytological examination. The functions of the main organs such as heart, lung, liver and kidney,and so on, were normal. The patients underwent an average of 8-course chemotherapy before the mobilization. Informed consents of all the patients were obtained.METHODS: MIT was intravenously injected at 10 mg/(m2·d)for 2 to 3 days, then Ara-C was also intravenously injected at 2 g/m2 every 12 hours for 1 to 2 days. When white blood cell (WBC) count recovered from the lowest value, 5 to 7.5 μg/ (kg·d)G-CSF was applied in 20 patients for 3 to 5 days successively. And 5 to 7.5 μg/ (kg·d)G-CSF and 5 to 7 μ g/(kg·d)GM-CSF were applied in another 22 patients at 6:00 in the morning and in the evening, respectively. PBSCs harvesting started when WBC > 2.5×109 L-1, especially when CD34+ cells≥ 1%,WBC was doubly increased. Autologous peripheral blood mononuclear cells (MNCs) were collected with CS3000 plus blood cell separator for detecting the level of CD34+ cells and T lymphocyte subsets. CFU-GM assays were performed in a methyl-cellulose-based clonogenic assay.① MNCs mixed with FITC-labeled CD34+, CD3 and CD8 monoclonal antibodies as well as CD4 PE-labeled CD monoclonal antibody at 4 ℃ for 30 minutes. 5×105 cells were determined, and CD3 and CD34+ levels, CD4/CD8 were determined by flow cytometer.Colony forming unit-granulocyte macrophage (CFU-GM) was determined with methyl cellulose. ② Related adverse reactions were observed after operation. ③ Aiming to different types of diseases,autologous PBSCs were back infused 36 to 48 hours after pre-disposal treatment. MNCs count and trypan-blue drying were done. Levels of CFU-GM and CD34+ cells were determined after unfreezing.MATN OUTCOME MEASURES: ① Changes in CD34+ cells and T lymphocyte subsets before and after mobilization. ② Postoperative related adverse reactions. ③ Back perfusion volume of autologous PBSCs (MNCs count, the number of CFU-GM and CD34+ cells).RESULTS: Forty-two involved patients participated in the final analysis. ① Changes in CD34+ cells and T lymphocyte subsets before and after mobilization: Without using hematopoietic growth factors (HGF), the percentage of CD34+ cells in peripheral blood of the patients was (0.054±0.032)%. After using G-CSF/GM-CSF treatment, it was (1.82±0.76)%,which was obviously increased compared with that of without using HGF (P < 0.001). The CD34+ cells and CFU-GM yields of 22 patients in C-CSF plus GM-CSF combination group [(8.76±3.39)×106/kg, (3.52±1.33)×105/kg, respectively]were significantly higher than those of 20 patients in G-CSF alone group [(6.12±2.11)×106/kg, (2.03±1.07)×105/kg,respectively (P < 0.05)]. There were no obvious changes of T lymphocyte subsets in the patients when using G-CSF/GM-CSF for some days except that CD34+ cells increased gradually (P > 0.05). ② Postoperative related adverse reactions: Ⅱ to Ⅲ degree hair-loss was seen in all the patients. Blood platelets dropped to (54.43±26.14)×109 L-1 at different degrees. Infective fevers (37.8 ℃ to 41.0 ℃) occurred in 21 patients. But they were controlled in short term after antibiotics treatment. All the side effects of G-CSF and GM-CSF were mild and reversible, easily controlled with paracetamol or steroids. Bone pain (mainly in lumbosacral region) occurred in 13 patients when WBC went up quickly. ③ Back perfusion volume of autologous PBSCs: PBSCs were cryopreserved at -80 ℃ without program control for 2.0 to 6.5 months. The cell recovery rate was (88.7±7.4) %. Trypan blue exclusion rate was (92.1±5.5) %. The back perfusion volume of MNCs, CD34+ cells and CFU-GM yields were (5.21±2.44)×108/kg, (6.89±3.55)×106/kg, (2.58±2.33)×105/kg,respectively. ④Circulation blood volume were 10 to 16 L (end-point separation blood volume were all above trebling TBV). Hematopoiesis was well reconstituted in 40 patients received autologous PBSCT.CONCLUSTON: MIT and high-dose Ara-C chemotherapy combined with both G-CSF alone and G-CSF plus GM-CSF can safely and effectively mobilize autologous PBSCs, while G-CSF plus GM-CSF is superior to G-CSF alone.Large-volume leukapheresis is an important method to enhance the productive rate of stem cells and decrease the times of harvesting.

9.
Clinical Medicine of China ; (12): 173-174, 2001.
Article in Chinese | WPRIM | ID: wpr-402144

ABSTRACT

Objective To investigate the early diagnosis of sudden cardiac death so as to increase the resuscitating success rate.Methods 24 cases with sudden cardiac death were analyzed.Results In 24 cases,there were 10 cases resuscitated successfully,8 cases with heartbeats recovered died in 1~3 days,5 of which had average blood glucose of 21.6 mmol/L and blood sodium of 156 mmol/L,and 6 cases resuscitated unsuccessfully.Conclusion 70.83% was caused by AMI in 24 cases.The early diagnosis of AMI is mainly based on clinical manifestation.Althogh early ECG did not show it,AMI should not be denied and misdiagnosed.Resuscitation should be performed on the spot as soon as cardiac arrest occurs.Naloxone and NaHCO3 should be used rationally.Meanwhile,be sure to prevent and treat reperfusion-injury.The prognosis would be extremely poor if the patient showed hyperglycemia and hypernatremia during resuscitation.

10.
Article in Chinese | WPRIM | ID: wpr-675784

ABSTRACT

Objective To explore whether acute graft versus host disease (aGVHD) could be alleviated by syngeneic bone marrow mixed with granulocyte colony stimulating factor (G CSF) mobilized H 2 haploidentical marrow grafting. Methods Female BALB/c and neonatal BALB/c mice were recipients and male (BALB/c?C 57 BL/6) F 1 (BCF 1) mice were donor mice respectively. Donor mice were injected subcutaneously with G CSF daily at 0.01 ?g/g body weight or saline for 6 days, and splenocytes were harvested on day 6. Spleen index(SI) represented GVHD in neonatal mice after the intraperitoneal injection of mixed spleen cells.Lethally irradiated ( 60 Co, 8.5 Gy) adult mice were transplanted with a mixture of syngenetic plus G CSF mobilized (control diluents) H 2 haploidentical marrow cells.Survival time and survival rate of the recipients were observed after mixed marrow transplantation (MBMT). GVHD was assessed by observing signs of weight loss, ruffled fur, diarrhea and histological change of skin, liver and small intestines. Enzyme linked immunosorbemt assay (ELISA) method was used to detect cytokines (IL 2, IL 4, IFN ?). Fluorescence activated cell sorting (FACS) analysis was used to detect T cell phenotype. Results (1) The neonatal mice subject to injection of 2∶1 and 1∶1 mixed spleen cells and H 2 haploidentical spleen cells all suffered from aGVHD. The severity of aGVHD in recipient mice receiving G CSF mobilized splenocytes was dramatically reduced. (2) The aGVHD signs and histological change were observed in most mice of 2∶1 and 1∶1 MBMT groups. However, the survival time of G CSF mobilized MBMT groups was longer than in control groups ( P

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