OBJECTIVE: To explore the impact of upper airway expansion surgery on the blood pressure of hypertension patients with upper airway obstruction. METHOD: Clinical data of 45 cases of hypertension patients with nasal obstruction who were underwent upper airway expansion surgery. The morning sitting blood pressure of patients were recorded. All of the patients were underwent nasal expansion surgery and 11 cases with obstructive sleep apnea hypopnea syndrome were combined with H-uvulopalatopharyngoplasty. RESULT: Blood pressure of all 45 cases were apparently reduced in 12 months after the operations [systolic blood pressure (157.38 +/- 15.15) mmHg to (132.18 +/- 8.43) mmHg, diastolic blood pressure (95.42 +/- 9.28) mmHg to (82.31 +/- 5.88)mmHg], in which 9 cases with type I hypertension were lowered to normal pressure. Results had statistical difference between pre-operation and post-operation (P < 0.01). CONCLUSION Upper airway expansion surgery (including nasal and pharynx cav ity) can help lower blood pressure of hypertensive patients with nasal obstruction. Post-operative results were better than H-uvulopalatopharyngoplasty only.
Adult ; Aged ; Aged, 80 and over ; Blood Pressure ; physiology ; Female ; Humans ; Hypertension ; physiopathology ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; Sleep Apnea, Obstructive ; complications ; physiopathology ; surgery

Objective To observe the function of immature CD8α+ dentritic cells (DCs) in vitro. Methods The bone marrow and spleen of C57BL/6(H-2b) and Balb/c (H-2d) mice were got to prepare immature CD8α+ DCs and spleen lymphocytes,and treated by mytomycin. MTT test was used.MLR group, MLR plus variable density syngeneic CD8α+ DC group, MLR plus variable density allogeneic CD8α+ DC group,MLR plus variable density CD8α+ DC supernatant group,CD8α+ DC plus syngeneic T cell group and negative control group were established. MLR group was set up by responder cell ratio of 0.2,0.5,0.8,1.0,to build the MLR plus syngeneic and allogeneic CD8α+ DC experimental groups. Culture supernatant from different density (1 × 105/ml - 5 × 106/ml) of CD8α+DCs was added into MLR to build CD8α+ DC supernatant group. CD8α+ DCs were co-cultured with syngeneic T cells to build CD8α+ DCs plus syngeneic T cells group. 2 × 105/well responder cells served as the negative control group. ELISA was used to detect the concentrations of IFN-γ and IL-10 in the DCs could both suppress MLR (P＜0. 05), and the difference was not statistically significant (P＞0. 05). When CD8α+ DCs were increased, the suppressive effect was enhanced. When CD8α+ DC/responder cell ratio ＞0. 2, the inhibitory effect could be observed, and this effect reached the peak when the ratio was 1.0. The CD8α+ DCs had weak ability to stimulate syngeneic lymphocyte proliferation in vitro, and certain stimulating effect could be seen only when CD8α+ DC/responder cell ratio ＞2 (P＜0. 05). Its culture supernatant also showed suppressive effect (P＜0. 05), and the supernatant with a cell density of 5 × 105/ml showed the maximum effect. IL-10 concentration in the concentration was 1.0 ± 1.2 pg/ml. Conclusion The in vitro function of immature CD8α+ DCs was immunosuppression/tolerance,and they could secret high level of IL-10. The CD8α+ DCs and their culture supernatant could suppress MLR in vitro.

[Objective] To establish an effective and stable method to induce hematopoietic cells from embryonic stem(ES) cells,the phenotype and function of ES-derived hematopoietic cells induced by stromal cell conditioned medium (SCCM) of yolk sac (YS),fetal liver (FL) or bone marrow (BM) were analyzed and compared.[Methods] 10% of YS-SCCM,FL-SCCM or BM-SCCM was added to culture system for differentiation of ES cells.Flow cytometric analysis was used to identify expression of Flk1,Integrin α4,Sca-1,and CD34.Colony analysis was used to identify the quantity of high proliferative potential colony-forming cells (HPP-CFC) in differentiated ES cells.The yield of CFU-S (colony-forming unit-spleen) was also analyzed by transplanting ES cell derivatives into lethally irradiated mice.[Results] Expression of Flk1,Integrin α4,Sca-1,and CD34 could be tested on induced EB cells.The percentage of Flk-1+,Integrin α4+ and Sca-1+ cells induced by were 3.03%,2.9%,and 13.74%,respectively,which are greater than other groups.The percentage of CD34+ cells induced by BMSC-CM was 1.07% which was greater than other groups.The yields of HPP-CFC from hematopoietic cells induced by FLSC-CM or BMSC-CM were 7.4 /105 cells (P ＜ 0.01) and 5.8 /105 cells (P ＜ 0.05) which were greater than the yields of control group.The yields of CFU-S from hematopoietic cells induced by FLSC-CM or BMSC-CM were 8.5/5 × 105 cells and 6.75/5 × 105 cells which were also greater than the yields of control group (P ＜ 0.001).[Conclusion] Both YS-SCCM,FL-SCCM,and BM-SCCM could promote hematopoietic differentiation of ESE14.1 cells.Hematopoietic differentiation induced by FL-SCCM or BM-SCCM is more effective,which generates hematopoietic progenitor cells with normal function.Application of FL-SCCM generates more primitive hematopoietic progenitor cells than that of BM-SCCM.

AIM: To establish hybrid mouse embryonic stem (ES) cell line from blastocysts of the (C57BL/6J × 129/J) F1 mouse. METHODS: 3.5 days post- coitus (d.p.c.) blastocysts were cultured on mouse embryonic fibroblasts (MEFs) in the medium, after 3 - 4 days, Inner cell mass were picked up and disaggregated, then reseeded. After the ES - like colonies appeared, passaged them to give permanent ES cell lines. The pluripotent propertes of ES cells obtained were analyzed by alkaline phosphatase (AKP) activity, expression of SSEA- 1 and Oct-4, and their capacity to form teratoma. RESULTS: Two hybrid ES cell lines, SC1001, SC1002 were obtained from blastocysts of the (C57BL/6J × 129/J) F1 genotype. Most of these ES cells had a normal karyotype and an XY sex chromosome composition. The pluripotent properties of the cell lines were analyzed on the basis of their alkaline phosphatase activity, expression of SSEA - 1 and Oct - 4, and their capacity to form teratoma in severe combined immunodeficiency (SCID) mice. CONCLUSION: Two hybrid mouse ES cell lines having pluripotent properties and capacity for long - term self renewal were generated from blastocysts of the ( C57BL/6J × 129/J) F1 genotype.

OBJECTIVETo prevent Graft-versus-host disease (GVHD) in rat model, we evaluated the feasibility of mesenchymal stem cells (MSCs) as a gene transfer target and studied the efficiency of recombinant adenovirus mediated gene therapy.

METHODSWe constructed the recombinant adenovirus containing CTLA4Ig gene. Rat MSCs of passages 3-5 were infected by the adenovirus, and the transfection efficiency was monitored by GFP markers. We performed flow cytometric analysis, immunohistochemical and Western blotting analysis to identify the CTLA4Ig expression. The gene transferred MSCs were tested for their ability to inhibit the allogeneic lymphocyte response in vitro and to prevent GVHD in a rat model.

RESULTSRecombinant adenovirus pAd-CTLA4Ig was correctly constructed and confirmed. After MSCs were infected by the adenovirus, the CTLA4Ig protein was detected not only in transgenic MSCs, but also in the culture medium. In a mixed lymphocytes response (MLR) test, the transgenic MSCs could significantly inhibit the allogeneic lymphocyte response compared with the control groups (P < 0.05). A model of GVHD was developed by transplanting bone marrow cells and spleen lymphocytes of F344 rats to lethally irradiated SD rats. The onset of GVHD could be ameliorated or prevented by co-administration of transgenic MSCs. All the rats in the control groups suffered severe acute GVHD. CTLA4Ig expression was observed in the liver, intestine, kidney and spleen 30 days post-transplantation.

CONCLUSIONSOur results indicate that adenoviral vectors could efficiently transfer CTLA4Ig gene into MSCs and sustain long-term stable expression in vitro and in vivo.

Abatacept ; Adenoviridae ; Animals ; Feasibility Studies ; Gene Transfer Techniques ; Genetic Vectors ; Graft vs Host Disease ; prevention & control ; Immunoconjugates ; genetics ; Mesenchymal Stromal Cells ; Rats ; Rats, Inbred F344 ; Rats, Sprague-Dawley ; Recombination, Genetic

AIM:To investigate the differentiation from human mesenchymal stem cells(hMSC) into neuron-like cells.METHODS:hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate with DMEM/monothioglycerol or DMEM/β-mercaptoethanol, respectively. Neuron-specific enolase(NSE), neurofilament(NF), and glial fibrillary acidic protein(GFAP) were detected by immunohistochemistry. RESULTS:hMSC were expanded as undifferentiated cells in culture for more than 5 passages. When treated with monothioglycerol or β-mercaptoethanol for 5 hours, hMSC exhibited neuronal phenotype. The expression of NSE and NF in the neuron-like cells was positive, but the glial astrocyte marker GFAP didn't express. CONCLUSION:hMSC can be induced to differentiate into neurous.

【Objective】To observe the efficacy and side effects of hematopietic stem cell transplantation with combination of umbilical cord blood(UCB) and neonatal peripheral blood(NPB) in the treatment of β-thalassemia major.【Methods】28 mL NPB was drawn from a HLA identical neonate within 5 hours after his birth to complement stem cell of the UCB he donated for transplantation to his sibling with β-thalassemia major.Various items of hematopoiesis reconstruction were detected in UCB and NPB respectively.After conditioning with chemotherapy by using busulfan 20 mg/kg,cyclophosphamide 200 mg/kg,melphalan 90 mg/m2 and antithymocyte globulin(ATG) 90 mg/kg,the patient received the 53 mL UCB and 28 mL NPB,achieving 5.7×107/kg nucleated cells(NC),93×105/kg CFU-GM and 3.1×105/kg CD34+CD38- cells from his HLA-identical sibling.【Results】Absolute nucleated cell(ANC) reached 0.5×109/L on 14th day post transplant,and platelets reached 20×109/L on 34th day after transplant.The heterozygosity of β-654 mutation point was detected by the PCR-RDB.The sexual chromosome changed from XX pretransplant to XY posttransplant.The patient was free red blood cell transfusion from 14th day post transplant.Her hemoglobin rose progressively from 86 g/L to 110 g/L.The patient survived for 197 days free from disease after transplantation.Following up for 9 months, the donor grew and developed normally.【Conclusion】The NPB contains a lot of stem cells.The transplantation with combination of suitable NPB and UCB is an effective tactics when the UCB cells are deficient.

Objective Study on producing human monoclonal antibodies against Rhesus (D) antigen that was suitable for use as blood group typing reagent. Methods B lymphocytes from a Rh negative woman, which can produce anti D antibodies were transformed by Epstein Barr virus(EBV). Antibody secreting cells were enriched by RhD + group O erythrocytes and cloned by limited dilute method. By using one step emzymatic method on microplates, one thousand normal blood donors with a common Rh phenotype were tested with the supernatant of cell culture medium as well as a polyclonal human anti D and a commercial monoclonal anti D serum. Results Three human B lymphocyte lines secreting monoclonal antibodies to Rh (D) were established. One of them produced lgM antibody. The titer of the monoclonal antibodies was 64～128. Study on 1000 blood donors, the results did not show any discrepancy among the three different anti Rh(D) serum. Conclusion These monoclonal antibodies against D antigen could be used in Rh(D) typing.

【Objective】 To explore the complication and engra ftment of human cord blood hematopoietic stem cells in utero transplantation thr o ugh abdominal cavity of fetal rats , and to establish an animal model for clini cal application. 【Methods】　Human cord blood (MNC) cells were transplanted into th e abdominal cavity of fetal rats, the complications and the outcome of pregn ancy were observed. The condition of engraftment was detected by flow cytometr y and immunohistochemistry methods after the fetus were born. 【Results】 Huma n CD3 cells were detected in rats and the engraftment rate was 64%. At 1 and 2 months of age, the mean value of human CD3 cells were 0.28%±0.05% and 0.41 %± 0.05% respectively （P＜0.05）.Human CD3 、CD20及 CD+34 ce lls were also detected in liver、spleen and thymus of rats at 2 months of age. The i ncidence of complication was significantly different between transplanted grou p and non-transplanted group. 【Conclusion】　Human cord blood cells transfused into the abdominal cavity of fetal rats were engrafted . There were some complication s occurred during operations which affected the outcome of pregnancy.

Objective　To obtain mammalian cell expression vector of human CD154 gene. Methods　A 820 bp cDNA fragment was amplified by RT-PCR method from total RNA of human peripheral blood mononuclear cell（PBMC） activated with 10 ng／mL PMA and 1 μg／mL PHA for 8 hours. The fragment was cloned into pcDNA3.1（＋） plasmids.The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes BamH Ⅰ and EcoR Ⅰ and sequenced by Sangers-dideory-mediated chain termination. Results　This cDNA fragment included 820 bp entire coding region and a part of the 3 non-coding region. The recombinant mammalian cell expression vector of pcDNA3.1（＋）／hCD154 was constructed， the sequence of the insert was identical to the published sequence encoding human CD154 antigen. Conclusion　The recombinant mammalian cell expression vector of pcDNA3.1（＋）／hCD154 was successfully constructed.