1.Autologous renal tumor lysate-loaded dendritic cell induce generation of antigen specific CTLs
Yuxia WAN ; Yuxian CHEN ; Jianhui MA
Chinese Journal of Urology 2001;0(09):-
Objective To make use of the characteristics of presenting and processing tumor antigen of dendritic cells to enhance killing capability of CTLs against autologous renal cell carcinoma. Methods Autologous dendritic cells were obtained by culturing bone marrow cell from patient with RCC in the presence of granulocyte macrophage colony stimulating factor(GM CSF) and IL 4. Dendritic cells were loaded with tumor cell lysate and co cultured with autologous PBMCs from patient to induce generation of tumor specific cytotoxic T cells(CTL). Killing activity and cytokine release of the CTL and the population of CTL were measured by cytotoxic assay and ELISA and FACS analyses. Results Immune response of DC Tuly induced CTL was demonstrated by the following facts:(1)the growth expansion of CTL enhanced 43 folds on day 16;(2)up regulation of the CD3 + and CD8 + population in CTL;(3)the cytotoxicity of specific CTL against autologous RCC was highly enhanced as compared with allogenic RCC and heterogenous tumor( P
2.Establishment and biological characteristics of six human renal cell carcinoma cell lines
Yunxia WAN ; Jianhui MA ; Yuxian CHEN
Chinese Journal of Urology 2001;0(09):-
Objective To establish human renal cell carcinoma (RCC) cell lines,and to investigate the cell phenotypes and expression of tumor associated antigens. Methods Fresh tumor tissues from pathologically proven human RCCs were primary cultured and passed generation to generation until a stably grow in vitro.The cell cloning form,chromosome and graft into nude mice in vivo were examined for cell lines.Immunofluorescent stain and flow cytometric analysis were carried out for cell's phenotype,RT PCR examination for MN/CA9 gene expression. Results Six human RCC cell lines have been established,including four of the cell lines derived from clear cells,one from mixed clear granular cells and one from papillary cells.All the cell lines showed the characteristics of malignant cells.All the lines highly expressed HLA ClassⅠ and HER2/neu.Three lines weakly expressed HLA ClassⅡ and one cell line highly expressed CD86 but all the lines did not express CD80.RT PCR showed that three cell lines have the expression of MN/CA9 gene. Conclusions These newly established RCC cell lines would provide a useful in vitro model for studies related to biological characteristics,tumor associated antigen,immunogenity and immunotherapy of human RCC.
3.Expression on dhfr-CHO cells and activity study of CD137 (human 4-1BB)
Bing WAN ; Lining ZHANG ; Chunhong MA
Chinese Journal of Immunology 2001;0(10):-
Objective:To set up a eukaryotic system for high expressing human CD137 and to investigate the role of CD137 and CD137L on signals transduction of cells.Methods:The pCDNA3 plasmid containing full length of human CD137 cDNA sequence(CMV ILA SEN,CIS) and pSV2 dhfr plasmid were cotransfected into dhfr CHO cells by lipoid mediating method. The positive clone was selected with G418. Expression of CD137 on dhfr CHO cells were induced by MTX and detected by RT PCR, immunocytochemistry and flow cytometry.It's activity study was done by method of incorporating 3H TdR.Results:CD137 expressed on the surface of dhfr CHO, expression rate was 96.07%, it's activity study indicated that CD137 increase the proliferation of PBMC stimulated by anti CD3 monoclonal antibody.Conclusion:dhfr CHO cells that highly express CD137 were established. CD137 can increase the proliferation of PBMC stimulated by anti CD3 monoclonal antibody.
4.CORRELATION ANALYSIS OF MYCOPLASMA PNEUMONIAE INFECTION AND ASTHMA
Yanfang MA ; Rui WANG ; Xin WAN
Modern Hospital 2015;(6):14-16
Objective To explore the relationship between Mycoplasma pneumoniae infection and bronchial asthma, and provide a theoretical basis for the diagnosis and treatment of bronchial asthma .Methods 54 cases of children with bronchial asthma were selected as the observation group , treatment of children with herpes infection of the upper respiratory tract of 54 cases as the control group .Two groups of children were assayed by eosinophil count , interleukin-13 (IL-13), immunoglobulin IgE (MP-IgM) and Mycoplasma pneumoniae antibody detection .The positive rate of MP-IgM test in two groups were compared .According to the observation group , MP-IgM detection results were divided into MP -IgM positive group and MP -IgM negative group .Positive contrast group , MP-IgM MP-IgM negative group and a control group of children with eosinophil count , IL-13 and IgE were detected .Re-sults The positive rate of MP-IgM in children with observation group detection was 75.93%;The positive rate of MP-IgM in children with control group of detection was 27.78%;The positive rate of MP-IgM detection in chil-dren with the observation group was significantly higher than the control group ( p<0.05); The MP-IgM positive group, MP-IgM negative groups eosinophil count , the levels of IL-13 and IgE levels were significantly higher than that of control group (all p<0.05); In MP-IgM positive group, Eosinophils count (538.18 ±41.63) ×106/L, IL-13 level (253.27 ±40.36) ng/L and IgE level (327.13 ±56.91) kU/L were significantly higher than in MP -IgM negative group (all p<0.05).Conclusion The incidence of asthma is associated with mycoplasma pneumonia infection , and we should strengthen the detection and anti -infection treatment of Mycoplasma pneumoniae in children with asthma .
5.Protection and mechanism of bushenhuoxue on retinal ganglian cells under the hypoxic condition in vitro
Li, WAN ; Xuejun, XIE ; Rong, MA
Chinese Journal of Experimental Ophthalmology 2014;32(3):211-215
Background The early disorder of diabetic retinopathy (DR) is the damage of retinal neural cells induced by high glucose and lack of oxygen.Previous studies show that bushenhuoxue serum can enhance the activity of glutamine synthetase (GS) in Müller cells under hypoxia,and glutamate-mediated retinal excitotoxicity also can be reduced by bushenhuoxue serum intervention.However,whether the concentration of glycine can be increased by bushenhuoxue serum is not clear.Objective This study was to investigate the protective effects of bushenhuoxue serum on retinal ganglion cells (RGCs) under hypoxia.Methods The Sprague Dawley (SD) rat serum containing bushenhuoxue was prepared.The RGCs of newborn SD rats were purified and identified by a twostep immunopanning procedure.After 72 hours,all RGCs were cultured in 96-well plates and divided into four groups:normal control group (cultured in adult SD rats normal serum),bushenhuoxue group (cultured in bushenhuoxue serum),hypoxia group (cultured in 1 mmol/L sodium dithionite); hypoxia + bushenhuoxue intervention group (cultured in bushenhuoxue serum+sodium dithionite).Glutamate and glycine contents in the extracellular fluid were detected by L-8800 automatic amino acid analyzer,and the content of lactate dehydrogenase (LDH) was assayed using LDH kits in 24,48 and 72 hours after culture.Results Cultured cells showed the green fluorescence under the immnofluorescence microscope.The contents of glutamate,glycine and LDH in the extracellular fluid were (0.0805±0.0010)mg/L,(0.0554±0.001 5)mg/L and (1 626.03 ±122.10)μmol/(min · L) in the normal control group in 24 hours after culture,and those in the hypoxia group were (0.022 5±0.001 1) mg/L,(0.014 6±0.001 1)mg/L and (1 458.68±94.23)μμmol/(min · L),showing significant reducing in the hypoxia group (q =-3.53,P =0.00 ; q =-2.45,P =0.00 ; q =-2.98,P =0.02).Compared with the normal control group,LDH and glycine contents in the extracellular fluid were significant raised in the hypoxia group 48 hours after culture (q =2.55,P =0.01 ;q =4.48,P =0.00).Seventy two hours after culture,the glutamate and glycine contents in the hypoxia group were higher than those of the normal control group (q =2.45,P =0.00 ;q =3.72,P =0.00).In 48 and 72 hours of culture,the contents of glycine were (0.017 4±0.001 5) and (0.019 2±0.001 2) mg/L in the hypoxia+bushenhuoxue intervention group,which were significantly higher than (0.016 0±0.001 2) and (0.018 0±0.000 8) mg/L in the hypoxiagroup (q=2.28,P=0.04;q=2.33,P=0.03),but the LDH level were (1 632.94±264.31) and (1 875.00±137.45)μmol/(min · L) in the hypoxia+ bushenhuoxue intervention group,which were lower than (1 688.49 ± 112.86) and (2 267.86 ± 175.21) μmol/(min · L) of the hypoxia group (q =-2.95,P =0.02 ; q =-2.35,P=0.00).No significant differences were seen in the glutamate content 24,48 and 72 hours after culture (P=0.55,0.28,0.46).A positive correlation was seen between the glutamate content and glycine content in the extracellular fluid (Kendall coefficient =0.519,Spearman coefficient =0.696,both at P =0.000).Conclusions The release levels of glutamate and glycine increase in the hypoxia RGCs,which probably is a compensatory response of RGCs.Bushenhuoxue serum can protect RGCs against injury by increasing the release of glycine and decreasing the LDH leakage from RGCs.
6.Current situation and development in treatment of laryngopharyngeal reflux.
Yichen WAN ; Yan YAN ; Furong MA
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2013;27(12):676-680
The mainstay of treatment of laryngopharyngeal reflux focuses on a concoction involving proton-pump inhibitor and modified beneficial behaviors, sometimes referring to medications like H2-R blocker etc. Therapeutic effect seems to be promising although may at times be challenged by conflict researches. Voice therapy and voice parameters emerge to provide a grand new perspective in treatment and evaluation of this disease.
Humans
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Laryngopharyngeal Reflux
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therapy
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Proton Pump Inhibitors
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therapeutic use
7.The very severe sensorineural deafness patients caused by rubella virus infection: two cases report.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(17):1567-1568
To explore the audiological features in children who were sever sensorineural hearing loss infected with rubella virus. There were two cases of rubella virus infection in children who were deaf, they conducted the distortion product otoacoustic emission, ABR and auditory steady-state evoked response (ASSR) examination, then analyzed the results comprehensively. Two patients' mothers were prompted to have infected rubella virus during the early three months pregnant period by history and laboratory tests. The two patients were not detected deafness gene mutation. Audiology results implied the two patients were very severe binaural sensorineural deafness, so they were recommended to equipped with hearing aids and cochlear implant surgery. Early pregnancy women infected with rubella virus can cause very severe offspring sensorineural deafness. The crowd whose mother were suspected to infect with rubella virus in early pregnancy, that should be tracked and detected hearing in order to achieve early detection, early intervention and early treatment.
Child
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Cochlear Implantation
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Cochlear Implants
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Evoked Potentials, Auditory
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Female
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Hearing Aids
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Hearing Loss, Sensorineural
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etiology
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virology
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Humans
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Otoacoustic Emissions, Spontaneous
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Pregnancy
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Rubella
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complications
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Rubella virus
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pathogenicity
8.A case of parapharyngeal space infection followed abdominal pain.
Cairong YANG ; Song MA ; Baoluo WAN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(1):90-91
Descending necrotizing mediastinitis that has an abdominal pain as a main clinical manifestation is seldom. Here one case is reported. At the beginning, the patient had pharyngalgia and his swallowing was not smooth. After that, abdominal pain became a main symptom. Pharyngalgia relieved . However CT showed mediastinal infection. Surgical drainage,antibiotics treatment and nutritional support were performed. The patient was cured.
Abdominal Pain
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etiology
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Deglutition
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Drainage
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Humans
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Infection
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Mediastinitis
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complications
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diagnosis
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therapy
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Necrosis
9.c-erbB-2 protein of human renal cell carcinoma can be recognized by cytotoxic T lymphocytes
Yunxia WAN ; Yuxian CHEN ; Jianhui MA
Chinese Journal of Urology 2001;0(04):-
ObjectiveTo provide experimental proof for the specific immunotherapy in patients with renal cell carcinoma (RCC).MethodsThe expression of c-erbB-2 and HLA-A2 molecules in RCC were examined by flow cytometery and HLA-A2 cDNA RT-PCR.Tumor lysatic antigens (Tuly) was used to load DCs (DC-Tuly) to induce the generation of c-erbB-2 specific CTLs derived from peripheral blood mononuclear cells of RCC patients in vitro and CTLs was used to kill RCC cells which express or not express c-erbB-2 and HLA-A2.The cytotoxic activity of induced CTLs was further studied by antibodies (anti-c-erbB-2 and anti-CD 8) blocking assay.Resultsc-erbB-2 protein is tumor associated antigens for RCC.DC-Tuly induced specific CTLs highly killed c-erbB-2+ HLA-A2+ autologous and allogenic RCC,but non-specific ones lowly killed c-erbB-2- HLA-A2+ or c-erbB-2+ HLA-A2-RCC cells. The cytotoxic activity against tumor cells was blocked by anti-c-erbB-2 and anti-CD 8 monoclonal antibodies.ConclusionsThese results suggest that this specific CTLs adoptive immunotherapy for RCC patients with c-erbB-2+ HLA-A2+ could be a novel approach for clinical use.
10.Post-operative deformity after posterior approach vertebral pedicle fixation in treatment of thorac-lumbar fracture
Orthopedic Journal of China 2006;0(14):-
[Objective]To analyze the factors of post-operative deformity after posterior approach vertebral pedicle fixation in treatment of dorso-lumbar spinal fractures.[Method]Between 2000.10~2005.11,32 patients of single dorso-lumbar spinal fracture were hospitalized.compressive type 5 cases,explosive type 27 cases.T11 12 cases,T12 11 cases,L1 5 cases,L2 3 cases,L3 1 case.The patients were devided into 3 groups according to the severity of related chondral plates injuly,group 1: without chondral fracture 11cases,group 2: with upper chondral plate fracture 15 cases,group 3: with both upper and lower chondral plate fractures 6 cases.By the formula to measure the average height of fractured vertebral body and its kyphotic Cobb's angle before operation,after operation and at time of follow-up,and calculate the result stastically.[Result]All 32 patients were followed-up for 10~56 months(average 23 months).The results indicated that the patient's preoperative condition was negatively correlated with the average vertebral height which is followed-up.The worse condition was,and the greater loss of postoperative vertebral height was.Furthermore,the patient's situation before operation had a positive correlation with the Cobb's angle of vertebral followed-up.When the patient's condition was worse,the postoperative angle of deformity was larger.[Conclusion]Posterior approach vertebral pedicle fixation in treatment of dorso-lumbar spinal fractures may result in reccurence of vertebral deformity,which is related to the severity of injury of vertebra and its chondral plates,so that to those patients related with chondral plates injuries,the vertebral pedicle fixation should be combined with anterior bone grafting,or with bone grafting into vertebral body after reduction.