1.SELDI-TOF-MS ProteinChip technology for screening of urine markers of bladder carcinoma
Denglong WU ; Yuanfang ZHANG ; Ming GUAN
Chinese Journal of Urology 2001;0(07):-
Objective To search for new tumor markers from urine of bladder transitional cell carcinomas. Methods Urine samples from 61 bladder cancer patients who were histologically diagnosed and 53 healthy volunteers and 42 cases with other urogenital diseases were analyzed using surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) IMAC-Cu-3 ProteinChip technology,which can specifically bind the metal-combining-proteins.Proteomic spectra were generated by mass spectrometry.A blinded test set was used to determine the sensitivity and specificity of the differentially expressed markers. Results Four differentialy expressed potential markers were identified.Their corresponding molecular weights were 3445,3703,3896 and 5932, respectively.The 3445 protein was identified as ?-defensin,with its sensitivity for diagnosing bladder cancer being 80% and specificity being 75%.The sensitivity of 3703 protein was 32%,while its specificity was 94%.97% of the patients who were positive for 3703 protein were with high grade and invasive bladder cancer.The sensitivity for 3896 and 5932 proteins were 78% and 55%;the specificity for them were 40% and 35%,respectively. Conclusions SELDI-TOF-MS ProteinChip technology is a quick,easy and practical,high throughput analytic method.It can screen out several relatively specific,potential markers from urine of bladder cancer patients,so it has better clinical feasibility.
2.Exploration on cultivation of clinical thinking for eight-year program medical students in during clinical internship
Xuedong WU ; Jianyun WEN ; Fuyu PEI ; Yuanfang JING
Chinese Journal of Medical Education Research 2012;11(6):579-581
Pediatrics internship is a bridge that brings medical theory into clinical reality.Cultivate systematic clinical thinking during internship is the key to the growth of clinical doctors.Good outcome has been achieved among these eight-year medical students by using teaching methods such as case discussions,lectures,teacher instructions,which can cultivate proper clinical thinking and make students more active.
3.Intraocular Pressure Sensor Based on a Contact Lens.
Xuhong GUO ; Weihua PET ; Zhaolin YAO ; Yuanfang CHEN ; Xiaohui HU ; Hongda CHEN ; Jingyuan ZHU ; Huijuan WU
Journal of Biomedical Engineering 2016;33(1):18-22
Intraocular pressure detection has a great significance for understanding the status of eye health, prevention and treatment of diseases such as glaucoma. Traditional intraocular pressure detection needs to be held in the hospital. It is not only time-consuming to doctors and patients, but also difficult to achieve 24 hour-continuous detection. Microminiaturization of the intraocular pressure sensor and wearing it as a contact lens, which is convenient, comfortable and noninvasive, can solve this problem because the soft contact lens with an embedded micro fabricated strain gauge allows the measurement of changes in corneal curvature to correlate to variations of intraocular pressure. We fabricated a strain gauge using micro-electron mechanical systems, and integrated with the contact lens made of polydimethylsiloxane (PDMS) using injection molding. The experimental results showed that the sensitivity was 100. 7 µV/µm. When attached to the corneal surface, the average sensitivity of sensor response of intraocular pressure can be 125.8 µV/mm Hg under the ideal condition.
Contact Lenses, Hydrophilic
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Dimethylpolysiloxanes
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Glaucoma
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Humans
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Intraocular Pressure
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Tonometry, Ocular
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instrumentation
4.Eukaryotic expression and preparation of polyclonal antibody for osteoclast inhibitory lectin related protein
Suxia WU ; Lihui CHAI ; Zhanzheng WANG ; Guangchao LIU ; Wenzhi TIAN ; Yuanfang MA
Chinese Journal of Immunology 2014;(10):1374-1377,1392
Objective:In order to get recombinant protein OCILRP 2-Fc and anti-OCILRP2 antibody for further study of OCILRP2.Methods:Eukaryotic expression vector pIg-CD5-OCILRP2 which fused with extracellular domain of OCILRP 2 and human IgG1 Fc fragment was constructed.G418 was used for stable expression cell strain after pIg-CD5-OCILRP2 transfected into CHO cells.Recombinant protein OCILRP 2-Fc purified from CHO cell supernatant was used to immunize rabbit and anti-OCILRP2 polyclonal antibody was purified from rabbit serum by using protein G column.Results: ELISA data showed that we got a high-titer anti-serum and anti-OCILRP2 antibody purified from the rabbit serum.Western blot indicated this antibody could specifically bind to OCILRP 2-Fc and OCILRP2 in NIH/3T3 lysate.OCILRP2 expression in murine bone marrow derived dendritic cells ( DC) was detected by this polyclonal antibody ,too.Compared to immature DC ,OCILRP2 expression was elevated in LPS induced-mature DC.Conclusion: This study has offered an available tool and provided a clue for further study of the roles of OCILRP 2 in immune response.
5.Study on the pharmacolinetics,tissue distridution and excretion of nerve growsth factof in mice
Caiguo HUANG ; Shanjian WEI ; Hua JIANG ; Shukui SONG ; Huiyuan LI ; Yuanfang WU
Academic Journal of Second Military Medical University 2001;22(1):65-67
Objective:To study the pharmacokinetics,thissue distribution and secretion of nerve growth factor(NGF)in mice.Methods:The conecntration of NGF in various body fluids and tissue were determined by isotope tracer combined SDSPAGE method.Results:The plasma concenmtration-time curve was in accordance with the two-compartment pharmacokinetic model.The elimation half-life(t1/2β)was 3.1.The half-life of distribution(t1/2ka)was 5min.Tpeak was 25 min.AUC was 72.4 mg·kg-1·h-1.The concentrations of NGF were high in thyroid,blood,submaxillary glands,superior cervical ganglion,adrenasl and kidneys.Conclusion:NGF has a wide distribution,high tissue concentrationa nd excrtet mainly through the urine.
6.Osteoclast inhibitory lectin related protein 2 protects mice from endotoxemia
Suxia WU ; Lihui CHAI ; Fei YANG ; Weihua LI ; Jingli GU ; Yuanfang MA
Chinese Journal of Microbiology and Immunology 2014;(9):657-661
Objecitv e To investigate the effects of a recombinant protein osteoclast inhibitory lectin related protein 2( OCILRP2)-Fc on LPS-induced endotoxemia by blocking OCILRP 2 signaling pathway and to in-vestigate the roles of OCILRP2 during inflammation.Methods Real-time PCR was used to detect OCILRP2 ex-pression at mRNA level in RAW264.7cells before and after in vitro stimulation with LPS.A mouse model of en-dotoxemia was established by intraperitoneal injection of BALB /c mice with a median lethal dose of LPS .Two hours prior to LPS treatment, mice were intraperitoneally injected with OCILRP2-Fc, human IgG or PBS, re-spectively .Several parameters including the survival rate of BALB/c mice with and without LPS treatment , spleen weight for arterial hyperemia analyzing , histopathological changes of lung and liver by HE staining , serum levels of inflammatory cytokines (IL-6, IL-12, TNF-αand IFN-γ)by ELISA , NF-κB activity by Western blot, were analyzed .Results Real-time PCR showed that LPS elevated in vitro OCILRP2 expression at mRNA level in macrophages (P<0.05).Upon the treatment of OCILRP2-Fc, BALB/c mice suffered from endotoxemia showed obviously increased survival rate , decreased spleen hyperemia , attenuated pathological injury of lung and liver, reduced levels of IL-6, IL-12, TNF-αand IFN-γin serum samples (P <0.05) as compared with mice treated with human IgG and PBS .LPS induced NF-κB p65 phosphorylation and IκB degradation were inhibited by OCILRP2-Fc treatment.Conclusion OCILRP2-Fc protects mice from endotoxemia by blocking OCILRP 2 signaling, which suggests that OCILRP2 plays an important role in LPS induced inflammation.
7.Expression of pigment epithelium-derived factor in bladder tumour is correlated with interleukin-8 yet not with interleukin-1α.
Chenchen, FENG ; Ming, GUAN ; Qiang, DING ; Yuanfang, ZHANG ; Haowen, JIANG ; Hui, WEN ; Paohsun, WANG ; Zhong, WU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(1):21-5
Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In this study the expression of PEDF, interleukin-1α (IL-1α) and -8 (IL-8) in bladder tumours was investigated. Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples. Expression change of the factors was compared with clinicopathological parameters. Correlations between PEDF, IL-1α and IL-8 were analyzed. None of the factors was in relation to gender, tumour occurrence, and size or onset pattern. PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression. PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping. PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049, r=-0.578) as well as in tumour grouping (P=0.033, r=-0.276). Deranged expressional change of PEDF, IL-1α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.
8.Clinical study of PEI combined with HIFU for treating unresectable middle and advanced stages of primary liver cancer
Guangping WU ; Yuanfang PU ; Tian TIAN ; Yong PANG ; Ke CAO ; Yong YANG ; Bo XU ; Yuan LI ; Yan XIE ; Min HE
Chongqing Medicine 2015;(10):1367-1369
Objective To investigate whether the percutaneous ethanol injection (PEI)under sedation and analgesia can in-crease the energy deposition and curative efficiency of the high intensity focused ultrsound(HIFU)in treating unresectable middle and advanced stages of primary liver cancer.Methods Thirty-six cases of clinically diagnosed unresectable middle and advanced sta-ges of primary liver cancer were randomly divided into the PEI+ HIFU group(combination group,n = 23)and the simple HIFU group (HIFU group,n=13);10mL of the mixture of 99.7% ethanol and iodized oil (9:1)was given by intratumoral injection at 30 min before ablation in the PEI+HIFU group,while 0.9% physiological saline 10mL was replaced in the simple HIFU group.The ablation energy efficiency factor(EEF)and irradiation time were compared between the two groups.Results The ablation EEF in the PEI+HIFU group and the simple HIFU group were (13.82+4.26)J/mm3 and (25.63+6.31)J/mm3 respectively,the PEI+HIFU group was significantly lower than the simple HIFU group (P <0.05);the irradiation time were (1 468.28+253.21)s and (2 352.56+463.34)s respectively;which in the PEI+ HIFU group was significantly shortened (P <0.05).Conclusion PEI can enhance the HIFU ablation energy deposition and improve the efficiency of HIFU for treating unresectable primary liver cancer.
9.Treatment of Gaucher disease with allogeneic hematopoietic stem cell transplantation: report of three cases and review of literatures.
Xiangfeng TANG ; Zuo LUAN ; Nanhai WU ; Bo ZHANG ; Yuanfang JING ; Hong DU ; Wei LU ; Shixia XU
Chinese Journal of Pediatrics 2015;53(11):810-816
OBJECTIVETo explore the efficacy of unrelated umbilical cord blood transplantation (UCBT) in the treatment of Gaucher disease.
METHODThe clinical characteristics of three children with Gaucher disease underwent UCBT in our hospital between April 2013 and September 2014 were retrospectively analyzed. Literature on allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of Gaucher disease was searched at Wanfang and Pubmed databases between 1983 and 2015 and was reviewed and summaried.
RESULTThree children with Gaucher disease, all were female, received UCBT. These patients' age at receiving transplantation was 3.8 years, 7.1 years and 2.6 years, respectively. The second case received the second transplantation. The first and third case received splenectomy before UCBT. The pretreatment regimen was busulfan (Bu)/fludarabine (Flu)/cyclophosphamide (CTX)/antithymocyte globulin (ATG), and for the patient received the second transplantation melphalan was added to the myeloablative conditioning regimen of Bu/Flu/CTX/ATG. Cyclosporine and mycophenolate mofetil (MMF) wee used for prophylaxis of acute graft versus host disease (aGVHD). The dose of cord blood stem cell nucleated cell counts was 9.7 × 10⁷ /kg,11.9 × 10⁷ /kg and 7.6 × 10⁷/kg respectively. The dose of cord blood stem cell CD34⁺ cell counts was 5.4 × 10⁵/kg , 3.5 × 10⁵/kg and 3.2 × 10⁵/kg respectively. The day of granulocytes exceeding 0.5 × 10⁹/L was day 11, 12 and 19 after transplantation, respectively. The day of platelets exceeding 20 × 10⁹/L was day 14, 33 and 74 after transplantation, respectively. At one month after transplantation the rate of chimerism was over 95% and all patients got donor complete chimerism. The level of β-glucocerebrosidase recovered to normal at one month after transplantation. During transplantation, all patients developed cytomegalovirus (CMV) and Epstein-Barr virus (EBV) viremia. In case 1 immune thrombocytopenia occurred at five month after transplantation unresponding to steroids and mesenchymal stem cells infusion was administered and his platelet in routine blood test recovered to normal. But the patient died because she was infected with varicella-zoster virus out of hospital at nine month after transplantation and the level of β-glucocerebrosidase was normal before death and chronic GVHD (cGVHD) was not found. The case 2 is now in 19th month after transplantation and his level of β-glucocerebrosidase was normal. cGVHD was not found. The patient is currently free of disease. The case 3 was in 9th month after transplantation and his level of β-glucocerebrosidase was normal. cGVHD was found at 112 day after transplantation and was localized and could be controlled by hormonal therapy. The patient is currently free of disease. Three patients' size of liver was significantly reduced after their level of β-glucocerebrosidase ecovered. There were 50 cases with Gaucher disease who were treated with allo-HSCT in the literature and none of them were reported from China. Disease-free survival rate of patients treated with allo-HSCT for Gaucher disease was 85%. In all reports, there were 31 cases who had information of typing of Gaucher disease, of whom 22 cases had type 1 and 9 cases had type 3. Twenty-nine cases had information of survival, of whom 24 cases survived and 5 cases died of infection. Fifteen cases had data of engraftment, 2 of whom had graft failure and one had late graft failure.Glucocerebrosidase recovered to normal in 25 of 31 cases who had relevant data, in one of whom with late graft failure the enzyme recovered to normal 3 month after transplantation, but his enzyme decreased to the initial level 9 month after transplantation. Along with enzyme level's recovery to normal, in a part of cases bone pain and hepatomegaly were relieved and growth delay was improved.
CONCLUSIONThe unrelated UCBT may be a form of treatment that offers the potential of permanent cure and a procedure with possible long-term benefits in patients with Gaucher disease.
Antilymphocyte Serum ; therapeutic use ; Busulfan ; therapeutic use ; Child ; Child, Preschool ; China ; Cyclophosphamide ; therapeutic use ; Cyclosporine ; therapeutic use ; Disease-Free Survival ; Female ; Gaucher Disease ; therapy ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Mycophenolic Acid ; analogs & derivatives ; therapeutic use ; Retrospective Studies ; Transplantation Conditioning ; Vidarabine ; analogs & derivatives ; therapeutic use
10.Molecular epidemiology of noroviruses from infantile diarrhea at Suzhou and Nanjing area in Jiangsu province in 2010
Jianguang FU ; Bin WU ; Hong JI ; Liang LI ; Xian QI ; Yuanfang QIN ; Shenjiao WANG ; Fei DENG ; Zhifeng LI ; Fenyang TANG ; Rongqiang ZU ; Changjun BAO
Chinese Journal of Microbiology and Immunology 2011;31(11):989-993
ObjectiveTo investigate norovirus infection status and indentify its epidemiological characteristics and genotype distribution in infantile viral diarrhea in Jiangsu.MethodsFour hundred and ninety-eight fecal specimens of infantile virus diarrhea cases were collected from Suzhou Children's hospital and Nanjing Children's hospital in 2010.Norovirus genegroup were detected by real-time RT-PCR,and genetype were determined by sequence analysis.Results Among all fecal specimens,2 (0.4%) cases were positive for norovirus G Ⅰ,and 190 (38%) cases for G Ⅱ.Nucleotide sequence analysis revealed that in the 2 samples for G Ⅰ,one strain was G Ⅰ 1 and another was G Ⅰ 3.Twenty-one strains were belonged to G Ⅱ 4 and 2 strains were G Ⅱ 3 in the 23 samples for G Ⅱ.ConclusionAs one of the most important pathogens causing infantile viral diarrhea in Jiangsu province,subtype G Ⅱ 4 was the main epidemic strain of norovirus,meanwhile other genotypes also existed.