2.Comparative study on different methods for quantitative analysis of oral common microorganisms
ming-zhu, ZHANG ; chao-lun, LI ; yun-tao, JIANG ; wei, JIANG ; jing-ping, LIANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(02):-
flow cytometry.A strong linear relationship was observed in the standard curve of real-time PCR of each bacteria. Conclusion These three non-culture methods can be used in the quantitative analysis of oral microorganisms.Real-time PCR and laser scanning confocal microscopy are better than the traditional culture-based CFU count,and real-time PCR is the most sensitive method.
3.Differential gene expression between Streptococcus sobrinus 6715 and its fluoride-resistant strains
wei, JIANG ; jing-ping, LIANG ; chao-lun, LI ; yun-tao, JIANG ; ming-zhu, ZHANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(02):-
Objective To detect the differential gene expression between Streptococcus sobrinus(S.sobrinus) 6715 and its fluoride-resistant strains. Methods The fluoride-resistant strains of S.sobrinus 6715 was induced by increasing the concentration of fluoride step by step.Total RNA of both S.sobrinus 6715 and its fluoride-resistant strains was extracted,mRNA was separated and purificated,and then cDNA was obtained by reversed transcription.Suppression subtractive hybridization(SSH) technology was used to detect the differential gene expression between them.The differential gene expression fragments were cloned and compared with the GenBank by BLAST.Results After comparing with the GenBank by BLAST,it was identified that there were two differential gene expression fragments,fruA and SMU.438c. Conclusion The cDNA subtractive lib of differential gene expression between S.sobrinus 6715 and its fluoride-resistant strains was successfully constructed through SSH,which paves a way for the further study of fluoride-resistant mechanism.
4.Quantitative analysis of cariogenic strains in dental plaque of caries-susceptibile children by real-time PCR
jing-ping, LIANG ; yun-tao, JIANG ; chao-lun, LI ; wei, JIANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(02):-
8).The amount of the targeted microorganisms(Streptococcus mutans,Streptococcus sobrinus,Actinomyces viscosus and Actinomyces naeslundii) and the total number of bacterial cells were determined by real-time PCR based on SYBR-Green I fluorescence.Results The percentages of the four targeted bacteria in high-caries group were significantly higher than those in caries-free group(P
5.Combination of EMPs and BMSCs in promoting regeneration of periodontal tissue
zhong-chen, SONG ; rong, SHU ; yu-feng, XIE ; xiu-li, ZHANG ; bin, ZHANG ; ai-mei, SONG ; chao-lun, LI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(06):-
Objective To evaluate the feasibility of reconstructing horizontal periodontal bone defects by tissue engineering based on bone marrow stromal cells(BMSCs)as seed cells and enamel matrix proteins(EMPs)as growth factors. Methods Two healthy rhesus monkeys were selected, and BMSCs were isolated from iliac marrow and serial subcultivation was conducted. The cells of induced BMSCs at passage 3 were harvested and mixed with Bio-oss collagen. The models of horizontal periodontal bone defects were established surgically in each buccal side of the posterior teeth, and were divided into four groups (blank control group, material group, cells/material group and cells/material/EMPs group). The histological and Micro-CT observation were carried out 8 weeks later. Results In the blank control group, the defects were filled with fibrous connective tissue. There was newly-formed alveolar bone in the material group. In the cells/material group, periodontal regeneration could be observed, while the newly-formed cementum was irregular and less in quantity. In the cells/material/EMPs group, the amount of newly-formed alveolar bone was larger, and the newly-formed cementum was continuous and regular. Conclusion The tissue engineering technique of BMSCs as seed cells in combination with EMPs induction can significantly promote the regeneration of periodontal tissue.
6.Detection of Brain Neurotransmitters in Patients with Vasovagal Syncope with Encephalofluctuogram Technology
Jun XIAO ; Lin WANG ; Chaoying ZHU ; Hongmei LI ; Lun SHU ; Hongling XU ; Yusong GUO ; Chao HU ; Junxian XUE ; Xin CHEN
Chinese Journal of Rehabilitation Theory and Practice 2017;23(3):349-351
Objective To observe the changes of brain neurotransmitters in patients with vasovagal syncope (VVS) with encephalofluc-tuogram technology (ET). Methods From August, 2015 to December, 2016, 30 patients with VVS were selected as case group, 30 controls matched with sex and age were selected from the outpatients without syncope. They were detected the function of gamma aminobutyric acid (GABA), glutamate (Glu), 5-hydroxytryptamine (5-HT), acetylcholine (Ach), norepinephrine (NE) and dopamine (DA) with ET. Results There was no significant difference between the two groups in the values of GABA, Glu, 5-HT, Ach and DA (t<1.680, P>0.05), while the values of NE was higher in the case group than in the control group (t=-3.552, P<0.001). Conclusion VVS may be related to the high level of activity of NE in the brain.
8.Culturing Schwann cells by explants and modified enzyme digestion technique
Hui WANG ; Lun LUO ; Wen-Sheng LI ; Chao-Feng LIANG ; Hai-Yong HE ; Ying GUO
Chinese Journal of Neuromedicine 2011;10(5):461-463
Objective To introduce an efficient method for culturing and purifying Schwann cells (SCs) in vitro. Methods Sciatic nerves were harvested from neonatal SD rats and the epineuria were removed. Single enzyme digestion combined with explans and double enzyme digestion were employed, respectively, to digest the nerve tissues following trituration. The proliferation of SCs and degree of purification were evaluated by viable count method and the combination of S-100 labeled monoclonal antibody and SCs. Results Proximally 3.5 × 106 cells were harvested with 96% survival rate and a purity of Schwann cells over 94% by single enzyme digestion combined with explans, however only 3.0×106 cells were gained with the purity being 90% and survival rate being 92% by double enzyme digestion. Conclusion This method yields large amount of viable Schwann cells with high purity and survival rate.
9.Study of anti-myeloma activity of interleukin-2 activated bone marrow in vitro.
Wei LIU ; Shi-Lun CHEN ; Jin-Wei LIU ; Xin LI ; Cheng-Qing XIA ; Man QI
Chinese Journal of Hematology 2005;26(4):205-208
OBJECTIVETo study the anti-myeloma activity of interleukin-2 activated bone marrow (ABM).
METHODSBone marrow mononuclear cells (BMMNC) from multiple myeloma and iron-deficiency anemia patients were cultured in the presence of rIL-2. The anti-myeloma activity of ABM against U266 cells, cells expressing surface CD45, CD38, CD138, the levels of TNF-alpha and IFN-gamma in ABM culture supernatant were measured with MTT method, flow cytometry and ELISA method respectively after bone marrow was activated with rIL-2 for 24 and 72 hours.
RESULTSThe tumor-killing activities against U266 cells of ABM were significantly increased compared with that of non-activated bone marrow (NBM) at 72 hours [(69.70 +/- 26.57)% vs (43.20 +/- 12.39)%, P < 0.05] and 24 hours [(34.25 +/- 11.93)% vs (26.53 +/- 5.48)%]. The CD45(-)CD38(+)CD138(+) cells of ABM from myeloma group at 72 hours were decreased from (8.46 +/- 3.66)% to (4.79 +/- 1.56)% (P < 0.05). TNF-alpha and IFN-gamma were detectable after cultured for 24 hours in both normal control group and myeloma group and went higher at 72 hours. The level of TNF-alpha and IFN-gamma were significantly increased in ABM compared with that in NBM (P < 0.05). Meanwhile, there was a positive relationship between the level of TNF-alpha, IFN-gamma and cytotoxicity of ABM from normal control group at 24 hours and 72 hours (P < 0.05), and was a negative relationship between TNF-alpha and IFN-gamma levels and the CD45(-)CD38(+)CD138(+) cells in myeloma group at 72 hours (P < 0.05).
CONCLUSIONNormal BMMNCs activated with rIL-2 have tumor-killing activities against U266 cells. Myeloma cells and tumor burden were decreased in myeloma bone marrow after the marrow was activated with rIL-2. Production of TNF-alpha and IFN-gamma from bone marrow cells including T cells, monocyte-macrophages and NK cells activated with rIL-2 might be involved in anti-myeloma activity of ABM.
Adult ; Aged ; Bone Marrow Cells ; drug effects ; immunology ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Cytotoxicity, Immunologic ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Humans ; Interferon-gamma ; biosynthesis ; Interleukin-2 ; immunology ; pharmacology ; Male ; Middle Aged ; Multiple Myeloma ; blood ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; biosynthesis
10.Evidence for determining the safe surgical margin for pleomorphic adenoma of parotid gland.
Yi-quan XU ; Chao LI ; Jin-chuan FAN ; Bing ZHANG ; Jian-chao CHEN ; Zhao-hui WANG ; Ying WANG ; Sheng QIN ; Xiao-yan QING ; Jun-feng WAN ; Lun LI ; Mei TANG ; Hong YANG ; Bo LIANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(2):137-141
OBJECTIVETo compare the treatment outcomes, complications and histopathologic features between conventional parotidectomy and functional regional parotidectomy in the treatment for pleomorphic adenoma of parotid gland and to provide clinical, and pathological evidence for determining the safe surgical margin.
METHODSOf 109 patients, 60 patients received conventional parotidectomy and 49 patients received functional regional parotidectomy. The rates of tumor recurrence and complications were compared between the groups of patients.
RESULTSThere was no significant difference in the incidence of tumor recurrence, the facial paralysis and sialosyrinx between two groups. The rates of Frey's syndrome, numbness of auricular region, and facial asymmetry were 30.0%, 61.7%, and 38.3% in the patients with conventional parotidectomy respectively, while the rates were 6.1%, 30.6%, and 8.2% in the patients with functional regional parotidectomy, with significant statistically difference, respectively (P < 0.05). Of 109 patients, 33 with incomplete capsule, 29 with capsule penetration, 25 with pseudopodia, and 13 with satellite nodules. There was no significant difference in the depth of tumor infiltration between two groups of patients. For the tumor smaller than 2 cm, the depth of infiltration in conventional group was from 0.061 to 1.122 mm, functional group was from 0.442 to 3.127 mm (Z = -1.093, P = 0.057); for the tumors between 2 - 4 cm, the depth in conventional group was from 0.081 to 7.908 mm, functional group was from 0.082 to 6.632 mm (Z = -0.214, P = 0.831); for the tumor larger than 4 cm, the depth of infiltration was from 0.340 to 8.476 mm.
CONCLUSIONSCompared with conventional parotidectomy, functional regional parotidectomy has good outcomes and less complications. The surgical margins of pleomorphic adenomas of the parotid gland should be determined by the size of tumor. The 1 cm-surgical margins are safe for the tumors less than 4 cm, and the tumors more than 4 cm should be treated with superficial parotidectomy.
Adenoma, Pleomorphic ; pathology ; surgery ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Parotid Neoplasms ; pathology ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult