1.The study of isolation and culture in vitro of human umbilical cord mesenchymal stem cells and their biological properties
Hai BAI ; Ke YANG ; Jianfeng OU
Chongqing Medicine 2016;45(7):876-879
Objective To identify a detailed biological characterization of mesenchymal stem cells (MSCs) isolated from hu‐man umbilical cord(UC) tissue regarding their morphology ,immunophenotype ,purity and proliferative capacity and establish a rea‐sonably cultured and amplified system .Methods After stripping off arteries and veins ,the remaining parts of umbilical cord were cut into 1 mm3 small sections and cultured with DMEM/F12 containing 10% fetal bovine serum .Adhere cells were obtained and the morphology of the cells was observed under inverted phase contrast microscope .The growth curves of them were drawn by CCK‐8 and the cell cycle and surface antigens (CD29 ,CD73 ,CD90 ,CD105 ,CD31 ,CD14 ,CD34 ,CD45 ,CD11b ,HLA‐DR) were detected by flow cytometry .Results Seven to ten days after primary culture ,adhere cells came out of fragments .The MSCs harvested were a high purity and mainly presented as a fibroblast‐like morphology .UC‐MSCs had a strong ability of proliferation through the cell growth curve .The special surface antigens CD29 ,CD73 ,CD90 ,CD105 were positive expression ,while CD31 ,CD14 ,CD34 ,CD45 , CD11b ,HLA‐DR were negative .More than 80% cells of MSCs were found at G0/G1 phase .Conclusion Human UC‐MSCs could be cultured and proliferated in vitro .
3.Diagnostic Significance of Detecting Peripheral Blood Granulocyte CD55/CD59 and FLAER in Anemia and PNH
Ke YANG ; Xiaoyu GUO ; Jianfeng OU ; Hai BAI ; Yaozhu PAN
Journal of Modern Laboratory Medicine 2017;32(3):6-10
Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019<0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163~17.890,P=0.000~0.038<0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ~ 6.308,P=0.000 ~0.016< 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.
4.Immunocompetent cells and immune regulator improve apopotosis of mesenchymal stem cells by down-regulating heme oxygenase-1
Ling ZHANG ; Hai BAI ; Cunbang WANG ; Jianfeng OU ; Guorong YANG
Journal of Third Military Medical University 2003;0(21):-
Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P
5.Effect of cobalt chloride-induced hypoxia on proliferation of human umbilical cord-derived mesenchymal stem cells and related gene and protein expressions
Xiao HAN ; Hai BAI ; Jiaojiao YIN ; Ke YANG ; Yanxia HAN ; Jianfeng OU ; Cunbang WANG
Chinese Journal of Tissue Engineering Research 2015;(45):7268-7273
BACKGROUND:Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cels (hUC-MSCs) in a time- and concentration-dependent manner, and meanwhile, CoCl2 can regulate the expression of genes and proteins in hUC-MSCs. OBJECTIVE:To explore the effects of CoCl2 induced-hypoxia on the proliferation of hUC-MSCs and gene and protein expressions in hUC-MSCs, thereby establishing an effective method for MSCs culture and amplificationin vitro. METHODS: hUC-MSCs were extracted using tissue explant method. Under hypoxia conditions induced by CoCl2 (0, 100, 150, 200, 250 μmol/L) for different periods (0, 1, 2, 3, 4 days), flow cytometry was used to identify cel surface-associated antigens; cel counting kit-8 was used to detect cel proliferation; RT-PCR was used to determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cel-derived factor-1, interleukin-6, transforming growth factor-β mRNA; western blot assay was used to detect protein expression of hypoxia inducible factor-1α. RESULTS AND CONCLUSION:The cels were positive for CD29, CD73, CD90, CD105, while negative for CD31, CD14, CD34, CD45, CD11b, HLA-DR. Moreover, the antigen expression was not affected by CoCl2 induced-hypoxia. CoCl2 induced-chemical hypoxia could promote the proliferation of hUC-MSCs in a time- and concentration-dependent manner. RT-PCR results showed thatunder hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cel-derived factor-1 mRNA expressions were significantly up-regulated, but interleukin-6 and transforming growth factor-β mRNA expressions were down-regulated significantly (P < 0.05). Additionaly, the protein expression of hypoxia inducible factor 1α was increased under hypoxia conditions. These findings indicate that CoCl2 induced-hypoxia environment may promote the proliferation of hUC-MSCs and the optimal concentration of CoCl2 is 200μmol/L. However, a higher concentration of CoCl2 (≥ 250μmol/L) inhibits the proliferation of hUC-MSCs, and the mechanism may be related to the increase of hypoxia inducible factor-1α at protein and mRNA levels.
6.Suggestion of clinical laboratory biosafety administration
Junwei DENG ; Ou SHE ; Xiaoliu SHI ; Jianzhen FANG ; Minneng LI ; Xinjie YANG ; Zhijie HAI ; Hua WANG ; Qun LONG
Chinese Journal of Medical Science Research Management 2008;21(3):143-144,186
Clinical laboratory biological safety is one of whole society safety. This paper introduced briefly the current situation of clinical medical laboratory biosafty in the hospital. and set forth common biological hazards specifically for whose characteristics. Combining the biosafety administration measures from abroad, the issue of laboratory biological safety administration was considered, and put forward some suggestions according to related law and regulation of national laboratory safety administration in order to strengthen clinical laboratory biosafety administration.
7.Vacuum-assisted biopsy and wire localization for the diagnosis of non-palpable breast lesions.
Kun-Lun SU ; Hai-Bin XU ; Zu-Jian HU ; Jun-Ling HE ; Ou-Ou YANG ; Wang-Hua HU
Chinese Journal of Oncology 2010;32(6):472-475
OBJECTIVETo compare the effectiveness and accuracy of the use of vacuum-assisted biopsy (VAB) versus wire localization (WL) in the diagnosis of non-palpable breast lesions (NPBL).
METHODSNinety-seven consecutive women with NPBL were randomized into VAB group and WL group. All specimens were identified by mammography. The patients were requested to score the cosmetic appearance of their breast after operation, and a numerical rating scale was used to measure pain on the first postoperative day. Underestimation rates for atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS) were recorded if open surgical biopsy revealed DCIS and invasive cancer, respectively. Clear margins were also recorded in the two groups.
RESULTSVAB and WL located all the NPBL successfully. In the VAB group, the specimen volume was smaller than that of the WL group (2.3 cm(3) vs. 18.4 cm(3), P = 0.03). Underestimation rates of ADH and DCIS in the VAB group were 16.7% and 11.1%, respectively. The diagnostic accordance rate of VAB was 97.9%, the false negative rate was 2.1%, and there was no false positive case. The means of the numerical rating pain scale were different in both groups (1.7 for VAB vs. 2.5 for WL, P = 0.02). When cosmetic results were taken into account, 40 VAB patients had excellent outcomes and 8 good outcomes, compared with 25 excellent and 24 good for the WL group. There were better cosmetic outcomes with the VAB procedure (P < 0.05).
CONCLUSIONVAB is highly reliable and may avoid diagnostic open surgery in the majority of patients with benign lesions. However, because of the underestimation of histologic diagnosis and tumor margin involvement, VAB can not be used to completely substitute wire localization.
Adult ; Biopsy, Needle ; instrumentation ; methods ; Breast ; pathology ; Breast Neoplasms ; diagnosis ; pathology ; Carcinoma in Situ ; diagnosis ; pathology ; Carcinoma, Ductal, Breast ; diagnosis ; pathology ; Diagnostic Errors ; Female ; Fibroadenoma ; diagnosis ; pathology ; Humans ; Hyperplasia ; Middle Aged ; Precancerous Conditions ; diagnosis ; pathology ; Stereotaxic Techniques ; instrumentation ; Vacuum
8.Analysis on comprehensive noise control effects in an automobile engine manufacturing enterprise
Lin CHEN ; Youmei SHU ; Yang LIAO ; Hai ZHANG ; Kunpeng ZHONG ; Jun LIU ; Fengyi OU ; Zhi WANG
China Occupational Medicine 2024;51(2):168-171
ObjectiveTo analyze the measures and effects of comprehensive noise control in an automobile engine manufacturing enterprise. Methods An automobile engine manufacturing enterprise with noise exposure work-sites was selected as the research subject. The noise hazard status of the enterprise was investigated. The noise hazard exceeded the national standard in work-sites was comprehensively treated. The treatment effect was analyzed. Results A total of 395 work-sites exposed to noise were detected. Among them, 16 work-sites exceeded the national noise standard, with the rate of 4.1%. The work-sites with noise exceeding the national standard were mainly in the machine workshop and the casting workshop, with normalization of equivalent continuous A-weighted sound pressure level to a nominal eight hours working day (LEX,8h) ranging from 86.8 to 89.2 and 85.2 to 90.2 dB(A), respectively. In addition, the classification of occupational noise exposure of the mold protection group in the casting workshop was level Ⅱ, and classification of occupational noise exposure of other work-sites with excessive national standard was grade Ⅰ. After the enterprise implemented noise control measures such as process reform, equipment replacement, sound insulation, noise reduction, sound absorption, and vibration isolation, the LEX,8h of nine positions were lower than those before rectification [(86.7±1.1) vs (83.3±1.3) dB(A), P<0.01]. Conclusion The use of comprehensive control measures can achieve the purpose of reducing noise from the sources, and can be used for reference by relevant enterprises with noise hazards.
9.Features of the dual energy technique with dual-source computed tomography for anterior cruciate ligament injuries.
Rui BAI ; Shan-xing OU ; Hai-ling LIU ; Guo-qing QIAO ; Ping-yue LI ; Hua-yang HUANG
Acta Academiae Medicinae Sinicae 2010;32(6):663-665
OBJECTIVETo explore the diagnostic value of the dual-energy technique with dual-source computed tomography (DSCT) for anterior cruciate ligament injuries.
METHODSThe clinical data of 8 patients with arthroscopic results were retrospectively reviewed. All patients underwent two- and three-dimensional imaging by multiplanar reconstruction, volume rendering, and tendon mode on DSCT. Dual-energy characteristics were compared with arthroscopic results.
RESULTSSix patients who were arthroscopically diagnosed as anterior cruciate ligament injuries, all of them were also correctly diagnosed by DSCT. Two patients who were arthroscopically diagnosed as normal, one was also diagnosed as normal by DSCT and the other was misdiagnosed. The overall agreement rate was 87.5% (7/8) . Under the dual energy tendon mode, the dual energy staining of the injured anterior cruciate ligament was lower than that of the contralateral normal cruciate ligament of the patient.
CONCLUSIONThe staining diminution in DSCT imaging may be a new feature that can be used to effectively diagnose anterior cruciate ligament injury.
Adolescent ; Adult ; Anterior Cruciate Ligament Injuries ; Female ; Humans ; Knee Injuries ; diagnostic imaging ; Male ; Tomography, X-Ray Computed ; methods ; Young Adult
10.Evaluation of sealing ability of three kinds endodontic materials as root canal sealers.
Hai-jing GU ; Li-min LIU ; Qiong XU ; Yong OU-YANG ; Jun-qi LING
West China Journal of Stomatology 2007;25(3):257-259
OBJECTIVETo compare the apical microleakage of Vitapex (calcium hydroxide based paste) with that of AH-plus and zinc oxide eugenol sealer when used with laterally condensed gutta percha obturation technique.
METHODSOne hundred single rooted human anterior teeth were instrumented and randomly divided into three experimental groups (A, B, C) of 30 teeth each and two control groups (D, E) of 5 teeth each. Group A was filled with laterally condensed gutta-percha using Vitapex as sealer. Group B was filled with laterally condensed gutta-percha using AH-plus as sealer. Group C was filled with laterally condensed gutta-percha using zinc oxide eugenol as sealer. Group D was the positive control. Group E was the negative control, which were coated with nail polish to entire root surface. Teeth were then suspended in 2% methylene blue. After this, teeth were demineralized dehydrated and cleared. Linear dye penetration was determined under stereomicroscope with calibrated eye piece.
RESULTSThe mean dye penetration for group A, B, C were respectively (0.57 +/- 0.56) mm, (0.79 +/- 0.96) mm and (1.07 +/- 1.12) mm. Group D demonstrated maximum dye penetration. Group E showed no dye penetration. There was no statistically significant difference between group B and group C (P > 0.05). However, there was statistically significant difference between group A and group B, C (P < 0.01).
CONCLUSIONThis study showed that Vitapex used as endodontic sealer material are better than AH-plus sealer and zinc oxide eugenol sealer.
Calcium Hydroxide ; Dental Leakage ; Dental Pulp Cavity ; Gutta-Percha ; Humans ; Molar ; Root Canal Filling Materials ; Root Canal Obturation ; Silicones ; Zinc Oxide-Eugenol Cement