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 .

Accidents, Occupational ; Humans ; Work Schedule Tolerance

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.

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

Objective To explore the effects of exemestane combined with chemotherapy on the expressions of matrix metalloprotein-9 (MMP-9) and E-cadherin in breast cancer mice.Methods The SPF BALB/c female mice were selected,25 mice with breast cancer model were successfully established and randomly divided them into 3 groups:endocrine group with 8 mice and was given 25 mg of exemestane tablets by gavage once a day;chemotherapy group with 8 mice and was given a range of TP chemotherapy (paclitaxel,135 mg · m-2 and cisplatinum 20 mg · m-2);combined group with 9 mice and was given 25 mg of exemestane tablets by gavage once a day and a range of TP chemotherapy.The normal group contained 6 normal healthy mice and was given the same feeding without drugs.After feeding for 3 weeks,all mice were killed.The expression levels of MMP-9 and E-cadherin in mice right axillary subcutaneous tissue of the four groups were investigated by fluorescence quantitative PCR,immunohistochemistry and Western blot.Results The expression levels of MMP-9 protein in mice:Compared with normal group 0.01 ± 0.00,endocrine group,chemotherapy group,combined group were 0.75 ± 0.37,0.71 ± 0.25,0.33± 0.01 with increased significantly (all P ＜ 0.05);compared with combined group,endocrine group and chemotherapy group increased significantly (all P ＜ 0.01).The expression levels of E-cadherin protein in mice:Compared with normal group 0.64 ±0.28,endocrine group,chemotherapy group,combined group were 0.17 ± 0.08,0.20 ± 0.17,0.44 ± 0.28 with decreased significantly(all P ＜ 0.05);compared with combined group,endocrine group decreased significantly(all P ＜ 0.01).The expression levels of MMP-9 mRNA (MMP-9/β-actin) in mice:Compared with normal group 0.48 ± 0.04,endocrine group,chemotherapy group,combined group were 10.80 ± 0.04,10.54 ± 0.13,3.48 ± 0.04 with increased significantly (all P ＜0.05);compared with combined group,endocrine group decreased significantly(all P ＜ 0.01).The expression levels of E-cadherin mRNA (E-cadherin/β-actin) in mice:Compared with normal group 13.36 ± 0.06,endocrine group,chemotherapy group,combined group were 5.13 ±0.04,5.25 ±0.02,11.36-±0.06 with decreased significantly (all P ＜0.05);compared with combined group,endocrine group decreased significantly (P ＜ 0.01).Conclusion There are close relationships between the expression levels of MMP-9 and E-cadherin and breast cancer,and the combination of exemestane and chemotherapy could significantly improve the expression levels of MMP-9 and E-cadherin.

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.

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.

Objective To develop a ventilation system for the enhaned mobile biosafety level-2(BSL-2)laboratory to control the dispersed aerosol within the laboratory effectively.Methods A ventilation system for the enhanced mobile biosafety level-2 laboratory was designed with the concept of full fresh air ventilation,which realized air supply with 3-stage filtration and constant air volume control,and air exhaust with 1-stage high-efficiency filtration and variable air volume control.The buffer room had the layout of upside air supply and upside exhaust,with the air supply and exhaust equipment integrated into the ceiling;the experimental room had the layout of diagonally upside air supply and diagonally downside exhaust.Computa-tional fluid dynamics(CFD)software was used to analyze the airflow organization,temperature field and velocity field at 0°,30°,45°,60° and 90° air supply angles in order to verify the feasibility of the ventilation system design,and the distributions of aerosol concentration in the experimental room at different air supply angles were simulated to determine the optimal air supply angle.Results Investigations in terms of airflow organization,temperature field,velocity field and aerosol dispersion control showed in case of 60° air supply angel the enhanced mobile BSL-2 laboratory behaved the best and its core indexes all met the requirements of GB 27421-2015 Mobile laboratories—Generalrequirements for biosafety and T/CESC 662-2020 Architectural technical standard for BSL-2 laboratory in medical facilities including the number of air changes,static pressure difference,temperature and humidity,cleanliness and airflow direction.Conclusion Considerations have to be taken on the locations of supply and exhaust outlets,the facility layout and air supply angle when the enhanced mobile BSL-2 laboratory is designed so as to enhance the biosafety of the protective area.[Chinese Medical Equipment Journal,2024,45(7):29-34]

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

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