1.Functions of human periodontal myofibroblast in vitro.
West China Journal of Stomatology 2015;33(2):130-134
OBJECTIVETo investigate the functions of human periodontal myofibroblast (MFB) in vitro.
METHODSHuman periodontal fibroblast (hPDLFs) was cultured and induced to MFB by transforming growth factor-β1 (TGF-β1). MFB was denoted as the experimental group, whereas the hPDLFs was the control group. The groups were continuously cultured and harvested at 0, 12, 24, 48, and 72 h. The MFB marker α-smooth muscle actin (α-SMA) was examined by immunocytochemistry. The expression of fibronectin (FN) between MFB was examined by immunocytochemistry to detect the MFB contact relationship. The mRNA expression levels of α-SMA, collagen (Col) I, and Col III were measured by reverse transcription-polymerase chain reaction (RT-PCT) to analyze extracellular matrix secretion. The protein expression levels of α-SMA and Col I were also assessed by Western blot.
RESULTSThe experimental group had significantly higher α-SMA expression than the control group at 0 h (P < 0.001). A positive expression of FN was found between MFB. The experimental group had significantly higher expression levels of Col I and Col III than the control group at 24 h (P < 0.001).
CONCLUSIONHuman periodontal MFB presents a continuous, high expression of α-SMA. MFB could interact through FN. MFB is significantly capable of extracellular matrix secretion.
Actins ; Epithelial Cells ; Extracellular Matrix ; Fibroblasts ; Fibronectins ; Humans ; Jaw ; metabolism ; Myofibroblasts ; Transforming Growth Factor beta1
2.Isolation and identification of lung cancer stem cells from NCI-H1650 cell line
Man DING ; Anmei ZHANG ; Bo ZHU
Chongqing Medicine 2015;44(12):1588-1591
Objective To isolate and identify the lung cancestem like cellfrom the cell line NCI-H1650 .MethodThe NCI-H1650 cellwere continuously cultured in serum-free medium with human insulin ,recombinanepidermal grow th facto(EGF) ,recombinanbasifibroblasgrowth facto(bFGF) and bovine serum albumin(BSA) ,and paclitaxel waadopted to induce the stem cell.Flow cytometry and immunofluorescenstaining were used to analyze the expression levelof lung cancestem cell markeCD133 and CD326 .The expressionof stem cell related markeOct4 ,Nanog and Sox-2 were detected by RT-Pcand West-ern blo.ResultThe NCI-H1650 cellcontinuously cultured by serum-free medium and induced by combining paclitaxel induction showed the spherical suspension growth .Compared with NCI-H1650 cellcultured by serum ,CD133 and CD326 had significantly high expression and the expression levelof Oct4 ,Nanog and Sox-2 were significantly increased (P<0 .05) .Conclusion Adopting the serum-free culture in vitro combined with paclitaxel foinducing NCI-H1650 cellcan effectively isolate the stem cellamong them.
3.Clinical Curative Effect and Safety of MCT/LCT Fat Emulsions at Different Doses in Preterm and Low Birth Weight Infants
Jing YUAN ; Man HE ; Wujun DING
China Pharmacist 2015;(3):444-445
Objective:To analyze the effect of MCT/LCT fat emulsions at different doses on the growth and development of prema-ture and low birth weight infants. Methods: Totally 76 cases of preterm or low birth weight infants were divided into the treatment group (40 cases) and the control group (36 cases), both groups were given MCT/LCT fat emulsions, amino acid and glucose injec-tions in 12-24 h after the birth. The initial dose of MCT/LCT fat emulsions in the treatment group was 2. 0 g·kg-1 ·d-1 and the dose was gradually increased to 3. 0 g·kg-1 ·d-1 with 0. 5 g·kg-1 ·d-1 progressive increase. The initial dose of MCT/LCT fat emulsions in the control group was 0. 5 g·kg-1 ·d-1 , and the dose was increased to 3. 0 g·kg-1 ·d-1 with 0. 5 g·kg-1 ·d-1 progressive in-crease. The treatment course was 7 days. The neonatal weight, serum total bilirubin, direct bilirubin, alanine aminotransferase, total cholesterol, triacylglycerol and blood glucose were monitored in the two groups. Results:The born body mass recovery time of the ob-servation group was (4. 38+0. 93) d, which was significantly shorter than that in the control group [(6. 81+1. 90) d]. After the re-covery, the body weight growth rate of the observation group was (30. 41+1. 81) g·kg-1 ·d-1 , which was significantly higher than that of the control group [(18. 21 +2. 08) g·kg-1 ·d-1, P <0. 05]. After the 7-day treatment, the blood biochemical indices showed no statistically significant difference between the two groups (P>0. 05). Conclusion:Early application of MCT/LCT fat emul-sions at high dose is beneficial to the improvement of nutritional status in premature infants.
5.Effect of positive end expiratory pressure on pulmonary function of one-lung ventilation patient during perioperation
Peiyan DING ; Wenjun YAN ; Man HE ; Ruobin LIU ; Yabing MA
Chinese Journal of Postgraduates of Medicine 2011;34(18):1-3
Objective To explore the effect of positive end expiratory pressure (PEEP) on arterial oxygenation and intrapulmonary shunt during one-lung ventilation (OLV) and pulmonary function during perioperation. Methods Forty patients with normal pulmonary function,ASA I - II :scheduled for pulmonary lobectomy, were divided into control group and PEEP group by random digits table with 20 cases each. Patients were induced by double-lumen tubes under intravenous anesthesia and were received 10 ml/kg tidal volume, 12 frequents/min breathing rate during the two-lung ventilation (TLV), secondary reduced to 6 ml/kg tidal volume, 16-18 frequents/min breathing rate without PEEP (control group) or with 5 cm H2O cm H2O =0.098 kPa) PEEP (PEEP group) during OLV.Hemodynamics and respiratory mechanical parameters were continuously monitored, lung function before operation and at 72 h after operation was detected. Results Compared to before OLV,arterial oxygen tension (PaO2), arterial oxygen saturation (SpO2), oxygenation index (OI) were decreased and intrapulmonary shunt ratio (Qs/Qt) was increased in control group and PEEP group at 30 min after OLV (P < 0.01 or < 0.05). However,PaO2 and SpO2 and OI were higher and Qs/Qt was lower in PEEP group than that in control group at the same time point (P<0.05). In addition, FEV1%, FVC% and FEV1/FVC were (121.8 ± 25.0% ,(117.2 ± 24.3)% , (87.6 ± 15.7)%before operation and (84.9 ± 21.6)%, (77.2 ± 18.3)% , (70.5 ± 12.5)% at 72 h after operation respectively in control group, (116.9 ±24.5)% , (112.1 ±23.6)% , (85.3 ± 13.8)% before operation and (96.3 ± 20.4)%, (88.1 ± 19.8)% , (78.4 ± 10.2)% at 72 h after operation respectively in PEEP group. Although decreased in control group and PEEP group at 72 h after operation comparing with preoperation (P< 0.01 or < 0.05 ), FEV1%, FVC% and FEV1/FVC were higher in PEEP group than those in control group at 72 h after operation (P<0.05). Conclusion Appropriate PEEP increases arterial oxygenation,reduces Qs/Qt and improves pulmonary function during OLV,reduces the risk of hypoxernia and lung injury induced by OLV during perioperation.
6.Piggyback implantation using Toric intraocular lens in high myopia
Qiong-Yan, TANG ; Man-Qiang, PENG ; Ding, LIN
International Eye Science 2014;(9):1599-1601
To assess the postoperative outcomes of piggyback implantation using Acrysof Toric intraocular lens ( lOL ) in high myopia combined with corneal astigmatism.
●METHODS: Sixty patients who had phacoemulsification with lOL implantation due to high myopia, cataract and corneal astigmatism from January 2010 to June 2013 were randomly divided into observation group (piggyback Toric lOL implantation, both an Acrysoft lQ Toric lOL and a minus foldable acrylic three piece lOL were implanted in the capsular bag, n= 30) and control group (foldable lOL implantation, n = 30). Postoperative follow - up went on 6mo. lnformation collected included uncorrected visual acuity ( UCVA), lOL position, residual astigmatism and complications.
● RESULTS: The UCVA increased from 3. 52 ± 0. 03 preoperatively to 4. 78±0. 01 at 6mo postoperatively in the observation group, from 3. 51±0. 03 preoperatively to 4. 30± 0. 13 at 6mo postoperatively in the control group. The observation group's postoperative UCVA was better than that of the control group. There was statistically significant difference ( t = 3. 612, P < 0. 05 ). The preoperative corneal astigmatism was 2. 97 ± 0. 87 (1. 70 -4. 27) D in the observation group and 2. 92 ± 0. 97 (1. 50 -4. 90)D in the control group. The postoperative residual astigmatism was 0. 48 ± 0. 23 ( 0. 25 - 1. 00 ) D in the observation group and 2. 87 ± 1. 11 (1. 00 - 5. 20) D in the control group. There was statistically significant difference postoperatively (t = - 11. 995, P < 0. 05) between the two groups. No complications occurred.
●CONCLUSlON: Piggyback implantation using Toric lOL can help to solve the problem of no matching Toric lOL power for the high myopia combined with corneal astigmatism at the current stage. lt improves the UCVA and reduces the astigmatism after the cataract surgery.
7.Impacts of RNA interference targeting the polo-like kinase-1 gene on the invasion of and anoikis in human malignant melanoma cells
Keyun DING ; Juan XU ; Changfeng MAN ; Yu FAN
Chinese Journal of Dermatology 2014;47(6):413-416
Objective To investigate the effects of down-regulation of polo-like kinase-1 (PLK1) gene by RNA interference (RNAi) on the invasion of a human malignant melanoma cell line A375 and their possible mechanisms.Methods Cultured A375 cells were classified into several groups:blank control group receiving no treatment,liposome group transfected with lipofectamine only,and three siRNA groups transfected with three concentrations of a small interference RNA (siRNA) targeting PLK1 respectively.After additional culture,real time quantitative PCR and Western blot analysis were performed to quantify the expressions of PLK1 mRNA and protein in A375 cells respectively,Transwell invasion assay to evaluate the invasive capacity of A375 cells,agarose gel electrophoresis and terminal deoxynucleotidyl-transferase-mediated dUTP-biotin nick end labelling (TUNEL) to detect anoikis in A375 cells.The colony-forming capacity was also evaluated for A375 cells.Statistical analysis was carried out by one-factor analysis of variance.Results There was a significant decrease in PLK1 mRNA and protein expressions as well as in colony-forming units in the siRNA groups compared with the blank control group (all P < 0.05).The invasive capacity of A375 cells was significantly inhibited in the siRNA groups with the number of migrating cells in Transwell assay being 39 ± 5,19 ± 5 and 9 ± 3 in A375 cells transfected with 3.125,6.250 and 12.500 nmol/L siRNAs respectively,compared to 56 ± 5 in the blank control group (all P < 0.05).A characteristic DNA ladder was observed on agarose gel electrophoresis in the siRNA (6.250 nmol/L) group.Compared with the blank control group and liposome group,the three siRNA groups showed increased apoptotic index (3.86% ± 0.35% (3.125 nmol/L siRNA),7.35% ± 0.36% (6.250 nmol/L siRNA) and 17.56% ± 0.38% (12.500 nmol/L siRNA) vs.1.15% ± 0.25% (blank control group) and 1.18% ± 0.22% (liposome group),all P < 0.05).Conclusions PLK1 siRNA can inhibit the invasion of malignant melanoma cells,likely by inducing anoikis in these cells.
8.Establishment of a three-dimensional finite element model of transmission straight wire appliance
Rui DING ; Yunna MAN ; Peiling WU ; Wei WANG
Chinese Journal of Tissue Engineering Research 2013;(33):5942-5948
BACKGROUND:With the development of modern orthodontics, to invent an efficient appliance is the focus in
recent studies. Transmission straight wire appliance was born on this background. This appliance can accelerate occlusion and shorten treatment duration. The relationship between distal width and angulation of gable bends of main arch wire needs to study in depth.
OBJECTIVE:To establish three-dimensional finite element model of transmission straight wire appliance with better biological and mechanical similarity, and to obtain the relationship between distal width and angulation of gable bend of main arch wire.
METHODS:By using scanning of spiral CT with 64 rows, the sectional image data in DICOM format of maxil ary dentition and maxil ae of the volunteers (Class Ⅱ, division 1) were obtained. With the help of Ansys workbench 13.0, Mimics 10.01, Unigraphics NX and Geomagic Studio 8.0 softwares, the three-dimensional finite element model including transmission straight wire appliance, bend, Australian Orthodontic Wire, maxil ae, maxil ary tooth and periodontal ligament was established in Windows XP Service Pack 3 system.
RESULTS AND CONCLUSION:A three-dimensional finite element model of transmission straight wire appliance was established, which consisted of 250 929 elements and 657 766 nodes. Furthermore, three-dimensional finite element model had higher geometric similarity and mechanical similarity, as wel as the advantages of adding or subtracting components according to the requirement of the research. The model was conductive to analyze the mechanical system of transmission straight wire appliance and guide the clinical application and appliance modification.
9.DOT1L-long enhances breast cancer metastasis
kai Xiao DING ; kun Yin FU ; MOHAN MAN
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(10):1327-1331
Objective · To investigate the histone methyltransferase capability of DOT1L-long form and its role in breast cancer metastasis.Methods · The existence of DOT1L-long form was confirmed by PCR, and the mRNA level of DOT1L was tested by real-time PCR. In HEK293T cells in which DOT1L canonical and DOT1L-long were overexpressed respectively, Western blotting was used to test the expression level of DOT1L and the histone methyltransferase capability. In the MCF10A cell line with inducible expression of DOT1L-long, real-time PCR was used to detect the mRNA level of epithelial-mesenchymal transition (EMT) marker, and transwell assay was used to detect the migration of breast cancer cells in which the expression level of DOT1L is low or high. Results · PCR demonstrated the existence of DOT1L-long form, and real-time PCR showed it widely exists in HCT116, T98G, MCF10A cells, etc. Western blotting showed the expression of DOT1L-long form and its H3K79 methyltransferase activity. In MCF10A cells in which overexpressed canonical DOT1L and DOT1L-long, mRNA levels of N-cadherin and fibronectine increased. Transwell showed canonical DOT1L and DOT1L-long both substantially increased the migration of breast cancer cells. Conclusion · The existence of DOT1L-long was confirmed and investigated, which is 202 amino acids longer than the canonical DOT1L, and is coded by a new exon, located between exon 27 and 28. Further, the DOT1L-long has H3K79 methyltransferase activity, and is able to promote breast cancer metastasis.
10.Control Modes of Hospital Disinfection and Supply Rooms in Hubei Province
Min LIU ; Hua CAO ; Man WU ; Qiuling XIONG ; Chuanxin DING
Chinese Journal of Nosocomiology 2006;0(09):-
OBJECTIVE To survey the current situation of disinfection and supply rooms in 34 hospitals of Hubei Province under the Committee of Hubei Distinfection.METHODS On the basis of certain references,an questionaire was designed to survey on site.RESULTS All 34 hospitals were taken disperse control mode of disinfection;the education was not good enough;most of the disinfection and supply rooms covered limited space,without meeting the standards of Ministry of Health.The rate of equipement usage was low;the working spheres were narrow.CONCLUSIONS There is not any control mode of disinfection and supply rooms in these 34 hospitals;if working staff and equipment are improved,the working spheres can be broaden.it should be transferred to the centralized control mode of disinfection.