1.Passage of superparamagnetic iron oxide labeled rat bone marrow mesenchymal stem cells in vitro: Do iron particles change with passage?
Hua YANG ; Xiaoming ZHANG ; Hong JIANG ; Yang SHAO ; Nanlin ZENG
Chinese Journal of Tissue Engineering Research 2009;13(49):9705-9708
BACKGROUND: As magnetic resonance (MR) contrast, a large number of clinical and experimental researches have been done on superparamagnetic iron oxide (SPIO), while the report on the labeled cell passaged cells is rare.OBJECTIVE: MR imaging was performed to the labeled bone mesenchymal stem cells (BMSCs) and its passaged cells in vitro, in order to establish the base of monitoring magnetic labeled BMSCs with magnetic resonance imaging (MRI) in vivo.DESIGN, TIME AND SETTING: The cytological in vitro experiment was performed at the Imaging Research Institute and Institute of Rheumatology and Immunology of North Sichuan Medical College Hospital from June 2006 to January 2007. MATERIALS: Two clean female albino rats (Animal Center, North Sichuan Medical College), SPIO (Schering AG,Germany) were used in this study.METHODES: Bilateral femur and tibia bone marrow was extracted from rats. BMSCs were harvested and purified using the adherent method, and then labeled with 600 μL ferric oxide-polylysine compound (42 mg/L iron concentration) in vitro. MAIN OUTCOME MEASURES: Cell maker-positive rate and the MR signal intensity were respectively measured to the labeled cells and its passaged cells under the inverted microscope and magnetic resonance imaging. RESULTS: Following Prussian blue staining, labeling rate of SPIO labeled cells at the first passage was 100%. With increased passage, the labeling rate was reduced from the first to fifth passages. Compared with non-labeled PBS control, there was no significant difference in signal intensity in the first and second passages cells, but the signal intensity percentage was gradually decreased with signal intensity of increased cell passage from the third passage. Cell labeling rate was negatively correlated with T2~*WI signal intensity (r=-0.986 6, P <0.005). CONCLUSION: The iron particles in the magnetic labeled cells can be passaged to the offspring cells, and can be monitored in a certain period of time with MRI in vitro. These results firstly introduced that SPIO-labeled cell iron particles can decrease with cell passage.
2.Superparamagnetic iron oxide labeled mesenchymal stem cells and their magnetic resonance imaging in vitro
Hua YANG ; Xiaoming ZHANG ; Yang SHAO ; Hong JIANG ; Zhaohua ZHAI
Journal of Third Military Medical University 2003;0(17):-
Objective To explore the optimal situation of labeling bone mesenchymal stem cells (BMSCs) with superparamagnetic iron oxides (SPIO) mediated by poly-L-lysine (PLL), and determine the most optimal protocol of magnetic resonance imaging according to the patterns of MR in vitro. Methods BMSCs were isolated from white rat and purified, incubated with SPIO-PLL complexes at the range of concentrations (0, 4.2, 8.4, 21, 42, 84 ?g Fe per ml medium). The labeling ratio and distribution of SPIO particles in BMSCs, and the morphological evidence of abnormal visualization were evaluated by Prussian blue staining, fluorescent microscope and electron microscopy. MTT growth curves and magnetic resonance imagings were obtained at the range of concentrations. Trypan blue exclusion test was performed to elevate the viability of BMSCs labeled with PLL at the range of concentrations (0, 0.05, 0.25, 0.5, 1.0, 5.0 ?g PLL per ml medium). Results The cellular labeling ratio was strongly correlated to the concentrations of SPIO (P
3.The significance and the clinical analysis of calcification in thyroid node
Minghao LI ; Jiaxing YANG ; Yanru JIANG ; Benling SHAO
Chinese Journal of Postgraduates of Medicine 2008;31(14):33-35
Objective To investigate the significance of calcification in thyroid node for diagnosis of thyroid carcinoma.Method Retrospective analysis of 107 thyroid nodules' pre-operative ultrasonic and postoperative pathologic results.Results Total ultrasonic thyroid calcification ratio was 27.1%(29/107),which in benign samples was lower than Ihat in malignant samples(17.2%vs 70.0%,P<0.01).Micro-calci-fication ratio in benign samples Was lower than thai:in malignanl samples(8.0%vs 50.0%,P<0.01).Conclusion The ralio of thyroid carcinoma with calcification is higher,so the detection of thyroid carcinoma,especially micro-single-calcification should be significant.
4.Comparison of intraocular pressure measured by Icare rebound tonometer and Goldmann applanation tonometer
He-zheng, ZHOU ; Shao-yang, ZHANG ; Wen-shan, JIANG
Chinese Journal of Experimental Ophthalmology 2011;29(5):456-459
Background As a new tonometer,it is necessary to assess the clinical value of Icare rebound tonometer.Objective This study was to compare the intraocular pressure(IOP)values measured by Icare with that measured by GAT,and discuss the clinical value of leare rebound tonometer. Methods IOP measurement was performed on 152 eyes of 78 subjects with suspicious glaucoma,glaucoma,refractive error and normal examinnee by Icare and GAT respectively.The Icare IOP was measured firstly and then the GAT IOP was carried out with the 3-or 5-minute interval.The IOP values were compared between Ieare and GAT.This study was approved by Ethic Committee of Wuhan General Hospital of Chinese PLA.Written informed consent was obtained from each subject prior to this study. Results The mean IOP values of Icare and GAT were(19.16±5.03)mmHg and(18.41±4.52)mmHg respectively.The differences between Icare IOP and CAT IOP were less than or equal to 1 mmHg in 96 of 105 eyes(63.2%).The positive correlation was found between the Icare IOP and GAT IOP(r=0.940,P<0.01).The Ieare IOP was lower than that of GAT when IOPIcare<16 mmHg,however,the IOP of Icare were higher when IOPIcare≥6 mmHg;the IOP of Icare were higher than that of GAT in the total CCT range.The correlation coefficients of IOP of Icare or CAT with CCT were 0.341(P<0.01)and 0.333(P<0.01),respectively. Conclusion Compared with GAT,Icare is more feasible in clinic because it is practicable and reliable.
5.Effect of E-cadherin in epithelial-mesenchymal transition of neuroblastoma
Jingwei YANG ; Hui JIANG ; Zhimei GAO ; Jingbo SHAO
Chinese Journal of Applied Clinical Pediatrics 2021;36(5):369-373
Objective:To analyze the expression of E-cadherin in the epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1) in neuroblastoma.Methods:TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), was applied to SK-N-SH cells in vitro compared with the blank control group.EMT-related genes mRNA and protein expression were detected by carrying out real-time PCR assays and Western blot.A scratch test and migration assay were performed to verify the alteration of SK-N-SH cell migration capacity.Data collected from 18 cases of neuroblastoma patients were selected from the Department of Hematology Oncology, Shanghai Children′s Hospital from January 2008 to December 2012.The expression of E-cadherin in the tumor tissue of the neuroblastoma patients after operation was detected by immunohistochemistry.The clinical features and survival prognosis of these patients were analyzed. Results:Compared with the control group, after SK-N-SH cells were treated with TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), real-time PCR assays and Western blot revealed that the mRNA(0.603±0.081, 0.606±0.008, 0.716±0.166 vs.1.000) and protein expression levels(0.855±0.026, 0.600±0.017, 0.495±0.011 vs.1.000) of E-cadherin were significantly decreased ( F=8.144, P=0.040; F=74.810, P<0.001), while the mRNA(2.132±0.167, 3.494±0.017, 4.184±0.021 vs.1.000) and protein expression levels (1.175±0.053, 1.189±0.058, 1.225±0.106 vs.1.000)of α - smooth muscle actin were significantly increased ( F=547.300, P<0.001; F=68.810, P=0.007), suggesting that EMT changes occur in cells.Scratch test and Transwell migration assay revealed that the number of migrating cells increased obvious with the treatment of TGF-β1 (5 μg/L) ( t=16.070, P=0.040). The 10-year overall survival(OS) rates of neuroblastoma patients with E-cadherin strong positive expression, positive expression, weak positive expression and negative expression in the pathology were (77.78±13.86)%, (75.00±21.66)%, (25.00±21.65)% and 0, respectively ( F=8.160, P=0.040). Conclusions:TGF-β1 can induce the EMT in SK-N-SH cells and increase cell migration.The decrease expression of E-cadherin in neuroblastoma patients is closely associated with clinical progress and recurrence or metastasis of the disease.
6.Clinical analysis of 45 children with Langerhans cell histiocytosis
Xuelian LIAO ; Hui JIANG ; Zhenghua LU ; Jingbo SHAO ; Jingwei YANG ; Shayi JIANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(15):1145-1148
Objective To analyze the clinical data of children with Langerhans cell histiocytosis (LCH),to discuss the therapeutic effect,and to analyze the factors related to prognosis.Methods A total of 45 children diagnosed as LCH were divided into group A (18 cases with bone lesion only),group B(6 cases with soft tissue lesion),and group C (21 cases with viscera lesion) according to Shanghai Children's Hospital-LCH-2007 scheme [SCH-LCH-2007 (modified DAL-HX83/90) scheme].(1) Initial treatment:group A was treated with Prednisone (Pred) + Vincristine (VCR) for 28 weeks,and group B was treated with Pred + VCR + Etoposide (VP16) + Mercaptopurine (6MP) for 43 weeks,and group C was treated with Pred + VCR + VP16 + Methotrexate (MTX) +6MP for 52 weeks.(2) Re-treatment scheme after relapse included:①upgrading treatment,group A to group B,group B to group C.②Individual treatment for group C included VP16 modification,and maintained Thymosin and/or Ciclosporin etc.Results The total survival rate was 93.3% (42/45 cases) and recurrence rate was 26.7% (12/45 cases).Children in group A and B were all effective,while 2 patients in group C died,and 1 case missed follow-up.Multi-factor analysis showed that the factors like age,sex,group,skeleton,soft tissue,erythra,lymph gland,lung,mouth,ears,hypophysis pituitary had no statistical significance,but liver,spleen and blood involvement had statistical significance in disease relapse:liver (P=0.007 1),spleen (P=0.016 9),and blood (P=0.011 1).Conclusion LCH can affect several organs of children and relapse,and modified DAL-HX83/90 scheme is very effective.The liver,spleen and hematopoiesis system involvement is correlates with the relapse.
7.In vitro sensitivity to macrolides and mutation analysis in the 23S rRNA gene of Chlamydia trachomatis
Lili SHAO ; Yong JIANG ; Lina YANG ; Xiaojing YANG ; Yuanjun LIU ; Quanzhong LIU
Chinese Journal of Dermatology 2010;43(5):305-308
Objective To test the in vitro susceptibility to macrolides of urogenital Chlamydia trachomatis (Ct) isolates, screen resistant Ct strains, and to explore resistance mechanism at the molecular level. Methods A total of 42 Ct strains were isolated from cervical or urethral swab samples and propagated in McCoy cells until the infection rate reached more than 90%. Then, susceptibility test was performed to evaluate the activity of three macrolides. Reverse transcription PCR and PCR were used to amplify two macrolide-resistance related genes, i.e., 23S rRNA gene and L4 gene, respectively in 2 erythromycin-resistant Ct strains and 4 erythromycin-sensitive strains followed by direct sequencing. Results The minimal inhibitory concentration (MIC) varied from 0.5 to 2 mg/L for erythromycin, 0.008 to 0.032 mg/L for clarithromycin, 0.125 to 0.5 mg/Lfor azithromycin. Erythromycin resistance was found in 2 isolates with the MIC value being 2 mg/L. Two mutations, C2452A and T2611A/C (Escherichia coli numbering) in the 23S rRNA gene, were detected in the resistant strains only, while the other 2 mutations, Pro113Leu and Pro156 Ala in L4 gene, were observed in all the tested strains. Conclusions Erythromycin-resistant Ct strains have emerged in clinical settings. The low-level erythromycin resistance may be associated with C2452A and T261 1C mutations in the 23S rRNA gene, whereas the point mutations in L4 gene is unlikely related to erythromycin resistance.
8.Postoperative complications of complex proximal humeral fractures after treated with proximal humeral internal locking system plate fixation
Fuyuan MA ; Tieyi YANG ; Rui JIANG ; Yan ZHANG ; Yue LIU ; Jin SHAO
Chinese Journal of Tissue Engineering Research 2013;(48):8381-8387
BACKGROUND:Proximal humeral internal locking system fixation for complex humeral fractures via deltoid splitting approach provides good clinical results, but certain complications stil existed.
OBJECTIVE:To explore the postoperative complications and the related risk factors for displaced three-part and four-part fractures of proximal humerus treated with proximal humeral internal locking system fixation via deltoid-splitting approach, and to propose the corresponding countermeasures.
METHODS:106 cases with displaced three-part and four-part fractures of proximal humerus were retrospectively analyzed. The relationship between postoperative complications and the related risk factors was analyzed with Logistic regression analysis.
RESULTS AND CONCLUSION:A total of 81 patients were fol owed-up for 12 to 30 months. The mean Constant score at 12 months after operation was (76.57±4.70) points. The postoperative complications occurred in 31 patients (38.3%) of which impingement syndrome involved in 16 cases (19.8%), head-shaft angle loss in six cases (7.4%), head-shaft angle loss combined with screws cut-out in two cases (2.5%), pure screws cut-out in two cases (2.5%), humeral head necrosis in two cases (2.5%), fat liquefaction in five cases (6.2%). Single factor analysis showed that there were significant differences in the superiorly located greater tuberosity, superiorly located plate and Neer classification between impingement group and un-impinged group (P<0.05). There were statistical y significant differences in age, postoperative medial cortical defects and Neer classification between head-shaft angle loss group and un-loss group (P<0.05). By means of logistic regression analysis, the superiorly located greater tuberosity, superiorly located plate and Neer classification were the individual predictors for postoperative impingement syndrome;postoperative medial cortical defect and Neer classification were the individual predictors for postoperative head-shaft angle loss.
9."Double ""T"" tube drainage in hepatic echinococcosis which ruptured into the common bile duct"
Tiemin JIANG ; Deng YANG ; Bo RAN ; Qiang GUO ; Yingmei SHAO ; Erganaili.aji TU
Chinese Journal of Hepatobiliary Surgery 2017;23(8):539-541
Objective To investigate the double T tube drainage method in the treatment of hepatic echinococcosis which ruptured into the common bile duct.Methods A retrospective study was conducted on 86 patients who were treated surgically for hepatic echinococcosis which had ruptured into the common bile duct at the First Affiliated Hospital of Xinjiang Medical University from June 2012 to December 2014.The average postoperative hospitalization,postoperative complications (residual cavity bile leakage and residual cavity effusion,residual cavity infection) and biliary complications of biliary tract infection were analyzed.Results Significant differences were found on the postoperative residual cavity complications in group A:(2,7.1%) when compared with Group B:(9,15.5 %),and also on the postoperative hospitalization between the double T tube drainage group [group A:(7.1 ± 1.3) d] and the traditional T type tube decompression group [B group:(8.2 ± 1.5) d] (P < 0.05).Conclusions The doubleT tube drainage in the treatment of hepatic echinococcosis which had ruptured into the common bile duct was simple,safe and effective.This treatment could completely cure residual cavity bile leakage,and it had the advantage of avoiding occurrence of common bile duct related complications caused by the traditional suture method for bile leakage.
10.Effect of gefitinib on mucus hypersecretion in chronic obstructive pulmonary disease
Min ZHANG ; Shujuan JIANG ; Huaichen LI ; Lili SU ; Daowei LI ; Yang SHAO ; Weixia MA
Chinese Journal of Geriatrics 2013;32(7):778-781
Objective To investigate the effect of gefitinib on mucus hypersecretion by inhibiting epidermal growth factor receptor (EGFR) activity in chronic obstructive pulmonary disease (COPD).Methods Human airway epithelail cell lines 16HBE cells were exposed to cigarette smoke extraction (CSE) to establish the COPD model.EGFR activity was inhibited by tyrosine kinase inhibitor gefitinib.The mRNA expressions of EGFR and MUC5AC were detected by real-time PCR.EGFR,p-EGFR and MUC5AC protein levels were determined by Western blot and ELISA.Results EGFR mRNA level was increased by 12.7% in CSE and 8.6% in gefitinib group,but had no significant differences among CSE,gefitinib group and control group (all P> 0.05).MUC5AC mRNA levels were enhanced by 141.7%,26.4% in CSE group and gefitinib group respectively,and there were significant differences among CSE,gefitinib group and control group (all P<0.05).EGFR protein levels were (600.34±64.58) μg/mg,(632.58±72.94) μg/mg,(584.57±67.39) μg/mg,in control,CSE and gefitinib groups,respectively,and there were no significant differences between groups (all P>0.05).p-EGFR protein levels were (338.62±45.28) μg/mg,(679.43±78.23) μg/mg,(292.74±59.17) μg/mg in control,CSE and gefitinib groups,respectively.MUC5AC protein levels were(72.80±6.25)μg/mg,(187.00±±10.26)μg/mg,(92.57±8.32)μg/mg in control,CSE and gefitinib groups respectively.Compared with control group,p-EGFR and MUC5AC protein levels were increased significantly in CSE group (both P<0.05),and had no significant differences in p EGFR and MUC5AC protein levels between control group and gefitinib group.Conclusions CSE may lead to mucus hypersecretion through activating the EGFR-mediated signaling pathways.Gefitinib may inhibit mucus hypersecretion by inhibiting EGFR tyrosine kinanse activity.EGFR may serve as a potential target for COPD.