1.Homing of bone marrow hemopoietic stem cells in infarct area following myocardial infarction
Chinese Journal of Tissue Engineering Research 2009;13(27):5252-5256
BACKGROUND: Studies have shown that after bone marrow stem cell transplantation, there are juvenile cells with the episode of myocardial cells around the infarct areas, which was recognized as deriving from bone marrow stem cells.OBJECTIVE: To explore the homing situation of bone marrow stem cells by autologous mobilization around the infarct areas aftar myocardial infarction.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the China-Japan Clinical Pathology Center, Daliar Medical University from February 2005 to April 2006.MATERIALS: A total of 30 myocardial samples from the autopsy at the Department of Pathology and Forensic Medicine, Dalian Medical University between 2000 and 2005. Fifteen normal samples dyed from the mechanical injuries or craniocerebral injury,without abnormity following autopsy. Fifteen samples in the myocardial infarction group suffered from coronary atherosclerosis and coronary tube stenosis, and died within 10 days after myocardial infarction.METHODS: The autopsy was performed within 48 hours after the death and the tissues were obtained from antedor wall, lateral wall and posterior wall of the left ventricle, and made into tissue sections. We checked the expression of CD34, CD133 and transforming growth factor-β, in the normal area, infarct area and around the infarct area by immunohistochemistry.MAIN OUTCOME MEASURES: The expression of CD34, CD133 and transforming growth factor-β in different areas and the relationship between them.RESULTS: The positive expression of CD34 in infarct area was significantly stronger than that around the infarct area and in normal area (P < 0.01). The positive expression of transforming growth factor-β in infarct area was significantly stronger than that around the infarct area (P < 0.01), and the expression of transforming growth factor-β showed negative reactJon in normal area. The expression of CD34, CD133 and transforming growth factor-β, in infarct areas and around the infarct area was closely related (P < 0.01).CONCLUSION: There are the homing of bone marrow hemopoietic stem cells in infarct area and around the infarct ares after myocardial infarction. The increase in transforming growth factor-β in infarct area provides the possibility for the differentiation of the homing stem cells, the following angiogenesis and the regeneration of cardiomyocytes.
2.An experimental study of folic acid rivaling methylenetetrahydrofolate reductase (MTHFR) gene
Wenlin XIAO ; Bing SHI ; Qian ZHENG ; Yan WANG ; Lei HUANG
Journal of Practical Stomatology 2010;26(1):15-19
Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.
3.Scanning the SRSF2 gene mutations in acute myeloid leukemia by using high-resolution melting analysis
Dongming YAO ; Jing YANG ; Zhen QIAN ; Lei YANG ; Gaofei XIAO ; Jiang LIN ; Hong GUO ; Jun. QIAN
The Journal of Practical Medicine 2016;32(7):1169-1172
Objective To study the SRSF2 mutations in acute myeloid leukemia (AML) patients by using high-resolution melting analysis (HRMA). Methods PCR-HRMA analysis was performed to screen SRSF2 mutations in 140 cases with AML, and the direct DNA sequencing was used to confirm the HRMA results. Results Five percent (7/140) of AML patients were found with heterozygous SRSF2 mutations, including one case of P95R mutation, two case of P95L mutation, and four cases of P95H mutation, the above mutations were confirmed by direct DNA sequencing. The maximal sensitivity of HRMA in detecting SRSF2 mutation was close to 10%. There were no difference in gender, age and blood parameters among cases with or without SRSF2 mutations (P > 0.05). The overall survival (OS) of patients with SRSF2 mutations was inferior to those without SRSF2 mutations in AML patients (P=0.016). Conclusions HRMA analysis was a convenient, rapid, specific, high-throughput technique for scanning of SRSF2 gene mutations in AML patients. SRSF2 mutation may predict the adverse prognosis in AML patients.
4.Rapid Detection of Active Pharmaceutical Ingredients in Weak Active Pharmaceutical Ingredient Signal Drugs by Paper Substrate-Surface Enhanced Raman Scattering Spectrometry
Xiao LI ; Yun MENG ; Lei WANG ; Qian QIAN ; Fang FANG ; Li MIAO
Chinese Journal of Analytical Chemistry 2015;(11):1735-1742
A rapid detection method of active pharmaceutical ingredients( API) in weak API signal drugs by surface enhanced Raman scattering ( SERS) technology combined with paper substrate was established in this work. By soaking the filter paper in silver nanoparticles solution ( Ag NPs) to synthesize Ag NPs-paper as the substrate, and then the sample solution was dropping on the substrate with SERS detection. On the basis of strengthen ability of Ag NPs-paper, result of SERS detection and optimal preparation conditions, the fast identification method of weak API signal drugs was established. In this case, the SERS spectra of weak API signal drugs and their standards SERS spectra were obtained, where the correlation coefficient of weak API signal drug SERS spectra and its standard was more than 0. 9. The result showed that by this method, the low content API in weak API signal drugs could be well investigated, and the deficiencies of the normal Raman spectroscopy efficiently was also overcome. In conclusion, the synthesize method of Ag NPs-paper was simple, and the strengthen effect of this Ag NPs-paper on the intensity was obviously observed. Paper substrate-SERS method was simple, rapid and sensitive, and could be used to detect weak API signal drugs, presenting broad application prospects in the rapid detection of weak API signal drugs.
5.Impact of Antimicrobial Agents on Transcription of Gene hld in Forming Process of Staphylococcus epidermidis Biofilm
Qian-Guo HAN ; Xue-Zhong LEI ; Lei ZHANG ; Xiao-Ju LV ; Ru-Jia YU ;
Chinese Journal of Nosocomiology 2009;0(16):-
OBJECTIVE To investigate the transcription level of gene hld of Staphylococcus epidermidis in the biofilm forming and detaching under MIC antibiotic and explore the relationship between biofilm-related drug resistance and persistant infection.METHODS The transcription level of gene hld of S.epidermidis under MIC concentration of 4 antibiotics was compared with those of the control group without antibiotics by SYBR real-time fluorescent quantitative RT-PCR at the different time point of biofilm formation and detachment.RESULTS The transcription of gene hld decreased rapidly from initial adherence,and droped continuously for few hours.There was an increase from 24 hours to 72 hours in groups without antibiotics but not in antibiotics groups,the differenet was significant.CONCLUSIONS Antibiotics improve adherence at first and then prevent matrix decomposition water-conducting tube and detachment of cells by impact of transcription of gene hld,it can protect cells from killing by inhibiting the penetration of biotics and prevent them become planktonic cells after detachment from biofilm.
6.Effects of total flavonoid of chrysanthemum on Fas and Fasl expression in lacrimal gland cells of male castrated rabbits with dry eye
Xiao-Lei, YAO ; Qing-Hua, PENG ; Qi-Lei, CHEN ; Yong-Hua, TANG ; Qian, ZHONG
International Eye Science 2014;(10):1749-1754
AIM: To evaluate the effects of total flavonoid of chrysanthemum on the expressions of Fas and FasL in male rabbits with dry eye, and to investigate the therapeutic effects of the total flavonoid of chrysanthemum on dry eye.
METHODS: Totally 150 male Japanses white rabbits were divided into blank group ( group A ) , sham -operated group ( group B ) , model group ( group C ) , androgen control treatment group (group D), and total flavonoid of chrysanthemum treatment group ( group E ) . The dry eye model was established with orchiectomy on group C, D and E. Rabbits in group E were treated with total flavonoid of chrysanthemum. Rabbits in group D were treated with androgen intramuscular injection. Rabbits in the group A, group B, group C was treated with normal saline. All rabbits were detected with Schirmer's Ⅰ test and tear break-up time (BUT). Fas, FasL were checked on immunohistochemistry.
RESULTS:The Schirmer's I test values of group E was significantly higher than that of group C ( P<0. 01 ) and the BUT value of group E was significantly longer than that of group C ( P<0. 01 ). The quantity of positive expression of Fas in glandular tube cell and acinar epithelial celland apoptosis cells of group E after treatment at 1, 3, 5mo were significantly lower than that of group C, cell population of the positive expression of FasL was obviously higher than that of group C (P<0. 01).
CONCLUSION:The main component of chrysanthemum is flavonoid, which could significantly inhibit happening of dry eye in rabbit after androgen level lowered and lacrimal gland apoptosis and keep basic tears secretory volume and tear film stability.
7.Feasibility assessment of visual quality analyzer KR - 1 W guiding personalized aspheric lOL implantation
Xiao-Li, WANG ; Qian, LI ; Xiao-Lei, TANG ; Ying-Chun, LÜ ; Li, GUO
International Eye Science 2015;(1):43-45
Abstract?AlM:To discuss the feasibility of using the visual quality analyzer KR - 1W to guide the relatively personalized aspheric intraocular lens ( lOL ) implants to make the whole eye spherical aberration close to 0. 1μm.?METHODS: ln this prospective case series study, the corneal spherical aberration with 6mm aperture of 73 patients (100 eyes) was measured with KR-1W Visual Function Analyzer 1d before surgery. For the sake of the whole postoperative spherical aberration were close to 0. 1μm, 9 cases ( 16 eyes ) with corneal spherical aberration <0. 15μm were implanted Sofport Advanced Optic lOL, named AO group;45 cases ( 57 eyes ) with corneal spherical aberration 0. 25~0. 3μm were implanted AcrySof lQ lOL, named lQ group;19 cases (27 eyes) with corneal spherical aberration > 0. 35μm were implanted Tecnis ZA9003 lOL, named Tecnis group. Aspherical lOL was implanted after phacoemulsification through a cornea 2. 75mm incision without suture. Uncorrected visual acuity, beat corrected visual acuity, spherical aberration of the whole eye and jnternal optics (mainly lOL) at 6mm pupil diameter were examined at 3mo postoperatively. The relevant data were analyzed using t-test and variance analysis.?RESULTS: The whole ocular spherical aberration at 6mm pupil diameter in all postoperative were 0. 084 ± 0. 032μm;in Tecnis group, the data were 0. 091 ± 0. 021μm;in AO group, the data were 0. 0814-0. 013μm;lQ group were0. 093 ± 0. 042μm. There was no significantly different between the predicted value and actual value of ocular spherical aberration at 6 mm pupil diameter in all postoperative ( t = 1. 932, P = 0. 061 ) and in the three groups. The difference value in the predicted values of the preoperative spherical aberrations of the whole eye and the actual values after surgery was 0. 013±0. 041μm; there was no statistically significant difference ( F=2. 537, P=0. 091 ) . Respectively compared the uncorrected visual acuity and besta corrected visual acuity among three groups of postoperative, no significant difference were found (F=0. 897, P=0. 421;F=1. 423, P=0. 097).?CONCLUSlON: Personality selection of aspheric lOL based on preoperative corneal spherical aberration of patients is feasible and produces satisfactory target postoperative total spherical aberration.
8.Expression of Golgi glycoprotein 73 and secreted Clusterin in hepatocellular carcinoma
Qian CAO ; Gulibiye SHABIER ; Ying YANG ; Lei XIAO ; Rui MAO ; Ruili ZHANG ; Hua ZHANG ; Yongxing BAO
China Oncology 2013;(11):880-884
it was 4% (3/75). GP73-positive rate in HCC was higher than that of the normal liver tissues (χ2=73.60, P<0.05). sCLU-positive rate in HCC was also higher than that of the normal liver tissues (χ2=207.94, P<0.05). GP73 expression was positively correlated with sCLU expression in HCC (r=0.405, P<0.05). GP73 and sCLU were associated with clinicopathological features including tumor differentiation, TNM stage and vascular invasion (P<0.05); GP73 and sCLU had no correlation with the patient’s gender, age, HBsAg, cirrhosis, AFP value, portal vein thrombosis and tumor numbers (P>0.05). GP73 was associated with survival but not sCLU. Conclusion:GP73 and sCLU have higher positive rates in HCC and GP73 is positively correlated with sCLU. The expression of GP73 and sCLU are probably closely related with the invasion of HCC, which can help evaluate the prognosis of the patients.
9.Effects of CO_2 pneumoperitoneum on focal adhesion kinase of gastric cancer MKN-45 cells
Lin XUE ; Yan SHI ; Peiwu YU ; Feng QIAN ; Yongliang ZHAO ; Xiao LEI ; Bo TANG ; Huaxing LUO
Chinese Journal of Digestive Surgery 2009;8(5):347-349
Objective To investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.
10.Effects of methamphetamine acute exposure on neural damage
Lei JIANG ; Wenyi QIAN ; Jinsong ZHANG ; Jun WANG ; Xufeng CHEN ; Hao SUN ; Hang XIAO
Chinese Journal of Emergency Medicine 2016;25(11):1393-1399
Objective To explore the neural damage induced by acute exposure to methamphetamine (METH).Methods The mice were administrated with METH,then the stereotyped behavior of mice was evaluated,and spatial recognition memory was analyzed by Y-maze test.In addition,nitric oxide synthase (NOS) activity was detected by kit,and the apoptotic proteins including Bax,Bcl-2,Caspase-3 were assayed by using Western blot.The DNA injury induced by METH was observed by using the comet assay.Moreover,mitochondrial membrane potential was detected to assess the toxic effects of METH on mitochondria by JC-1.With the Western blot assay,the phosphorylation of MAPK signaling pathways were also investigated.Results Acute METH exposure significantly increased the stereotyped behavior in mice,and spatial recognition ability of mice was obviously decreased.On the molecular level,total nitric oxide synthase (TNOS) and induced nitric oxide synthase (iNOS) were increased,and the apoptotic proteins,such as Bax and cleaved caspase-3 were markedly enhanced.With the comet assay,it showed that METH exposure resulted in DNA damage.In parallel,mitochondrial membrane was damaged which manifested as mitochondrial membrane potential decreased.With the western blot,It was further found that METH enhanced the activation of MAPKs.However,p38 MAPK signahng pathway was demonstrated to be the only one factor involved in METH-induced neural damage.Conclusion METH induced neural damage,and MAPK signaling pathways might be involved in this process,since inhibition of p38 MAPK signaling pathway significantly ameliorated METH-induced neural damage.