Objective To observe the eotaxin expression of rat airway smooth muscle cells ( ASMCs) induced by serum from asthmatic rats, and explore the possible mechanism. Methods ASMCs isolated fromrat tracheas were cultured in vivo. Then they were treated with serum from asthmatic rats, or treated with serum and dexamethasone simultaneously. The level of eotaxin protein in supernatant and eotaxin mRNA in ASMCs were measured by ELISA and reverse transcription-polymerase chain reaction. The expression of cAMP in ASMCs was examined by radioimmunoassay. Results After the treatment with sensitized serum, the eotaxin level in supernatant and mRNA expression in ASMCs were significantly higher [ ( 107. 09 ±7. 12) ng/L vs. ( 0. 63 ±0. 56) ng/L, P lt; 0. 05; 1. 39 ±0. 04 vs. 0. 05 ±0. 01, P lt;0. 05] , and the level of cAMP in ASMCs was significantly lower compared with the control group [ ( 17. 58 ±3. 62) ng/L vs. ( 32. 39 ±3. 36) ng/L, P lt; 0. 05] . After intervened by the sensitized serum and dexamethasone simultaneously, the protein and mRNA expressions of eotaxin were lower compared with those intervened by sensitized serumalone [ ( 64. 18 ±4. 04) ng/L and 0. 77 ±0. 19] . The level of eotaxin in supernatant was negatively correlated with cAMP level in ASMCs ( r = - 0. 788, P lt; 0. 01) . Conclusions There is anautocrine function in ASMCs as inflammatory cells after stimulation with sensitized serum. Eotaxin may play an important roll in the pathogenesis of asthma via a cAMP-dependent pathway.
Objective To explore the effect of the platelet-rich plasma (PRP) on proliferation and osteogenic differentiation of the bone marrow mesenchymal stem cells (MSCs) in China goat in vitro. Methods MSCs from the bone marrow of China goat were cultured. The third passage of MSCs were treated with PRP in the PRP group (the experimental group), but the cells were cultured with only the fetal calf serum (FCS) in the FCS group (the control group). The morphology and proliferation of the cells were observed by an inverted phase contrast microscope. The effect of PRP on proliferation of MSCs was examined by the MTT assay at 2,4,6 and 8 days. Furthermore, MSCs were cultured withdexamethasone（DEX）or PRP; alkaline phosphatase (ALP) and the calcium stainingwere used to evaluate the effect of DEX or PRP on osteogenic differatiation of MSCs at 18 days. The results from the PRP group were compared with those from the FCS group. Results The time for the MSCs confluence in the PRP group was earlier than that in the FCS group when observed under the inverted phase contrast microscope. The MTT assay showed that at 2, 4, 6 and 8 days the mean absorbance values were 0.252±0.026, 0.747±0.042, 1.173±0.067, and 1.242±0.056 in the PRP group, but 0.137±0.019, 0.436±0.052, 0.939±0.036, and 1.105±0.070 in the FCS group. The mean absorbance value was significantly higher in the PRP group than in the FCS group at each observation time (P＜0.01). Compared with the FCS group, the positive-ALP cells and the calcium deposition were decreased in the PRP group; however, DEX could increase boththe number of the positiveALP cells and the calcium deposition. Conclusion The PRP can promote proliferation of the MSCs of China goats in vitro but inhibit osteogenic differentiation.
摘要:目的:探討PTTG的表達在非小細胞肺癌發生、發展中的作用及其與CMYC蛋白表達的關系。方法:應用免疫組化SP法檢測PTTG、CMYC二種蛋白在44例非小細胞肺癌、20例肺良性病變組織和12例正常支氣管粘膜上皮組織中的表達。結果:PTTG和CMYC蛋白在非小細胞肺癌組織中的表達明顯高于肺良性病變組及癌旁組織,在TNM分期、淋巴結轉移組間差別有統計學意義。非小細胞肺癌組織中PTTG與CMYC表達呈顯著正相關。結論:提示PTTG和CMYC可能參與了非小細胞肺癌的發生和發展,可作為反映其生物學行為的指標。Abstract: Objective: To investigate the expression of PTTG and its relationship with expressions of CMYC protein in human nonsmall cell lung cancer (NSCLC).Methods: Immunohistochemical methods were applied to detect the expression of PTTG,CMYCproteins in 44 surgical specimens from NSCLC patients,20 pneumonic benign lesion and 19 normal bronchial epithelium. Results:There were high erexpressions of PTTG,CMYC in NSCLC tissues than inadjacent tissues and benign lesions.There were statistical relationships between their expressions and TNM stage,lymphnode metastasis.The expression of PTTG was positively correlated with CMYC. Conclusion: Overexpression of PTTG,CMYC may be related to human NSCLC,PTTG and CMYC play a cooperative role inthe process of NSCLC,all of them may be used as important indices for biologic behavior of NSCLC.
OBJECTIVE: To study the effect of simvastatin on the expression of bone morphogenetic protein-2 (BMP-2) and alkaline phosphates (ALP) activity in the primary cultured bone marrow stromal cells, and to elucidate the mechanism of the anabolic osteogenetic effect of simvastatin. METHODS: Bone marrow stromal cells in femur and tibia of adult mouse were cultured in vitro. after treated with different concentrations of simvastatin (0, 0.1, 0.2, 0.5 and 1.0 mumol/L) or recombinant human BMP-2 for 72 hours, ALP activity of bone marrow stromal cells was determined. BMP-2 expression of bone marrow stromal cells was analyzed by using immunocytochemistry and Western blotting. RESULTS: After treated with simvastatin for 72 hours, BMP-2 expression increased, while little BMP-2 expression could be observed in the control group. ALP activity also increased in a dose-dependent manner; t-test showed that ALP activity in the group which concentrations of simvastatin were 0.5 mumol/L (t = 2.35, P = 0.041), 1.0 mumol/L (t = 2.348, P = 0.041) had significant difference when compared with control group. CONCLUSION: Simvastatin lead to high expression of BMP-2 in bone marrow stromal cells, via the increased auto- or para-crine of BMP-2, and ALP activity increased. These may be parts of the mechanism on the anabolic osteogenetic effect of simvastatin.
Objective To investigate the possible mechanism of arsenic trioxide (As2O3) inducing P16 gene demethylation and transcription regulation in the retinoblastoma (RB) Cell Line Y79． Methods The induced growth inhibition of Y79 cell was assayed by MTT; The DNA content of Y79 cell was analyzed by flow cytometry after being exposed to As2O3; the methylation status of the P16 gene in Y79 cell line before and after treatment with As2O3 was detected by the nestedmethylation specific PCR and DNA sequencing; the mRNA of P16,DNA methyltransferases (DNMT3A and 3B)gene were determined by RT-PCR． Results As2O3 was able to inhibit the growth of Y79 cell and increase the cell number in G0-G1 phase；P16 gene was not expressed in Y79 cell line and As2O3 can induce itrsquo;s mRNA expression；after 48 hour disposal of As2O3，the methylation levelof P16 gene was apparently attenuated in Y79 cell line,the expression of DNMT3A and DNMT3B was obviously down-regulated． Conclusions P16 gene is the hypermethylation in the retinoblastoma cell line Y79, and As2O3 can inhibite the methylation of P16 gene and upregulate the expression of p16 gene mRNA which inhibits the proliferation of Y79 cell by inducing the G0-G1 arrest, by inhibiting the expression of DNA methyltransferases.
Objective To investigate the effect of S-adenosyl-l-methionine (SAM) on oxidative stress and alveolar septal cell apoptosis in mice with emphysema after smoking cessation. Methods Twenty-two male SPF C57BL/6J mice aged 6 - 8 weeks were randomly divided into 4 groups, ie. a healthy control group, an emphysema group, a smoking cessation group, and a SAM intervention for 8 weeks after smoking cessation group, with 8 mice in each group. The mice model of emphysema was established by intraperitoneal injection of cigarette smoke extract (CSE) combined with cigarette smoke exposure. Smoking cessation started after the emphysema model was successfully constructed and lasted for 8 weeks. After smoking cessation, the mice in SAM intervention groups were intraperitoneally injected with SAM mg·kg–1·d–1 for 8 weeks. The right lung sections of the mice were taken for hematoxylin-eosin staining to observe pathological changes, and the mean linea rintercept (MLI) and mean alveola rnumber (MAN) of lungs were measured. The concentrations of malondialdehyde (MDA), superoxide-dismutase (SOD) and glutathione (GSH) in alveolar lavage fluid of left lung were detected by spectrophotometry. Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) technique was carried out to detect the alveolar septal cells apoptosis. Results MLI, apoptosis index of alveolar septal cell and MDA concentration in bronchoalveolar lavage fluid (BALF) increased significantly in the emphysema group compared with healthy controls, increased significantly in the smoking cessation group compared with the emphysema group, and decreased in the SAM intervention group compared with the smoking cessation group (all P<0.05). GSH concentration and SOD activity in BALF and MAN was significantly lower in the emphysema group compared with the healthy control group, significantly lower in the smoking cessation group compared with the emphysema group, and significantly higher in the SAM intervention group compared with the smoking cessation group (all P<0.05). Conclusions Lung histopathology and apoptosis of alveolar septal cells in emphysema mice progress continuously after smoking cessation. SAM may reduce oxidative stress and improve apoptosis of alveolar septal cells, so as to protect emphysema mice after smoking cessation.
ObjectiveTo observe the expression of interleukin (IL)-17, IL-4 and interferon γ (IFN-γ) in experimental autoimmune uveoretinitis (EAU).
MethodsC57BL/6 mice were immunized with interphotoreceptor retinoid-binding protein 1-20 to induce EAU. The inflammatory reaction before and on 7, 14, 21, 28 days after immunization were observed. The level of IL-17, IL-4 and IFN-γ in the serum were measured by enzyme-linked immune sorbent assay (ELISA). mRNA and protein expression of spleen and retina were analysed using quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot at the same time, respectively.
ResultsThe most serious inflammatory reaction occurred at the 14th day after immunization. The highest level of IFN-γ in serum, highest mRNA and protein expression of IFN-γ in spleen and retina of mice occurred at day 7 after being immunized. The highest level of IL-17, IL-4 in serum, highest mRNA and protein expression of IL-17, IL-4 in spleen and retina of mice occurred at day 14 after being immunized. The increase degree of IL-17 was more than IFN-γ and IL-4. At 7, 14 and 21 days after immunization, compared with the pre-immunization, the level of IL-17, IL-4, IFN-γ in serum of mice were significantly increased (F=1 817.346, 268.600, 164.621; P < 0.05). There was no difference in the levels of IL-17, IL-4, IFN-γin serum of mice between pre-and 28 days after immunization (P > 0.05). At 7, 14 and 21 days after immunization, compared with the pre-immunization, the protein expression of IL-17, IL-4, IFN-γ in spleen (F=312.67, 114.250, 216.220) and retina (F=271.504, 85.370, 80.722) of mice were significantly increased (P < 0.05). There was no difference in protein expression of IL-17, IL-4, IFN-γ in spleen and retina of mice between pre-and 28 days after immunization (P > 0.05).
ConclusionsThere were IL-17, IL-4 and IFN-γ expression in EAU. IL-17, IL-4 and IFN-γ play a key role in the occurrence and development of the EAU.
Objective To compare the short-term efficacy and long-term survival of patients with non-small cell lung cancer (NSCLC) treated by uniportal or three portal thoracoscopic radical resection. Methods A total of 388 patients who underwent uniportal or three portal thoracoscopic radical resection of lung cancer in the Department of Thoracic Surgery of Anhui Chest Hospital from 2015 to 2016 were analyzed retrospectively. The patients were divided into two groups including an uniportal group and a three portal group according to the procedure. The clinicopathological features, perioperative data and long-term survival of the two groups were compared. Results Finally, we included 205 patients with 105 males and 100 females at an average age of 58.73±10.93 years. There were 102 patients in the uniportal group and 103 patients in the three portal group. There was no statistical difference in clinicopathological features between the two groups (P>0.05). But compared with the three portal group, the uniportal group had less postoperative drainage, shorter postoperative catheterization time and postoperative hospital stay (P<0.05). There was no statistical difference in the number of lymph node dissection stations between the two groups (P=0.058). The pain score at 24 hours after operation in the uniportal group was significantly lower than that in the three portal group (P<0.001). There was no statistical difference in the total incidence of complications and the incidence of pulmonary complications between the two groups (P=0.161 and P=0.275). The median survival period and the 1st, 3rd, and 5th year survival rate in the uniportal group was 63.0 months and 95.0%, 75.2%, 51.5%, respectively. The median survival period and the 1st, 3rd, and 5th year survival rate in the three portal group was 61.0 months and 89.3%, 70.9%, 50.5%, respectively. There was no satistical difference in the survival results between the two groups (P=0.440). Conclusion Uniportal thoracoscopic radical resection of lung cancer is more minimally invasive and safe and effective in the treatment of NSCLC. It can make patients recover faster after operation.
Abstract: Objective To investigate the effect of autologous bone marrow mesenchymal stem cells (MSCs) transplantation on cardiac function and their proliferation and differentiation in the post-infarct myocardium in rabbits. Methods Twenty New Zealand rabbits were randomly divided into two groups, the autologous bone marrow mesenchymal stem cells group (MSCs group,n=10) and control group (n=10). Myocardial infarct model was set up by ligation of the left anterior descending (LAD), two weeks after establishment of the infarct model,either 400μl of cell suspension (total cells 1×106) labled by 1,1’-dioctadecyl3,3,3’,3’-tetramethyl indocarbocyanine perchlorate (Dil) or a comparable volume of L-DMEM medium were autologously transplanted into several different points of the periphery of the scar respectively. To evaluate the heart function, echocardiography were performed before modeling,two weeks after modeling, 2 and 4 weeks after the cells transplantation for asurements of left ventricular end systolic diameter (LVESD) and left ventricular end diastolic diameter (LVEDD), tocalculate left ventricular eject fraction(LVEF) and left ventricular fractional shortening (LVFS). Meanwhile the myocardial contrast echocardiography (MCE) were performed for evaluating the blood perfusion of the post-infarct myocardium. Eight weeks after the transplantation, the animalswere undergoing euthanasia, specimens were acquired for pathology. Results Echocardiography indicated that:The LVEF and LVFS between two groups were fundamentally the same before modeling,two weeks after modeling respectively (0.72±0.08 vs. 0.71±0.04,0.56±0.11 vs. 0.55±0.09; 0.35±0.06 vs. 0.35±0.04, 0.24±0.08 vs. 0.23±0.03, Pgt;0.05), but those were improved significantly in group MSCs when compared with control group at two weeks and four weeks after the cells transplantation(0.71±0.05 vs. 0.60±0.05，0.72±0.07 vs. 0.62±0.08 and 0.34±0.03 vs. 0.29±0.01， 0.35±0.06 vs. 0.27±0.05 respectively,Plt;0.05). There were no differences in LVESD and LVEDD between two groups in any time points(Pgt;0.05). MCE showed the blood perfusion of the infarct myocardium were improved two and four weeks after the cell transplantation. Pathology indicated that Dil positive cells were survived in MSCs transplanted hearts, stained positively for αsarcomeric actin and desmin eight weeks after cell transplantation, HE slides indicated that the capillary density in all the cells transplanted hearts were much higher when compared with control group (38.6±7.6/mm2 vs. 21.4±3.9/mm2，Plt;0.05). ConclusionMSCs can differentiate into cardiomyocytes, improve myocardial perfusion and cardiac function when transplanted into ischemic myocardium.
Objective To test the hypothesis that marrow stromal cells （MSCs）, when implanted into selfmyocardium in rabbits, can undergo milieu-dependent differentiation and express cardiomyogenic phenotypes and enhance cardiac function of ischemic hearts, through establish a clinically relevant model for autologous MSCs transplantation, Methods Thirteen New Zealand White rabbits were randomly divided into experimental group （n= 7） and control group （n= 6）. In experimental group, autotogous MSCs（3× 106 cells/30μl） labeled with Bromodeoxyuridine （BrdU） were respectively injected into superior, central and inferior sites in the periphery of the myocardial infarct region. Phosphate buffer saline （PBS） was injected into the scar of the control group hearts according to the same procedure used in the experimental group. Four weeks later, the transplanted labeled MSCs were detected by laser scanning confocal microscopy and the cardiac function were examined by echocardiogram and muhichannel physiologic recorder. Results After 4 weeks, transplanted MSCs were demonstrated myogenic differentiation with the expression of α-sarcomeric actin and connexin 43 located in intercalated disk. MSCs increased the number of vessels compared with controls in myocardial ischemia area. MSCs implantation resulted in markedly improved left ventricular contractility[left ventricular ejection fraction （LVEF）： 0. 51 ± 0.07 vs. 0. 43 ± 0.06 ,left ventricular lateral wall motion distance （LVLWMD） ：1. 75±0. 42mm vs. 1.09±0. 28mm, left ventricular systolic wall thickening ratio（LVAT） ：0. 19%±0.05% vs. 0. 11%±0.04%, left ventricular systolic pressure （LVSP）： 113. 1± 6.3mmHg vs. 99, 5 ± 5, lmmHg, left ventricular end diastolic pressure （LVEDP）： 11. 5±2. lmmHg vs, 14, 3 ±3. lmmHg, maximum rate of left ventricular pressure rise （＋dp/dtmax）：4 618. 3±365. 2 mmHg/s vs. 3 268. 1± 436.9 mmHg/s, maximum rate of left ventricular pressure fall （-dp/dtmax） ：3 008.8±346.7 mmHg/s vs. 2 536.9± 380.4 mmHg/s, P〈0.05]. Conclusion Transplanted autologous MSCs are able to undergo differentiation to form myocardial cells and improve the cardiac function of ischemia myocardium effectively. Autologous MSCs transplantation may have significant clinical potential in treatment myocardial ischemia.
