1.PI3K/GSK-3β signaling pathway mediates cardiotrophin-1 cardioprotection against cardiocyte hypoxia-reoxygenation injury
Juxiang LI ; Lei WAN ; Hao DING ; Zirong XIA ; Hai SU ; Sujuan YAN ; Yanqing WU ; Qinghua WU ; Xiaoshu CHENG
Chinese Journal of Emergency Medicine 2009;18(8):814-818
Objective To study the effect of Cardiotrophin-1 (CT-1) on cardiocyte hypoxia-reoxygenation injury,and to investigate the signaling pathways involved in the protective effect. Method This study was carried out in Key Lab of Molecular Medicine in Jiangxi Province. Cardiomyocytes from the hearts of 2-day-old Sprague-Dawley neonatal rats were prepared by a modified method. Five groups were included in the study. Group (ⅰ): control, Group (ⅱ): hypoxia/reoxygeuation, Group (ⅲ): hypoxia / reoxygenation + CT-1, Group (iv) : CT- 1 + hypoxia/ reoxygenation + LY294002 (PIK3/Akt inhibitor), Group (ⅴ): CT-1 + hypoxia / reoxygenation +DMSO. The concentration of CT-1 was 10 ng/mL. Myocytes survival rote was evaluated by MTS method, apopto-sis, mitochondrial permeability transition pore (△ψm) and reactive oxygen species(ROS) were detected by flow cy-tometer, phosphorylased GSK-3β and PI3K protein by western blotting. Analysis of variance and q test as statistical methods was used to analyze the data. Results Cardiomyocyte apoptosis and ROS increased markedly after hy-poxia/reoxygenation,but cardiomyocyte survival rate and the level of △ψm [(40.55±4.25) vs. (86.28±7.15), P < 0.01]decreased significantly. With CT-1 intervention, cardiomyocyte survival rate increased markedly (87%),apoptosis and ROS reduced significantly. The level of △ψm increased, the level of phosphorylased GSK-3β and phosphorylased PI3K protein obviously increased. The effect of CT-1 was inhibited by LY294002, but no significant effect was observed on ceils survival in DMSO group, which confirmed that LY294002 specifically in-volved blocking the protective effect of CT-1. Conclusions CT-1 can protect cardiac cells against hypoxia- reoxy-genation injury, these effects are dependent upon its ability to activate the PI3K/GSK-3β pathway.
2.Change of subunit of NADPH oxidation enzyme complex nox -1 protein in cardiocyte hypoxia - reoxygenation injury and the role of cardiotrophin -1
Lei WAN ; Juxiang LI ; Kui HONG ; Hao DING ; Zirong XIA ; Hai SU ; Sujuan YAN ; Yanqing WU ; Qinghua WU ; Xiaoshu CHENG
Chinese Journal of Pathophysiology 2009;25(11):2113-2117
AIM: To observe the change of subunit of NADPH oxidation enzyme complex nox - 1 protein in cardiocyte hypoxia - reoxygenation injury and the role of cardiotrophin -1.METHODS: Cardiomyocytes from the hearts of 1 -3 d old neonatal rats were prepared by a modified method. Five groups were included in the study: control; hypoxia/ reoxygenation; hypoxia/reoxygenation + CT - 1; CT - 1 + hypoxia/reoxygenation + LY294002 (PIK3/Akt inhibitor) ; CT -1 + hypoxia/reoxygenation + PD98059 (ERK inhibitor) ; CT - 1 + hypoxia/reoxygenation + DMSO. The concentration of CT -1 was 10 μg/L. The survival rate of myocytes was evaluated by MTS method. Apoptosis, mitochondrial permeability transition pore ( △ψm) and reactive oxygen species ( ROS) were detected by flow cytometry. Nox - 1 protein was determined by Western blotting. RESULTS: Apoptosis of cardiomyocytes and the level of ROS (19.7% ±1.4% vs 2.1% ± 0.5% , 14.07% ± 1.25% vs 3.54% ± 0.86% , P < 0.05 ) increased markedly after hypoxia/reoxygenation, but cardio-myocyte survival rate and the level of△ψm (40.55% ±4.25% vs 86.28% ±7.15% , P <0.01) decreased significantly. The expression of nox - 1 protein was upregulated markedly. With CT - 1 intervention, cardiomyocyte survival rate increased markedly, apoptosis, both ROS and expression of nox - 1 protein reduced significantly. The level of△ψm increased obviously. The effect of CT - 1 was inhibited by LY294002.No significant effect was observed on cells survival in DMSO group, which confirmed that LY294002 was specifically involved in blocking the protective effect of CT - 1.CONCLUSION : The expression of subunit of NADPH oxidation enzyme complex nox - 1 protein is upregulated markedly in cardiocyte hypoxia - reoxygenation injury.CT - 1 protects cardiac cells against hypoxia - reoxygenation injury by downregulating the expression of nox -1 protein to decrease the level of ROS.
3.Role of platelet-derived growth factor in directional migration of human meseuchymal stem cells toward gliomas
Yong-Fei DONG ; Chao-Shi NIU ; Ge GAO ; Wan-Hai DING ; Guo-Ping HE
Chinese Journal of Neuromedicine 2009;8(5):437-440
Objective To investigate the effect ofplatelet-derived growth factor (PDGF) and conditional medium of U87 glioma cells on the migration ability of human mesenchymal stem cells (hMSCs) to understand the possible role of PDGF in the directional migration of hMSCs toward gliomas. Methods hMSCs were isolated from the whole bone marrow by adherent culture, and the expression of PDGF receptor (PDGFR-α, β) in the cells was examined by RT-PCR. In vitro migration assay was performed using transwell inserts to observe the effect of PDGF (0, 5, 50, and 125 ng/mL) and the conditional medium on the directional migration ability ofhMSCs. The changes in the migration ability of hMSCs in response to addition of anti-PDGF antibody in the conditional medium were investigated. Results RT-PCR detected the expression of PDGFR- αand PDGFR-β mRNA in the isolated hMSCs. In the cell migration assay, both PDGF and the conditional medium induced directional migration of hMSCs (P<0.05), which was significantly suppressed by anti-PDGF antibody P<0.05). Conclusion With chemokine-like activities, PDGF concentration-dependently enhances the directional migration of hMSCs toward gliomas in vitro.
4.The effect of separating red blood cells from bone marrow graft in vitro by methylcellulose.
Miao ZHANG ; Bo-Long ZHANG ; Hai-Jie JIN ; Chun-Ji GAO ; Fang-Ding LOU ; Wan-Ming DA
Journal of Experimental Hematology 2002;10(6):561-563
It has need to separate red blood cells (RBC) from marrow graft in ABO group unmatched BMT and auto-BMT with purging tumor cells, the separating effect of methylcellulose was observed. The mixture of 0.5% methylcellulose and bone marrow was laid up in an open transfusion system, and then sedimentation of RBC was performed in the transfusion tube. The separating results of 18 marrow grafts showed that the recovery rates of mononuclear cells and CD34(+) cells were (83.8 +/- 55.2)% and (90.3 +/- 7.2)%, respectively. RBC residual rate was (4.3 +/- 1.5)%. The yield of CFU-GM was (60.8 +/- 22.4)/2 x 10(5) MNC, and there was no difference to [(69.8 +/- 23.4)/2 x 10(5) MNC] yielded from same marrow samples, separated by Ficoll-Hypaque separation. It is concluded that this method could be used for bone marrow transplantation.
Bone Marrow Transplantation
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methods
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Cell Separation
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methods
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Erythrocytes
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immunology
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Humans
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Methylcellulose
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pharmacology
5.Outcome analysis on drinking-water type endemic fluorosis in Qinghai in 2008
Sheng-ying, WEI ; Qing, LU ; Ping, DING ; Sheng-rong, DING ; Guan-glan, PU ; Ping, CHEN ; Duo-long, HE ; Hai-yan, ZHANG ; Wen-jiang, SI ; Qiu-xiang, LIU ; Xing, WANG ; Ma, WAN
Chinese Journal of Endemiology 2010;29(1):77-79
Objective To observe the state of endemic flurosis in Qinghai province in 2008 and to provide scientific basis for endemic flurosis control and prevention. Methods All the endemic fluorosis villages in Qinghai province was divided into slight,medium and heavy degree according to the water fluoride content before improving water,and 2,4,4 villages were collected from each degree,respectively. Water samples were collected from each direction of east,west,south,north and centre,and fluoride concentration were determined from each surveyed villages with unimproved-water. At the same time,3 copies of the peripheral water samples and a sample of source water were collected to determine fluoride concentration. In all the village children aged 8 to 12 were tested for dental fluorosis by Dean method. Six copies of the urinary fluoride were randomly sampled in different age groups. The fluorine content in urine was determined with F-ion selective electrode. The situation of clinical skeletal fluorosis of adults over 16 years of age was examined,and 20 adults in the villages of medium and heavy levels were filmed for skeletal fluorosis. Results The detectable rate of dental fluorosis of children aged 8-12 years old was 43.59% (245/562),that of skeletal flurosis of adults aged above 16 years was 50.32% (1582/3144) and detectable rate of X-ray was 29.45% (48/163). Fluoride content in urine was 1.52 mg/L. Conclusions Endemic fluorosis is still very serious in Qiaghai province in a rising trend,so the control measures need to be strengthen.
6.Comparing the effect of adjuvant chemotherapy by portal vein infusion with intraluminal chemotherapy for colorectal cancer.
Zhi-Zhong PAN ; De-Sen WAN ; Zhen-Hai LU ; Li-Ren LI ; Gong CHEN ; Zhi-Wei ZHOU ; Xiao-Jun WU ; Pei-Rong DING ; Fu-Long WANG
Chinese Journal of Surgery 2004;42(15):918-921
OBJECTIVETo compare the effect of 5-fluorouracil (5-FU) portal vein infusion (PVI) for 7 days after radical resection, with intraluminal chemotherapy during operation for eliminating liver metastasis and elevating long-term prognosis in colorectal cancer.
METHODS162 colorectal cancer patients with radical resection were divided into portal vein chemotherapy group (group A, 82 cases) and intraluminal chemotherapy group (group B, 80 cases) randomly. In group A, 5-fluorouracil were infused with 1g per day constantly for 7 days after operation through portal vein catheters, which placed into greater omental vein and fixed on the abdominal wall. In group B, intraluminal chemotherapy was given and 5-fluorouracil 0.5 g was injected into the greater omental vein during operation.
RESULTSThe short-term complications and long-term effect in the two groups were compared by statistical software SPSS 8.0. Group A had more operative complications, and no statistical differences was found in hospital time and survival rate of the two groups. The 5-year survival rate is 76.7% (group A: 74.3%, group B: 79.2%), and the liver metastasis rate is 19.8%. There is no significant difference between the two group-survival curves. Multiple variable analysis suggested that Dukes' stage was the prognosis factor (P < 0.05).
CONCLUSIONSThe present study demonstrated that the two chemotherapy methods play an important role in preventing liver metastasis and improving the survival rate, and the intraluminal chemotherapy would be easier and simpler. The result should be further improved by using combined chemotherapy.
Adult ; Aged ; Antimetabolites, Antineoplastic ; administration & dosage ; Chemotherapy, Adjuvant ; Chemotherapy, Cancer, Regional Perfusion ; methods ; Colorectal Neoplasms ; drug therapy ; mortality ; therapy ; Combined Modality Therapy ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Portal Vein ; Survival Rate ; Treatment Outcome
7.Hepatitis B x protein activated vascular endothelial growth factor expression through hypoxia inducible factor-1 pathway.
Hai-ping WANG ; Xiao-ping CHEN ; Lei DING ; Song-qing HE ; Muthanna ALI ; Dong-hua LI ; Wan-guang ZHANG
Chinese Journal of Oncology 2003;25(5):433-436
OBJECTIVETo investigate whether hepatitis B x protein (HBx) stimulates vascular endothelial growth factor (VEGF) through hypoxia inducible factor-1 (HIF-1 alpha) pathway.
METHODSTwo plasmids including pIRES-EGFP-HBx and pTK-Hyg were co-transfected to a hepatocellular carcinoma cell line SMMC-7721. With fluorescence-positive and fluorescence-negative hygromycin-resistant colonies selected, expressions of VEGF and HIF-1 alpha in protein or/and mRNA level were detected.
RESULTSFluorescence-positive cells were stably integrated with HBx, in which expression of HIF-1 alpha and VEGF were upregulated. Fluorescence-negative cells did not express HBx, VEGF or HIF-1 alpha.
CONCLUSIONHBx can activate VEGF through HIF-1 alpha pathway.
Cell Line, Tumor ; Gene Expression Regulation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; Trans-Activators ; physiology ; Transcription Factors ; genetics ; physiology ; Vascular Endothelial Growth Factor A ; genetics ; physiology
8.Complications and prevention of single-nostril transsphenoidal surgery for pituitary adenomas
Ying JI ; Chao-Shi NIU ; Wan-Hai DING ; Shi-Ying LING ; De-Jun BAO ; Xiao-Feng JIANG ; Xian-Ming FU ; Ye-Han WANG
Chinese Journal of Neuromedicine 2008;7(4):384-387
Objective To discuss the common complications of single-nostril transsphenoidal surgery for pituitary adenomas, and the prevention ofpostsurgical complications. Methods Clinical and follow-up (1-36 months) data of 241 patients with pituitary adenomas treated with single-nostril transsphenoidal surgery were collected and analyzed retrospectively by EXCEL software with regard to the patients' sex, age, tumor dimension and quality, postsurgical complications, and so on. Results Total removal of tumors was achieved in 171 (71%) cases, subtotal removal in 28 (11.6%), most removal in 26 (10.7%), and partial removal in 16 (6.6%). After the operation, 38 (15.8%) cases were found with transient diabetes insipidus, 12 (4.9%) cases with worse sight, 4 (1.6%) with cerebrospinal fluid rhinorrhea, 2 (0.8%) with unilateral oculomotor paralysis, and 2 (0.8%) with anterior pituitary insufficiency; during the operation, there were 2 (0.8%) cases with severe cavernous sinus hemorrhage, 2(0.8%) with nasoseptal perforation, no internal carotid artery injuries or death. Conclusions Though the damage of single-nostril transsphenoidal surgery for pituitary adenomas was small, there were a few complications. In order to prevent the complications and reduce the mortality, we should be familiar with the regional anatomy and better master the surgical skills, meanwhile, clinical experience is also very important in improving the therapeutic efficacy.
9.Characteristics of brainstem auditory evoked potential and preservation of facial and auditory nerves in patients with acoustic neuromas
Hui-Lin LIU ; Chao-Shi NIU ; Shi-Ying LING ; Ying JI ; Xiao-Feng JIANG ; Lian YU ; Wan-Hai DING ; Xian-Ming FU
Chinese Journal of Neuromedicine 2010;9(2):145-148
Objective To study the variational features of the brainstem auditory evoked potential (BAEP) and the preservation of the facial and auditory nerves during the microsurgery in patients with acoustic neuromas (AN). Methods We retrospectively analyzed the data of 76 patients with AN, admitted to our hospital fi'om May 2006 to May 2009: preoperative BAEP were examined; the peak latency (PL) of Ⅰ, Ⅲ and Ⅴ waves and their interpeak latency (IPL) were noted and compared. The tumors were microsurgically removed under intraoperative electrophysiologic monitoring to preserve the facial and auditory nerves. The relationships between the preservation of facial and auditory nerves and both the tumor size and the utilization of intraoperative monitoring were discussed. Results Before the operation, the 100% abnormal rate of BAEP was showed on the diseased side with the disappearance of Ⅲ and Ⅴ waves as its main features, while 82.9% (63/76) of that was noted on the contralateral side with the IPL ratio of Ⅲ-Ⅴ / Ⅰ-Ⅲ greater than or equal to 1 as its main features; the BAEP features between the 2 sides had statistical significance (P<0.05). Seventy patients (92.1%) were found abnormal of BAEP on the contralateral side during the operation; the BAEP features of the contralateral side had statistical significance between preoperation and intraoperation (P<0.05). The tumors were totally removed in 69 (90.8%), subtotally removed in 6 (7.9%) and partially removed in 2 (1.3%). The preservation of facial and auditory nerves increased obviously through monitoring the response of BAEP during the surgery and a positive correlation was observed between the preservation of facial and auditory nerves and the response of BAEP in the contralateral side(r=0.912, P=0.000; r=0.613, P=0.000). However, an inverse correlation was observed between the preservation of facial and auditory nerves and the size of AN (r=-0.869, P=0.000; r=-0.738, P=0.000). Conclusion Preoperative BAEP can help to make an exact diagnosis on patients with AN and the preservation of facial and auditory nerves can increase obviously through monitoring the normal response of BAEP during the surgery. The anatomy preservation of facial and auditory nerves depends on the size of AN.
10.Complications of large acoustic neuromas and their prevention
Ying JI ; Chao-Shi NIU ; Shi-Ying LING ; Chuan-Dong CHENG ; Wan-Hai DING ; Xian-Ming FU ; Ye-Han WANG
Chinese Journal of Neuromedicine 2011;10(7):693-696
Objective To discuss the common complications of large acoustic neuromas performed surgery via suboccipital retrosigmoid approach under neurophysiological monitoring, and the prevention of postsurgical complications. Methods One hundred and sixteen patients with large acoustic neuromas, admitted to our hospital from May 2006 to April 2010, were performed surgery via suboccipital retrosigmoid approach under neurophysiological monitoring; clinical data and follow-up data (3-12 months) of these patients were collected and analyzed retrospectively; the influence of tumor dimension on short- and long-term postoperative complications after the surgery were compared.Results Total tumor resection was achieved in 107 patients, subtotal resection in 7 patients, and partial resection in 2 patients. After the operation, the facial nerve was preserved anatomically in 101 patients (87.07%); and the functional valuation of facial nerve according to postoperative House-Brackman showed 89 patients (76.72%) in grade Ⅰ-Ⅱ, 13 patients (11.21%) in gradeⅢ-Ⅳ and 14 patients (12.07%) in grade Ⅴ-Ⅵ. Short-term postoperative complications included hoarse voice in 16 patients and herpes catarrhalis of injured side in 20 patients. Long-term postoperative complications included loss of hearing (n=83) and permanent facial paralysis (n=23). Patients with tumor dimension larger than 5 cm had a significantly higher incidence of short-term postoperative complications than those within 3-5 cm (P< 0.05). No patient died. Conclusion The keys to avoiding the complications are to master the clinical anatomy of the approach, accumulate surgical experiences, preoperatively study the individual imaging and clinical data, and use intraoperative nerve monitoring.