1.Combined endovascular and extravascular neurointerventional technique for treatment of acutely ruptured cerebral aneurysms.
Jinqing HU ; Dong LIN ; Huawei LING
Journal of Interventional Radiology 2003;0(S1):-
Objective To assess the initial clinical effect of combined endovascular and extravascular neurointerventional technique for treatment of acutely ruptured cerebral aneurysms.Methods Forty patients with acutely ruptured aneurysms were embolized with Guglielmi detachable coil (GDC) or Dendron electrically detachable coil (EDC) firstly. Immediately after the embolization, all patients were received lumbar puncture and over-the-wire microcatheters were catheterized into cisterna magna via the spinal subarachnoid space. Urokinase(100,000 IU in 2mL normal saline) was injected into cisterna magna via the microcatheter to dissolve clot 2 hours later. Urokinase injection was repeated according to the CT results. Results All the procedures of aneurysm embolization and catheterization of cisterna magna were successful, no technical complications were found. In all patients, CT scans of the day 3th to day 7th revealed complete disappearance of clot in basal cistern. One patient experienced transient sympotomatic cerebral vasospasm without permanent neurologic deficit. The other 39 patients didn't manifest symptomatic cerebral vasospasm. No aneurysmal rebleeding was detected. Conclusions Combined endovascular and extravascular neurointerventional technique can obliterate acutely ruptured cerebral aneurysms and remove the clot of basal cistern. It can prevent aneurysm rebleeding and cerebral vasospasm. Therefore it is a minimally invasive, safe and reasonable method for treatment of acutely ruptured cerebral aneurysms.
2.advanced aged patients with Bipolar artificial femoral head replacement in the treatment of advanced aged ;patients with femoral neck fracture
Huawei CHEN ; Yanlu CAI ; Wanan CHEN ; Ruizhong LIN
Chinese Journal of Primary Medicine and Pharmacy 2013;20(z2):18-19
Objective To explore the clinical curative effect of the bipolar artificial femoral head replace-ment in the treatment of advanced aged patients with femoral neck fracture .Methods 32 advanced aged patients with femoral neck fracture were treated with the bipolar artificial femoral head replacement ,the effect was observed . Results 32 patients were stage Ⅰincision healing ,and after two weeks all could ambulate .Followed up for 6 to 36 months,two cases of dislocation ,two cases of postoperative pain ,3 cases of deep venous thrombosis ,one case of hip limited functionality.Harris score function assessment:excellent 24 cases(74%),good in 5 cases(16%),3 cases (10%).Conclusion Bipolar artificial femoral head replacement is the effective treatment of advanced aged patients with femoral neck fracture,with shorter operation time,small trauma,early and activities,and reduce the complications and reduce mortality rate etc .
3.Changes of Th17 cytokine in human bronchial epithelial cells induced by coke oven emission
Hongli LI ; Qiuyan XIE ; Xiuling LIU ; Yong NIU ; Yufei DAI ; Yuxin ZHENG ; Lin YAO ; Huawei DUAN
Chinese Journal of Pharmacology and Toxicology 2014;(2):199-204
OBJECTIVE To find the infla mmation bio markers induced by coke oven e missions (COE),we investigated the changes of T helper 17 (Th17 )cytokines in hu man bronchial epithelial (16HBE)cells.METHODS 16HBE cells were exposed to organic extracts of COE collected fro m co-king plant at the concentrations of 5,10 and 20 mg·L -1 for 24 h or 5 d to establish short-term and long-term cell models,respectively.Cell viability was measured by MTT assay and infla mmatory da mage was assessed by lactate dehydrogenase assay (LDH).The cytokines in culture supernatant sa mples was detected by co mmercial hu man Th17 cytokine panel kit.RESULTS COE Can induce infla mmation in COE 20 mg·L -1 group and no expression on IL-17 F and IL-1 β.The concentration of IL-10 was 1 .25 ± 0.54,1 .39 ±0.13 and (1 .90 ±0.73)pg·mL -1 in COE 5,10 and 20 mg·L -1 group showing good con-centration-effect relationship (r=0.98,P <0.05 ).IL-23 expression was found only higher at 10 and 20 mg·L -1 and the concentrations were 3.38 ±3.90 and (1 .74 ±2.00 )pg·mL -1 ,respectively.In 16HBE cells treated by COE for 5 d,elevated expression of IL-17A was found in COE 5 and 10 mg·L -1 group,and there was statistically sigificant difference between COE 10 mg·L -1 and DMSO group (P<0.05).Elevated concentration of IL-17F of 10.2 ±1 1 .78 and (6.79 ±7.84)pg·mL -1 was found in COE 5 and 10 mg·L -1 group.The concentration of IL-10 was 1 .71 ±0.02,1 .49 ±0.25 and (2.82 ± 0.33)pg·mL -1 in COE 5,10 and 20 mg·L -1 group,respectively.We found increased IL-1 βexpression with concentration of 2.72 ±0.62,2.25 ±0.33 and (0.93 ±0.21 )pg·mL -1 in COE 5,10 and 20 mg·L -1 group with negative dose-response relationship.We also found more elevated TNF-αlevels in the 5 d than in the 24 h model with no COE specific relationship.CONCLUSION COE induces expression changes of Th17 cytokines profile in 16HBE cells,including IL-23 and IL-1 βfor early and long-term infla mmation,respectively.IL-10 may be a candidate marker for population study on COE induced infla mmatory injury.
4.Analyses of the changes of sphenoid sinus and related factors of sphenoid sinusitis after microscopic transsphenoidal pituitary adenoma resection
Yong QIN ; Huawei WU ; Shanwu WU ; Hongyan QU ; Kunzhe LIN ; Yehuang CHEN ; Shousen WANG
Cancer Research and Clinic 2021;33(1):48-52
Objective:To investigate the changes of sphenoid sinus and related risk factors of sphenoid sinusitis after microscopic transsphenoidal pituitary adenoma resection.Methods:The clinical and imaging data of 106 patients with large pituitary adenoma in 900 Hospital of the PLA Joint Logistics Team between August 2012 and March 2015 were continually collected. The changes of accumulated blood and fluid, inflammation, mucocele, mucosa remodeling in sphenoid sinus cavity at preoperative and postoperative different time points were observed through the analysis of magnetic resonance imaging (MRI). Binary logistic multiple factors regression model was used to analyze the independent risk factors for postoperative sphenoid sinusitis.Results:MRI results showed that the blood and fluid accumulated in sphenoid sinus cavity were absorbed and dissipated 3 months after the surgery, and the saddle bone windows were covered by new mucous membrane at this time, but they were not complete; the remaining tumors in the saddle all sank into the saddle to different degrees. The reconstruction of sphenoid sinus mucosa was basically complete 6 months after the surgery. There were 7 (6.6%) cases of mucocele in sphenoid sinus and 26 (24.5%) cases of sphenoid sinusitis 3 months after the surgery among 106 patients. The results of multivariate analysis showed that growth hormone adenoma ( OR = 2.981, 95% CI 1.480-26.207, P = 0.014), preoperative sphenoid sinusitis ( OR = 12.392, 95% CI 2.927-52.462, P = 0.001), frequency of multiple transsphenoidal surgery ( OR = 14.758, 95% CI 2.431-89.584, P = 0.003) and perioperative cerebrospinal fluid leakage ( OR = 11.644, 95% CI 2.175-62.344, P = 0.004) were independent risk factors for postoperative sphenoid sinusitis. Conclusions:The evolution of sphenoid sinus cavity contents has its own rules after microscopic transsphenoidal pituitary adenoma resection. Patients with growth-hormone pituitary adenoma, sinusitis before surgery, multiple transsphenoidal surgery and cerebrospinal fluid leakage during the surgery should receive enhanced anti-infection treatment and nasal care in perioperative period to reduce the possibility of sphenoid sinusitis after surgery. The staged time of reoperation for pituitary adenoma resection by transsphenoidal approach should be about 3 months after the previous operation.
5.Reform on diversified teaching of medical imaging
Huawei WU ; Qi FENG ; Linghua LIN ; Jiong ZHU ; Jiejun CHENG ; Jianrong XU
Chinese Journal of Medical Education Research 2013;(6):643-645
Discipline of medical imaging has developed very quickly,which play an increasingly important role in the diagnosis and treatment of clinical disease.According to the characteristics of medical imaging disciplines,we launched reform on teaching management,teaching methods,course content,teacher training and modes of teaching,examing and researching in an aim to improve medical imaging education standards.
6.Cloning and recombinant expression of human soluble TRAIL in Pichia pastoris.
Yingying LI ; Lin WAN ; Hao YANG ; Shan LIU ; Huawei CAI ; Xiaofeng LU
Journal of Biomedical Engineering 2010;27(6):1307-1326
TNF-related apoptosis-inducing ligand (TRAIL) is a member of factor TNF family, which could be potentially developed as novel antitumor agent due to its selective and efficient induction of apoptosis in tumor cells. Gene recombinant expression is an important tool for production of pharmaceutical protein. In this paper, the gene encoding human soluble TRAIL (114-281aa fragment) was cloned by PCR and then inserted into the Pichia Pastoris expression vector pPIC9K. The transformants were double-screened on plates containing neomycin G418 and many clones with high levels of G418-resistance were selected for further studies on protein expression. The recombinant human soluble TRAIL was secreted into the BMMY media under the condition of 3% methanol. And the recombinant protein was purified to homogeneity (-80% purity) by using Ni-agarose affinity chromatography. The yield of this protein is about 1-2 mg per liter culture. Cell viability assays demonstrated that human soluble TRAIL was cytotoxic in both leukemia cells Jurkat and lung cancer cells A549. After treatment with 0.05 microg/ml TRAIL, the survival rate of Jurkat cells was about 10%. The expressed TRAIL showed dose-dependent cytotoxicity in A549 cells within the range of 0.1-1 microg/ml. When the protein concentration reached 1 microg/ml, the survival rates of A549 cells were about 30%. However, the recombinant human soluble TRAIL did not show obvious cytotoxicity in human skin fibroblast cells (HSF) at concentrations tested. There results demonstrate that human soluble TRAIL is selectively cytotoxic in tumor cells. The expression system constructed in this experiment might contribute to further production of soluble TRAIL and TRAIL-based novel fusion proteins in large quantities.
Antineoplastic Agents
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pharmacology
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Cell Line, Tumor
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Cloning, Molecular
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Cytotoxicity Tests, Immunologic
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methods
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Genetic Vectors
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genetics
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Humans
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Pichia
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genetics
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metabolism
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Recombinant Proteins
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biosynthesis
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genetics
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pharmacology
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TNF-Related Apoptosis-Inducing Ligand
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biosynthesis
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genetics
7.The correlation in transcriptional expression with promoter hypermethylation of AKAP12 gene in bladder transitional cell carcinoma
Weiwei LIU ; Ming GUAN ; Huawei ZHANG ; Haowen JIANG ; Zehua YANG ; Quhao WEI ; Nan CHEN ; Yong LIN ; Yuan Lü
Chinese Journal of Laboratory Medicine 2008;31(8):929-932
Objective To investigate transcriptional expression and promoter CpG methylation status of A-kinase anchoring protein 12 (AKAP12) gene and analyze their correlation with clinical pathological stage in bladder transitional cell carcinoma. Methods AKAP12 mRNA expression level and promoter CpG metbylation status was measured by fluorescent quantitative RT-PCR (FQ-RT-PCR) and methylation specific PCR (MSP) in 30 bladder transitional cell carcinoma and adjacent normal tissues. The products of PCR were cloned and bisulfite sequenced. Results Decreased AKAP12 mRNA expression was demonstrated in 22 carcinomas (73. 3% ) and was significantly associated with turnout grade (P =0. 02).The frequency of promoter methylation of AKAP12 gene was 53. 3 % (16/30) and correlated with the tumor stage(r =0.52,Pn =0.03)and grade(r =0.61,Pn =0.01). Conclusion Aberrant promoter methylation of AKAP12 can result in the loss of gene expression and may association with bladder transitional cell carcinoma.
8.Enhancement of cytotoxicity of cantionic antimicrobial peptide in tumor cells by conjugation to cell-penetrating peptide.
Shan LIU ; Hao YANG ; Huawei CAI ; Lin WAN ; Xiaofeng LU
Journal of Biomedical Engineering 2011;28(1):110-114
Due to their lower risk for induction of resistance, membrane-active antimicrobial peptides with anticancer effect are attractive in cancer therapy. Because cell binding contributes to the cytotoxicity of peptide, it is possible to enhance the cytotoxicity of antimicrobial peptide in tumor cells by conjugation to a cell-penetrating peptide (CPP). In this paper, a fusion peptide MPGA by conjugation of antimicrobial peptide MP to CPP Antp at its N-terminus was constructed. After compared the cytotoxicity of unconjugated MP with that of the fusion peptide, it was found that MPGA showed higher cytotoxicity than that of unconjugated MP. And the fusion peptide MPGA induced cell death in tumor cells by membrane disruption. These results demonstrated that the cytotoxicity of antimicrobial peptide can be significantly enhanced by conjugation to CPP, which might be an effective way to develop novel anticancer drugs.
Antimicrobial Cationic Peptides
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pharmacokinetics
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pharmacology
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Antineoplastic Agents
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pharmacokinetics
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pharmacology
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Cell Line, Tumor
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Cell Membrane Permeability
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drug effects
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Humans
9.Expression and purification of recombinant human cytochrome C in Escherichia coli.
Huawei CAI ; Lin WAN ; Hao YANG ; Lihong CHEN ; Xiaofeng LU
Journal of Biomedical Engineering 2007;24(3):620-625
Cytochrome C plays important roles in electron transferring, oxidative stress and apoptosis. In this study, soluble cytochrome C was accumulated in cytoplasm of E. coli by utilizing the co-expression of human cytochrome c and yeast heme lyase from a single plasmid. After ultrasonic disruption of the bacteria, a lot of contaminated proteins were discarded by addition of 350 g/L ammonium sulfate into the supernatant. Then the recombinant human cytochrome C was purified to 80% homogeneity after two times cation exchange chromatography on SP-Sepharose Fast Flow. Yields of cytochrome C greater than 10 mg per liter culture were attained. This efficient system for producing human cytochrome C is helpful for us to understand the roles of this protein in biological processes and therapy of human diseases relevant to apoptosis and oxidative stress.
Cytochromes c
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biosynthesis
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genetics
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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Humans
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Recombinant Proteins
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biosynthesis
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genetics
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isolation & purification
10.Influence of specifically knocking out AMP-activated protein kinase α1 subunit gene in excitatory neurons in brain energy metabolism and cognitive function in mice
Weiwei JIA ; Bingbing LIN ; Lewen CHEN ; Yaling DAI ; Huawei LIN ; Xiaojun HE ; Shengxiang LIANG ; Zhifu WANG ; Weilin LIU
Chinese Journal of Neuromedicine 2021;20(5):433-439
Objective:To investigate the changes of brain energy metabolism and cognitive function in mice with specifically knocking out AMP-activated protein kinase α1 subunit ( AMPKα1) gene in the excitatory neurons by Cre-loxP recombination system. Methods:Sixteen 6-month-old mice with genotype AMPKα1 flox/flox/Camk2a-Cre/ERT2 obtained by hybrid breeding were randomly divided into AMPKα1 knockout group ( n=8) and AMPKα1 wild-type group ( n=8). Mice in the AMPKα1 knockout group were intraperitoneally injected 0.1 mL tamoxifen (20 mg/mL, dissolved in corn oil) daily for a consecutive 5 d to control AMPKα1 gene knockout in the excitatory neurons; and mice in the AMPKα1 wild-type group were intraperitoneally injected 0.1 mL corn oil daily for a consecutive 5 d. Seven d after that, Morris water maze and T maze experiments were employed to detect the spatial learning and memory abilities and spatial working memory of these mice; chemical exchange saturation transfer imaging (CEST) was used to observe the glucose metabolism in the hippocampus and cortex surrounding the hippocampus; Western blotting was used to detect the AMPKα1 and glutamate receptor 1 (GluR1) protein expressions in the hippocampus and cortex surrounding hippocampus of two groups. Results:(1) Morris water maze showed that, as compared with those in the AMPKα1 wild-type group, mice in the AMPKα1 knockout group had significantly prolonged escape latency ([13.90±3.72] s vs. [22.40±6.28] s; [11.95±3.86] s vs. [22.39±9.77] s]) on the 3 rd and 4 th d of experiment, statistically decreased times crossing the platform ([5.25±1.83] times vs. [1.75±1.28] times, P<0.05). (2) T-maze experiment showed that as compared with that of the AMPKα1 wild-type group, the free alternation rate in mice of the AMPKα1 knockout group was significantly decreased ([73.21±9.16]% vs. [48.21±11.29]%, P<0.05). (3) CEST showed that the glucose metabolism levels in the hippocampus and cortex surrounding the hippocampus of AMPKα1 knockout group were significantly lower than those in AMPKα1 wild-type group (1.51±0.81 vs. 2.77±0.67; 1.31±0.83 vs. 2.42±0.95, P<0.05). (4) Western blotting showed that the AMPKα1 and GluR1 protein expressions in the hippocampus and cortex surrounding the hippocampus of the AMPKα1 wild-type group were significantly higher than those of the AMPKα1 knockout group (AMPKα1: 0.70±0.05 vs. 0.49±0.03, 0.98±0.04 vs. 0.64±0.06; GluR1: 1.22±0.18 vs. 0.60±0.11, 0.96±0.08 vs. 0.79±0.04, P<0.05). Conclusion:Specifically knocking out AMPKα1 in excitatory neurons can result in abnormal glucose metabolism in the brain of mice, and thus cause cognitive dysfunction, whose mechanism may be related to excitatory synaptic disorder caused by energy metabolism disorder.