1.The cloning,expression and purification of cDNA in E.coli
Xiaodong WU ; Lijuan XUE ; Shaojuan YANG ; Hui ZHU ; Weizhong WANG
Chinese Journal of Immunology 1986;0(04):-
Objective:The human heparin-binding growth factor Midkine(hMK) gene was constructed and expressed in E.coli, and the activity of MK protein was tested.Methods:The MK gene was amplificated from the tissue of human embryo kidney by RT-PCR techniques. The MK fragment without signal peptide was inserted into plasmid pET30a. The gene was expressed in E.coli, the protein was purificated by affinity chromatography, and the activity was tested by ~3H-TdR method.Results:The sequence of hMK in our experiment was as same as the sequence published in Genebank. The activity assay was shown high activity of MK protein in our experiment.Conclusion:The expression carrier of hMK was established successful and it can expression high activity hMK protein in E.coli.
2.Docetaxel combined with cisplatin and fluorouracil in the treatment of advanced gastric cancer
Hui LI ; Wenhui SUN ; Bei XUE ; Changping WU
China Oncology 2006;0(10):-
Background and purpose:Gastric cancer is one of the leading causes of cancer death throughout the world.When compared with optimal supportive care alone,combination chemotherapy yields a significant advantage in the management of advanced gastric cancer.However,no single regimen has emerged or been accepted as being clearly superior over another.Here we investigated the efficacy and safety of "docetaxel+ cisplatin + fluorouracil" 5-day combination chemotherapy as treatment in patients with advanced gastric cancer.Methods:Between 2004 January and 2005 July,we enrolled 30 patients [males 22,median age 53 years(range 28-72)] with advanced gastric cancer.The regimen consisted of docetaxel 75 mg/m~(2) on day 1,cisplatin 15 mg/ m~(2) on days 1-5,and fluorouracil 500 mg/ m~(2) on days 1-5,every 3 weeks.All patients received over two cycles of this regimen.Results:A total of 99 cycles were administered.Mean cycle number per patient was 3.3.The administered dose intensity of docetaxel was 23.5 mg/m~(2)/week,fluorouracil 735 mg/ m~(2)/week and cisplatin 14.7 mg/ m~(2)/week,which corresponded to 94%,97.5% and 92.3% of planned doses.Of these patients,16(53.3%) achieved a partial response,8(26.6%) stable disease,and 6 patients(20%) showed progressive disease.The median time to progression was 5.1 months.(95% CI 4.1-6.0 months).Median overall survival was 9.9 months.(95% CI 8.1-11.6 months).Leukopenia occurred during 36.7% of cycles(36 of 99 cycles);18.2% grade Ⅰ,10.1% grade Ⅱ,5.0% grade Ⅲ and 3.0% grade Ⅳ.Anemia occurred in 12.1%(12 of 99 cycles);7.1% grade Ⅰ and 5.0% grade Ⅱ.Thrombocytopenia was 15.2%(15 of 99 cycles) and all were grade Ⅰ or Ⅱ.Diarrhea,stomatitis and hypersensitivity occurred in 16.7%(5 out of 30 patients),respectively.Neutropenic fever occurred in two patients(6.7%) and myalgia in nine(30%).Conclusions:Docetaxel combined with fluorouracil and cisplatin is an active and tolerable regimen for the treatment of patients with advanced gastric cancer.
3.AdH1-siRNA-Survivin inhibits tumor growth and metastasis of mouse melanoma
Zhiyong YU ; Weigan SHEN ; Qingyu XUE ; Hui WU
Journal of Third Military Medical University 2003;0(18):-
Objective To evaluate the effect of adenovirus-mediated siRNA targeting Survivin on tumor growth and metastasis of mouse melanoma.Methods A replication-defective recombinant adenovirus AdH1-siRNA-Survivin targeting mouse Survivin was constructed using the homologous recombination technique.The expression of Survivin protein in B16BL6 cells after infection was detected by Western blotting.Mouse melanoma models were established by subcutaneous injection of B16BL6 cells.AdH1-siRNA-Survivin of 2?109 pfu every week by multi-spot intratumoral or peritumoral injection was performed to every mouse(totally three times).The tumor volume was measured every five days after tumor cell implantation,and the microvessel density(MVD) of tumor was counted by immunohistochemistry under the microscope.For the experimental metastasis assay,B16BL6 cells infected with AdH1-siRNA-Survivin were injected into the tail veins of mice,then the visible black nodules on the lung surface were counted macroscopically at 21th day after tumor cell injection.Results AdH1-siRNA-Survivin could obviously decrease the expression of Survivin protein in B16BL6 cells.When compared with the blank control and AdH1-empty control,the tumor growth was inhibited and MVD was less in the mice injected with AdH1-siRNA-Survivin(P
5.Progress of potential therapeutic targets and small molecule ligands of anti-colorectal cancer pathogenic Fusobacterium nucleatum
Xue-xin BAI ; Ya-hui HUANG ; Chun-quan SHENG ; Shan-chao WU
Acta Pharmaceutica Sinica 2023;58(3):639-645
The composition of intestinal microflora is closely related to the occurrence and development of colorectal cancer (CRC). Among them,
6.Effect of astragaloside IV on SDF-1 α and CXCR4 secretion of human umbilical vein endothelial cells damaged by high glucose
Xue BAI ; Hui XIAO ; Yicheng YU ; Hongwei LAN ; Tingting WANG ; Chenhong ZHU ; Ajian PENG ; Wu XIONG
Journal of Chinese Physician 2021;23(1):24-28
Objective:To investigate the effect of astragaloside IV (AS-IV) on the secretion of stromal cell-derived factor-1α (SDF-1α) and CXC chemokine receptor 4 (CXCR4) by high glucose injured human umbilical vein endothelial cells (HUVECs), so as to lay a foundation for further study on AS-IV improving angiogenesis by regulating SDF-1 α/CXCR4 axis of endothelial cells.Methods:HUVECs were isolated and cultured from the umbilical vein of full-term healthy newborns and identified by von Willebrand factor (vWF) combined with 4-diamino-2-phenylindole (DAPI) nuclear staining. The obtained HUVECs was cultured in EGM-2 medium with 30 mmol/L glucose for 120 h to obtain high glucose damaged HUVECs. After intervention with different concentration gradients (25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, 400 mg/L) AS-IV for 72 hours, the contents of SDF-1α and CXCR4 were detected by enzyme linked immunosorbent assay (ELISA) method to determine the best concentration of AS-IV. The supernatant of damaged HUVECs were collected at 6, 12, 24, 48 and 72 hours after intervention with the best concentration of AS-IV, and the contents of SDF-1α and CXCR4 were detected by ELISA method to determine the best action time of AS-IV. The damaged HUVECs was randomly divided into experimental group and control group, and the blank group was set up at the same time. The experimental group was treated with the best concentration of AS-IV and the best time, the control group and the blank group were treated with the same volume of phosphate buffered saline (PBS) solution, and the contents of SDF-1α and CXCR4 in each group were detected by ELISA method.Results:The vWF factor on the cell membrane was green fluorescence, and the nucleus was blue after DAPI staining. When the fusion image showed green fluorescence, HUVECs were identified by blue fluorescence. The expression of SDF-1α in damaged HUVECs was the best when treated with AS-IV of 100 mg/L for 24 hours (1 642.87 pg/ml), and the expression of CXCR4 in damaged HUVECs was the best when treated with AS-IV of 50 mg/L for 48 hours (8.44 ng/ml). Compared with the control group, the contents of SDF-1α and CXCR4 in the experimental group were significantly increased, and the difference was statistically significant ( P<0.05). While the contents of SDF-1α and CXCR4 in the experiment group were slightly less than those in the blank group and there was no statistically significant difference ( P>0.05). Conclusions:AS-IV can promote the expression of SDF-1α and CXCR4 in HUVECs damaged by high glucose to return to normal physiological level, so as to play the role of vascular repair and neovascularization.
7.Histopathological observation of acquired immunity in skin of hamsters elicited by Necator americanus third-stage infective larvae
Jian, GUO ; Jian, XUE ; Hui-qing, QIANG ; Jia-tong, WU ; Shu-hua, XIAO
Chinese Journal of Endemiology 2008;27(6):613-616
Objective To obtain acquired immunity evidence in hamsters elicited by third stage hookworm larvae of Necator americanas(NaL3).morphology changes of NaL3 and inflammatory responses in the skin and undedying subcutaneous tissue and muscles of hamsters were observed.Methods Hamsters were immunized subcutaneously with one dose of 150 NaL3 at 2 weeks earlier,and then challenged pereutaneously with 900 NaL3.Skins were excised from post-challenge hamsters at 6,24,72 hours and 1,2 weeks,and then examined under light microscopy.Non-immunized hamsters served as negative controls.Results In non-immunized hamsters the number of NaL3 were 15,33,11.0 and 0 at 6,24,72 hours and 1,2 weeks post-infection.No damaged or dead NaL3 section was observed.All NaL3 exhibited no structural damage and infihrating inflammatory cells were absent from the sunDunding tissues.There were no cutaneous changes.In contrast.the total number of Nak sections in the skin of immunized hamsters were 25,53,15,5 and 4 at 6,24,72 hours and 1,2 weeks post-challenge.Among these NaL3 sections,damaged and dead section number were 0,24,6,0,0 and 0,0,7,5,4.At 24 hours post-challenge the Nak exhibited cutieular swelling and damage.By 72 hours post-challenge pyknosis of the somatic cells nuclei and sparseness or loss of definition in the internal structures of NaL3 were seen.One or two weeks after chanenge,the NaL3 showed severe damage or even dead with remnants.Inflammatory responses including macrophages,epithelioid cells and eosinophils infiltrating and granulomata forming were mainly seen around the NaL3 sections in the skin of immunized hamsters.Conclusions Hamsters initially immunized with NaL3 exhibited obvious acquired immunity protection against percutaneously challenged infection as evidenced by vigorous inflammatory responses in the skin and underlying subcutaneous tissue and muscle.
8.Analysis of external quality control results of iodine deficiency disorders network laboratory in Ningxia from 2002 to 2011
Xue-qin, WANG ; Hui-zhong, WU ; Ya-jun, ZHANG ; Jun, ZHAN
Chinese Journal of Endemiology 2013;32(4):461-463
Objective To analyze the examination results of external quality assessment (EQA),at all levels of iodine deficiency disorders (IDD) network laboratories in Ningxia Province and to further standardize and improve the laboratory,and to provide a reliable laboratory quality assurance for surveillance and control of IDD.Methods The examination results of EQA at all levels of IDD laboratories in Ningxia Province were statistically analyzed in accordance with the National Reference Laboratory (NRL) of IDD (2002-2011).Results Laboratory hardware equipment and technology at all levels met the testing requirements,and qualified rate of quality control increased year by year.Both of the response rate and qualification rate of urine iodine laboratories at provincial level were 100% in the past decade.From 2005 on,the response rate of city laboratories had been 100%,and the qualification rate had been 100% since 2007.The response rate and qualification rate of salt iodine laboratories at both the provincial level and the city level were 100% in the past decade.The response rate of salt iodine laboratories at county level had been 100% since 2004,and the qualification rate had been 100% since 2009.Salt iodine and urinary iodine levels were fully qualified for the past three years at provincial,municipal and county levels.Conclusions All levels of IDD network laboratory in Ningxia Province runs good,EQA is fully qualified,and is able to provide a reliable laboratory quality assurance for surveillance and control of IDD.
9.Surgical treatment of primary pulmonary artery sarcoma.
Chinese Medical Journal 2011;124(3):461-463
BACKGROUNDPulmonary artery sarcoma originating from the pulmonary artery is a rare disease and its prognosis is extremely poor. Most patients are initially thought to have pulmonary embolism.
METHODSFive patients (3 males and 2 females, mean age was (42.0 ± 8.5) years) of primary pulmonary artery sarcomas underwent surgical procedures. Sarcoma resection and right ventricle outflow tract reconstruction were performed.
RESULTSThere was no death 30 days post-operation except one patient. The function of heart was improved to NYHA II.
CONCLUSIONSGiven the significant morbidity and mortality of pulmonary artery sarcomas, an aggressive strategy for resection is indicated. Early diagnosis and complete surgical resection is perhaps the best way to improve patients' survival with pulmonary artery sarcoma.
Adult ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Artery ; pathology ; surgery ; Sarcoma ; surgery ; Treatment Outcome