1.Expression,Purification and Bioactivity Identification of IFN-tau
Jia-Lei FU ; Chang-Zheng SONG ; Geng-Lin ZHANG ;
China Biotechnology 2006;0(03):-
Interferon-tau (IFN-tau) is a newly discovered IFN of type I. It was originally found for its role as a pregnancy recognition hormone in ruminant animals such as sheep and cows. Like the other type I IFNs, IFN-tau have the same biological activities including antiviral, antiproliferative and immunomodulatory effects. In order to clearly identify the function of IFN-tau, the coding sequence of IFN-tau was amplified by PCR from IFN-tau cDNA, this fragment digested by EcoR I and BamH I and was inserted into pBV220 to form the recombinant plasmid pBV220/IFN-tau which was then transformed into E.coli BL21. It was found that pBV220/IFN-tau was highly expressed as inclusion body in BL21. Next, the expressed protein was purified on S-100 High Resolution and the purified product was confirmed by amino acid sequence analysis. Moreover, the standard antiviral activity test indicated that the activity of IFN-tau was about 2.09?106IU/ml.
2.The diagnosis and therapy of intestinal lymphangiectasia,report of 15 cases
Wande GENG ; Wenbin SHEN ; Yuguang SUN ; Song XIA ; Kun CHANG
Chinese Journal of General Surgery 2008;23(5):332-335
Objective To evaluate the diagnosis and therapy of intestinal lymphangiectasia.Methods In this study 15 patients were admitted in our hospital during recent 7 years.Clinical manifestations included hypoalbuminemia,symmetrical edema,emaciation,diarrhea and lymphopenia.Lymphangiography,lympanscintigraphy and biopsy were performed for diagnosis.Therapy conducted included conservative therapy,low-fat and medium-chain triglycerides(MCT)diet,albumin infusions,diuretics,total parenteral nutrition and octreotide.Surgical therapy ineluded thoracic duct-vein anastomasis and segmental resection.Results In this group 8 patients receiving conservative therapy were followed-up from 1.5 to 7 years(average 2.5 years).Symptoms were alleviated in 6 patients.Seven patients underwent operative therapy,among them,4 patients received thoracic duct-exterior jugular vein anastomasis and followed-up from 1 to 5 years,with symptoms mitigated in 2 patients.3 patients underwent local intestinal resection,follow-up from 1 to 3 years found one patient was cured,one was improved,and 1 patient died 3 months afterthe operation. Conclusion Intestinal lymphangiectasia is rather rare and there was no definite and effective therapy.
3.Rhizosecretion of HIV-1 Recombinant Capsid Protein from Transgenic Lycium barbarum L.Hairy Roots
Dan-Ru LIU ; Chang-Zheng SONG ; Geng-Lin ZHANG ; Guo-Li DU ;
China Biotechnology 2006;0(02):-
HIV p24 core protein can induce both cellular and neutralizing antibody responses.HIV-1 CA-virus-like particles(VLPs)vaccines provide a promising approach for the development of an effective vaccination strategy against HIV infection.Rhizosecreion of the recombinant proteins provides a new manufacturing platform that can simplify the extraction and purification procedure.Lycium barbarum L.was transformed by Agrobacterium tumefaciens EHA105 harboring the plant expression vector pCAMBIA1305.2-MA4-CA with a GRP signal peptide and MA4-CA fusion gene.Transgenic hairy roots were induced and cultivated in hydroponic culture.Western blotting indicated that the recombinant CA proteins were present in two forms,a glycosylated monomer(37 kDa)and a dimer(50 kDa)in the roots and hydroponic medium.It appeared from the present immunohistochemical data that the recombinant CA proteins fused with GRP signal peptide were confined to the cytoplasm,cell wall and intercellular space,indicating targeting into the secretory pathway.It demonstrated for the first time the rhizosecretion of HIV-1 recombinant capsid protein in Lycium barbarum L.hairy roots,and may offer a novel method for expressing HIV-1 CA-VLPs vaccines in plants.
4.Concomitant precise hemihepatectomy for the treatment of hilar cholangiocarcinoma
Lin WANG ; Zhimin GENG ; Xiaogang SONG ; Yaling ZHAO ; Yi Lü ; Yingmin YAO ; Chang LIU ; Qingguang LIU
Chinese Journal of Digestive Surgery 2013;(3):204-209
Objective To investigate the efficacy of concomitant precise hemihepateetomy for the treatment of hilar cholangiocarcinoma.Methods The clinical data of 38 patients with hilar cholangiocarcinoma who received concomitant precise hemihepatectomy at the First Affiliated Hospital of Xi'an Jiaotong University from January 2009 to October 2012 were retrospectively analyzed.All patients were examined by B ultrasonography,computed tomography (CT),magnetic resonance cholangiopancreatography (MRCP) and CT angiography (CTA)preoperatively.The hepatic function was tested before operation.Of the 7 patients with obstructive jaundice,5 received percutaneous transhepatic cholangial drainage,and 2 received endoscopic nosalbiliary drainage.Surgical procedures were determined according to the results of imaging examination.The resection of hilar cholangiocarcinoma,postoperative histopathological examination,pre-and postoperative hepatic function and prognostic indicators were analyzed.The count data and measurement data were analyzed using the chi-square test and t test,respectively; the survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed using the Log-rank test.COX proportion hazards model was used for multivariate analysis.Results The positive rates of B ultrasonography,CT and MRCP were 65.8% (25/38),71.1% (27/38) and 89.5% (34/38),respectively.The results of 5 patients who received CTA were positive.Concomitant left hemihepatectomy was performed on 28 patients,concomitant right hemihepatectomy on 10 patients; concomitant caudate lobectomy on 22 patients,concomitant resection and reconstruction of portal vein on 4 patients (including 1 patient who received left hepatic vein repair),concomitant hepatic artery resection on 12 patients (including 3 patients who received hepatic artery reconstruction).Of the 38 patients,R0 resection was performed on 32 patients,R1 resection on 4 patients,R2 resection on 2 patients.Hepatic function indicators including total bilirubin,direct bilirubin,alkaline phosphatase,gamma-glutamyl-transferase,alanine aminotransferase and aspartate aminotransferase were significantly decreased after operation (t =7.799,8.445,5.697,6.633,4.469,4.140,P < 0.05).Two patients died perioperatively,with the mortality rate of 5.3% (2/38).The main postoperative complications included bile leakage and hepatic function insufficiency,with the incidences of 28.9% (11/38) and 21.1% (8/38),respectively.Postoperative histopathological findings included 31 patients with invasive adenocarcinoma,5 patients with nodular adenocarcinoma,1 patient with mucinous adenocarcinoma and 1 patient with adenosquamous carcinoma.The overall 1-,2-,3-year survival rates were 66%,37% and 21%,and the median survival time was 22.0 months.There were significant differences in the survival rates between patients who received R0 resection and those with R1/R2 resection,and between patients with N0 and N1/N2 stage (x2 =4.516,10.397,P < 0.05).The results of multivariate analysis showed that positive margin and lymph node metastasis were prognostic indicators.Conclusions Concomitant precise hemihepatectomy has significantly improved the radical resection rate and the efficacy of treatment for hilar cholangiocarcinoma.Comprehensive preoperative imaging examination and hepatic function test are important for the assessment for resectability of hilar cholangiocarcinoma.Selective preoperative biliary drainage are key points to decrease postoperative morbidity and morality.
5.MR conventional sagittal-axial plane and coronal iterative decomposition of water and fat with echo asymmetry and least-squares estimation technique in diagnosis of extraforaminal lumbar disc herniation
Junzu GENG ; Xulin LIU ; Qing CHANG ; Changzheng FAN ; Xiaolei SONG ; Hongri ZHAO
Chinese Journal of Medical Imaging Technology 2018;34(2):288-292
Objective To compare the diagnostic value of MR conventional sagittal-axial plane and coronal iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL) technique in diagnosis of extraforaminal lumbar disc herniation (ELDH).Methods Totally 32 patients with ELDH confirmed by surgery underwent preoperative MR sagittal-axial plane and coronal IDEAL scanning.Disc herniation,nerve involvement,nerve angle,nerve deep impression,nerve thinning or truncation,nerve adhesion unsmooth and nerve swelling were analyzed between the two methods.Results Disc herniation,nerve involvement and number of reduction or disappearance of surrounding fat had no statistically significant differences between the two methods (all P>0.05),while nerve angle,nerve deep impression,nerve thinning or truncation,nerve adhesion unsmooth and nerve swelling were significantly different between the two methods (all P<0.05).Surgical operation confirmed that 32 patients with nerve compression were consistent with findings by IDEAL.Conclusion MR sagittal-axial plane and coronal IDEAL both can be used to diagnose ELDH,but their display ability of nerve compression is different.Coronal IDEAL is superior to conventional sagittal-axial plane and is an effective imaging method,being able to display nerve compression more intuitively,clearly and comprehensively.
6.Apoptosis-inducing effect of annexin A2 on multiple myeloma cells and its related mechanisms.
Hong-Yu BAO ; Jian-Ning WANG ; Yan-Qiu HOU ; Ming SONG ; Liu-Bo ZHANG ; Qing-Qi MENG ; Chang-Geng RUAN
Journal of Experimental Hematology 2012;20(6):1384-1388
This study was purposed to investigate the apoptosis-inducing effect of Annexin A2 gene (AnxA2) on multiple myeloma (MM) cells and its mechanisms. The human MM cell lines U266 and RPMI8226 were transfected by using siRNA targeting at AnxA2; the expressions of AnxA2 mRNA and protein in the siRNA-transfected cells were detected by real-time PCR and Western blot, respectively; the cell apoptosis was assayed by flow cytometry. The results showed that silencing AnxA2 gene by siRNA resulted in decreased expressions of AnxA2 gene and protein, increased apoptosis of U266 and RPMI8226 cell lines (P < 0.05), at the same time resulted in down-regulation of apoptosis-related gene expressions including p65NF-κB, IL-2, IL-6 (P < 0.05), and up-regulation of P53 gene expression (P < 0.05). It is concluded that the AnxA2 silence plays a promoting role in apoptosis of MM cell lines U266 and RPMI8226.
Annexin A2
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genetics
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Apoptosis
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genetics
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Cell Line, Tumor
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Humans
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Multiple Myeloma
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genetics
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pathology
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RNA, Small Interfering
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genetics
7.The diagnosis and therapy of chylous ascites.
Wen-bin SHEN ; Yu-guang SUN ; Song XIA ; Guo-fu WU ; Wan-de GENG ; Ying-xian SUN ; Kun CHANG
Chinese Journal of Surgery 2005;43(1):25-28
OBJECTIVETo discuss the diagnosis and therapy of chylous ascites.
METHODSTo diagnose 40 patients of chylous ascite with regular test and quantitative analysis of chyle, direct lymphangiography, CT (immediately after direct lymphangiography), lymphangioscintigraphy, MRI. Twenty-two patients received conservative therapy, 18 patients received retroperitoneal lymphangiectomy and (or) lymph-vein shunting.
RESULTSLymphatic dysplasia and chylous reflux were found in almost every patient, total parenteral nutrition showed good results. Followed up from 1 month to 5 years, in conservative therapy group, 9 patients were controlled well clinically, the condition of 6 patients was improved better. Seven patients showed no effect. In operation group, 11 patients were controlled well clinically. Four patients got mitigated. Total 7 patients died, although 4 of them ameliorated temporarily.
CONCLUSIONSDirect lymphangiography, CT (immediately after direct lymphangiography) are the most important diagnosis methods. The influence of the therapy to the malformed lymphatic system of patients should be well considered. Lymph-vein shunting, such as thoracic duct-left external jugular vein anastomosis, gastroenteral or retroperitoneal lymphatics-testicular or ovarian vein anastomosis, could improve the circulation of lymph and chyle of patients. Lymphatic microsurgery will play more and more important roles in the treatment of chylous diseases.
Adolescent ; Adult ; Aged ; Child ; Chylous Ascites ; diagnosis ; therapy ; Female ; Follow-Up Studies ; Humans ; Infant ; Lymphatic Vessels ; surgery ; Lymphography ; Magnetic Resonance Imaging ; Male ; Microsurgery ; Middle Aged ; Parenteral Nutrition, Total ; Tomography, X-Ray Computed
9.The role of Toll-like receptor 4 on inflammation and Aβ formation in cortex astrocytes.
Chang-Yin GONG ; ; Ai-Ling ZHOU ; Jia-Hui MAO ; Ya-E HU ; Jin-Song GENG
Acta Physiologica Sinica 2014;66(6):631-638
To investigate the role and possible molecular mechanism of astrocytes in inflammation and amyloid β-protein (Aβ) formation, in this research, by using LPS to stimulate cultured rat astrocytes in vitro with or without anti-Toll-like receptor 4 (TLR4) antibody pretreatment, we first detected the TLR4, TNF-α, IL-1β, β-amyloid precursor protein (β-APP) and β-site APP clearing enzyme 1 (BACE1) mRNA with real-time PCR, and TLR4, NF-κB/P65 protein in cultured astrocytes by Western blot, and then further probed the translocation of NF-κB/P65 using immunofluorescence and the contents of TNF-α, IL-1β and Aβ in culture supernatant through ELISA. We found that all of these indexes increased at different degrees after LPS-stimulation. However, if pretreatment with anti- TLR4 antibody, such stimulating effects of LPS on the nuclear translocation of NF-κB/P65 and TNF-α, IL-1β, Aβ contents in astrocytic culture supernatant were reduced significantly or disappeared in comparison with the group with only LPS-administration. Our results suggest that TLR4 in astrocytes might play an important role in the inflammation and Aβ formation through the TLR4/NF-κB signaling pathway, thus providing new knowledge and understanding of the inflammatory hypothesis of AD pathogenesis.
Amyloid Precursor Protein Secretases
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metabolism
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Amyloid beta-Protein Precursor
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metabolism
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Animals
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Aspartic Acid Endopeptidases
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metabolism
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Astrocytes
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metabolism
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Cells, Cultured
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Cerebral Cortex
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cytology
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Inflammation
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metabolism
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Interleukin-1beta
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metabolism
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RNA, Messenger
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Rats
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Real-Time Polymerase Chain Reaction
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Signal Transduction
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Toll-Like Receptor 4
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metabolism
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Transcription Factor RelA
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metabolism
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Tumor Necrosis Factor-alpha
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metabolism
10.Intravenous Tenecteplase for Acute Ischemic Stroke Within 4.5–24 Hours of Onset (ROSE-TNK): A Phase 2, Randomized, Multicenter Study
Lu WANG ; Ying-Jie DAI ; Yu CUI ; Hong ZHANG ; Chang-Hao JIANG ; Ying-Jie DUAN ; Yong ZHAO ; Ye-Fang FENG ; Shi-Mei GENG ; Zai-Hui ZHANG ; Jiang LU ; Ping ZHANG ; Li-Wei ZHAO ; Hang ZHAO ; Yu-Tong MA ; Cheng-Guang SONG ; Yi ZHANG ; Hui-Sheng CHEN
Journal of Stroke 2023;25(3):371-377
Background:
and Purpose Intravenous tenecteplase (TNK) efficacy has not been well demonstrated in acute ischemic stroke (AIS) beyond 4.5 hours after onset. This study aimed to determine the effect of intravenous TNK for AIS within 4.5 to 24 hours of onset.
Methods:
In this pilot trial, eligible AIS patients with diffusion-weighted imaging (DWI)-fluid attenuated inversion recovery (FLAIR) mismatch were randomly allocated to intravenous TNK (0.25 mg/kg) or standard care within 4.5–24 hours of onset. The primary endpoint was excellent functional outcome at 90 days (modified Rankin Scale [mRS] score of 0–1). The primary safety endpoint was symptomatic intracranial hemorrhage (sICH).
Results:
Of the randomly assigned 80 patients, the primary endpoint occurred in 52.5% (21/40) of TNK group and 50.0% (20/40) of control group, with no significant difference (unadjusted odds ratio, 1.11; 95% confidence interval 0.46–2.66; P=0.82). More early neurological improvement occurred in TNK group than in control group (11 vs. 3, P=0.03), but no significant differences were found in other secondary endpoints, such as mRS 0–2 at 90 days, shift analysis of mRS at 90 days, and change in National Institutes of Health Stroke Scale score at 24 hours and 7 days. There were no cases of sICH in this trial; however, asymptomatic intracranial hemorrhage occurred in 3 of the 40 patients (7.5%) in the TNK group.
Conclusion
This phase 2, randomized, multicenter study suggests that intravenous TNK within 4.5–24 hours of onset may be safe and feasible in AIS patients with a DWI-FLAIR mismatch.