1.Cloning of asparagine synthetase gene and preparation,identification of the monoclonal antibody against asparagine synthetase
Ling YI ; Qing-Yun ZHANG ; Wei-Ben YONG ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To clone human asparagine synthetase(ASNS)gene,express the MS2- ASNS fusion protein through gene engineering and use the purified target protein to immune BALB/C mice, prepare and identify the monoclonal antibody(McAb),which forms the base for studying mechanism of L- asparaginase used as salvage regimen in midline NK/T cell lymphoma nasal type.Methods ASNS gene fragment was amplified by RT-PCR from HepG2 cell line and constructed into prokaryocytic expression vector.Fusion-protein of MS2-ASNS was expressed and used for immunizing BALB/C mice to prepare McAbs against ASNS.Identified the McAb and detected the expression of ASNS in tumor.Results Part of the ASNS reading frame(NCBI,M27396:179-1 420 bp)was cloned and product length of RT-PCR was 1 263 bp.Molecular weight of MS2-ASNS was about 54 700 Da.Two strains of hybridoma secreting ASNS McAbs were obtained.The subtype of the ASNS McAb was IgG2a.Western-blot showed that the McAbs could specifically react with MS2-ASNS fusion protein and ASNS protein in tumor cell lines.ASNS expression was detected by immunocytochemistry and Immunohistochemisrry.Conclusion We have cloned human ASNS gene,obtained the anti-ASNS McAb and examined the expression of ASNS in tumor.
2.Protective effects of curcumin or dexamethasone on ischemia-reperfusion injury of rat lung grafts
Jia-Yuan SUN ; Wei-Gang GUO ; Yong BEN ;
Chinese Journal of Organ Transplantation 2005;0(07):-
Objective To investigate the curative effects of curcumin(CUR)or dexamethasone (DXM)on ischemia-reperfusion injury(IRI)of rat lung grafts.Methods Male SD rats were randomly divided 4 groups:CUR group(CUR was administered intraperitoneally to both donors and recipients at 3 h prior to operation);DXM group(DXM was administered intraperitoneally at 30 min prior to operation);vehicle group(Animals were injected with the DMSO to both donors and recipients at 3 h prior to operation);sham group(Time-matched control animals underwent the same surgery,except that no graft was implanted).Six animals were sacrificed at different reperfusion periods of 2 h and 24 h,respectively.Oxygenation indexes(PO_2/FiO_2),lung injury scores,wet/dry ratio(W/D)and myeloperoxidase(MPO)activity in the transplanted lung were measured.Malondialdehyde(MDA), total anti-oxidative capacity(TAOC),tumor necrosis factor(TNF)-?and interleukin(IL)-6 in the transplanted lung and serum were determined.Results The levels of LPV PO_2/FiO_2 were significant- ly higher in the CUR and DXM groups than in the vehicle groups both 2 and 24 h after reperfusion,re- spectively(P
3.Development of Tripterygium glycosides nano-carries based on "nanoemulsion-gels" and its pharmacodynamics.
Qing GU ; Ben-Ming YOU ; Di-Shun YANG ; Yong-Hua PAN ; Wei SHU ; Ji-Yong LIU
China Journal of Chinese Materia Medica 2015;40(1):73-78
The aim of this study is to develop the Tripterygium glycosides nanoemulsion gels and investigate its pharmacodynamics. Oleic acid was used as oil phase, polyoxyethylene castor oil as surfaetant, and 1,2-propanediol as cosurfactant to screen the formula of Tripterygium glycoside nanoemulsion using the pseudo-temary phase diagrams. Then the nanoemulsion gels was prepared. The ICR mouse ears were sensitazated by 7% DNCB, and then were excited by 0.3% DNCB to stimulate the model of mouse chronic dermatitis and eczema. The concentrations of IFN-γ, IL-4 and IL-8 in mouse blood were determined by ELISA. The results showed that Tripterygium glycosides nanoemulsion gels could significantly inhibit the swelling of mouse ears(P < 0.01) and ameliorate the edama and erythema of model mouse ears skin. Also it could significantly decrease the expression of IFN-γ and IL-4 in model mouse blood. Tripterygium glycosides nanoemulsion gels had a good therapeutic effect on mouse model of dermatitis and eczema. It was expected to provide a new and long-acting exterernal preparation for the treatment of dermatitis and eczema.
Animals
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Chemistry, Pharmaceutical
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instrumentation
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methods
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Dermatitis
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drug therapy
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immunology
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Drug Carriers
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chemistry
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Drugs, Chinese Herbal
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chemistry
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pharmacokinetics
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Emulsions
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chemistry
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Female
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Glycosides
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chemistry
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pharmacokinetics
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Humans
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Interleukin-4
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immunology
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Interleukin-8
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immunology
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Mice
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Mice, Inbred ICR
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Nanoparticles
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chemistry
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Tripterygium
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chemistry
4.Thoracoscopy combined laparoscopic combined surgery: the value of esophageal cancer
Zhaohui GUO ; Shuliang ZHANG ; Ben ZHENG ; Wei ZHENG ; Yong ZHU ; Guobing XU ; Chun CHEN
Chinese Journal of Thoracic and Cardiovascular Surgery 2015;31(3):164-167
Objective The incidence of complications after esophageal cancer surgery is relatively high.The purpose of this paper was to explore the feasibility of combined thoraco-laparoscopy surgical treatment for senile esophageal cancer(over 70 years old).Methods A total of 526 esophageal cancer cases were retrospectively analyzed in this study.The operative procedures was esophageal carcinoma resection surgery and thoracic and abdominal lymphadenectomy which were operated through video-assisted by thoracoscope and laparoscopic From January 2010 to June 2014 in the Union Hospital of Fujian Medical University,divided into aged groups(≥70 years old,n =132) and non-aged group(< 70 years old,n =394).Statistical analysis was carried on the rate of two groups of preoperative risk factors(hypertension,diabetes,cardiac insufficiency,pulmonary insufficiency,cerebral infarction),surgical trauma and the number of lymph node dissection (laryngeal recurrent nerve injury,operation time,intraoperative blood loss),postoperative complications(cardiac complications,chylothorax,pulmonary complications,anastomotic fistula,poor healing of the incision,intrathoracic stomach atony),postoperative recovery and early postoperative mortality.Results Preoperative risk factors:the prevalence of the aged group with hypertension,cerebral infarction,cardiac insufficiency,and pulmonary insufficiency was significantly higher than the non-aged group (P < 0.05) but diabetes (P > 0.05).Surgical trauma and the number of lymph node dissection:no significant difference (P > 0.05).Postoperative complications:the aged group was higher in the incidence of pulmonary complications and cardiac complications than the other (P < 0.05) ; but the chylothorax,anastomotic fistula and poor healing of incision,intrathoracic stomach atony in the two groups seemed no significant difference(P > 0.05).Furthermore,pulmonary complications were highest in the all complications,significantly higher than the others in both groups.Postoperative recovery:significant difference existed in postoperative hospitalization days,the aged group was obviously longer than the non-age groups.Early postoperative mortality rates in the two groups had no significant difference.Conclusion The aged patiences had a higher prevalence in the preoperative risk factors and so was the cardiac complications as well as pulmonary complications among the postoperative complications.However the combined thoraco-laparoscopy in the surgical treatment of elderly patients with esophageal cancer is safe and feasible.
5.Survey of adult carotid atherosclerosis in water-related endemic fluorosis areas in Heilongjiang province in 2008
Jia-yong, LIU ; Hui, LIU ; Wei, DONG ; Ben, GAO ; Yun-qi, LIU ; Dian-jun, SUN
Chinese Journal of Endemiology 2010;29(6):634-636
Objective To investigate the disease situation of adult carotid atherosclerosis in water-related endemic fluorosis areas in Heilongjiang province in 2008 so as to explore the relationship between water-related endemic fluorosis and incidence of carotid atherosclerosis disease. Methods A total of 266 participants over the age of 40 from four villages in Zhaozhou county with water fluoride ≥ 1.0 mg/L in a fluorosis area and 283 residents over the age of 40 from four villages in Tailai county with water fluoride < 1.0 mg/L in a nonfluorosis area were investigated. A portable-type B mode color ultrasound was used to examine the left carotid artery of all participants.The carotid atherosclerosis was diagnosed and graded through the ultrasonograms. Results The prevalence rates of carotid atherosclerosis in the 4 fluorosis villages(Xinfeng, Taipingshan, Baochan and Houzheng villages) were 47.3% (35/74), 63.5%(40/63), 73.3%(33/45) and 60.7%(51/84), respectively, and in the 4 nonfluorosis villages(Hala,Qianxing, Sanjia and Ailin villages) were 32.7% (17/52), 32.9% (24/73), 39.2% (31/79) and 30.4% (24/79),respectively. The prevalence rates of carotid atherosclerosis in every villages standardized by age were 47.3% ,63.5%,73.3% ,60.7% and 34.7% ,36.3% ,43.0% ,41.3%, respectively. Statistic method used was Wilcoxon two sample test and the differences were significant(T = 26, P < 0.05 ). Total carotid atherosclerosis positive rate standardized by age in the 4 fluorosis villages was significantly higher than that in the 4 nonfluorosis villages[57.5%(153/266) ,37.8% (107/283), x2 = 21.36, P < 0.01 ]. After standardized by age, the severity of carotid atherosclerosis was significantly different between fluorosis villages and nonfluorosis villages (x2 = 36.15, P < 0.01 ). Conclusion The prevalence rate of carotid atherosclerosis in endemic fluorosis area is higher than that in nonfluorosis area.
6.Clinical analysis of 38 elderly patients with early double primary cancers.
Chang-hao CAI ; Ben-yan WU ; Dao-hong WU ; Yong SHAO ; Meng-wei WANG
Chinese Journal of Oncology 2004;26(7):440-442
OBJECTIVETo study the clinical features and proper treatment of 38 elderly patients with early double primary cancers.
METHODSThirty-eight elderly patients with early double primary cancers treated from January 1980 to March 2003 were retrospectively reviewed for involved organs, treatment and prognosis.
RESULTSDigestive tract was the most frequently involved, followed by urogenital system and lung. Long-term results of endoscopic mucosal resection (EMR), operation and radiotherapy were superior to other methods. The prognosis of gastrointestinal carcinoma was better than that of prostate carcinoma and hematopoietic system. The operation rate decreased with increasing age. The 5-year survival rates of EMR, operation and radiotherapy were 85.7%, 71.1% and 75.0%, respectively. The medium survival time was 120 months in first cancer and 39 months in the second primary cancer. The 5-year survival rates of the first cancer and second primary cancer were 88.6% and 53.8%.
CONCLUSIONYearly follow-up for elderly patients with endoscopy, beta ultrasonic scan and X-ray contribute to finding of early double primary cancers. Operation is the best treatment of early double primary cancers. Endoscopic mucosal resection is especially suitable for old patients with digestive tract and bladder cancer.
Aged ; Aged, 80 and over ; Colorectal Neoplasms ; diagnosis ; radiotherapy ; surgery ; Endoscopy, Digestive System ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; radiotherapy ; surgery ; Male ; Middle Aged ; Neoplasms, Multiple Primary ; Prostatic Neoplasms ; radiotherapy ; surgery ; Retrospective Studies ; Stomach Neoplasms ; radiotherapy ; surgery ; Survival Rate
7.The clinical value of sixteen-detector row computed tomography angiography for the assessment of coronary artery bypass graft.
Wei SONG ; Shu-xuan JIN ; Yong-ping DU ; Jian-ping LIU ; Ben HE ; Bin-yao WANG
Chinese Journal of Cardiology 2005;33(8):704-707
OBJECTIVEThe aim of this study was to evaluate the value of sixteen-detector row computed tomography angiography (CTA) for the assessment of coronary artery bypass graft (CABG).
METHODSSixty-two consecutive patients undergoing coronary artery bypass grafting were recruited. Among them, 6 patients were excluded from the study due to unfavorable control of heart rate. A total of 56 patients with 152 coronary artery bypass grafts (internal mammary artery, n = 48; saphenous venous grafts, n = 104) were examined by computed tomography angiography (CTA) with sixteen-detector row CT and by conventional invasive coronary angiography (CAG). All CT procedures were performed with retrospective electrocardiogram gating method. The patients' mean heart rate was 58 +/- 6 beats/minute. 120 ml of Visipaque 320 were continuously injected with the rate of 4.0 ml/sec during the procedure. The patency and the stenosis of coronary artery bypass grafts were evaluated by two experienced readers.
RESULTSAll the coronary artery bypass grafts were visualized by CTA, and all the proximal bypass anastomoses and 71% of the distal bypass anastomoses were also visualized by CTA. Furthermore, 29 occlusions and 13 significant stenoses of coronary artery bypass grafts were detected by CTA. The comparison of the results between CTA and CAG showed that among all the 42 occluded and stenosed coronary artery bypass grafts detected by CTA, 34 were confirmed by CAG; among all the 110 normal coronary artery bypass grafts detected by CTA, 108 were confirmed by CAG. There were 8 false positive and 2 false negative findings, resulting in a sensitivity of 94%, a specificity of 95%, a positive predictive value of 86%, and a negative predictive value of 99%.
CONCLUSIONSixteen-detector row CTA technology may provide a reliable visualization and higher diagnostic accuracy of coronary artery bypass grafts lesions. This technique can be used as a noninvasive procedure for the diagnosis of suspected coronary artery bypass grafts dysfunction.
Aged ; Coronary Angiography ; methods ; Coronary Artery Bypass ; Coronary Artery Disease ; diagnostic imaging ; surgery ; Coronary Restenosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed
8.Interaction between p38 mitogen-activated protein kinase signal transduction pathway and NF-kappaB/IkappaB system on the proinflammatory cytokines release after burn trauma.
Xu-lin CHEN ; Zhao-fan XIA ; Duo WEI ; Dao-feng BEN ; Yong-jie WANG ; Nian-qing DENG
Chinese Journal of Surgery 2006;44(7):492-495
OBJECTIVETo investigate the interaction between p38 mitogen-activated protein kinase signal transduction pathway and nuclear factor (NF)-kappaB/IkappaB system on the proinflammatory cytokines release after burn trauma.
METHODSHuman monocyte line THP-1 were incubated with serum from eight healthy controls, burn sera, burn sera pretreatment with SB203580, and burn sera pretreatment with pyrrolidine dithiocarbamate (PDTC). After 24 hours incubation with serum, tumor necrosis factor (TNF)-alpha and interleukin-1beta (IL-1beta) levels in THP-1 culture supernatants were measured by ELISA. The activities of p38 MAPK and expressions of IkappaBalpha in THP-1 were measured by Western blot analysis. The EMSA method was used to characterize the binding activities of NF-kappaB and activating protein (AP)-1 in THP-1.
RESULTSIn comparison with normal controls, burn sera resulted in a significant higher level release of TNF-alpha and IL-1beta in THP-1 [(7.30 +/- 0.84) ng/ml vs (2.20 +/- 0.28) ng/ml, P < 0.05; (2.88 +/- 0.38) ng/ml vs (0.81 +/- 0.14) ng/ml, P < 0.05], which were significantly inhibited by pretreatment with SB203580 or PDTC. Burn sera showed increased activities of p38 MAPK and AP-1 in THP-1 (4728 +/- 582 vs 1291 +/- 163, P < 0.05; 946 +/- 137 vs 361 +/- 40, P < 0.05), which were abolished by pretreatment with SB203580 but not PDTC. The expression of IkappaBalpha in THP-1 incubated with burn sera was significantly decreased than those incubated with control sera (1211 +/- 115 vs 2658 +/- 318, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580. Burn sera also leaded to an increased activity of NF-kappaB in THP-1 (1636 +/- 170 vs 317 +/- 32, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580.
CONCLUSIONSThere are no direct interaction between p38 MAPK signal transduction pathway and NF-kappaB/IkappaB pathway. These two pathways, which regulate the production of TNF-alpha and IL-1beta in monocyte following burn trauma, are parallel and independent.
Adolescent ; Adult ; Burns ; immunology ; physiopathology ; Female ; Humans ; I-kappa B Proteins ; physiology ; Immune Sera ; pharmacology ; In Vitro Techniques ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Monocytes ; drug effects ; physiology ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; physiology ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism ; physiology
9.Role of adjuvant chemoradiotherapy in treatment of resectable esophageal carcinoma: a meta-analysis.
Bin ZHENG ; Wei ZHENG ; Yong ZHU ; Xiao-Yan LIN ; Ben-Hua XU ; Chun CHEN
Chinese Medical Journal 2013;126(6):1178-1182
BACKGROUNDThe effectiveness and influence of surgery followed by adjuvant chemoradiotherapy (CRT) on the survival of patients with resectable esophageal carcinoma are still under debate. The outcomes of clinical trials have not been consistent. This study aimed to perform a meta-analysis of eligible published clinical trials to compare CRT with surgery without adjuvant chemoradiotherapy (non-CRT) for resectable esophageal carcinoma.
METHODSComputerized bibliographic and manual searches were undertaken to identify all eligible literature between 1990 and 2012. PubMed, EMBASE, Chinese National Knowledge Infrastructure, and Wanfang databases were our primary sources for published clinical trials. The quality of the methodology and reliability of the data from all of the clinical trials were assessed. All data were extracted by three independent researchers.
RESULTSSeven studies that included a total of 523 patients were selected. It was found that CRT significantly improved survival. The odds ratio (OR) in comparing CRT and non-CRT groups was 1.75 (95% confidence intervals (CI): 1.17 - 2.60, P = 0.006) for 1-year survival, 2.07 (95%CI: 1.45 - 2.96, P < 0.0001) for 3-year survival, and 2.17 (95%CI: 1.45 - 3.26, P = 0.0002) for 5-year survival. There have been no CRT treatment-related deaths reported in the literature. The incidence of related complications was high in the cases with CRT. Patients treated with CRT had a lower incidence of local-regional cancer recurrence (OR: 0.49, 95%CI: 0.31 - 0.76, P = 0.002) and a similar incidence of distant cancer recurrence (OR: 0.90, 95%CI: 0.60 - 1.34, P = 0.60).
CONCLUSIONSIt was found that patients with resectable esophageal carcinoma could gain a survival benefit from CRT. However, CRT was associated with a high incidence of related complications.
Carcinoma ; surgery ; therapy ; Chemoradiotherapy, Adjuvant ; methods ; Esophageal Neoplasms ; surgery ; therapy ; Humans
10.Role of p38MAPK signal transduction pathway in Kupffer cells production of TNF-alpha and IL-1beta in severely burned rats.
Xu-Lin CHEN ; Zhao-Fan XIA ; Duo WEI ; Dao-Feng BEN ; Yong-Jie WANG
Chinese Journal of Surgery 2005;43(3):185-188
OBJECTIVETo investigate the role of p38 mitogen-activated protein kinase (MAPK) signal transduction pathway in the Kupffer cells production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in severely burns rats.
METHODSMale health adult Sprague-Dawley rats were randomized into four groups: sham burn rats given vehicle, sham burn rats given the p38 MAP kinase inhibitor SB203580, rats given a 30% total body surface area (TBSA) full-thickness burn and fluid resuscitation plus vehicle, and burn rats given injury and fluid resuscitation plus SB203580. Rats from each group were killed at 24 h after burn or sham burn and Kupffer cells (KCs) were isolated. After 18 h incubation, KCs next were stimulated with 50 ng/ml of LPS for 18 h. After stimulation, supernatants were removed for analysis of TNF-alpha and IL-1beta levels by ELISA. The TNF-alpha and IL-1beta mRNA expressions (by quantitative real-time RT-PCR) and the activities of p38 MAPK and JNK (by Western blot analysis) in KCs were examined.
RESULTSEighteen hours after 50 ng/ml LPS stimulation, KCs from burn rats released significantly higher levels of TNF-alpha and IL-1beta than did shams. The mRNA levels of TNF-alpha and IL-1beta in KCs increased significantly postburn. Western blot analysis suggested that expression of phosphorylated p38 MAPK and JNK were increased in KCs harvested from burn group after stimulation with LPS compared with those from sham group. In vivo administration of SB203580 markedly suppressed both the release of TNF-alpha and IL-1beta and the mRNA expressions of TNF-alpha and IL-1beta in KCs from both sham and burn rats. p38 MAPK activity in KCs was abolished by administration with SB203580, whereas JNK was not.
CONCLUSIONSp38 MAPK signal transduction pathway mediates KCs production of proinflammatory cytokines TNF-alpha and IL-1beta in severely burned rats.
Animals ; Blotting, Western ; Burns ; enzymology ; physiopathology ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Interleukin-1 ; genetics ; metabolism ; Kupffer Cells ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism ; physiology