1.A New Method for Accurate Determination of Peroxidase Activity Based on Fluorescence Decrease of Guaiacol
Yang ZHAO ; Yongsheng LI ; Xiufeng GAO
Chinese Journal of Analytical Chemistry 2015;(7):1040-1046
At present the most common colorimetry for the activity determination of the peroxidase ( POD) is based on the detection of the absorbance of product at 470 nm in a reaction system of the H2 O2/POD/guaiacol ( GA) , but the shortcoming of the method is that the formed product is not stable and there is the serious adsorption phenomenon on the cell. To solve this problem, a new method was established for accurate determination of POD activity based on the fluorescent feature of GA. By using standard solutions of horseradish peroxidase as the test samples and under these optimum conditions such as 0. 5 mmol/L of GA, 0. 5 mmol/L of H2 O2 , pH 6, 0. 05 mol/L of phosphate buffer solution and the reaction temperature of 20℃, the sample volume was only consumed 20 microlitre at a time, the linear response range was 500-60000 U/L ( r=0 . 9993 ) , the detection limit was 385 U/L and relative standard deviation was ≤2 . 4% ( n=11 ) . The comparisons for the determination results of the POD activity in the white radish’s extraction solutions were conducted among our method (9714±132 U/L) and the colorimetric method (9926±352 U/L) as well as the recirculating-catalytic flow analysis ( 9608±456 U/L ) . The results showed that the mutual consistency is better.
2.Studies and prospects of the candidate virulence gene of familial medullary thyroid carcinoma
Ming GAO ; Yang YU ; Yongsheng JIA
Chinese Journal of Clinical Oncology 2014;(3):204-206
Familial non-medullary thyroid carcinoma (FNMTC) is de fi ned as the presence of two or more affected fi rst-degree relatives with non-medullary thyroid cancers without other known familial syndromes. FNMTC is one of the most inheritable forms of all cancers, with a high risk of a first-degree relative developing the disease. Compared with sporadic non-medullary thyroid carcinoma (NMTC), FNMTC presents at a younger age and is associated with a higher incidence of multifocal disease and metastasis. This in-creased aggressiveness has been hypothesized to translate into higher recurrence rates and decreased survival of patients with FNMTC. The genes involved in the pathogenesis of FNMTC are yet to be elucidated, although some recent studies identified several predisposi-tion loci with a high degree of genetic heterogeneity. Since 2005, next-generation sequencing (NGS) technologies have been developing as rapid, high-throughput, and cost-effective approaches to fulfill medical sciences and research demands. With the use of NGS, the un-derlying causative genes can be directly distinguished via systematic filtering, through which the identified gene variants are verified for novelty and functionality.
3.Clinical evidence-based study on new-generation drug-eluting stent:everolimus-eluting stents(Xience Ⅴ)
Wenjun GAO ; Wenjie LIU ; Yongsheng KE
Chinese Journal of Clinical Pharmacology and Therapeutics 2002;0(05):-
Percutaneous coronary intervention(PCI) has been a mainstay in the management of coronary artery disease since its introduction in the late 1970s.Bare-metal stents and,more recently,first-generation drug-eluting stents(DES),such as sirolimus-eluting(Cypher) and paclitaxel-eluting stents(Taxus),have further improved results of percutaneous coronary intervention by improving early results and reducing the risk of restenosis.There are currently debates on the safety of these first-generation DES,given the potential for late stent thrombosis which is a first-generation drug-eluting stent of the largest security issue,especially after discontinuation of dual antiplatelet therapy.Next-generation DES such as everolimus-eluting stents(Xience V) holds the promise of superior anti-restenosis efficacy as well as long-term safety.This review makes a presentation of the evidence-based clinical research according to everolimus-eluting stents(Xience Ⅴ).
4.Relation of DNA ploidy and proliferative activity with the clinical and pathologic features in colorectal cancer
Fei GAO ; Yongsheng GUO ; Yu WANG
Chinese Journal of Rehabilitation Theory and Practice 2003;9(5):290-292
ObjectiveTo study the relationship between DNA aneuploidy and proliferative activity and the clinical and pathologic features.MethodsDNA ploidy and cell cycle analysis were analyzed in 119 colorectal fresh cancer apecimens using flow Cytometry and prospectively compared with the CEA in serum,tumor size,tumor morphology,lymph node metastases,Dukes' Classfication,histologic type and grade in colorectal cancer.ResultsThere was no relation between serum CEA and DNA aneuploidy and proliferative activity. Aneuploidy was detected at 56.5% in ulcerating carcinoma, which was significantly higher than 14.7% in bulge carcinoma (P<0.01). Aneuploidy was detected at 64.7% in middle and lower grade carcinoma, which was significantly higher than 36.5% of high grade carcinoma (P<0.01). No significant differences in aneuploidy were observed with respect to tumor size, lymph node metastases, Dukes' classfication,tumor histologic type.ConclusionsDNA aneuploidy of cancer cell can express the degree of malignancy of colorectal cancer. But proliferative activity does not relate to all the clinical and pathologic features. CEA in serum does not relate to DNA aneuploidy and proliferative activity.
5.Role of the Wnt/β-catenin signaling pathway in regulating the phenotypic transformation of aortic valvular myofibroblasts to osteoblast-like cells
Yingnian SHEN ; Hongjie WANG ; Di CHEN ; Gao XU ; Yongsheng LI
Chinese Journal of Geriatrics 2016;35(2):209-213
Objective To elucidate the role of the Wnt/-catenin signaling pathway in regulating the phenotypic transformation of aortic valvular myofibroblasts to osteoblast-like cells.Methods Cultured primary valvular myofibroblastes isolated from porcine aortic valve leaflets were treated with oxidized low-density lipoprotein (ox-LDL) for different lengths of time:24 h,48 h and 72 h.The Wnt signaling pathway inhibitor Dickkopf-1 (DDK-1) was co-incubated with ox-LDL for 72 h.After cells harvest,the expression of myofibroblastic or osteoblast-like phenotype related markers,a-smooth muscle actin (α-SMA),bone morphogenetic protein 2 (BMP2),alkaline phosphatase (ALP) and corebinding factora-1 (Cbfα 1),was detected by Western blotting.The expression and sub cellular localization of β3-catenin was assessed by immunocytochemistry.Changes of the Wnt/β-catenin signaling pathway and the transformation of aortic valvular myofibroblast to osteoblast-like cells were monitored.Results BMP2,ALP and Cbfa 1 protein expression was not or barely detectable in the control group.However,after ox-LDL treatment,the expression of α SMA,BMP2,ALP and Cbfa 1 increased significantly (each P<0.01) in a time-dependent manner (each P<0.05).Besides,ox-LDL was also able to up-regulate the protein expression of β-catenin in a time-dependent manner (P<0.05) and promoted its nuclear translocation.After DKK-1 treatment,the protein expression of β3 catenin and osteogenesis related markers was down regulated (P<0.05).Conclusions The Wnt/β-catenin signaling pathway may play a crucial role in regulating the transformation of aortic valvular myofibroblasts to an osteoblast like phenotype.
6.Understanding of Clinical Application of Modified Xiaochaihu Decoction in Artificial Total Knee Arthroplasty
Weitao GE ; Zhenzhu LIU ; Yongsheng SUN ; Yun GAO
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(8):111-112
Xiaochaihu Decoction comes from Shang Han Lun, which is a main formula for Shaoyang diseases. According to the principle of prescriptions corresponding to syndromes, the clinical application of modified Xiaochaihu Decoction for the treatment of swelling, pain, nausea, vomiting, bloating, and constipation after artificial total knee arthroplasty can achieve good efficacy.
7.Investigation of the correlation between a group of metastasis-associated proteins and metastatic potency in breast carcinoma
Aiju FANG ; Yongsheng GAO ; Shengmei LI ; Wenjun LIU ; Bin MENG
Chinese Journal of General Surgery 2000;0(11):-
Objective To investigate the expression and the significance of a group of metastasis-associated proteins in invasive ductal breast carcinoma(IDC).Methods Tissue microarray containing 247 IDC specimens was constructed.The expressions of ?-B crystallin,CD44v6,MMP-2 and TIMP-2 were detected by immunohistochemistry,and the relation between the expression of these proteins and the clinicopathologic character was analyzed.Results(1) The expression rates of ?-B crystallin,CD44v6,MMP-2 and TIMP-2 in IDC were 70.0%,61.5%,57.5% and 57.1% respectively,and significantly higher than those of normal breast tissues(P
8.Curative effect and safety of percutaneous closure of patent foramen ovale on 23 migraineurs
Yanling DENG ; Kangding LIU ; Xiujuan WU ; Juan WANG ; Yi YANG ; Yongsheng GAO ; Yingqi XING
Chinese Journal of Neurology 2013;(3):180-184
Objective To evaluate the clinical effect and safety of percutaneous closure of patent foramen ovalein treatment of migraine.Methods We studied 23 patients with migraine who were tested positive in Contrast Transcranial Doppler (TCDc) at the Department of Neurology in Bethune First Hospital of Jilin University between 2010 May and 2011 November.Procedures of transcatheterclosure of patent foramen ovale were performed at Department of Cardiovascular Surgery.At varied time points post-surgery,the patients were re-examined by TCDc and transthoracic echocardiograph (TTE) and followed up by phone interviewed with Headache Impact Test-6 (HIT-6) questionnaire to elevate improvement of migraine.The data was analyzed statistically.Results Percutaneous closure was successful in all patients without major long-term side-effects.At follow-ups at 48-542 days ((247 ± 145) days) post-surgery,the scores of HIT-6 were reduced to 38-70 (51 ± 9),compared to the pre-operative scores of 42-76 (61 ± 8).Disability by migraine had significantly improved (t =4.80,P < 0.05).In 6 patients with migraine with aura,the scores of HIT-6 were reduced to 46-56 (51 ± 3),from preoperative scores of 63-76 (69 ± 4) and their disability affected by migraine had significantly improved (t =6.80,P <0.05).In 17 patients with migraine without aura,the scores of HIT-6 after surgery were reduced to 36-70 (52-10) from the preoperative scores of 42-69 (60 ± 8) and disability affected by migraine in this group had significantly improved (t =3.77,P <0.05).Among 18 patients having MRI or CT examination before the operation,8 patients had neurological events (such as cerebral infarction,cerebral ischemia) with average age of (44 ± 7) years old.In this group,disability affected by migraine had significantly improved (the HIT-6 scores:49 ± 7 vs 58 ± 10,t =2.49,P < 0.05).Conclusion Percutaneous closure of PFO showed significant improvement in migraine treatment and it is a safe procedure.
9.Clinical significance of plasma fibrinogen level in patients with lung cancer
Wen TIAN ; Jinghua GAO ; Yongsheng LI ; Jinghua ZHANG ; Jijun LI ; Chunling LIU
Clinical Medicine of China 2015;31(10):871-875
Objective To investigate the relationship among the plasma levels of fibrinogen and the lung cancer,and its clinical significance.Methods From 2011 to 2013,121 cases newly diagnosed lung cancer patients(lung cancer group) and 37 cases healthy individuals(control group) were evaluated.The patients had no history of coagulation system disorders or anticoagulant therapy.Plasma prothrombin time (PT), activated partial thromboplastin time(APTT), thrombin time (TT), fibrin original (FIB), platelet (PLT) of the patients were obtained.The relationship between the plasma levels of fibrinogen and clinical characteristics, therapy modalities (surgery, chemotherapy and radiotherapy), therapy outcomes and survival durations of the patients were analyzed.Results (1) Serum levels of fibrinogen at stage Ⅱ A, stage Ⅱ B, stage Ⅲ A, stage Ⅲ B, stage Ⅳ were (2.001±0.813) g/L, (2.191±0.827) g/L, (3.121 ±2.016) g/L, (4.174±0.595) g/L, (4.332 ± 1.534) g/L, a significant difference was observed between the fibrinogen levels of patients with stage Ⅱ A and those with stage Ⅳ disease (P<0.001), and there were no significant differences among other stages (P>0.05).(2)The mean fibrinogen level was significantly higher in the patient group with ECOG performance status 2 than in the other groups(r=0.613,P<0.05).The mean fibrinogen level was (3.780±1.731) g/L (95%CI,3.122-4.439,P<0.001) in the group with ECOG 0, (4.182 ± 1.661) g/L(95%CI 3.583-4.781 ,P<0.001) in the group with ECOG 1 ,and (4.725±2.153) g/L(95%CI,4.007-5.443,P<0.001) in the group with ECOG 2.(3) The treatment responses of 81 patients who received chemotherapy and/or radiotherapy without surgical intervention were evaluated, serum levels of fibrinogen in 39 patients with partial remission (PR) were (4.005 ±1.177) g,/L,42 patients with stable disease(SD) were (3.192±0.479) g/L, 17 patients with progressive disease(PD) were (7.530± 1.885) g/L,fibrinogen levels were found to be significantly higher in cases with progression,and the difference was significant(P =0.015).(4)Correlation analysis on fibrinogen and chnical indicator: clinical stage (r =0.529, P =0.008), ECOG score (r =0.273, P =0.031), therapy response (r =0.529, P=0.012) were positively correlated with fibrinogen levels.(5)Fibrinogen levels in patients with lung cancer were higher than those of the control group ((2.891 ± 0.484) g/L vs.(3.586± 1.692) g/L, t =-4.620, P <0.05), and the difference was statistically significant (P<0.01).(6)The survival duration was significantly longer in patients with lower fibrinogen levels(321 d vs.435 d,P<0.05).The mean fibrinogen concentration in patients who were still alive at the end of a 2-year follow-up was (3.531 ± 1.482) g/L, whereas the meanfibrinogen level of patients who died was (3.725± 2.063) g/L, and the difference between the two groups was significant(P =0.015).Conclusion The results suggest that Fibrinogen plasma levels might be useful to predict the clinical outcome and survival of patients with lung cancer.
10.Effect of phosphorylated-P38 MAPK on caspase-3 expression in substantia nigra of the MPTP mouse model of Parkinson's disease
Zifeng WEI ; Yongsheng WANG ; Qian WANG ; Liren MA ; Zuofeng ZHANG ; Junling GAO ; Yuxin ZHANG
Basic & Clinical Medicine 2010;30(1):54-58
Objective To investegate the effect of phosphorylated-P38 MAPK(mitogen-activated protein kinase) on the expression of caspase-3 in the substania nigra (SN) of MPTP-induced mouse model of(PD). Methods Mice were randomly divided into MPTP model group, which were treated with MPTP and inhibitor group. Once a day for 5 days; control group was treated with saline and DMSO as much as the model group received per day for 5 days. The behavioral were observed, immunohistochemistry and Western blot for TH, caspase-3 and phosphorylation of P38 MAPK were used to observe the change of positive cell number and the expression level in the SN of midbrain. Results Compared with the mice in control group, the model group showed typical symtoms of PD with decreased numbers of TH-positive neurons and the protein level of TH in SN of the midbrain by about 60% and 65% respec-tively(P<0.01) , the numbers of caspase-3 and phosphorylation of P38 MAPK immunoreactive cells and their protein level in the SN of the midbrain increased markedly (P<0.01). After giving SB203580, the above changes were reduced obviously (P <0. 01). Conclusion In the mouse model of subacute Parkinson's disease induced by MPTP, phosphorylated-P38 MAPK regulated caspase-3 in the SN of midbrain, the specific P38 MAPK inhibitor SB203580 is neurologically oprotective to the mouse model.