1.Neuroprotective effect of inhibition of soluble epoxide hydrolase on focal cerebral ischemia/reperfusion in Rats
International Journal of Cerebrovascular Diseases 2013;(2):108-113
Objective To investigate the neuroprotective effect of soluble epoxide hydrolase (sEH) inhibitor 12-(3-adamantan-l-yl-ureido) dodecanoic acid (AUDA) on focal cerebral ischemia/reperfusion in rats and its mechanisms.Methods Sixty male Sprague-Dawley rats were randomly divided into sham operation and saline control groups,as well as low-dose (0.157 ml/kg),medium-dose (0.235 ml/kg) and high-dose (0.314 ml/kg) AUDA groups (n =12 in each group).Four rats in each group were selected for infarct volume,cell apoptosis and p-Akt immunohistochemistry detection.A model of middle cerebral artery ischemia/ reperfusion was induced by the suture method.The corresponding dose AUDA or equal volume of saline was injected intraperitoneally before reperfusion in each AUDA group and the saline control group.Neurological deficit scores were performed at 24 h of reperfusion.2,3,5 triphenyltetrazolium chloride (TTC) staining was used to detect infarct volume.TdT-mediated dUTP nick end labeling (TUNEL) was used to detect apoptotic cells of brain tissue in the periinfarction area.Immunohistochemical method was used to detect p-Akt expression of brain tissue in the peri-infarction area.Results TTC staining showed no infarction was observed in the sham operation group.The infarction volumes in the saline control group as well as the low-dose,medlum-dose and high-dose AUDA groups were 254.146 ± 25.481,212.679 ± 7.514,150.188 ± 33.997,and 99.563 ± 3.415 mm3,respectively.There were significant differences (F =39.637,P =0.000).The each dose AUDA group was significant less than the control group (all P=0.000).The medium-dose AUDA group was significantly less than the low-dose AUDA group (P=0.002),and the high-dose AUDA group was also significantly less than the low-dose AUDA group (P =0.000) and medium-dose AUDA group (P =0.006).TUNEL staining showed that a small number of apoptotic cells (6.400 ± 1.477/high-power field) were observed in the sham operation group.The numbers of apoptotic cells in the saline control group as well as in the low-dose,medium-dose and high-dose AUDA groups were 57.550 ± 13.067,47.030 ± 8.423,34.530 ± 4.393 and 26.400 ± 2.683/high power field,respectively.Each dose AUDA group was significantly less than the saline control group (all P <0.01).The medium-dose and high-dose AUDA groups were significantly less than the low-dose AUDA group (P < 0.01),and the high-dose AUDA group was also significantly less than the medium-dose AUDA group (P <0.01).Immunohistochemistry showed that only a few p-Akt-positive cells (3.325 ± 1.438/high power field) were observed in the sham operation group.The numbers of p-Akt-positive cells in the saline control group as well as the low-dose,medium-dose and high-dose AUDA groups were 9.450 ±2.531,16.400 ± 3.865,22.875 ± 7.974,and 29.300 ± 3.203/high-power field,respectively.Each dose AUDA group was significantly more than the saline control group (all P <0.01).The medium-dose and high-dose AUDA groups were significantly more than the low-dose AUDA group (all P <0.01).The high-dose AUDA group was also significantly more than the medium-dose AUDA group (P < 0.01).Conclusions The inhibition of sEH may decrease neuronal apoptosis and reduce infarct volume in the peri-infarction area by upregulating the PI3K/Akt pathway.It has a neuroprotective effect for focal cerebral ischemia/reperfusion in rats.
2.Analysis of SOD and HCY in acute cerebral infarction and acute cerebral hemorrhage patients
Famei QI ; Xiumei YUAN ; Yulan DONG
International Journal of Laboratory Medicine 2015;(10):1323-1324
Objective To investigate the significance of superoxide dismutase (SOD) and homocysteine (HCY) in patients with acute cerebral infarction and acute cerebral hemorrhage .Methods Serum levels of SOD and HCY were detected in 251 acute cere‐bral infarction patients ,204 acute cerebral hemorrhage patients and 485 healthy subjects .Results Compared with healthy subjects , serum levels of SOD in acute cerebral infarction patients and acute cerebral hemorrhage patients were lower ,but serum levels of HCY were higher (P<0 .05) .There were significant differences of serum SOD and HCY levels among different types of cerebral infarction patients (P<0 .05) .Conclusion There could be important value for determine of SOD and HCY in patients with acute cerebral infarction and acute cerebral hemorrhage .
3.Research on PCR determining of type b Haemophilus influenzae strains and immunogenicity of polysaccharide conjugates
Yulan YUAN ; Jingrong GUO ; Jiwei ZHOU ; Hua GAO
Chinese Journal of Microbiology and Immunology 2012;32(4):346-350
Objective To determine Haemophilus influenzae type b strains in molecular level using PCR,and to study the immunogenicity of capsular polysaccharide conjugates in mice.Methods Extracting genomes using bacterial DNA extract kit from Hoemophilus influenzae type b strains,and PCR for determining the strains through serotyping-specific and capsular genotyping primers respectively.Various capsular polysaccharides conjugated TT respectively,and the conjugates were administered subcutaneously to mice through dilution.After vaccination with two doses,blood samples were collected for the detection of antibody levels to polyribosylribitol phosphate ( PRP),the capsular polysaccharide of Hib.Results All five Haemophilus influenzae type b strains contain type-specific(482 bp) and capsular type (343 bp)DNA fragment through PCR detecting.The DNA fragments were sequenced.BLAST show that these sequences are 100% homology comparing the above strains respectively,and are 99% and 100% homology comparing the GenBank X78559.1 and M19995.1 respectively.The immunogenicity of mice from various capsular polysaccharide conjugates (PRP-TT) was not significantly different by ELISA detecting.Conclusion Through PCR,Haemophilus influenzae type b strain can be determined in molecular level.The immunogenicity of mice from purified capsular polysaccharide conjugates was not different.The study provides a detection means for the features and heredity stability of Haemophilus influenzae type b strain and reference data for the immunogenicity of different polysaccharide conjugates in vaccine research and development and production.
4.Dissolution testing combined with computer simulation technology to evaluate the bioequivalence of domestic amoxicillin capsule.
Ruixue PAN ; Yuan GAO ; Wanli CHEN ; Yulan LI ; Changqin HU
Acta Pharmaceutica Sinica 2014;49(8):1155-61
Re-evaluation of bioequivalence of generic drugs is one of the key research focus currently. As a means to ensure consistency of the therapeutic effectiveness of drug products, clinical bioequivalence has been widely accepted as a gold standard test. In vitro dissolution testing based on the theory of the BCS is the best alternative to in vivo bioequivalence study. In this article, the conventional dissolution method and flow-through cell method were used to investigate the dissolution profiles of domestic amoxicillin capsules in different dissolution media, and the absorption behavior of the drugs with different release rates (t85% = 15-180 min) in the gastrointestinal tract was predicted by Gastro Plus. The flow-through cell method was thought better to reflect the release characteristics in vivo, and amoxicillin capsules with regard to the release rates up to 45 min (t85% = 45 min) were having a satisfied bioequivalence with the oral solution according to the C(max) and AUC. Although two different dissolution profiles of domestic amoxicillin capsules were found by flow-through cell methods, prediction results revealed that domestic capsules were probably bioequivalent to each other.
5.Comparison of emergency percutaneous coronary intervention and intravenous thrombolysis combined with emergency percutaneous coronary intervent for acute myocardial infarction
Xiangguang YUAN ; Yun YANG ; Zhiming WANG ; Yulan CHEN
Chinese Journal of Primary Medicine and Pharmacy 2005;0(02):-
Objective To evaluate the efficacy and safety of intravenous thrombolysiswith urokinase combined with emergency interventional therapy acute myocardial infarction(AMI).Methods 128 patients with AMI were randomized to thrombolysis plus PCI group and primary PCI group,the patency rate of infarct related artery(IRA) before intervention,the procedural success rate,the incidence of bleeding complications and acute ischemic events during hospitalization and the left ventricular ejection function(LVEF) measured by echocardiography before discharge were compared.Results The IRA patency rate in the thombolysis plus PCI group(65.7%) was significantly higher than that in the primary PCI group(24.3%)(P
6.The vitro anticancer activity analysis of mouse IL-28 recombinant adenovirus vector
Yulan YAN ; Jin ZHANG ; Yang LIU ; Lixue YUAN ; Taofeng ZHU ; Jinxu ZHENG ; Xuefeng BU
Chinese Journal of Microbiology and Immunology 2011;31(7):638-642
Objective To transfect the recombinant mIL-28A adenovirus vector into lung adenocarcinoma cell line LA795 and research its anticancer activity. Methods Transfected the constructed mouse IL-28(mIL-28) recombinant adenovirus vector into LA795 cell line, detected with PCR, immunocytal fluorescence, Tunel, Annexin V and MTT. Results Transfected with rAd-mlL-28A into the LA795 cells, mIL-28A gene expression products mRNA increased obviously, IL-28 expression was detected in cells obviously,apoptosis cell number increased, and the growth of LA795 cells transfected with rAd-mIL-28A were inhibited obviously. Conclusion The recombinant miL-28A adenovirus vector we have constructed, which expresses IL-28 when transfected to lung adenocarcinoma cell line LA795, inhibits growth of carcinoma cell to some extent, and may work by promoting the apoptosis of cancer cells.
7.Application of modified Z-stent in interventional therapy of Budd-Chiari syndrome
Fengji ZHANG ; Xuan ZHANG ; Shuang QIN ; Dejing ZHANG ; Wenyong YUAN ; Guoqiang SU ; Yulan LIANG
Journal of Xinxiang Medical College 2003;20(5):305-307
Objective To evaluate the utility of modified Z-stent in treatment for Budd-Chiarisyndrome (BCS). Methods A retrospective study was used in twelve BCS patients treated in twohospitals. The Doppler examination was carried out in all patients preoperatively, so as to confirm thenature of the lesion and chose correct type of endovascular modified Z-stent. Under DSA monitoring formembranotomy and dilation of the inferior vena cava(IVC), after that, insert the marked modified Z-stent to IVC correctly, and put the non-stent part to hepatic vein orifice. The cavography and hepaticvenography should confirm the position of the Z-stent. All of 12 patients with membranous obstructionof the IVC(MOVC) or segmental obstruction of the IVC (SOVC)were underwent modified Z-stentplacement. Results The IVC pressure (IVCP) before smd after membranotomy (dilation) were 27.33± 4.12cmH2O and 18.67 ± 5.07cmH2O (P<0.01). Comparing with dilation and putting stent group,the IVCP decreased from 18.67 ± 5.07cmH2O to 11.42 ± 2.11 cmH2O ( P < 0.01 ). The modffied Z-stent could avoid hepatic vein orifice getting compression and resist the retraction of IVC throughly infollowing-up period of 2.5 years. Conclusion s The endovascular treatment of BCS with modified Z-stent is more effective and safer to prevent thrombosis. Further study will be required to observe theirlong term effects.
8.Construction and practice of the diabetes management program with the goal of behavior changes
Yulan XU ; Hongdi YUAN ; Qingqing LOU ; Guoxing WANG ; Li WANG ; Linyan YAO
Chinese Journal of Practical Nursing 2010;26(8):27-29
objective To explore the effect of diabetes management program with the goal of behavior changes on behavior change and metabolic index of patients with type 2 diabetes mellitus.Methods By setting up the diabetes management team,establishing personalized management file,carrying out education program,setting the goal of behavior changes and evaluating goal's trace,One-year management was carried out for 56 cases of type 2 diabetic patients.Patients' behavior changes were evaluated after six months management and one year later the metabolic indexes were contrasted.Results After 6 months management,the rate of goal achievement was 96%,the rate of one year behavior stabilization was 92%,the body mass index (BMI),fasting and postprandial blood glucose,glycosylated hemoglobin (HbAlc) one year later were decreased dramatically.Conclusions There is important significance of diabetes management program with the goal of behavior changes for behavior changes and metabolic indexes control in type 2 diabetic patients.
9.Neuronal damage and neurite change in cell model of intractable epilepsy
Yuan WU ; Xiuying LIU ; Xuefeng WANG ; Yulan TANG ; Lu YU ; Jie SU ; Yuejuan WU ; Meigang MA
Chinese Journal of Neurology 2011;44(3):196-199
Objective To establish the cell model of intractable epilepsy and to observe its neuronal damage and morphologic change of neurites.Methods The model was established by exposing hippocampal neurons to Mg2+ -free media for 3 hours on days 10 of culture.Expression of lactic acid dehydrogenase (LDH) in supernatant was measured as an index of neuronal damage.The morphologic change of neurons and neurites was observed by optical microscope and scanning electron microscope (SEM).Results Compared to the control group, level of LDH (U/L) was significantly increased in the model group at different time points (3 hours: 4.26 ± 1.28, 6 hours: 6.56 ±2.34 and 24 hours: 16.67 ±3.57, P <0.05).With time prolonging, release of LDH in the model group was gradually increased (F = 39.316,P <0.05).Under optical microscope, neurons of model group migrated closely to each other and neurite connections appeared to be gradually "reticulated" after Mg2+ -free media treatment for 24 hours; and the "reticulated" neurites connections become more obvious after 72 hours.Under SEM, neuronal membrane was rough and had several small depressions, neurites were interlaced in cluster.Conclusions Neuronal damage and morphologic change of neurites are verified in the cell model of intractable epilepsy.
10.Stable expression of mouse IFN-λ2 in CHO cells and its biological activity analysis
Yulan YAN ; Lixue YUAN ; Yang LIU ; Wenyan CAO ; Xuefeng BU ; Zhigao BU ; Jinxu ZHENG
Chinese Journal of Microbiology and Immunology 2010;30(2):104-109
Objective To express mouse IFN-λ2 stably and study its biological activity. Methods Full-length of mIFN-λ2 cDNA was obtained by using RT-PCR from cells of mouse spleen stimulated by ve-sicular stomatitis virus(VSV) and then subcloned to eukaryotic expressing vector PCAGG-EGFP. The recom-binant was transfected into CHO cells. VSV * GFP-B16 system was used to measure the antivirus activity. The constructed cell line MDBK-Mxp-Luc was used to study the character of Mx1 protein induced by the mIFN-λ2. Results The recombinant pMD18-T-mIFN-λ2 was digested by two kinds of enzyme, Sac I and Xho I, to produce the fragment was of 582 bp, and of which the sequence analysis of sequence shows it was entirely consistent with the nucleotide sequences reported in GenBank. PCAGG-EGFP-mIFN-λ2 eukaryotic expressing vector was constructed successfully and expressed stably in CHO cells, and the mRNA of mIFN-λ2 was verified expressing in CHO-PCAGG-EGFP-mIFN-λ2 cell line by RT-PCR. The antivirus activity of in the supernatant secreted by the CHO-PCAGG-EGFP-mIFN-λ2 cell line was 10~4 AU/ml. The mIFN-λ2 pro-tein can could induce the expression of the antivirus protein Mx1, and the expression of Mx1 protein induced by mIFN-λ2 enhanced with time going, 9 to 12 hours achieved the peak, 24 hours vanished. Conclusion Gene cloning of mIFN-λ2 was successful. The eukaryotic expressing vector of mIFN-λ2 was constructed suc-cessfully and expressed stably in CHO cells, and its product has obvious antivirus activity in vitro. And the antivirus activity of the product was closely correlated with inducing expression of antivirus protein Mx1.