1.MicroRNAs and neuronal apoptosis after cerebral ischemia
Lu XIAO ; Xianxian ZHANG ; Yingzhu CHEN
International Journal of Cerebrovascular Diseases 2012;20(9):712-716
MicroRNAs (miRNAs) are a class of highly conserved non-coding small RNA molecules.They regulate gene expression by inhibiting transcription or translation.Ischemic stroke is one of the major diseases of resulting in death and disability worldwide.The final outcome of its pathological process is neuronal death.Neuronal apoptosis is mainly seen in ischemic penumbra,and saving the neurons in penumbra are the key to the treatment of ischemic stroke.This article reviews the roles of miRNAs in the neuronal apoptosis during cerebral ischemia.
2.Applied research of Doppler tissue imaging technique in evaluating accessory pathways in pre-excitation syndrome
Yu WANG ; Yingzhu LU ; Tao GUO
Chinese Journal of Ultrasonography 1997;0(06):-
Objective To evaluate the accuracy,practical value and methods of Doppler tissue imaging(DTI) in localizing the ventricular emergence site of accessory atrioventricular pathways.Methods Forty patients with pre-excitation syndrome were examined by DTI before intracardiac electrophysiologic studies(EP) and radiofrequency catheter ablation(RFCA).The pre-excitation zone was determined as a zone of maximum acceleration with “DTI acceleration mode” at the time of the delta wave or before the onset of the QRS complex.At the same time,the contraction velocity was measured from subendocardium of corresponding myocardial segment by “pulsed wave Doppler imaging”.Results Assuming the results of EP and RFCA as a gold standard,the correct localization by DTI was 80% but was only 47.5% and 52.5% by eletrocardiographic algorithms( P
3.Anticoagulant therapy in prevention and treatment of ischemic stroke
Xianxian ZHANG ; Lu XIAO ; Gang LIU ; Yingzhu CHEN
International Journal of Cerebrovascular Diseases 2011;19(11):829-835
The activation of coagulation system,especially in the occurrence and development of cardiogenic cerebral embolism,plays an important role.As one of the major preventive measures in ischemic stroke,the anticoagulant therapy is getting more and more attention.At the same time,the studies of anticoagulant drugs aiming to the intervention of different links in coagulation pathway have also made significant progress.
4.Prokaryotic expression and characterization of receptor binding domain protein of the Middle East respiratory syndrome coronavirus
Shuai LU ; Jiaming LAN ; Yingzhu CHEN ; Jianfang ZHOU ; Kun QIN ; Yongliang LOU ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2016;36(2):98-102
Objective To express the receptor binding domain (RBD) protein of the Middle East respiratory syndrome coronavirus (MERS-CoV) and to characterize the antigenicity of the purified recombi-nant protein. Methods The codon-optimized gene encoding the RBD protein of MERS-CoV was synthesized and then cloned into the pET30a ( +) vector to construct the recombinant expression plasmid. The trans-formed E. coli BL21 (DE3) strains carrying expression plasmid were induced by IPTG under different condi-tions. The expressed products were purified by using nickel affinity chromatography and further analyzed by SDS-PAGE and Western blot assay. Indirect ELISA was performed to analyze the antigenicity and specificity of RBD proteins expressed in prokaryotic expression systems in human serological test. Results The recom-binant RBD proteins were mainly expressed as conclusion body in an optimal induction condition of 37℃ and 0. 5 mmol/ L IPTG for 4 h. The high purified recombinant RBD proteins were obtained through denaturation and renaturation with a relative molecular mass of about 29×103 . Results of the Western blot assay showed that the recombinant RBD proteins could have specific reaction with the serum samples collected form mice with MERS-CoV infection. Indirect ELISA revealed that the RBD proteins expressed in the prokaryotic ex-pression system showed better sensitivity and specificity in the detection of antibodies against MERS-CoV in human serum samples. Conclusion This study reported the prokaryotic expression and purification of RBD protein of MERS-CoV for the first time, which might pave the way for further investigation on immunological detection of MERS-CoV and development of vaccines against MERS-CoV infection.
5.Effects of 1-3-n-Butylphthalide on the blood-brain barrier following whole brain irradiation in rats
Yingzhu CHEN ; Xianxian ZHANG ; Lu XIAO ; Yanhong QI ; Pu YANG ; Ye TIAN ; Shirao BAO
Chinese Journal of Radiation Oncology 2012;21(4):392-395
ObjectiveTo investigate the effects of 1-3-n-Butylphthalide on the blood-brain barrier (BBB) following whole brain irradiation in rats.Methods144 male Sprague Dawley rats were randomly divided into sham-irradiation group,irradiation group,1-3-n-Butylphthalide group,and irradiation plus 1-3-n-Butylphthalide group.Whole-brain irradiation was given as a single-dose of 10 Gy using 4 MV X-ray.The rats were injected intraperitoneally with 1-3-n-Butylphthalide at 0.3 mg/kg,1.0 mg/kg,3.0 mg/kg once per day.The changes of the BBB were assessed by Evans blue (EB) assay.The expression of vascular endothelial growth factor (VEGF) in the brain tissue was determined by immunohistochemistry. The circulating endothelial cells (CECs) isolated from right ventricular blood were counted.MRI was evaluated with the T1-weighted images,T2-weighted images and MRI enhancement images induced by Gd-DTPA.The data were compared among the groups through Student-Newman-Keuls test.ResultsCompared with the sham-irradiation group,the EB content,the expression of VEGF in the brain tissue and the CECs were significantly increased in the irradiation group (2.81∶ 7.82,P =0.002;5.83∶ 10.26,P=0.003;3.16∶6.14,P =0.002).The signal intensity of T1-weighted images was significantly decreased while T2-weighted images and the enhancement rate significantly increased in the irradiation group (P =0.004 -0.018 ).Compared with irradiation group,the EB content,the expression of VEGF and the CECs were decreased significantly in the irradiation plus 1-3-n-Butylphthalide group ( 7.80∶ 3.86,P =0.007 ; 10.83 ∶ 5.26,P =0.008 ;6.36∶ 3.64,P =0.009 ).However,the changes in the MRI were significantly attenuated ( P =0.008-0.026,and 0.006 -0.038,respectively).Conclusions Following whole brain irradiation,1-3-n-Butylphthalide can decrease the permeability of the BBB in rats via decreasing VEGF expression and decreasing the CECs.
6.Protective effect of DL-3-n-Butylphthalide on radiation injury of rat brain tissue
Yingzhu CHEN ; Xianxian ZHANG ; Lu XIAO ; Yanhong QI ; Pu YANG ; Jinzhong HUANG ; Shiyao BAO
Chinese Journal of Radiological Medicine and Protection 2012;32(3):255-258
Objective To investigate the protective effect and its mechanism of DL-3-n-Butylphthalide on the brain damage in rats following whole brain irradiation.Methods A total of 120 male Sprague Dawley rats were randomly divided into sham-irradiation group,irradiatien group and DL-3-n-Butylphthalide group.The model of whole-brain irradiatien was established by exposuring rat brain to 4 MeV X-rays with a single-dose of 10 Gy.The rats were intraperitoneally injected with DL-3-n-Butylphthalide at the dosages of 0.3,1.0,and 3.0 mg/kg once a day.The contents of malondialdchyde and super oxide dismutase activity were measured,while the expressions of apoptosis-associated genes and the ultrastructural changes in hippocampus were examined by immunohistnchemisty staining and electron microscope,respectively.Results After irradiation,the content of malondialdehyde and the expression of apoptosis gene bax in rat brain tissue increased while the activity of super oxide dismutase(SOD) and the expression of anti-apoptosis gene bcl-2 decreased.Apoptosis was also observed in the neurons of hippocampus CA1.Compared with irradiation group,the content of malondialdehyde and the expression of bax gene in the DL-3-n-Butylphthalide group wen significantly reduced ( t =-3.89--1.96,2.72-3.48,P < 0.05 ),while the activity of SOD and bcl-2 gene were significantly elevated ( t =2.94-3.76,-3.18--2.08,P < 0.05),and the injury degree of neuron structure in the DL-3-n-Butylphthalide group was slighter than that in the irradiation group.Conclusions DL-3-n-Butylphthalide executes protective effects in a dose-dependent manner againest the radiation injury in rats brain by reducing the induction of malondialdehyde,raising the activity of SOD and inhibiting the generation of apoptosis.
7.Changes of plasma fibrinogen level among acute ischemic stroke subtypes according to TOAST criteria and effects of Songling Xuemaikang.
Degang YANG ; Nianfang LU ; Xiaojie LIU ; Gang LIU ; Yao XU ; Jinzhong HUANG ; Yingzhu CHEN
China Journal of Chinese Materia Medica 2010;35(10):1339-1341
OBJECTIVETo investigate the changes of plasma fibrinogen level among acute ischemic stroke (ACI) subtypes according to Trial of Org10172 in Acute Stroke Treatment (TOAST) criteria and effects of Songling Xuemaikang.
METHODThe 160 patients with acute ischemic stroke were divided into two groups randomly: treatment group 85 cases (Songling Xuemaikang + Shuxuetong + Aspirin enterie coated tablets), control group 75 cases (Shuxuetong + Aspirin enterie ccoated tablets). The plasma fibrinogen was detected before and after treatment.
RESULTCompared with OC subtype, Fbg was higher in LAA, CE and SAO subtypes (P < 0.05). Compared with UE subtype, Fbg was higher in LAA, CE and SAO subtypes (P < 0.05). There was a significantly difference between LAA and SAO (P < 0.05). In LAA, SAO, CE of treatment group,the Fbg level were lowered significantly at the 15th day compared with pretherapy (P < 0.05). There was a significantly difference of Fbg between treatment group and control group In LAA, SAO and CE subtypes at the 15th day (P < 0.05).
CONCLUSIONFbg produces a marked effect at the pathomechanism of LAA, SAO and CE subtypes. Songling Xuemaikang can depress the plasma fibrinogen level of ACI, and be an effective adjunctive therapy on ACI.
Aged ; Aged, 80 and over ; Aspirin ; therapeutic use ; Brain Ischemia ; drug therapy ; metabolism ; pathology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fibrinogen ; metabolism ; Humans ; Male ; Middle Aged ; Stroke ; drug therapy ; metabolism ; pathology ; Treatment Outcome
8. Generation and preliminary characterization of monoclonal antibodies against neuraminidase of H7N9 subtype influenza A virus
Yueyang YU ; Yiran XIE ; Yingzhu CHEN ; Ying SUN ; Heng ZHANG ; Jian LU ; Liqi LIU ; Dayan WANG ; Yuelong SHU ; Kun QIN ; Jianfang ZHOU
Chinese Journal of Experimental and Clinical Virology 2017;31(1):66-70
Objective:
To develop the monoclonal antibody (mAb) against neuraminidase of H7N9 subtype influenza A virus and identify its biological function.
Methods:
Female 8 week-old BALB/c mice were immunized and the splenocytes of the mice were fused with Sp2/0 myeloma cells. Indirect ELISA was used to screen hybridoma and the positive clones were subject to be subcloned. Positive clones were identified and the monoclonal antibodies(mAbs) were obtained by purifying the ascetic fluid of mice injected with the hybridoma. The NA-binding as well as neuraminidase-inhibition activity of these mAbs were determined.
Results:
Three mAbs against neuraminidase of H7N9 subtype influenza A virus, 1G8, 3C4 and 4E8, were obtained. They demonstrated different epitop-recognizing. 3C4 and 4E8 exhibited neuraminidase inhibitory activity, with a IC50 of 1.45 μg/ml and 8.65 μg/ml, respectively.
Conclusions
The results suggested that mAbs specific to neuraminidase of H7N9 subtype influenza A virus were developed, providing an useful tool in control and preventing the novel H7N9 influenza A virus.
9.Rapid screening of SARS-CoV-2 inhibitors via ratiometric fluorescence of RBD-ACE2 complexes in living cells by competitive binding.
Lu MIAO ; Wei ZHOU ; Chunyu YAN ; Yuebin ZHANG ; Qinglong QIAO ; Xuelian ZHOU ; Yingzhu CHEN ; Guangying WANG ; Zhendong GUO ; Jun LIU ; Hailong PIAO ; Xia PAN ; Mengxue YAN ; Weijie ZHAO ; Guohui LI ; Yueqing LI ; Zhaochao XU
Acta Pharmaceutica Sinica B 2022;12(9):3739-3742