1.A brief review on the progress of mouse models of liver cancer
Shun LI ; Lixiang CHEN ; Xiuhua PENG ; Jiangming ZHU ; Xiaohui ZHOU
Acta Laboratorium Animalis Scientia Sinica 2016;24(2):213-216
Liver cancer remains one of the leading cause of cancer death in the world.Animal models, especially mouse models, are important tools for studying the biological characteristics, pathogenesis, new drug screening and therapy of liver cancer.Up to now, although the development of various animal models accelerates the research of liver cancer, all the existing models have their own disadvantages.Lacking of economical and applicable animal models that can mimic the human liver cancer seriously restrict the further study of liver cancer.With the development of genetically modified technologies, it provides a fast, easy and reliable method to establish liver cancer models.In this review, we describe the different types of mouse models used in liver cancer research, with emphasis on genetically engineered mice used in this field, which may open an avenue for functional cancer genomics and generation of liver cancer models by using gene editing technologies.
2.Establishment of a flow cytometry-based test for the detection of pyroptosis of BMDM
Xue LIU ; Lixiang CHEN ; Shun LI ; Xiaohui ZHOU
Chinese Journal of Microbiology and Immunology 2016;36(2):149-154
Objective To establish a flow cytometry-based test for the detection of pyroptosis of murine bone marrow-derived macrophages (BMDM). Methods Bone marrow cells were isolated from wild type (WT) C57BL/ 6 mice and/ or caspase-1-/ - C57BL/ 6 mice and then stimulated with macrophage colony-stimulating factor (M-CSF) to differentiate into murine BMDM. PBS, LPS and LPS+adenosine triphosphate (ATP) were respectively used to stimulate the BMDM. Western blot assay was performed to detect the cleav-age of IL-1β and caspase-1. The levels of IL-1β in the supernatants of cell culture were measured by ELISA. Lactate dehydrogenase (LDH) released in the culture media was detected by using LDH kit. The pyroptosis of murine BMDM was detected by using flow cytometry analysis after double-staining with TMR red+caspase-1, AnnexinⅤ+caspase-1 and propidium iodide (PI)+caspase-1. Results IL-1β was detected in the culture medium of BMDM treated with LPS+ATP and the cleavage of IL-1β and caspase-1 was confirmed by Western blot assay, which indicated that the NLRP3 inflammasome was activated by LPS+ATP treatment. Compared with the caspase-1-/ - mice group, higher levels of LDH were detected in the culture medium of BMDM isolated form the WT mice. Results of the flow cytometry analysis after staining BMDM with caspase-1 plus AnnexinⅤ or PI showed that more cells undergoing pyroptosis were detected in the LPS+ATP treat-ment group than that in LPS or PBS treatment group, which were consistent with the results of the reported flow cytometry with caspase-1+TMR red staining. Conclusion The flow cytometry-based test with double-staining of caspase-1 plus AnnexinⅤ or PI could be used for the detection of pyroptosis of murine BMDM.
3.Effect of quercetin on proliferation of rat glioma C6 cells
Lixiang ZHOU ; Yinan LUO ; Shuanglin FU ; Pengfei GE ; Hanting ZHUANG
Journal of Jilin University(Medicine Edition) 2006;0(02):-
Objective To study the effects of quercetin(QUE) on proliferation of rat glioma C6 cell line in vitro.Methods The cells were divided into 5 treatment groups(10,25,50,75 and 100 ?mol?L~(-1) QUE),blank control and menstruum control group.The rat C6 cells were cultivated to 1?10~6?mL~(-1) in the RPMI 1640 medium,then added into 96 holes board with various doses of QUE by 3 holes per group,and MTT assay was used to observe the proliferation of the cells treated for 24,48 and 72 h.The change of cell cycle was also observed by flow cytometry(FCM) after the cells were treated with 50 and 100 ?mol?L~(-1) QUE for 48 h.The changes of the protein P53 and Bcl-2 of C6 cells treated with 50 ?mol?L~(-1) QUE for 48 h were detected by immunocytochemical methods.(Results With) the augmentation of QUE and the extension of the treated time,the C6 cell growth was inhibited,the A values decreased and the cell number in G_0/G_l phase was increased,the cell numbers in S and G_2/M phases were cut down,and the decreased expression of Bcl-2 protein and the increased expression of P53 protein were also observed after treatment with QUE.Conclusion Inhibitory effect of QUE on C6 cell line is proved to be dependent on the treated time of the drug and the dose of QUE,and the induced apoptosis of C6 cells is implemented by the means of up-regulation of P53 protein expression and down-regulation of Bcl-2 protein expression.
4.Value of postoperative indocyanine green retention rate at 15 minutes combined with standard remnant liver volume in predicting liver dysfunction after hepatectomy
Lixiang MEI ; Dong WANG ; Huanni LI ; Zengbo LI ; Ledu ZHOU
Journal of Central South University(Medical Sciences) 2017;42(6):635-640
Objective:To investigate the value of indocyanine green retention rate at 15 minutes (ICG R15) on postoperative day 3 combined with standard remnant liver volume (SRLV) in predicting the occurrence of liver dysfunction after hepatectomyin hepatocellular carcinoma (HCC).Methods:The clinical data of 61 HCC patients undergone hepatectomy in Xiangya Hospital of Central South University from January 2015 to February 2016 were collected and analyzed.The patients were divided into 2 groups:a normal liver function group (n=40) and a liver dysfunction group (n=21).Univariate analysis was used to evaluate the risk factors for postoperative liver dysfunction.Logistic regression was used to assess the independent risk factors for postoperative liver dysfunction,and the regression equation between independent risk factors and postoperative liver dysfunction was established,The receiver operating characteristic (ROC) curve was used to examine the regression equation and compare the value difference in predicting postoperative liver dysfunction between single and combined independent risk factors.Results:Postoperative liver dysfunction occurred in 21 of the 61 patients,with an incidence rate at 34.4%.There was no significant difference in the time of operation,time of hepatic portal occlusion,volume of tumor and volume of resected liver between the 2 groups (all P>0.05),but there were significant differences in the ICG R15 on postoperative day 3,intraoperative blood loss and SRLV between the 2 groups (all P<0.05).The ICG R15 on postoperative day 3,intraoperative blood loss,SRLV were the risk factors for postoperative liver dysfunction.Logistic regression analysis showed ICG R15 on postoperative day 3 and SRLV were the independent risk factors for postoperative liver dysfunction,and the regression equation between independent risk factors and postoperative liver dysfunction was as follows:logit(P)=1.277+0.140×ICG R15 on postoperative day 3-5.125×SRLV.The area under the ROC curve ofICG R15 on postoperative day 3 combined with SRLV was more than that of single ICG R15 and single SRLV.Conclusion:ICG R15 on postoperative day 3 and SRLV are the independent risk factors for postoperative liver dysfunction.The regression equation,which is established by combination of ICG R15 with SRLV,can predict the occurrence of postoperative liver dysfunction.The accuracy of ICG R15 on postoperative day 3 combined with SRLV is better than that of single ICG R15 or single SRLV.
5.Effects of combined use of heparinoid,aspirin,and losartan on the experimental vascular intima hyperplasia in rats
Liyan PANG ; Xubin ZHOU ; Lixiang WANG ; Qingfen ZHU ; Hongyan XU
Chinese Pharmacological Bulletin 1986;0(05):-
AIM To observe the effects of the combination of heparinoid, aspirin and losartan on the experimental vascular intima hyperplasia induced by squeezing carotid artery in rats. METHODS Rats except the normal control group were under operation after two days oral administration once a day. At the second hour after injury, blood coagulation time(CT), time of arterial thrombus formation by electrical stimulus(ATFTES) and the level of TXB_2 in plasma were measured. At the fourteenth day after the surgery, the following indexes were examined: intima/media ratio, intima area ratio, PDGF-B and PCNA immunohistochemistry in carotid artery. RESULTS CT, TATFES increased and the level of TXB_2 decreased in the groups of Hep, Asp and Hep+Asp. The PDGF-B content, the positive ratio of PCNA and the inti- ma/media ratio also decreased in the groups of Hep, Asp, Los, Hep+Asp, Hep+Los and Asp+Los. Compared with single drug or two drugs combination, PDGF-B, positive ratio of PCNA, intima/media ratio and intima area ratio decreased significantly in the group of three drugs combination( P
6.CT and MRI manifestations of intracranial melanomas
Pengfei GE ; Yinan LUO ; Shuanglin FU ; Haiyan HUANG ; Lixiang ZHOU
Chinese Journal of Radiology 1994;0(06):-
Objective To explore the CT and MR imaging characteristics of intracranial melanomas. Methods CT and MRI characteristics in five patients admitted to our hospital from June 1993 to June 2000 and diagnosed as intracranial melanomas were retrospectively analyzed. Results There were two cases of primary melanoma and three cases of secondary melanoma. All the cases were examined by CT. The lesions presented as high density in 4 cases, and low density in only 1 case. Four cases were examined by MRI. Short T 1 and short T 2 signals were found in 3 cases, and slightly long T 1 and short T 2 signal was found in 1 case. Conclusion There are some special characteristics of melanomas on the MR imaging, which are helpful to ensure the diagnosis and distinguish the primary melanomas from secondary melanomas.
7.The establishment of the uPA inducible expression system
Lixiang CHEN ; Xiaojing ZHOU ; Wenwen LIU ; Wenjiang ZHOU ; Xiaonan REN ; Shiyan YU ; Xiaohui ZHOU
Chinese Journal of Comparative Medicine 2015;(1):1-8
Objective To establish uPA inducible expression system using recombinant retroviral system for the further construction of inducible uPA-SCID animal model .Methods The Inducible expression system need to construct two plasmids:pLNHXO1O2-Alb-GLUC-FMN2A -rtTA and pLNHXO5O6-TRE2-uPA-IRES-ZsGreen respectively. Both plasmids were based on retroviral vector pLNHX , Albumin promoter gene ( Alb) and rtTA gene or uPA gene and ZsGreen were obtained by PCR reaction and inserted into pLNHX .The Gaussia enzyme fluorescent element ( GLUC) was used to monitor rtTA expression in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA, and the ZsGreen for uPA expression monitoring in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen.The correct constructed plasmids were transfected into packaging cell line GP 2-293 to gain recombinant viral particles .NIH/3T3 cells were infected with these viral particles and selected with G 418.Gene expression in the surviving cells was confirmed by the PCR method .Results The recombinant retroviral vectors harbouring target genes were successfully cloned .The rtTA gene in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA was expressed, and uPA can be induced to express in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen by doxycycline (Dox) when the plasmid transfected into the HepG-Tet-on cell.The constructed recombinant two retroviral vectors were transfected into GP 2-293 packaging cells respectively to gain infectious viral particles .Then,NIH/3T3 cells were infected with these viral particles and single-cell clones which stably expressed the transgenes were successfully established .Conclusion This study primarily established uPA inducible expression system , it laid a foundation for the murine model of inducible liver damage , and provided a novel technical platform for further building the liver humanised murine models for viral hepatitis studying .
8.MRI study of cerebral lesions and analysis of laboratory examination and prognosis of patients with neuropsychiatric systemic lupus erythematosus
Guangyu ZHOU ; Lixiang ZHOU ; Ye LI ; Ling JIN ; Wenlong ZHANG ; Min YIN
Chinese Journal of Rheumatology 2012;16(10):697-700
Objective To investigate the magnetic resonance imaging (MRI) characteristics of cerebral lesions,laboratory data and prognosis of patients with neuropsychiatric systemic lupus erythematosus (NPSLE) and evaluate the correlation of MRI findings,autoantibodies and prognosis of NPSLE.Methods Thirty-one patients with NPSLE admitted during Jan 1998 to Oct 2011 were enrolled into this study,including 2 males and 29 females.MRI characteristics of cerebral lesions,laboratory data and clinical outcomes were retrospectively analyzed.x2-test and Spearman's correlation were used for statistical analysis.Results Cranial MRI scans on admission were abnormal in 27 (87%) of the 31 patients,among which 17 cases showed diffuse manifestations,9 cases showed focal lesions in brain and 1 case showed brain atrophy.The percentage of cases with focal lesions in the dead group (86%) was significantly higher than that of the survival group (13%) (P<0.01),while the percentage of diffuse lesion cases was significantly lower in dead group (P<0.01).Although the mean lesion numbers in the dead group were fewer compared with the survival group,lesion numbers in brainstem (18%) (P<0.01) or basal ganglia (29%) (P<0.05) were much more in the dead group.The positive rate of anti-ribosomal P protein antibodies shown in the dead group (43%) was markedly higher than that in the survival group (29%) (P<0.05).The positive rates of ACL in cases with diffuse lesions and cases with focal lesion were higher compared with cases showing normal MR images (P<0.01).Furthermore,the positive rate of anticardiolipin antibodies (ACL) in cases with focal lesions (56%) were higher than cases with diffuse lesion (35%) (P<0.05).There was no correlation between the seven types of autoantibodies and lesion locations such as basal ganglia,subcortical white matter,anterior and posterior horn of lateral ventricle,semiovale center,cerebral cortex,brainstem and cerebellum.Conclusion MRI findings and autoantibodies are correlated in patients with NPSLE and both of them may be used as prognostic markers for NPSLE.Focal lesions,location of brainstem and basal ganglia on cranial MR images,positive anti-ribosomal P protein antibody may predict an unfavorable clinical outcome of NPSLE; however,the correlation between ACL and prognosis could not be determined.
9.Correlation of M-type phospholipase A2 receptor genetic polymorphism with idiopathic membranous nephropathy
Guangyu ZHOU ; Yanxia SUN ; Lixiang ZHOU ; Jing YU ; Ying GUO ; Min YIN
Chinese Journal of Nephrology 2013;(1):1-5
Objective To investigate the correlation of M-type phospholipase A2 receptor (PLA2R) genetic polymorphism in two single nucleotide polymorphisms (SNPs) with idiopathic membranous nephropathy (IMN) of Chinese Han population in Northeast China.Methods A total of 327 individuals were enrolled in the study including 95 adult patients with biopsy-proved IMN (IMN group) followed up for (25.4±11.6) months and 232 healthy people identified by healthy examination in China-Japan Union Hospital of Jilin University (HC group).Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotype and allele frequency of rs35771982 and rs3828323 site in PLA2R gene.The x2 test was performed to compare the distribution difference of allelic frequency and genotype frequency of the two sites in PLA2R gene between two groups.Unconditional Logistic regression analysis was used to determine the risk factor of IMN.Results IMN and HC group were matched in male predominance and body mass index (BMI).Patients with IMN were older than the healthy controls and had higher Scr,serum total cholesterol (TC),24-hour urine protein level and lower serum albumin (Alb) level,lower estimated glomerular filtration rate (eGFR) than the healthy controls (all P < 0.01).The CC genotype frequency and the C allele frequency at SNP rs35771982 site of PLA2R gene in IMN group were significantly higher than those in HC group (x2=13.658,P=0.001;x2=15.315,P=9.10× 10-5),whereas there was no distribution difference of genotype and allele frequency at rs3828323 site between two groups (x2=2.844,P =0.241; x2 =2.959,P=0.085).The CC genotype at rs35771982 site in patients with IMN was not related to,age,gender,BMI,blood pressure and several laboratory indexes such as Alb,TC,Scr,eGFR and 24-hour urine protein level (all P > 0.05).Unconditional Logistic regression analysis revealed that the genotype at rs35771982,age,TC,Scr and eGFR were correlated with IMN occurrence.The CC genotype at rs35771982 was the risk factor of IMN (OR =4.408,95%CI 1.488-13.058).Conclusions The CC genotype and C allele at rs35771982 site in PLA2R may be associated with the susceptibility to IMN,whereas the correlation between gene polymorphism at rs3828323 site and IMN is not demonstrated.The CC genotype at rs35771982 is the independent risk factor of IMN in Chinese Han population in Northeast China.
10.Chinese consensus of cardiopulmonary resuscitation guides prevention, treatment and rescue of cardiac arrest in pregnancy
Yuhong MI ; Feihu ZHOU ; Lixiang WANG ; Yinping LI ; Qingyi MENG ; Jun ZHANG ; Xinyu ZHANG
Chinese Critical Care Medicine 2023;35(1):5-22
Pregnant women are a group of people in a special period, once sudden cardiac arrest (CA) occurs, it will threaten the life of both mother and child. It has become a great challenge for hospital, doctors and nurses to minimize maternal mortality during pregnancy. All the efforts should ensure the safety of both mother and child throughout the perinatal period. Because difference of the cardiopulmonary resuscitation strategies for common CA patients of the same age, the resuscitation strategies for CA patients during pregnancy need consider the patient's gestational age and fetal condition. Different resuscitation techniques, such as manual left uterine displacement (MLUD), will involve perimortem cesarean delivery (PMCD). At the same time, drugs should be reasonably used for different causes of CA during pregnancy, such as hypoxemia, hypovolemia, hyperkalemia or hypokalemia and other electrolyte disorders and hypothermia in 4Hs, as well as thrombosis, pericardial tamponade, tension pneumothorax and toxicosis in 4Ts. In view of the fact that many causes of CA in pregnancy are preventable, it is more necessary to introduce guidelines for CA in pregnancy in line with our national conditions for clinical guidance. This paper systematically reviewed the pathophysiological characteristics of CA during pregnancy, the high-risk factors of CA during pregnancy, and identified the correct resuscitation methods and prevention and treatment strategies of CA during pregnancy.