1.Expression of Toll-like receptor 2 in glioma and its correlation
Yuhua WANG ; Qikai SUN ; Xin GENG ; Zefu LI ; Linxin MA
Chinese Journal of Postgraduates of Medicine 2017;40(7):645-649
Objective To investigate the expression of Toll like receptor 2 (TLR2) in glioma and its relationship with the malignant biological behavior of glioma, so as to provide a new therapeutic target for glioma immunotherapy. Methods The tumor tissues of 90 glioma patients undergoing surgical excision were collected, of which WHO gradingⅠ-Ⅱgrade 39 cases,Ⅲgrade 24 cases,Ⅳgrade 24 cases. In addition, the normal brain tissues of 12 patients undergoing routine intracranial decompression were selected. The human glioma cell lines U87-MG, U251-MG and human astrocyte cell line HA were cultured. The expression levels of TLR2 mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western Blot test. The correlation between TLR2 protein and different clinical pathological parameters was analyzed. Results The expression levels of TLR2 mRNA and protein in the glioma tissuesⅠ-Ⅱgrade,Ⅲgrade andⅣgrade were significantly higher than those in normal brain tissues (0.27 ± 0.09, 0.57 ± 0.12 and 0.96 ± 0.18 vs. 0.11 ± 0.05; 0.31 ± 0.05, 0.44 ± 0.05 and 0.71 ± 0.09 vs. 0.02 ± 0.01), there were statistical differences (P<0.05). In different grades of glioma tissues, there were statistical differences in the expression levels of TLR2 mRNA and protein (F = 205.9 and 194.9, P<0.05). The expression levels of TLR2 mRNA and protein in human malignant glioma cell lines U87-MG and U251-MG were significantly higher than those in human astrocyte cell line HA (1.000 ± 0.100 and 0.356 ± 0.060 vs. 0.245 ± 0.030, 0.720 ± 0.100 and 1.800 ± 0.150 vs. 0.004 ± 0.000), there were statistical differences (P<0.05). The expression of TLR2 protein was independent of age, sex and location(P>0.05), but was related to tumor diameter and WHO grading (P < 0.01). Conclusions TLR2 in different grade glioma tissues and glioma cell lines are expressed, and its expression level is associated with the malignant degree of glioma; TLR2 protein not only can be used as a biomarker of gliomas and prognosis, but also provide a new target for the treatment of glioma.
2.Decreased number and impaired function of endothelial progenitor cells in gld.apoE-/-C57BL/6 mice
Shiying WANG ; Linyu GENG ; Xia LI ; Xuebing FENG ; Lingyun SUN
Chinese Journal of Rheumatology 2013;17(8):513-517,后插1
Objective To detect the changes of number and function of endothelial progenitor cells (EPCs) in gld.apoE-/-C57BL/6 mice,and to investigate whether premature atherosclerosis of gld.apoE-/-C57BL/6 mice was mediated by the dysfunction of these EPCs.Methods EPCs were isolated from peripheral blood and bone marrow of four types of C57BL/6 female mice:gld+/+apoE+/+,gld,apoE-/-and gld.apoE-/-.The percentage of EPCs was analyzed by FACS as CD11b-Sca-1+CD309+ cells.The attached cells were stained with DiI labeled acetylated low-density lipoprotein (DiI-ac-LDL) and FITC-labeled Ulex Europaeus agglutinin 1 (FITC-UEA-1) double staining to determine their phagocytic function.The adherent function of EPCs was determined by calculating the number of re-cultured EPCs.The capacity of EPCs to form the cavity structure was detected by calculating the number of the formed vascular-like structure.Results The percentage of circulating EPCs was significantly decreased in the gld.apoE-/-group (0.012±0.002)% compared to the gld+/+ apoE+/+ group [(0.039±0.005)%,P<0.01],the gld group [(0.025±0.001)%,P<0.05],and the apoE-/-group [(0.028±0.002)%,P<0.01].The percentage of bone marrow derived EPCs was decreased in the gld.apoE-/-group (0.12±0.01)% compared to the gld+/+apoE+/+ group [(0.42±0.05)%,P<0.05].The percentage of DiI-acLDL and FITC-UEA-1 double positive cells was decreased in the gld.apoE-/-group [(59.2±2.1)%] compared to the gld+/+apoE+/+ group [(87.5±3.0)%,P<0.01],and the gld group [(84.2±6.0)%,P<0.01] ; the adherent function of EPCs was impaired in the gld.apoE-/-group [(50.0±5.3)%] compared to the gld+/+ apoE+/+ group [(86.0±7.3)%,P<0.01],the gld group [(73.0±1.0)%,P<0.01],and the apoE-/-group [(65.0±4.0)%,P<0.05] respectively.The capacity of EPCs to form the cavity structure was decreased in the gld.apoE-/-group (4.0±0.3) compared to the gld+/+apoE+/+ group (12.0±1.4,P<0.01).Conclusion The number of EPCs is decreased in the gld apoE-/-C57BL/6 mice,the adhesion,phagocytosis and angiogenic function of EPCs in the bone marrow are impaired,which may be one of the causes of lupus with atherosclerosis.
3.Prevention and treatment of acute graft-versus-host disease in mice by bortezomib
Haiying SUN ; Huayun GENG ; Lingyu ZENG ; Zhenyu LI ; Kailin XU
Chinese Journal of Organ Transplantation 2011;32(1):11-15
Objective To observe the effect of bortezomib on acute graft-versus-host disease (aGVHD) in an aGVHD model of mice and investigate the related mechanism. Methods Male C57BL/6( H-2Kb)mice were used as donors and female Balb/c (H-2Kd) mice used as recipients. Balb/c mice received total body irradiation (TBI) by 7.0 Gy X-radiation, and randomly divided into five groups. normal (group A), TBI (group B), TBI + bortezomib (group C), TBI + bone marrow cells (BMC) + spleen cells (SC) (group D) and TBI + bortezomib + BMC + SC (group E). The physical signs and the pathological damage of aGVHD, mean survival time, and chimerism were observed in recipients. The NF-κB p65 levels in nuclei of the liver and small intestine tissues of groups A,B and C were analyzed by Western blot. Results ( 1 ) The clinical aGVHD score in group D was (7.37±0. 32), significantly higher than in group E (5.85 ± 0.40) (P<0. 05). Histopathology of the gut, liver and skin illuminated that the Ⅲ-Ⅳ degree GVHD occurred in group D. The occurrence of aGVHD in group E was later than in group D. The symptoms and the pathological damage of aGVHD in group E were milder than in group D. The average survival time in group E was significantly longer than that in group D (P<0.05). The percentage of donor-derived cells in recipient mice was above 90% at day 12 after transplantation; (2) NF-κB p65 levels in nuclei of the liver and small intestine tissues in group B was significantly higher than in group C on the day 1,3 and 5 (P<0. 05). Conclusion Bortezomib can inhibit the activation and expression of NF-κB,which may be the underlying mechanism for it to relieve aGVHD.
4.Changes of cholinergic neurons in the hippocampus of vascular dementia rats after neural stem cell transplantation
Yuhua SUN ; Lijiao GENG ; Jingya ZHAO ; Weiya HE ; Baoping LI
Chinese Journal of Tissue Engineering Research 2015;(50):8126-8131
each group were sacrificed, respectively. Distributions of BrdU positive cels and ChAT positive cels were detected by S-P immunohistochemical method. The learning and memory abilities of rats were detected by Morris water maze system.
RESULTS AND CONCLUSION:BrdU positive cels were mainly distributed in the cortex and hippocampus, especialy around the blood vessels, and there was the presence of focal aggregation. A smal amount of BrdU positive cels were observed in the basal ganglia and thalamus as wel as in the ependyma. BrdU positive cels were counted at different time after operation. The number of BrdU positive cels decreased with time, and only a smal number of BrdU positive cels were observed at 60 days after transplantation. The number of ChAT positive cels at different time after transplantation was ranked as folows: neural stem cel transplantation group > model group > sham operated group (P < 0.05). Compared with the model group, the time for searching the platform was significantly lower in the neural stem transplantation group and sham operated group, but the number of crossing the platform was significantly higher in the neural stem cel transplantation group and sham operated group (P < 0.05). The results show that neural stem cels could be transplanted into the rats with vascular dementia, and the cels could survive and migrate in the brain of rats and significantly improve the learning and memory ability. This mechanism may be related to the differentiation and growth of cholinergic neurons in the hippocampus.
5.Effect of ropivacaine administered intrathecally on pulmonary function after caesarean section
Wenhui LI ; Guiqi GENG ; Xingfeng SUN ; Shaoqiang HUANG
Chinese Journal of Anesthesiology 2012;32(2):164-166
ObjectiveTo investigate the effect of ropivacaine administered intrathecally on the pulmonary function after caesarean section.MethodsThirty-six ASA Ⅰ or Ⅱ parturients,aged 22-34 yr,with body mass index ≤35 kg/m2,undergoing elective cesarean section,were randomly divided into 2 groups (n=18 each):bupivacaine group (group B) and ropivacaine group (group R).Spinal anesthesia was performed at L3,4 interspace and 0.5% bupivacaine 1.8 ml (in cerebrospinal fluid 3 ml) and 1.0% ropivacaine 1.4 ml (in cerebrospinal fluid 3 ml) were injected into the subarachnoid space over 10 s in groups B and R respectively.Forced vital capacity (FVC),forced expiratory volume first second (FEV1) and peak expiratory flow (PEF) were measured by spirometry before surgery (T0),and at 1,1.5 and 2 h after intrathecal injection (T1-3).The motor block score was recorded at T1-3.It is regarded as the criteria for respiratory dysfunction that the measured values of FVC and FEV1 are lower than their 80 % and 70 % predicted levels respectively.ResultsFVC and FEV1 were significantly decreased at T1 compared with the baseline value at T0 in both groups (P<0.05).Compared with that at T1,PEF was significantly increased at T2,3 in group R (P <0.05).Compared with group B,PEF at T1 and motor block score atT3 were significantly decreased in group R (P<0.05).The respiratory dysfunction was not found in both groups.ConclusionSpinal anesthesia with ropivacaine and bupivacaine can inhibit the parameter level of carly pulmonary function afrer caesarean section in parturient without respiratory complications,but the pulmonary function still remains normal.
6.The roles of ABCD2 score in the evaluation of the transient ischemic attack
Xuan LIU ; Xiaopei SUN ; Hua CAO ; Geng CHANG ; Ming LI
International Journal of Cerebrovascular Diseases 2012;20(5):351-355
Part of the patients with transient ischemic attack (TIA) may develop ischemic stroke.Some of them may die because of complicating cardiovascular disease.Studies in recent years have shown that the ABCD2 score has an important value in the evaluation of the prognosis of TIA.This article summarizes the source and application of the ABCD2 score,and focuses on the roles of the score in the evaluation of the prognosis of TIA.
7.Clinical Research of Stereotactic Biopsy in Diagnosis of Multiple Intracranial Lesions
Zhenguang ZHANG ; Zongfang LI ; Hualin YU ; Tao SUN ; Xin GENG
Journal of Kunming Medical University 2013;(11):80-83
Objective To investigate the clinical value of stereotactic biopsy in the diagnosis of the multiple intracranial lesions, and explore the operation methods, technical points and clinical experiences to reduce surgical complications. Methods Twenty-three patients in the first affiliated hospital of Kunming Medical University underwent stereotactic biopsy from January 2009 to June 2013 were analyzed retrospectively. The patients were aged between 11 and 73 years (the mean age of 34.6 years) . There were 12 males and 11 females. Operations were performed by thin thickness of spiral CT scan, ASA-602S and Leksell-Frame-G stereotactic frame, Sedan side-cutting needle, Backlund side-cutting needle and neuroendoscope of STORZ. Routine histopathological examinations of specimens were conducted. The immunohistochemical staining of the histopathological section of specimens was performed if necessary. Results The diagnostic yield was 91.3% (21 cases) . The result of pathological diagnosis was inflammatory granuloma in 1, inflammatory lesion in 1, calcification in 1, hyperplasia of colloid cells in 6, astrocytoma in 1 (WHOⅠ-Ⅱ), astrocytoma in 2 (WHOⅡ), astrocytoma in 1 (WHOⅡ-Ⅲ), mixed oligoastrocytoma in 1 (WHOⅠ-Ⅱ), glioblastoma multiforme in 3 (WHOⅣ), germinoma in 1, brain metastases in 1, diffuse large B-cell lymphoma in 1, intracranial granulomatousarteritis in 1 and negative in 2. There were no serious complications, such as coma, hemiparalysis, infection and intracranial hematoma. Conclusion Stereotactic biopsy is an important method in the diagnose of multiple intracranial lesions. It has the advantages of precise location, less damage, safe performance, and reducing the complication effectively. It is worth promoting.
8.D ifferent cytotoxic effects of hydroxycamptothecin on human lung cancer cells and human embryo lung fibroblast cells
Zimu LI ; Min WANG ; Zhilan XU ; Di GENG ; Zhenxiao SUN
Chinese Journal of Pharmacology and Toxicology 2014;(3):315-320
OBJECTIVE Toinvestigatethedifferenceofcytotoxiceffectsofhydroxycamptothecin(HCPT)onhuman lungcancercelsA549andhumanembryolungfibroblastcelsMRC-5.METHODS A549celsandMRC-5celswere treated with HCPT 20-200 μmol·L-1 for 24,48 and 72 h,or pulse treated with HCPT 50-400 μmol·L-1 for 24 h along with 5 d release.cellsurvival was detected by MTT assay.Morphological changes for both types of cells were observed under an inverted phase-contrast microscope.cellcycle and apoptosis in both cells treated with HCPT 50 μmol·L-1 for 48 h weredeterminedbyflowcytometry.RESULTS HCPT20-200μmol·L-1inhibitedthesurvivalofbothcelsinaconcen-tration-dependent manner and more cytotoxicity was observed in A549 cells for 48 h.The concentration-effect correlation coefficient(r)of HCPT in A549 and MRC-5 cells for 48 h was 0.898 (P=0.015)and 0.996 (P=2.56E-5)respectively. The inhibition rates were significantly different between A549 and MRC-5 cells with treatment of HCPT 20,50,80,1 00, 1 60 and 200 μmol·L-1 for 48 h (P<0.05).The IC50 of HCPT on A549 and MRC-5 cells was (24.00 ±0.69)μmol·L-1 and (1 23.63 ±3.89)μmol·L-1 respectively,indicating that A549 cells were 5-fold more sensitive to HCPT than MRC-5 cells at 48 h.After exposure to HCPT 50 μmol·L-1 for 48 h,some A549 cells were rounded up and shrank dramatical y, and some cells underwent membrane blebbing or lysing while MRC-5 cells had no obvious changes.cellcycle and apop-tosis analysis showed that A549 cells were arrested at both S and G2/M phases and apoptosis occurred but MRC-5 cells were just arrested at S phase.In the recovery growth curve,the growth of A549 cells was inhibited to a larger extent than MRC-5 cells and the growth retardation stil existed for 24 h in both cells.The survival of MRC-5 cells was faster than that ofA549cels,althoughtherewasnocompleterecoveryineithercel.CONCLUSION A549celsaremoresensitiveto HCPT than MRC-5 cells due to the fact that HCPT induces cellcycle arrest at both S and G2/M phases and apoptosis in A549 cells,but only triggers S phase arrest in the MRC-5 cells.
9.Glycoconjugate glycan and its association with breast cancer
Xingquan SUN ; Jing LI ; Meiyu GENG ; Huashi GUAN
Chinese Pharmacological Bulletin 1986;0(04):-
Breast cancer has the highest incidence of gynecological tumors. Investigating into pathogenesis of breast cancer resulting from the change of glycoconjugate glycan has become the hotspot in disease glycomics. The diversified situation of glycoconjugate glycan and its association with breast cancer are reviewed and some issues are also discussed.
10.N-terminal pro-brain natriuretic peptide but not high-sensitivity C-reactive protein is related to severity of coronary artery stenosis in patients with acute coronary syndrome
Shijun LI ; Zhijun SUN ; Dandan LI ; Geng QIAN ; Tingshu YANG ; Xiaoying LI
Journal of Geriatric Cardiology 2010;07(2):101-105
Objective Biochemical indicators such as N-terminal pro-brain type natriuretic peptide(NT pro-BNP)and high-sensitivity Creactive protein(hsCRP)predict mortality in acute coronary syndrome(ACS).However,little is known about the relationship of these factors with severity of coronary artery stenosis in patients with.Methods Three hundred and thirty-one subjects including 246 unstable angina pectoris patients and 85 myocardial infarction patients were recruited and classified into two groups:single-vessel disease group(1-vessel disease,n=93)and multiple-vessel disease group(≥2-vessels disease,n=238)according to selective coronary angiography.Plasma levels of NT pro-BNP and hsCRP were measured and severity of coronary stenosis was determined by Gensini score.Results NT pro-BNP but not hsCRP level was higher in patients with myocardial infarction than in patients with unstable angina pectoris.The patients with multiple-vessel disease had significantly higher NT pro-BNP level but not hsCRP compared with those with single-vessel disease.NT pro-BNP levels increased significantly as left ventricle(LV)function decreased,and only NT proBNP but not hsCRP level was related to Gensini score of severity of coronary stenosis in ACS.Conclusion NT proBNP but not hsCRP level is related to severity of coronary artery stenosis in patients in ACS.