1.Parathyroid Hormone-Related Protein Promotes the Proliferation of Patient-Derived Glioblastoma Stem Cells via Activating cAMP/PKA Signaling Pathway
Zhenyu GUO ; Tingqin HUANG ; Yingfei LIU ; Chongxiao LIU
International Journal of Stem Cells 2023;16(3):315-325
Background and Objectives:
Glioblastoma (GBM) is an aggressive primary brain tumor characterized by its hetero-geneity and high recurrence and lethality rates. Glioblastoma stem cells (GSCs) play a crucial role in therapy resistance and tumor recurrence. Therefore, targeting GSCs is a key objective in developing effective treatments for GBM. The role of Parathyroid hormone-related peptide (PTHrP) in GBM and its impact on GSCs remains unclear. This study aimed to investigate the effect of PTHrP on GSCs and its potential as a therapeutic target for GBM.
Methods:
and Results: Using the Cancer Genome Atlas (TCGA) database, we found higher expression of PTHrP in GBM, which correlated inversely with survival. GSCs were established from three human GBM samples obtained after surgical resection. Exposure to recombinant human PTHrP protein (rPTHrP) at different concentrations significantly enhanced GSCs viability. Knockdown of PTHrP using target-specific siRNA (siPTHrP) inhibited tumorsphere formation and reduced the number of BrdU-positive cells. In an orthotopic xenograft mouse model, suppression of PTHrP expression led to significant inhibition of tumor growth. The addition of rPTHrP in the growth medium counteracted the antiproliferative effect of siPTHrP. Further investigation revealed that PTHrP increased cAMP concentration and activated the PKA signaling pathway. Treatment with forskolin, an adenylyl cyclase activator, nullified the antiproliferative effect of siPTHrP.
Conclusions
Our findings demonstrate that PTHrP promotes the proliferation of patient-derived GSCs by activating the cAMP/PKA signaling pathway. These results uncover a novel role for PTHrP and suggest its potential as a therapeutic target for GBM treatment.
2.Nam Dinh virus was detected and isolated in arbovirus investigation in Shanxi, China
Shenghui LIU ; Xiaodong TIAN ; Weijia ZHANG ; Hongmei ZHENG ; Junying ZHAO ; Chongxiao XU ; Yali ZHANG ; Shihong FU ; Kai NIE ; Fan LI ; Peifang DAI ; Qikai YIN ; Ying HE ; Jingxia CHENG ; Huanyu WANG
Chinese Journal of Experimental and Clinical Virology 2023;37(1):25-29
Objective:In this study, the collected mosquito samples were subjected to viral isolation to identify the species and branch characteristics of arboviruses in five regions of Shanxi Province.Methods:Eight arboviruses in mosquito samples collected from July to September 2020 were detected by real-time fluorescent quantitative PCR, and virus isolation was carried out through cell culture. Virus isolates were identified and analyzed by molecular biology and bioinformatics method.Results:We detected 1 batch of positive samples of Japanese encephalitis virus, 2 batches of positive samples of Culex flavivirus and 8 batches of positive samples of Nam Dinh virus among 121 batches of mosquito samples. Seven virus isolates were isolated, numbered: SX-YJ-Cxp-4、SX-YJ-Ars-2、SX-YJ-Cxp-1、SX-LY-Cxp-10、SX-GP-Ars-5、SX-GP-Cxp-2、SX-GP-Cxp-4, all of which were identified as Nam Dinh virus, and the whole genome sequencing was performed on one of them, and the result showed that Shanxi Nam Dinh virus isolate and Yunnan Nam Dinh virus isolate belonged to the same evolutionary branch.Conclusions:Nam Dinh virus was isolated and identified on the specimen from Shanxi province for the first time.
3.Laboratory identification and evaluation of national standard strains of Japanese encephalitis virus G1/G3/G5
Shenghui LIU ; Mengnan JIANG ; Weijia ZHANG ; Shihong FU ; Jingdong SONG ; Chongxiao XU ; Kai NIE ; Qikai YIN ; Ying HE ; Fan LI ; Songtao XU ; Guodong LIANG ; Qiang WEI ; Huanyu WANG
Chinese Journal of Experimental and Clinical Virology 2023;37(3):273-279
Objective:To determine the evaluation indexes of national standard strains of genotypes 1, 3 and 5 of Japanese encephalitis virus (JEV) and evaluate the national standard JEV strains.Methods:According to the national standard strain evaluation technical standards of pathogenic microbial bacteria (virus) species, based on the application of Japanese encephalitis virus research, and according to the morphological characteristics, biological characteristics, molecular biological characteristics and other research data to identify the characteristics of G1, G3 and G5 genotypes of JEV.Results:Spherical virus particles with a diameter of about 60 nm were visible under electron microscope of the three Japanese encephalitis virus strains. The cytopathic effect was mainly characterized by cell shrinkage and exfoliation in BHK-21 and Vero cell lines, cell fusion and exfoliation were shown after infection with C6/36 cell line; the virus titer was 10 5-10 7 PFU/ml, and the plaque size was different by genotype. The median lethal dose of intrabitoneal challenge in G1, G3 and G5 JEV in three weeks-old mice was 50.51 PFU, 6.98 PFU, and 8.13 PFU, and the median lethal dose of intracranial challenge in five weeks mice was 3 PFU, 0.3 PFU, 1.35 PFU. The whole genome length of G1, G3 and G5 JEV was 10 967 bp, 10 976 bp and 10 983 bp, respectively. Conclusions:Three genotypic national standard strains of JE V were identified and evaluated by electron microscopy, cell, animal and genome laboratory indexes, which provided reference for the identification and evaluation of other national standard strains of JEV.
4.Catalpol improves non-alcoholic fatty liver and lipid-lowering mechanism through endoplasmic reticulum stress-mediated PERK-eIF2α signaling pathway
Lu LIU ; Chongxiao LIU ; Qingyang LENG ; Yi ZHU ; Hongli ZHANG ; Xiaohua LI
Chinese Journal of Endocrinology and Metabolism 2022;38(3):231-238
Objective:To investigate effect and underlying lipid-lowering mechanisms of catalpol in non-alcoholic fatty liver disease(NAFLD).Methods:In vivo model of NAFLD was established with high-fat diet-fed ICR mice for 8 weeks. Low(50 mg/kg), medium(150 mg/kg), and high(300 mg/kg) doses of catalpol were administered, and the body weight, liver weight, hepatic index, and biochemical parameters of the mice were analyzed. Free fatty acid-induced LO2 in human hepatocytes to establish NAFLD cell model. Quantitative realtime PCR reaction to detect fatty acid synthesis-related gene levels. Western blotting assay was adopted to analyze proteins in the endoplasmic reticulum stress(ERS)-mediated protein kinase RNA-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α) signaling pathway. Results:Compared with model mice, body weight [(39.43±1.84)g, (34.01±1.83)g, (32.28±1.11)g vs(42.17±1.37)g, all P<0.001], liver weight [(1.03±0.06)g, (0.79±0.05)g, (0.64±0.04)g vs(1.30±0.13)g, P<0.01 or P<0.001], and liver index [(2.60±0.09)%, (2.32±0.09)%, (1.99±0.11)% vs(3.07±0.30)%, P<0.05 or P<0.001] were reduced in low, medium, and high doses of catapol model. Medium and high doses of catalpol diminished total cholesterol, triglyceride, low density lipoprotein-cholesterol, aspartate aminotransferase, and alanine aminotransferase( P<0.01 or P<0.001), increased high density lipoprotein-cholesterol( P<0.01 or P<0.001). In the cell model, elevated levels of both fatty acid synthesis genes and PERK-eIF2α pathway proteins were attenuated by catalase, and this attenuation was reversed by signaling pathway agonists. Conclusion:The Chinese herb catalpol may play a role in improving NALFD by regulating the ERS-mediated PERK-eIF2α signaling pathway.
5.Electromagnetic navigation technique in removal of hematoma under neuroendoscope
Zhenyu GU ; Tingqin HUANG ; Ren ZHOU ; Wei SHI ; Yahong ZHANG ; Qin SONG ; Chongxiao LIU
Chinese Journal of Neuromedicine 2020;19(5):477-482
Objective:To explore the application value of electromagnetic navigation technique in the removal of hematoma under neuroendoscope.Methods:Forty-three patients with supratentorial spontaneous cerebral hemorrhage, accepted removal of hematoma under neuroendoscope in our hospital from October 2015 to February 2019, were chosen in our study; 22 patients (navigation group) were performed the removal under the guide of real-time electromagnetic navigation, and 21 (non-navigation group) were performed the removal under neuroendoscope only. The amount of cerebral hemorrhage, operation time, residual amount of hematoma and hematoma clearance rate were compared between the two groups. Fugl-Meyer Assessment (FMA) was used to evaluate the motor function of the affected limbs two weeks after surgery. The anisotropy fraction (FA) values of fibers of affected pyramidal tracts and contralateral pyramidal tracts were examined by diffusion tensor imaging (DTI), and the relative FA (rFA) value (FA values of affected side/contralateral side) was calculated. Barthel index was used to evaluate the basic daily activities of the patients 6 months after surgery.Results:There was no significant difference between the navigation group and the non-navigation group in the amount of cerebral hemorrhage before surgery, amount of residual hematoma after surgery, hematoma clearance rate, and operation time ( P>0.05). FMA scores of upper and lower limbs, FA and rFA values of the affected side in the navigation group were significantly higher than those in the non-navigation group two weeks after surgery ( P<0.05). Barthel index of patients in the navigation group was statistically higher than that of the non-navigation group 6 months after surgery ( P<0.05). Conclusion:Electromagnetic navigation technique can guide endoscopy to effectively clear the supratentorial hypertensive cerebral hemorrhage without obviously increasing the operation time; effective protection of pyramidal fibers can improve the prognoses of patients.
6.PredictionofthetextureofpituitaryadenomasbyMRIsignalintensity
Siping TONG ; Qiujuan ZHANG ; Adeem N ZAHID ; Chongxiao LIU ; Xiaohui LI
Journal of Practical Radiology 2019;35(3):353-356
Objective ToevaluatethepredictivevalueofpreoperativeMRIsignalintensityindifferenttexturesofpituitaryadenoma (PA).Methods Accordingtotheinclusioncriteria,47casesofPA werecollected,including37softcasesand10firmcases.Different texturesofPAonMRIsignalintensityanddynamicenhancementcharacteristicswereanalyzed.Results T2WIintensitybetweentwo texturegroups(T2PA/T2WM、T2PA/T2GM、T2PA/T2CSFandSER)hadsignificantstatisticaldifferences(P<0.05).T1WIintensity betweentwotexturegroups(T1WIPA/T1WIPN )hadnosignificantstatisticaldifference(P>0.05).TheT2signalintensity(T2PA/T2WM、T2PA/T2GM、T2PA/T2CSF)andSERinsoftgroupwashigherthanthatinthefirmgroup.Predictivevalueofthetextureof PAbyusingtheROCcurveofT2PA/T2WM,T2PA/T2GM,T2PA/T2CSF,T1PA/T1PNandSER wereobtained,andtheareasunder theROCcurvewere0.857,0.835,0.856,0.630and0.781respectively.ThelargestareaundertheROCcurvewasT2PA/T2WM (P<0.05).Conclusion MRIT2signalintensityisrelatedtothetextureofPAandthoseindexescanbeusedtojudgethetextureofPA.
7.Features of mammographic mass edge in digital X-ray three dimensional tomography and their correlations with pathological parameters in breast cancer
Caixian YANG ; Hongguang ZHAO ; Hui LIU ; Chongxiao QU
Cancer Research and Clinic 2019;31(1):16-21
Objective To analyze the diagnostic value of features of mammographic mass edge in digital three dimensional tomography, and to discuss the correlations between the mammographic mass edge features and pathological features and molecular biological indicators, in order to provide evidence for early and accurate diagnosis and treatment of breast cancer and prognosis evaluation. Methods A retrospective analysis was made in 392 cases of breast cancer confirmed by operation and pathology in the People ' s Hospital of Shanxi Province from August 2017 to June 2018. These patients were examined by digital breast tomography (DBT) and full-field digital mammography (FFDM) before operation. Postoperative specimens were stained by immunohistochemical SP method to measure the expression of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and Ki-67. The correlation between the mammographic mass edge features and pathological features and molecular biological indicators were analyzed by usingχ2 test and Fisher exact probability method. Results In 392 patients, 352 cases (89.80%) were invasive ductal carcinoma, 6 cases (1.53%) were ductal carcinoma in situ with microinvasion, 7 cases (1.79%) were invasive lobular carcinoma, 17 cases (4.34%) were papillary carcinoma, 8 cases (2.04%) were mucinous carcinoma, 1 case (0.26%) was medullary carcinoma, and 1 case (0.26%) was metaplastic carcinoma. DBT were significantly better than FFDM in the detection rate [93.6% (367/392) vs. 77.8% (305/392)] and the diagnostic coincidence rate [88.2%(345/392) vs. 76.8%(278/392)] for breast cancer and judging the marginal features of tumor. The DBT imaging features of mucinous carcinomas were mostly regular in shape and clear in margin. The DBT imaging features of invasive ductal carcinomas were marginal burr sign or burr with lobulation sign. Immunohistochemical detection displayed the positive expression rates of ER and PR were highest, the positive expression rates of HER2 and Ki-67 were the lowest in marginal burr masses;the positive expression rates of ER and PR were high, the positive expression rates of HER2 and Ki-67 were low in marginal lobulated masses; the positive expression rate of HER2 was highest in borderline masses or partial borderline masses; the positive expression rates of HER2 and Ki-67 were highest in clear margin masses. Conclusions Breast DBT can reduce or eliminate the tissue overlap in FFDM examination, and improve the detection rate, the diagnostic coincidence rate and the specificity of breast cancer. It can also indirectly provide evidence for preoperative judgment of biological behavior of tumors, guide clinical treatment and assess the prognosis through the morphological and marginal features of breast masses. Breast DBT is worthy of a wide clinical application in breast examination.
8.On the effects of NLRP3 inflammasome on mice pancreatic β-cell damage induced by advanced glycation end products
Xiang KONG ; Qing SU ; Hongmei ZHANG ; Ning LIN ; Xiaoyong LI ; Zhen YANG ; Ruyuan DENG ; Chongxiao LIU ; Jie JIN ; Guangxun MENG
Chinese Journal of Endocrinology and Metabolism 2018;34(8):690-695
Objective To explore the role of the pyrin domain-containing 3 ( NLRP3) inflammasome in advanced glycation end products ( AGEs )-induced mice pancreatic β-cell damage. Methods AGEs were administered intraperitoneally for 6 weeks in NLRP3 knockout mice or C57BL/6J mice. Intraperitoneal glucose tolerance test and insulin releasing test were performed. Pancreatic sections were stained with haematoxylin and eosin, or with F4/80 and NLRP3 antibodies. Insulin and pancreatic tissue monocyte chemotactic protein 1 ( MCP-1) as well as interleukin-1β( IL-1β) levels were measured with ELISA kits. Expression of MCP-1 protein was determined by western blot. MIN6 cells and mouse peritoneal macrophages cells were treated with AGEs and different interventions (antioxidant NAC, adenovirus NLRP3 shRNA or NLRP3 knockout). Reactive oxygen species production, NLRP3 mRNA expression, IL-1β secretion, caspase 1 activity, apoptosis and glucose stimulated insulin release were determined. Results Injection of AGEs induced an abnormal response to glucose, enhanced the insulitis score, and increased the levels of pancreatic tissue MCP-1 and IL-1β, as well as raised the expression of NLRP3 and F4/80 in pancreatic islet. Remarkably, co-localization of NLRP3 and macrophage marker F4/80 was observed in islet. The damages were improved in NLRP3 knockout mice. After incubation with AGEs, reactive oxygen species production and cell apoptosis was enhanced, NLRP3 inflammasome activated, with glucose-stimulated insulin release impaired in MIN6 cells. NAC treatment alliviated the above damages, but NLRP3 gene silencing had no effect on ROS level, apoptosis, and insulin secretion. Finally NAC treatment and NLRP3 gene knockout inhibited activation of NLRP3 inflammasome induced by AGEs in mouse peritoneal macrophages cells. Conclusion NLRP3 knockout ameliorates the islet β-cell damage induced by AGEs. These effects were associated with AGEs-induced islets macrophage infiltrating by up-regulation of MCP-1 expression, and AGEs-induced activation of NLRP3 inflammasome in macrophage through ROS pathway, which results in the release of active IL-1βand leads to the lesions of β-cell.
9.Clinicopathological characteristics of ovarian seromucinous borderline tumor
Yanyan DONG ; Yunxiao LIU ; Chongxiao QU
Cancer Research and Clinic 2018;30(9):603-606
Objective To explore the clinicopathological characteristics,immunohistochemical phenotype and differential diagnosis of ovarian seromucinous borderline tumor.Methods Fifteen cases of ovarian seromucinous borderline tumor in Shanxi Provincial People's Hospital from January 2013 to May 2017 were collected.The surgical specimens were observed after HE and immunohistochemical staining,the patients were followed-up,and the relevant literature was reviewed.Results The age of 15 cases of ovarian seromucinous borderline tumor ranged from 26 to 56 years(mean 37 years).Eight cases occurred in the right,5 cases occurred in the left,only 2 cases were bilateral tumors.The complaint of most patients was abdominal distention,3 cases was ascites.The maximum diameter of these tumors ranged from 4 to 13 cm(mean 9.3 cm).Grossly,15 cases mainly showed cystic performance,varying amounts of papillae inside wall of the cysts.Small region of 2 cases were solid.Microscopically,9 tumors were composed of endocervical-like mucinous epithelium,4 tumors were endocervical-like mucinous epithelium and serous epithelium.2 cases were accompanied with endometriosis.Tumor cells mainly expressed estrogen receptor(ER),progesterone receptor(PR),paired box gene protein 8(PAX-8),cytokeratin 7(CK7),these markers were immunophenotypes of Mullerian tumors.Followed up for 3 to 24 months(mean 16.7 months),2 cases showed bilateral tumors,1 case was peritoneal implantation.No tumor recurrence was found in the remaining 12 cases.Conclusions Different from mucinous borderline tumor,ovarian seromucinous borderline tumor possesses relative special clinicopathological features,morphological and immunohistochemical phenotypes,with better prognosis.Combination of immunohistochemical markers ER,PR,PAX-8,CK7,CK20,Vimentin,CDX-2 and WT-1 can make an accurate diagnosis of this tumor.
10.Application of navigation assisted neurosurgery operation system in clinical practice teaching of clinical medicine specialty
Zhenyu GUO ; Chongxiao LIU ; Ren ZHOU ; Gang WANG ; Wei SHI ; Yahong ZHANG
Chinese Journal of Medical Education Research 2016;15(11):1175-1179
Objective To explore the effect of navigation assisted surgery system in the medical teaching in the department of neurosurgery. Method From May 4 of 2015 to June 3 of 2015, 51 medical undergraduates of clinical medicine in the Second Affiliated Hospital of Xi'an Jiaotong University were ran-domly divided into experiment group (navigation assisted surgical technique system teaching, n=25) and traditional group (traditional teaching, n=26). Practical effect of the different modes was evaluated by ques-tionnaire and examination results. Data were analyzed by SPSS 20.0. Enumeration data were compared between groups using chi square test or t test. Result After the teaching, the theoretical results of the experimental group and the control group were (83.05 ± 6.03) and (74.32 ± 7.12), and the difference was statistically significant (t=4.96, P=0.005). Clinical skills scores were (89.43 ± 5.12) and (81.11 ± 8.02), and the difference was statistically significant (t=2.91, P=0.029). The questionnaire showed that the experimental group students'!satisfaction degree to their own teaching method was better than that of control group (P<0.05). Conclusion Compared with the traditional teaching, the navigation assisted neurosurgical operation has obvious advantages. It can improve students'!enthusiasm for learning the professional knowledge and skills in neurosurgery, stimulate students'!learning interest and improve students'!test scores.

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