Parathyroid Hormone-Related Protein Promotes the Proliferation of Patient-Derived Glioblastoma Stem Cells via Activating cAMP/PKA Signaling Pathway

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.

Catalpol improves non-alcoholic fatty liver and lipid-lowering mechanism through endoplasmic reticulum stress-mediated PERK-eIF2α signaling pathway / 中华内分泌代谢杂志

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.

Electromagnetic navigation technique in removal of hematoma under neuroendoscope / 中华神经医学杂志

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.

PredictionofthetextureofpituitaryadenomasbyMRIsignalintensity / 实用放射学杂志

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.

On the effects of NLRP3 inflammasome on mice pancreatic β-cell damage induced by advanced glycation end products / 中华内分泌代谢杂志

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.

Application of navigation assisted neurosurgery operation system in clinical practice teaching of clinical medicine specialty / 中华医学教育探索杂志

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.

Effects of maternal high-fat diet on liver fatty acid metabolism-related gene expression in the early life of rat offspring / 临床儿科杂志

Objective To study the impact of maternal high-fat diet during pregnancy and lactation on hepatic steatosis in the early life of offspring rats and its possible mechanism. Methods Female Sprague-Dawley rats were fed either a high fat diet (HF) or control (C) diet for 8 weeks before mating and throughout gestation and ifrst 3 weeks of lactation. The expressions of hepatic fatty acid catabolism related genes, including peroxisome proliferator-activated receptor alpha (PPARα), acyl-CoA syn-thease long-chain family member3 (ACSL3), carnitine palmitoyltransferase-1α(CPT-1α) and 3-hydroxyacyl CoA dehydrogenase (Ehhadh) were determined in offspring liver tissue. The liver pathology was examined in offspring rats at 3 weeks of age. Results Pathohistological ifndings at 3 weeks of age showed that there were diffuse vacuolar degeneration in cytoplasm of hepatocytes and spot necrosis in hepatic lobular in the HF offspring liver. The mRNA expressions of PPARαand Ehhadh genes were markedly increased in the HF offspring as compared to the control group (P<0.05). The mRNA expression of CPT-1αgene was also higher in the HF offspring than that in control group (P=0.19). The level of ACSL3 gene expression, however, was markedly decreased (P<0.05). Conclusions Maternal high fat diet during pregnancy and lactation could result in an increased expression of genes related to hepatic fatty acidβ-oxidation, including PPARα, CPT1αand Ehhadh, but the liver steatosis cannot be reversed in the early life of offspring.

Increased risk of metabolic disease in offspring rats due to maternal high fat diet / 临床儿科杂志

Objectives To examine the association of the maternal high-fat (HF) diet with increased susceptibility to obe-sity and the development of metabolic diseases in their offspring, and observe difference in the effect of maternal vs. acquired high fat diet on metabolic state in their offspring. Methods A total of 15 SD female rats were divided into HF diet group (group H, n=9) and control diet group (group C, n=6). After fed on different diet for seven weeks, they were mated at the age of ten weeks and became pregnant. Their offspring were then divided to groups CH and HH fed HF diet and groups CC and HC fed control diet. At the age of 3 and 8 weeks, the metabolic markers and the liver pathohistological evidences of their offspring were obtained. Results The body weight, area under curve (AUC) of glucose tolerance, cholesterol and triglyceride were all higher in group H than those in group C (P0.05). At the age of 8 weeks, there was no difference in fasting glucose and insulin levels among the four offspring groups. The AUC and body weight were higher in group H than in group C (main effect of maternal diet, P=0.024, P=0.013). The AUCs were also higher in groups CH and HH than groups CC and HC respectively (main effect of acquired diet, P=0.041). The levels of total cholesterol, triglycerides and LDL at the age of 8 weeks were all higher in HH and CH groups than those in HC and CC groups (main effect of acquired diet, P=0.008, 0.007, 0.000, respectively). Their histological analysis at 8 weeks showed different degrees of fatty liver in HH, HC and CH groups, and normal liver in CC group. Conclusions Maternal HF diet may result in increased body weight, fatty liver and impaired glucose tolerance in their adult offspring, and thus increase the risk of developing metabolic diseases at their later age. .

Glutamate enhances the expression of vascular endothelial growth factor in cultured SD rat astrocytes / 药物分析学报

Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups: control group (C), glutamate group (G), QA group (Q), DCG-IV group (D), L-AP4 group (L) and glutanmte-FMCPG gronp (G+M). Cells were cultured under nomoxic condition (95% air, 5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes, respectively. G+ M group was preincubated with lmM MCPG for 30 min prior to the stimulation with glutamate. There were 7 time points at 0,4,8,12,16,24 and 48 h in each group except G+M group. Results The expression of VEGF mRNA and protein did not differ significantly among D group, L group and C group. Different from that in C group, the expression of VEGF mRNA and protein could be enhanced both in a dose-dependent and time-dependent manner in G group and Q group. Meanwhile, the enhanced expression of VEGF mRNA and protein in G group was completely suppressed by MCPG after 24 h. Conclusion Glutamate can increase the expression of VEGF mRNA and protein in cultured astrocytes, which may be due to the activation of group I metabotropic glutamate receptors in astrocytes.

The development of an OxyHb animal model in mice and the study on OxyHb-induced apoptosis of mouse brain cells in vivo / 药物分析学报

Objective On the basis of developing a new animal model for oxyhemoglobin (OxyHb) injection into subarachnoid space in mice, this research was to explore the temporal dependence and spatial distribution of OxyHb- induced apoptosis in the mouse brain cells in vivo and the mechanism of neurocyte injury induced by OxyHb. Methods The animal model for OxyHb injection into subarachnoid space in mice was developed. Mice were divided randomly into the experimental group (n=40) and the control group (n= 35). The control group received saline injection (50 μL ) and the experimental group received OxyHb injection (50 μL ), both into the subarachnoid space. The mice of the two groups were subdivided according to different postoperative time (3 h, 6 h, 12 h, 24 h and 48 h). The apoptosis or necrosis of cells was distinguished with microscopy (HE staining), transmission electron microscopy and TUNEL method. Results The distribution of apoptosis was mainly in the ipsilateral neocortex and bilateral hippocampal gyrus. The apoptotic mouse brain cells showed morphological changes in the experimental group by HE staining and transmission electron microscopy. The count of TUNEL-positive cells showed substantial increase in the experimental group, and there was a significant difference between the control and experimental groups, and the number of OxyHb- induced apoptotic cells decreased with time. Conclusion OxyHb in subarachnoid space in mice can induce apoptasis, but not necrosis of mouse brain cells in viro. The apoptotic brain cells show the pattern of temporal dependence and spatial distribution. It is suggested that the early treatment should be the method of first choice for treating the hemorrhagic brain injury.

Expression of CyclinD1 protein in cerebral glioma in the evaluation of prognosis / 中国组织工程研究

BACKGROUND: Research has showed that the abnormal expression of some proteins closely relates to the occurrence and development of cerebral glioma. However, the relationship between the abnormal expression of CyelinD1protein and the occurrence, development and prognosis of glioma is still uncertain which needs further study.OBJECTIVE: To study the expression of CyclinD1 protein in cerebral glioma and relationship between it and the impact of tumor DESIGN: Control study based on pathological specimens.SETTING: Neurosurgery department of a university hospital.PARTICIPANTS: Totally 48 glioma specimens of different malignaut degree were collected from the patients who accepted surgery treatment in Neurosurgery Department of Second Hospital of Xi' an Jiaotong University during January 1990 and December 1995. Twelve normal cerebral specimens were from the non-tumor patients who were conducted intracranial pressure reducing in Neurosurgery Department of Second Hospital of Xi' an Jiaotong University during January 1990 and December 1995.METHODS: S-P immunohistochemical technique was used to detect the abnormal expression of Cyclin D1 protein. Simultaneously, the dyeing results and clinical characters of patients were associated in order to conduct comparison.RESULTS: The positive expression rate of CyclinD1 protein in human cerebral glioma was 54. 12% while in normal cerebral tissue it was about 8.33%. There was significant difference between them(x2 =8. 148 1,P = 0. 004 3 ) . And the positive expression rate in cerebral glioma of low malignancy was 37.04% while in specimens of high malignancy it was 76.19%, there was significant difference (x2 = 7. 294 0, P = 0. 006 9). The positive expression rate of CyclinD1 protein in specimens of patients with long survival period and short survival period after surgery was 70. 37% and 33.33% respectively with significant difference between them (x2 = 6. 5268,P =0.010 6).CONCLUSION: CyclinD1 is closely related to the occurrence and development of human cerebral glioma. It has provided experimental evidence for the prevention to the occurrence of glioma and the estimation of its prognosis by studying the abnormal proliferation of glioma cells targeted on CyclinD1.

RXPERIMENTAL AND CLINICAL STUDY OF CREATINE KINASE BB ACTIVITY FOR TH E DIAGNOSIS ON BRAIN DAMAGE / 药物分析学报

Objective To study the differential diagnosis o n cerebral concussion and mild cerebral contusion value of the brain type creati n e kinase isoenzyme(CK-BB),and evaluate the seriousness of brain damage and prog nosis of the patients with acute head injury.Methods Chromatographic separating and fluorometric quant ifying technique was used to detect the CK-BB activity in the cerebrospinal flu id(CSF) of 117 patients with acute head injury and 12 patients with increased in tracranial pressure and 20 normal people.Results The CSF-CK-BB activity of the patients with acu te head injury was remarkably higher than that of the normal people and the CSF -CK-BB activity increased with the seriousness of brain damage.There was a clo se relationship between CSF-CK-BB activity and prognosis,and higher activity o f CSF-CK-BB indicated poor prognosis.Conclusion CSF-CK -BB activity could be used as a new index to diagnose brain damage and evaluate the seriousness of brain damage and prognosis.