ABSTRACT
Objective To investigate the effect of lncRNA GIHCG on the radiosensitivity of glioma cells and its mechanism.Methods The expression levels of GIHCG and miR-146a-3p in human brain normal glial cells HEB and glioma cell lines U251,A172,SHG139 and U87 were quantitatively measured by qRT-PCR assay.U251 and SHG139 cells were used for subsequent experiment.After silencing the expression of GIHCG or overexpressing miR-146a-3p in U251 and SHG139 cells,cell proliferation was detected by MTI assay,cell apoptosis was detected by flow cytometry,cell radiosensitivity was detected by colony formation assay and the expression levels of CDK1,CyclinD1,Bcl-2 and Bax proteins were measured by Western blot.The bioinformatics software predicted the presence of a binding site for GIHCG and miR-146a-3p.Dual luciferase reporter gene assay and qRT-PCR assay were adopted to verify the targeting relationship between GIHCG and miR-146a-3p.Results Compared with HEB cells,the expression of GIHCG was significantly up-regulated in glioma U87,U251,A172 and SHG139 cells (all P＜0.05),whereas that of miR-146a-3p was remarkably down-regulated (P＜0.05).Silencing GIHCG expression or overexpression of miR-146a-3p significantly decreased the U251 and SHG139 cell survival rate,survival fraction and the expression of CDK1,CyclinDl and Bcl-2 proteins (all P＜0.05),whereas considerably increased the apoptotic rate and expression of Bax protein (both P＜0.05).GIHCG performed targeted negative regulation of miR-146a-3p expression in U251 and SHG139 cells and inhibition of miR-146a-3p expression reversed the effect of silencing GIHCG on proliferation,apoptosis and radiosensitivity of glioma cells.Conclusion Silencing GIHCG expression up-regulates the expression of miR-146a-3p,thereby enhancing the radiosensitivity of glioma cells.
ABSTRACT
Objective:To investigate the effect of long non-coding (lnc) RNA HCP5 on the radiation sensitivity of glioma cells and underlying mechanism.Methods:The glioma cells U251 and U87 were irradiated with 0, 2, 4, 6, and 8 Gy rays as different doses.si-Con, si-HCP5, pcDNA, and pcDNA-HCP5 were transfected into cells U251 and U87, recorded as si-con group, si-HCP5 group, pcDNA group, and pcDNA-HCP5 group.si-Con and si-HCP5 were transfected into cells U251 and U87, and then irradiated with 4 Gy rays, respectively, recorded as IR+ si-con group and IR+ si-HCP5 group, the cells only irradiated with 4 Gy rays were recorded as IR group.After si-HCP5 with anti-miR-con and anti-miR-508-3p was co-transfected into cell U251 and U87, respectively, irradiated with 4 Gy rays, recorded as IR+ si-HCP5+ anti-miR-con group and IR+ si-HCP5+ anti-miR-508-3p group, respectively, the transfection was performed by liposome method.RT-qPCR was used to detect the expression of miR-508-3p and HCP5.Cell clone formation assay was used to detect the radiosensitivity of glioma cells.Flow cytometry was used to detect apoptosis, dual luciferase Reporter gene detection experiments detects fluorescence activity.Results:HCP5 was highly expressed in radiation-treated glioma cells, and miR-508-3p was lowly expressed.After silenced HCP5, U251 and U87 cells had enhanced radiosensitivity and apoptotic rate((16.67±1.68) vs (3.58±0.62), t=21.929, P<0.05; (12.32±1.08) vs (4.48±0.71), t=18.198, P<0.05) was increased, and γ-H2AX( (0.45±0.04) vs (0.23±0.05), t=10.307, P<0.05; (0.38±0.04) vs (0.24±0.03), t=8.400, P<0.05), Cleaved caspase-3((0.37±0.04) vs (0.16±0.03), t=12.600, P<0.05; (0.38±0.04) vs (0.22±0.03), t=9.600, P<0.05) expressions were increased.Compared with silencing HCP5 or radiation treatment alone, silencing HCP5 and radiation treatment of U251 cells simultaneously, the apoptosis rate ((25.34±1.54) vs (16.67±1.68), t=11.413, P<0.05; (25.34±1.54) vs (11.13±1.06), t=22.802, P<0.05) was significantly increased, and γ-H2AX((0.69±0.05) vs (0.45±0.04), t=11.245, P<0.05; (0.69±0.05) vs (0.31±0.04), t=17.804, P<0.05), Cleaved caspase-3 ((0.52±0.06/0.37±0.04, t=6.240, P<0.05) (0.52±0.06/0.34±0.04, t=7.488, P<0.05) expressions were increased.The expressions of p-PI3K ((0.21±0.02) vs (0.52±0.04), t=20.795, P<0.05; (0.26±0.23 ), ( 0.67±0.07), t=5.116, P<0.05), p- AKT ((0.22±0.03) vs (0.66±0.07), t=17.332, P<0.05; (0.23±0.04) vs (0.71±0.03), t=28.800, P<0.05) in U251 and U87 cells were decreased.HCP5 can target the regulation of miR-508-3p expression; interfering with miR-508-3p reversed the effects of silent HCP5 and radiation on the radiation sensitization and apoptosis of U251 and U87 cells.It reduced the expression levels of reducing γ-H2AX and Cleaved caspase-3, while increased the expression levels of p-PI3K and p-AKT. Conclusion:Silencing lncRNA HCP5 can enhance the radiation sensitivity of glioma cells and promote apoptosis.The mechanism may be related with the miR-508-3p and PI3K/Akt signaling pathway, which will provide new targets and new ideas for glioma treatment.
ABSTRACT
Objective@#To investigate the effect of lncRNA GIHCG on the radiosensitivity of glioma cells and its mechanism.@*Methods@#The expression levels of GIHCG and miR-146a-3p in human brain normal glial cells HEB and glioma cell lines U251, A172, SHG139 and U87 were quantitatively measured by qRT-PCR assay. U251 and SHG139 cells were used for subsequent experiment. After silencing the expression of GIHCG or overexpressing miR-146a-3p in U251 and SHG139 cells, cell proliferation was detected by MTT assay, cell apoptosis was detected by flow cytometry, cell radiosensitivity was detected by colony formation assay and the expression levels of CDK1, CyclinD1, Bcl-2 and Bax proteins were measured by Western blot. The bioinformatics software predicted the presence of a binding site for GIHCG and miR-146a-3p. Dual luciferase reporter gene assay and qRT-PCR assay were adopted to verify the targeting relationship between GIHCG and miR-146a-3p.@*Results@#Compared with HEB cells, the expression of GIHCG was significantly up-regulated in glioma U87, U251, A172 and SHG139 cells (all P<0.05), whereas that of miR-146a-3p was remarkably down-regulated (P<0.05). Silencing GIHCG expression or overexpression of miR-146a-3p significantly decreased the U251 and SHG139 cell survival rate, survival fraction and the expression of CDK1, CyclinD1 and Bcl-2 proteins (all P<0.05), whereas considerably increased the apoptotic rate and expression of Bax protein (both P<0.05). GIHCG performed targeted negative regulation of miR-146a-3p expression in U251 and SHG139 cells and inhibition of miR-146a-3p expression reversed the effect of silencing GIHCG on proliferation, apoptosis and radiosensitivity of glioma cells.@*Conclusion@#Silencing GIHCG expression up-regulates the expression of miR-146a-3p, thereby enhancing the radiosensitivity of glioma cells.
ABSTRACT
Objective To evaluate the risk of intracranial infection in patients treated with neuroendoscopy optic nerve tube decompression, and compare the risk of intracranial infection between surgery treatment and methylprednisolone treatment. Methods The clinical data of 105 patients with traumatic optic neuropathy from January 2013 to December 2017 in the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed. Among them, 74 patients were treated with transsphenoidal approach neuroendoscopy optic nerve tube decompression (operation group), and cured with antibiotics during perioperative period; 31 patients were treated with intravenous methylprednisolone sodium succinate (hormone group), and combined with mouse nerve growth factor etc. Meanwhile, broad-spectrum antibiotics were used to prevent the potential infection during the first 3 d of hospitalization. The incidence of intracranial infection was compared between 2 groups. Results The incidence of intracranial infection in operation group was 4.05%(3/74), that in hormone group was 3.23%(1/31), and there was no statistical difference between 2 groups (χ2=0.127, P＞0.05). Conclusions Compared with conservative treatment, the neuroendoscopy optic nerve tube decompression has less or equivalent chance to increase the risk of intracranial infection in patients with traumatic optic neuropathy. Accordingly, it could be viewed as the first-line treatment.
ABSTRACT
Objective To discuss the feasibility of neuroendoscopic third ventriculostomy for chronic posttraumatic hydrocephalus (PTH).Methods Nineteen cases of chronic PTH treated with neuroendoscopic third ventriculostomy between October 2010 and October 2014 were analyzed retrospectively.There were 13 males and 6 females, aged 11-57 years (mean, 36.3 years).Trauma resulted from traffic accidents in 14 cases, falls in 4 cases and blunt object hitting in 1 case.Of the 19 cases analyzed, 5 had Glasgow Coma Scale (GCS) score of 13-15, 5 had score of 9-12 and 9 had score of 5-8 at admission.Results of operation were assessed with the Canada multicenter evaluation criteria.Prognosis was analyzed with the Glasgow Outcome Scale (GOS).Results All cases were followed up for mean 13.6 months (range, 6-26 months).Improvement of symptoms was achieved in 17 cases, but was not seen in 2 cases.Of the 2 cases, one required ventriculoperitoneal shunt two weeks after ineffective ventriculostomy, and one required second ventriculostomy one month after the presence of stoma blockage.No serious complications occurred.At follow-up, 9 cases had GOS score of 5, 8 cases had score of 4 and 2 cases had score of 3.Conclusions Neuroendoscopic third ventriculostomy is in line with the physical characteristics in cerebrospinal fluid circulation, which implies no shunt implantation, less operative trauma and less complications.The procedure is an effective approach for chronic PTH.
ABSTRACT
AIM:To study the urinary pharmacokinetics of five anthraquinones after oral administration of Xiexin Decoction(Radix et Rhizoma rhei,Rhizoma coptidis and Radix scutellariae) in rats.METHODS:A high-performance liquid chromatographic method with fluorescence detection(HPLC-FLD) was established and validated the quantification for five anthraquinones(aloe-emodin,rhein,emodin,chrysophanol and physcion) in rat urine.SD rats were given 12 g/kg of Xiexin Decoction.Urine was collected before and after perfusion.Anthraquinones components in urine were measured by HPLC-FLD.Urinary pharmacokinetic parameters were determined according to urinary output-time data.RESULTS:After oral administration of Xiexin Decoction all the five anthraquinones were excreted from the urine.The excretion T_ 1/2 of aloe-emodin,rhein,emodin,chrysophanol and physcion were 3.46?1.18,3.24?0.60,4.69?1.99,4.49?1.63,5.65?1.74 h,respectively.The amounts of aloe-emodin,rhein,emodin,chrysophanol and physcion excreted from urine during 0～48 h were(11.28?4.30)?g,(116.73?17.46)?g,(5.48?2.92)?g,(9.53?2.67)?g,(0.41?0.20)?g,respectively.CONCLUSION:After oral administration of Xiexin Decoction five anthraquinones were excreted from urine and a small quantity of five anthraquinones excreted from urine in rats is less than 10% of oral dose.
ABSTRACT
AIM: To study the pharmacokinetics of flavonoids in mice after ig administration of Xiexin Decoction (Radix et Rhizoma rhei, Rhizoma coptidis, Radix Scutellariae). METHODS: Mice were given a single ig dose of Xiexin Decoction 4.5, 9.0 or 18.0 g/kg. Flavonoids in plasma were analysed by HPLC and plasma concentration of baibalin was determined. Pharmacokinetic parameters were calculated from the plasma concentration-time data with the DAS software package. RESULTS: After ig administration of Xiexin Decoction in mice, baicalin, baicalein and another flavonoid were detected in plasma and baicalin concentration was the highest of the three kinds of flavonoids in plasma. After a single ig dose of Xiexin Decoction 4.5, 9.0 or 18.0 g/kg, the pharmacokinetic parameters of baicalin were as follows:T_ 1/2 =2.77、5.69、6.20 h,AUC_ 0-∞ =9.09、23.49、39.57 ?g?h/mL,CL= 12.52 、 6.962 、 11.50 L?h/kg,V_d= 50.11 、 79.56 、 102.95 L/kg,C_ max1 =1.89、3.32、4.79 ?g/mL(T_ p1 = 0.08 h ), C_ max2 =1.46、2.57、4.16 ?g/mL(T_ p2 =3 h), respectively. CONCLUSION: Three kinds of flavonoids can be absorbed after ig administration of Xiexin Decoction in mice, of which baicalin is the major component.