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【Objective】 To investigate the effects of total flavone of oldenlandia diffusa (FOD) on the proliferation and apoptosis of hepatocellular carcinoma (HCC) stem cells sorted from Huh7. 【Methods】 Human HCC cell lines Huh7 was cultured in vitro; CD133 positive (CD133+) stem cells in Huh7 cell line were sorted by flow cytometry, and stem cell markers such as Nanog, Oct4 and Sox2 were tested by Western blotting. CD133+-Huh7 was stimulated by different concentrations (0 μg/mL, 50 μg/mL, 100 μg/mL and 400 μg/mL) of FOD for different time (24 h, 48 h, 72 h and 96 h). CCK8 and plate cell cloning assay were used to detect the effect of FOD on CD133+-Huh7 proliferation while Annexin V-PE/7-AAD was used to detect the effect of FOD on CD133+-Huh7 apoptosis. Western blotting was used to detect the protein expressions of protein 53 (P53), factor associated suicide-Fas-associating protein with a novel death domain (Fas-FADD), B-cell lymphoma-2 (Bcl-2), Cleaved-Caspase3, and Bcl-2 associated X protein (Bax). 【Results】 More than 95% of stem cells were purified for further experiments. Cell proliferation of CD133+-Huh7 was significantly inhibited by FOD, with the significant suppression at the concentration of 100 μg/mL for 72 h compared with negative control group (P<0.05). The apoptosis rate was significantly upregulated than that in the negative control group (P<0.05). The protein expression of Bcl2 decreased while Bax and Cleaved-Caspae3 increased via FAS/FADDD and P53 axis. 【Conclusion】 FOD can significantly inhibit the proliferation and promote the apoptosis of CD133+-Huh7.
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【Objective】 To investigate the effect of total flavone of oldenlandia diffusa(FOD) on the stemness, proliferation and apoptosis of breast cancer(BC) stem cells sorted from MDA-MB-231. 【Methods】 Human BC cell lines MDA-MB-231 was cultured in vitro; MDA-MB-231 was stimulated by different concentrations(0 μg/mL, 100 μg/mL, 200 μg/mL and 400 μg/mL) of FOD for different time (24 h, 48 h and 72 h). CCK8 and plate cell cloning assay were used to detect the effect of FOD on MDA-MB-231 proliferation; CD44+/CD24-MDA-MB-231 cell line were tested by flow cytometry and stem cell markers such as Nanog, Oct4 and Sox2 were tested by Western blotting; Annexin V-PE/7-AAD was used to detect the effect of FOD on MDA-MB-231 apoptosis and Bcl2, cleaved-caspase3 and Bax were tested by Western blotting. 【Results】 Cell proliferation of MDA-MB-231 was significantly inhibited by FOD, with the significant suppression at concentrations of 400 μg/mL for 72 h compared with negative control group(P<0.05). The apoptosis rate was significantly upregulated than the negative control group (P<0.05). The protein expression of Bcl2 decreased while Bax and cleaved-caspae3 increased, and stemness markers such as Nanog, Sox2 and Oct4 decreased in FOD-treated cells. Moverover, Akt-GSK3β-β-catenin axis was inhibited in FOD-treated cells. 【Conclusion】 FOD could significantly inhibit the stemness and proliferation and promote the apoptosis of MDA-MB-231.
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Mutations and polymorphisms of genes associated with pancreatic function are involved in the development and progression of pan-creatitis. In addition,systemic inflammatory response also affects the severity of pancreatitis,while mutations and polymorphisms of genes which encode inflammatory factors and cytokines can regulate the expression of such factors and thus affect the progression of systemic inflam-matory response. This article reviews the association between pancreatitis and polymorphisms of genes related to cationic trypsinogen,pancre-atic secretory trypsin inhibitor,cystic fibrosis conductance regulator,inflammatory factors,and cytokines.
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Objective To analyze the etiology of cerebral ischemic complications occurring in perioperative period of interventional therapy for intracranial aneurysms,and to discuss its prevention and treatment.Methods From January 2011 to March 2015,a total of 1106 patients with intracranial aneurysms underwent interventional therapy at the First Affiliated Hospital of Zhengzhou University,China.Among the 1106 patients,48 patients developed cerebral ischemic complications;their clinical data and the imaging materials were retrospectively analyzed.The factors associated with complications,the effective therapeutic scheme and the clinical preventive methods were discussed.Three months after the treatment,the recovery of neurological function was assessed with modified Rankin Scale (mRS).Results The causes of ischemic complications were,in order of decreasing frequency,cerebral vasospasm/unable elimination of microemboli (n=19),thrombus (n=16),impaired blood flow of adjacent perforating or distal vascular branches caused by embolization (n=6),intra-procedural hypotension (n=4) and cerebral edema (n=3).Combined treatments,including integrated anticoagulation,anti-platelet aggregation,anti-vasospasm,medication of elevating blood pressure,endovascular intervention,etc.were adopted.The patients were followed up for 3 months.Thirty-four patients (70.8％) obtained a mRS of ≤2 (no serious sequelae),and mRS＞2 (poor prognosis) was seen in 14 patients (29.2％).Conclusion The etiology of cerebral ischemia occurring during interventional treatment of intracranial aneurysms is complicated.It is necessary to take useful measures such as individualized anticoagulation,anti-platelet aggregation,anti-vasospasm,etc.during perioperative period of interventional therapy.The procedure-related complications can be effectively treated with interventional therapy and reliably prevented by careful and skilled surgical manipulation.
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ABSTRACT:Objective To investigate the effects of total flavones of oldenlandia diffusa (FOD)on epithelial-mesenchymal transition in hepatocellular cancer cell line MHCC97-H.Methods TGF-β1 induced EMT in routinely cultured liver cancer cell line MHCC97-H;then MHCC97-H cell was divided into 5 groups:normal control group, TGF-β1 group,TGF-β1 + FOD group,TGF-β1 + 5-FU group,and TGF-β1 + FOD + 5-FU group.After 48 h of treatment,the invasion ability of MHCC97-H cell was detected by Transwell;the proteins of E-cadherin and vimentin were determined by Western blot.Results Compared with the normal form of MHCC97-H cell line,the cell had obvious long fusiform after TGF-β1 induction,and the invasion ability enhanced (P = 0.02 ).But after treatment,the invasion ability of MHCC97-H cell decreased in FOD group and 5-FU group compared with that in TGF-β1 group (P = 0.03,P = 0.02 ),and decreased more significantly in FOD + 5-FU group (P = 0.01 ).The expression of E-cadherin at the protein level decreased significantly (P = 0.01 )in TGF-β1 group,which was abolished in FOD group (P =0.03 )and 5-FU group (P = 0.02 ).The expression of vimentin at the protein level increased significantly (P =0.01)in TGF-β1 group,which was abolished in FOD group (P =0.04)and 5-FU group (P =0.03)and more obviously in FOD+5-FU group (P =0.01).Conclusion FOD can reverse the invasion of MHCC97-H cells in EMT induced by TGF-β1 through decreasing the expression of E-cadherin protein and inhibiting the epithelial-mesenchymal transition of MHCC97-H cell.
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ObjectiveTo investigate the effect of intrathecal (IT) Ro25-6981 (a selective NR2B receptor antagonist) on focal cerebral ischemia-reperfusion (I/R) injury in rats.MethodsSixty healthy male SD rats weighing 280-320 g in which IT catheter was successfully implanted without complication were randomly divided into 4 groups ( n ＝ 15 each):group sham operation (group S) ; group focal cerebral I/R(group I/R) ; group I/R + Ro25-6981 (group Ro) and group I/R+ normal saline (group NS).The right middle cerebral artery was occluded for 2 h with a nylon thread with rounded tip which was inserted into right internal carotid artery and advanced cranially until resistance was met in groups I/R,Ro and NS.In group Ro Ro25-6981 100 μg/10 μl was injected IT at 0,2,23 h of reperfusion,while in group NS NS was injected IT instead of Ro25-6981.Neurological dificit was assessed and scored (0 ＝ no deficit,3 ＝ severe deficit) at 24 h of reperfusion.The infarct size was determined by 2,3,5-triphenyl tetrazolium chloride staining.ResultsIT administration of Ro25-6981 significantly reduced cerebral I/R-induced infarct sized and neurological deficit score in group Ro as compared with groups I/R and NS.ConclusionRo25-6981 injected IT can protect the brain against local cerebral I/R injury.
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Aim To investigate the effect of intrathecal injection of fluorocitrate(Fc)on mechanical and thermal hyperalgesia induced by complete Freund's adjuvant(CFA)injection in rats.Methods The mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were measured before and after CFA or Fc treatment.The changes of glial fibrillary acidic protein(GFAP)and OX-42(a microglial marker)expression in the spinal cord dorsal horn were evaluated by immunohistochemistry analysis.Results Rats with CFA-induced arthritis showed mechanical allodynia and thermal hyperalgesia,which was correlated with the increased GFAP and OX-42 expression in the spinal cord dorsal horn.Intrathecal injection of Fc markedly suppressed CFA-induced thermal hyperalgesia and mechanical allodynia.Fc significantly attenuated the activation of GFAP and OX-42 in the spinal cord dorsal horn.Conclusions The glia activation in spinal cord is closely related to the progress of CFA-induced peripheral hyperalgesia.Fc may exert antihyperalgesic effect by inhibiting the activation of astrocyte and microglia.
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Objective To evaluate the role of gliocytes in the spinal cord in the development of inflammatory pain (IP) in rats. Methods Adult male SD rats weighing 180-220 g were used in this experiment. A catheter was implanted in the subarachnoid space according to the method described by Yang. Animals with abnormal motor function of the hindlimb after intrathecal (IT) catheter implantation were excluded. IP was induced by subcutaneous (sc) injection of complete Freund's adjuvant (CFA) 50 μl at the lateral side of the ankle joint of the right hindpaw. Sixty-five rats were randomly divided into 5 groups ( n = 13 each): group I IP control normal saline (NS) 50μl was injected sc instead of CFA; group II IP; group IE PC (IT) + IP control fluorinated citric acid (FC, a gliocyte metabolism inhibitor) 1 nmol/10μl was injected IT at 15 min before NS 50 μl sc injection; group IV NS (IT) + IP and group V FC (IT) + IP. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured 2 d before induction of IP (T_0, baseline) .before and at 2, 4, 6, 8, 10, 12, 24 and 26 h (T_(1-9)) after sc NS or CFA injection. Five enimals in each group were killed at T_5 (8 h after sc NS/CFA injection) and the lumbar segment (L_(4,5)) was removed for determination of glial fibrillary acidic protein ( CFAP) and OX-42 expression by immuno-histochemistry. Results In group Ⅱ and Ⅳ sc CFA significantly decreased MWT and TWL. Mechanical and thermal hyperalgegia induced by sc CFA was significantly suppressed by intrathecal FC in group V . IP significantly increased GFAP and OX-42 expression in the spinal cord. Intrathecal FC significantly attenuated IP-induced up-regulation of GFAP and OX-42 expression in the spinal cord. Conclusion The activation of gliocytes in the spinal cord is involved in the development of CFA-induced hyperalgesia in rats.
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Objective To investigate the effect of propentofylline on nerve growth factor (NGF) and IL-1βrelease from rat cerebral cortical astrocytes. Methods Primary cultured rat astrocytes from SD rats (1-3 d,weighing 6-8 g) after 4 passages were randomly divided into 8 groups ( n = 6 wells each): group Ⅰ control (group C); group Ⅱ , Ⅲ, Ⅳ the astrocytes were exposed to propentofylline 10, 100 and 1000 μmol/L respectively (group P1, P2, P3 ); group Ⅴ the astrocytes were exposed to LPS 1 μg/ml and group Ⅵ, Ⅶ, Ⅷ the astrocytes were exposed to propentofylline 10, 100 and 1000 μmol/L in addition to LPS 1 μg/ml (group P1 + LPS, P2 + LPS,P3 + LPS). The astrocytes were then incubated for 3 days in all 8 groups. The concentrations of IL-1β and NGF in the supernatant were detected at 1 and 3 days of incubation using ELISA. Results LPS activated astrocytes resulting in decrease in NGF release and increase in IL-1β release. Propentofylline significantly increased NGF release and decreased IL-1β release from astrocytes incubated alone or with LPS by suppressing activation of astrocytes. Conclusion Propentofylline can enhance NGF release and inhibit IL-1β release from rat cerebral cortical astrocytes.
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Objective To investigate the role of P2Y1 purinergic receptors in the spinal cord in a rat model of bone cancer pain. Methods Ninety female SD rats weighing 150-180 g were randomly divided into 5 groups (n = 18 each): Ⅰ group sham operation; Ⅱ group bone cancer pain; Ⅲ group sham operation + MRS2179 (a specific P2Y1 purinergic receptor antagonist); Ⅳ group BCP + vehicle (group NS); Ⅴ group BCP+ MRS2179.Bone cancer pain was induced by inoculating Walker 256 mammary gland carcinoma cells into medullary cavity of tibia. In group Ⅲ, Ⅳ, Ⅴ MRS2179 100 pmol/10 μl or NS 10 μl was injected intrathecally once a day for 3 days starting from the 7th day after operation. Mechanical pain threshold to von Frey stimuli was measured before and every other day after operation. The anirnals were sacrificed on the 9th day after operation. The L4-6 segment of the spinal cord was removed for detection of expression of P2Y1 receptor and phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in the spinal dorsal horn. Results P2Y1 receptors and p-ERK1/2 coexisted in spinal dorsal horn. Inoculation of cancer cells into tibia significantly decreased mechanical pain threshold at postoperative day 6-18 and increased the expression of P2Y1 receptor and p-ERK1/2 on the 9th day after operation in group Ⅱ and Ⅳ as compared with group Ⅰ . Intrathecal MRS 2179 significantly increased pain threshold and decreased expression of P2Y1 receptor and p-ERK1/2 in group Ⅴ compared with group Ⅱ and Ⅳ. Conclusion P2Y1 receptors in the spinal cord are involved in the development of bone cancer pain, which may be related to the activation of ERK1/2.
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Objective: To establish the standard for the quality control of Dougenguanshitong granule.Methods: The TCL method was used to identify the Radix Ootoginseng and Dioscorea Bulbifera in the granule and the content of matrine in the granule was determined by the HPLC method.Results: The linear relationship of matrine was the range of 0.68-10.2?g(r=0.9997,n=6).The average recovery was 100.23% and RSD was 1.54%(n=6).Conclusion:The method is convenient,rapid,accurate and suitable for the quality control of Dougenguanshitong granule.
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OBJECTIVE：Probe into the feasibility of preparation of diclofenac sodium with β-cyclodextrins in 1∶1 and 2∶1 inclusion compounds ,and find out the affection in solubility of diclofenac sodium. METHODS:The 1∶1 and 2∶1 melusion compounds were prepared by liquid-phase an alysis the content and determination them solubility.RESULT:The 1 ∶1 and 2∶1 inclusion compounds were prepared by different ratio of reactants and etermined them mp were 299～300.5℃ and 302～303℃ respectively. CONCLUSION：Of us is that solubilits of the 1∶1 and 2∶1 inclusion compounds increase 0.65 and 1 times respetively comparing with declofenac sodium.
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Objective:To evaluate the psychosomatic effects of relaxation and the correlation between the effects and the personality and behavior pattern of the subject.Methods:68 medical students were involved in the study. All the subjects received relaxation training for 2 months. They were tested with Self-Rating Anxiety Scale (SAS) and Pittsburgh Sleep Quality Index (PSQI) before and at the end of the study. They were also tested electroencephalogram (EEG), electromyography (EMG) and skin temperature at the above two time-points. The correlation between the personality and the extents of alteration in index of EEG were also studied.Results:After training, the scores of SAS and PSQI in the subjects were significantly lower than those before training. The indexes of EEG, EMG and skin temperature were improved. The extents of alteration in index of EEG were correlated positively with the N-scores of Eysenck Personality Questionnaire (EPQ), the total scores and TH-scores of Type A Behavior Pattern (TABP).Conclusion:Relaxation can improve psychosomatic states of undergraduates. The extents of alteration in index of EEG were correlated with the personality and behavior pattern of the subject.