BACKGROUND: Basic fibroblast growth factor (bFGF) possesses multiple functions such as promoting neuronal survival and growth of cell processes in vitro and antagonizing the toxicity of excitatory amino acids,thereby playing import roles in functional recovery of the central nervous system (CNS). But whether bFGF offers neuroprotection on ischemic brain tissues by modulating intracellular free calcium content remains unknown.OBJECTIVE: To explore the effect of bFGF on intracellular free Ca2+ in the neural cells in the event of focal cerebral ischemia-reperfusion (IR)injury.DESIGN: Randomized controlled study.SETTING: Department of Neurology of Second Hospital Affiliated to Zhengzhou University.MATERIALS: This study was conducted in the Laboratory of the Department of Neurology, Second Hospital Affiliated to Zhengzhou University between August and December 2003. Totally 24 SD were randomized into sham operation group, ischemic group, IR group and bFGF exposure group with 8 rats in each group.METHODS: Middle cerebral artery occlusion (MCAO) model was established in rats in IR group and bFGF exposure group by inducing arterial thrombosis with thread, which was not preformed in rats in the sham operation group. Rats in bFGF exposure group received intraperitoneal injection of 10 μg/kg bFGF immediately after ischemia,which was replaced by the same volume of physical saline in the other two groups. Free Ca2+ in brain cells was detected at 24 hours of IR.MAIN OUTCOME MEASURES: Free Ca2+ in the brain cells at 24hours of IR.RESULTS: All the 24 rats survived the experiment. Free Ca2+ in IR group was significantly higher than that of the sham operation group [(673.46±18.44) vs (224.71±10.58) nmol/L, F=1 329.06, P ＜ 0.01], and also significantly higher in bFGF exposure group [(378.37±21.08) nmol/L,F=1 329.06, P ＜ 0.01].CONCLUSION: Intracellular free calcium can be obviously depressed by bFGF following IR injury, which benefits cell membrane stability and help prevent intracellular Ca2+ overload.

BACKGROUND:Increasing evidence has shown that lovastatin with less toxicity to normal cels has crucial effects on proliferation, apoptosis and differentiation of various cancer cels. However, its roles in glioma stem cels remain unclear.
OBJECTIVE:To explore the effect of lovastatin on proliferation and apoptosis of glioma stem cels.
METHODS: Flow cytometric sorting was used to separate glioma stem cels from human glioblastoma cel line U87. Effects of lovastatin on the proliferation and apoptosis of glioma stem cels were determined by MTT and flow cytometry, respectively. Furthermore, expression levels of Ki67, Bax and Bcl-2 in glioma stem cels treated with lovastatin were detected using western blot analysis.
RESULTS AND CONCLUSION: The CD133-positive glioma stem cels were sorted from human glioblastoma cel line U87 with a positive percentage of 85%. MTT assay showed that lovastatin inhibited the proliferation of glioma stem cels in dose (5, 10, 20 μmol/L)- and time (24, 48, 72, 96 hours)-dependent manners. Flow cytometry analysis showed that 10 μmol/L lovastatin (48 hours) induced apoptosis in glioma stem cels. In addition, the expression level of Ki67 was decreased by lovastatin treatment in a dose-dependent manner, and the Bcl-2 and Bax expression levels were reduced and increased by 10 μmol/L lovastatin treatment, respectively. In conclusion, lovastatin can inhibit cel proliferation and induce apoptosis of glioma stem cels, and lovastatin may be a potential drug for treatment of brain tumors.

BACKGROUND:Several studies have reported that quercetin can inhibit the proliferation and migration but promotes apoptosis of tumor cel s. OBJECTIVE:To explore the effect of quercetin on the proliferation and apoptosis of glioma stem cel s and the signal pathway involved. METHODS:Glioma stem cel s were isolated by immunomagnetic beads and treated in culture medium containingdifferent concentrations of quercetin (0, 25, 50, 100μmol/L). Cel proliferation was measured by MTT and apoptosis measured by flow cytometry at 48 hours after culture. The expression levels of Bcl-2, Bax and survivin, which are related to apoptosis, were detected by western blot. The expression of proliferating cel nuclear antigen (PCNA), which is related to proliferation, was also detected by western blot. The expression of STAT3 and p-STAT3 was also determined by western blot. RESULTS AND CONCLUSION:(1) Compared with the control (0μmol/L) group, quercetin inhibited the proliferation of glioma stem cel s in a dose-depended manner (P<0.05). With the increase of the concentration of quercetin, the expression of PCNA was increased (P<0.05). (2) Quercetin induced apoptosis of glioma stem cel s dose-dependently (P<0.05). With the increase of the concentration of quercetin, the expression of Bcl-2 and survivin was decreased, while the expression Bax was increased. (3) Quercetin could also inhibit phosphorylation of STAT3 dose-dependently, but the level of STAT3 was not changed. To conclude, these results show that quercetin could inhibit the proliferation of glioma stem cel s and promote apoptosis via the STAT pathway.

Objective To explore the expression of microRNA506 in brain tissue of mice with cere-bral ischemia reperfusion injury and its effect on proliferation and apoptosis of SH-SY5Y cells. Method-s Middle cerebral artery occlusion(MCAO)was used to establish the model of cerebral ischemia reperfu-sion in mice,and RT-PCR(reverse transcription polymerase chain reaction)was used to detect the expres-sion level of microRNA506 in the brain tissue of mice. MicroRNA506 overexpression plasmids and interfer-ence plasmids were constructed and transfected into SH-SY5Y cells respectively.Cell apoptosis was detected by flow cytometry,and cell proliferation was detected by MTT assay. Results The level of microRNA506 in brain tissue of cerebral ischemia reperfusion model mice was significantly higher than that of sham operation group((51.16±1.64)vs(12.82±1.66),P=0.008). The siRNA targeting microRNA506 significantly re-duced the growth of SH-SY5Y cells(P=0.0072),and induced the apoptosis of SH-SY5Y cells(P=0.0073). Transfection of microRNA506 enhanced the proliferation rate(P=0.020)of SH-SY5Y cells,and reduced the apoptosis of SH-SY5Y cells(P=0.017).Conclusions The expression level of microRNA506 in brain tissue of mice with cerebral ischemia reperfusion is increased.Overexpression of microRNA506 promotes the cell proliferation and decreases the apoptosis rate of SH-SY5Y cell while the low expression of microRNA506 de-creases the growth rate and induce cell apoptosis of SH-SY5Y cell.

Objective:To investigate the effects of ketogenic diet on seizures, electroencephalogram(EEG) changes and neurobehavioral development in children diagnosed with epilepsy.Methods:A total of 122 children diagnosed with spastic epilepsy in Nanyang Central Hospital from March 2016 to March 2019 were enrolled.The patients were divided into the observation group and the control group, by the computerized random number table method with 61 cases in each group.The children in the control group were treated with conventional therapy, and the children in the observation group were combined with the ketogenic diet on the basis of conventional treatment.The Gesell developmental schedules scale scores were compared between the two groups to evaluate seizure control and EEG improvement.Results:The seizure control in the observation group was significantly better than that in the control group [78.69%(48/61 cases) vs.54.10%(33/61 cases)], and the difference was statistically significant ( χ2 = 12.114, P <0.05). The EEG improvement in the observation group was significantly better than that in the control group [81.97%(50/61 cases) vs.55.74%(34/61 cases)], and the difference was statistically significant ( χ2=13.623, P<0.05). After 12 months of treatment, the children in the observation group had significantly higher fitness, gross motor, fine motor, language, and personal social, and total development quotient scores than the control group [(56.64±13.29) scores vs.(46.04±12.86) scores, (54.84±12.18) scores vs.(47.62±11.91) scores, (54.44±10.70) scores vs.(44.31±11.56) scores, (51.48±12.99) scores vs.(42.04±11.18) scores, (57.88±11.04) scores vs.(47.42±13.16) scores, (275.28±54.71) scores vs.(227.42±55.79) scores], the differences were statistically significant ( t=5.997, 5.887, 6.003, 5.889, 6.007, 6.010, all P<0.05). Conclusion:The ketogenic diet can significantly reduce seizures, improve EEG and neurobehavioral development in children with epilepsy.

Objective To investigate the effects of Atorvastatin on reducing the levels of inflammatory factors and on improving the quality of life in elderly patients with chronic kidney disease complicated with hyperlipidemia.Methods One hundred patients with chronic kidney disease accompanied with hyperlipidemia were enrolled from March 2016 to March 2017,and randomized into two groups:a control group undergoing conventional treatment with anticoagulant,diet control and standard glucocorticosteroid,and an observation group receiving Atorvastatin as add-on therapy to conventional treatment.Improvements of serum levels of inflammatory factors and blood lipid indices as well as total curative effect were compared between the two groups.Results The total effective rate was significantly higher in the observation group[94.0％ (47/50)]than in the control group (76.0％,38/50)(x2 =6.352,P＜0.05).After treatment,the improvements of blood lipid indices(TC,TG,LDL-C,HDL-C)were more in the observation group than in the control group(t =71.804,10.544,15.266,3.499,all P＜0.05).Before treatment,the serum levels of TNF-alpha,CRP and the 24h urine protein had no statistically significant differences between control and observation groups(P =0.406 or 0.678,respectively),but the levels in post-vs.pre-treatment were statistically significant lower in TNF-alpha[(1.1 ± 0.2) vs.(1.8 ± 0.3) μg/L],CRP [(8.2 ± 0.1) vs.(9.3 ± 0.1) mg/L] and 24h urine protein[(1.1±0.7)vs.(2.3±1.1)g/24 h](all P＜0.05).Conclusions Atorvastatin as add-on treatment to conventional therapy of chronic kidney diseases complicated with hyperlipidemia in the elderly has significant effects on controlling dyslipidemia and reducing inflammatory reactions so as to improve patients' quality of life.

Objective:To analyze the clinical effect of TREVO stent thrombectomy combined with tirofiban on patients with acute large-artery occlusion of the anterior circulation.Methods:Seventy-two patients with acute large-artery occlusion of the anterior circulation accepted thrombectomy in our hospital from November 2016 to May 2020 were divided into two groups according to different treatment methods: 35 patients in the control group were treated with TREVO stent thrombectomy, and 37 patients in the treatment group were treated with TREVO stent thrombectomy combined with tirofiban via intra-variceal injection. The success rate of recanalization, specific conditions of thrombolysis, improvement degrees of nerve defect, coagulation function, prognoses 90 d after thrombectomy, and complications were compared between the two groups.Results:The success rate of postoperative vascular recanalization in the treatment group and control group was 91.89% (34/37) and 88.57% (31/35), respectively, without statistically significant difference ( P>0.05). The time and times of thrombotomy in the treatment group were significantly shorter/smaller than those in the control group (P<0.05). The National Institute of Health stroke scale (NIHSS) scores of patients from the treatment group 14 d after thrombectomy were significantly lower than those of the control group ( P<0.05). The postoperative thrombin time, prothrombin time, and activated partial thrombin time of the treatment group were significantly longer than those of the control group ( P<0.05). The good prognosis rate of patients in the treatment group and control group 90 d after thrombectomy was 86.49% (32/37) and 60.0% (21/35), with significant differences ( P<0.05); and the incidence of complications was 8.11% (3/37) and 14.29% (5/35), without significant differences ( P>0.05). Conclusion:TREVO stent thrombectomy combined with tirofiban has a significant effect on treatment of acute large-artery occlusion of the anterior circulation, enjoying high safety.

Objective:To explore the correlation of plasma kallikrein and bradykinin receptor 1 with prognosis of cerebral infarction in elderly rats after ischemic stroke.Methods:A total of 40 male Sprague-Dawley rats were randomly divided into 4 groups: the stroke model group(intraperitoneally injected with 150 μl 0.9% saline, n=10), the DX-88 group(intravenously injected with kallikrein inhibitor DX-88 30 μl/time, n=10), the R-954 group(intravenously injected with bradykinin B1 receptor antagonist R-954 30 μl/time, n=10)and the DX-88 combined with R-954 treatment group(intravenously injected with DX-88 and R-954, n=10). Protein expression levels of plasma kallikrein and bradykinin receptor 1 were determined by Western blotting.mRNA expression levels of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α)and caspase-3 were analyzed by using qRT-PCR.A neurologic function scale was used to score cerebral nerve injury and calculate the middle cerebral infarction area and cerebral swelling in experimental rats.Cerebral blood-brain barrier permeability was assessed by the cerebral infarction area.Results:Neurological injury scores decreased in the DX-88, R-954 and DX-88 + R-954 groups compared with the stroke model group(5.35±1.35, 6.49±1.16, 4.92±0.92 vs.11.17±2.18, F=15.589, P=0.022). Compared with the stroke model group, the cerebral infarction area was reduced in the DX-88, R-954 and DX-88+ R-954 groups[(4.35±1.05) mm 2, (5.43±0.26) mm 2, (3.88±0.13) mm 2vs.(8.26±1.24) mm 2, F=13.476, P=0.034)]. The extent of brain swelling was smaller in the DX-88, R-954 and DX-88+ R-954 groups than in the stroke model group[(31.28±7.45) %, (35.19±8.57) %, (19.68±3.14) % vs.(74.26±15.66) %, F=16.587, P=0.026)]. Plasma kallikrein protein levels were lower in the DX-88 and DX-88+ R-954 groups than in the stroke model group( P<0.05). The expression of bradykinin-1 receptor mRNA was lower in the R-954 and DX-88+ R-954 groups than in the stroke model group( P<0.05). The above results indicated that antagonism of plasma kallikrein and bradykinin receptor 1 played an important role.mRNA transcription levels of IL-1β, TNF-α and caspase-3 were higher in the stroke model group than in the DX-88, R-954 and DX-88+ R-954 groups( F=12.665、14.574 and 13.665, P=0.021、0.015 and 0.003). Conclusions:Inhibiting plasma kallikrein and bradykinin receptor 1 may provide protection against cerebral nerve injury in cerebral ischemia, and improve the prognosis of cerebral infarction.