Efficacy and Safety of Teriflunomide in Chinese Patients with Relapsing Forms of Multiple Sclerosis: A Subgroup Analysis of the Phase 3 TOWER Study / 中华医学杂志(英文版)

Background@#Disease-modifying therapy is the standard treatment for patients with multiple sclerosis (MS) in remission. The primary objective of the current analysis was to assess the efficacy and safety of two teriflunomide doses (7 mg and 14 mg) in the subgroup of Chinese patients with relapsing MS included in the TOWER study.@*Methods@#TOWER was a multicenter, multinational, randomized, double-blind, parallel-group (three groups), placebo-controlled study. This subgroup analysis includes 148 Chinese patients randomized to receive either teriflunomide 7 mg (n = 51), teriflunomide 14 mg (n = 43), or placebo (n = 54).@*Results@#Of the 148 patients in the intent-to-treat population, adjusted annualized relapse rates were 0.63 (95% confidence interval [CI]: 0.44, 0.92) in the placebo group, 0.48 (95% CI: 0.33, 0.70) in the teriflunomide 7 mg group, and 0.18 (95% CI: 0.09, 0.36) in the teriflunomide 14 mg group; this corresponded to a significant relative risk reduction in the teriflunomide 14 mg group versus placebo (-71.2%, P = 0.0012). Teriflunomide 14 mg also tended to reduce 12-week confirmed disability worsening by 68.1% compared with placebo (hazard ratio: 0.319, P = 0.1194). There were no differences across all treatment groups in the proportion of patients with treatment-emergent adverse events (TEAEs; 72.2% in the placebo group, 74.5% in the teriflunomide 7 mg group, and 69.8% in the teriflunomide 14 mg group); corresponding proportions for serious adverse events were 11.1%, 3.9%, and 11.6%, respectively. The most frequently reported TEAEs with teriflunomide versus placebo were neutropenia, increased alanine aminotransferase, and hair thinning.@*Conclusions@#Teriflunomide was as effective and safe in the Chinese subpopulation as it was in the overall population of patients in the TOWER trial. Teriflunomide has the potential to meet unmet medical needs for MS patients in China.@*Trial Registration@#ClinicalTrials.gov, NCT00751881; https://clinicaltrials.gov/ct2/show/NCT00751881?term=NCT00751881&rank=1.

Effects of nerve growth factor-polybutylcyanoacrylate-nanoparticles on in vitro cellular models of Alzheimer's disease / 中华神经医学杂志

Objective To evaluate the effects of nerve growth factor (NGF)polybutylcyanoacrylate (PBCA)- nanoparticles (NP) on PC12 cells induced by Aβ1-40 Methods The emulsion polymerization method was used to prepare the NGF-PBCA-NP. Morphology of the NGF-PBCA-NP was observed by transmission electron microscopy. The entrapment efficiency and loading efficiency of NGF-PBCA-NP were measured by means of ultraviolet spectrophotometry. The effects of NGF-PBCA-NP on proliferation and cell apoptosis of β1-40-PC12 cells were observed by MTT and FCM methods. Results The NGF-PBCA-NPs were uniform spheres with drug entrapment efficiency of 80.87％ and loading efficiency of 19.88％.Compared with the cells in the NGF group,cells in the NGF-PBCA-NP group exhibited a significantly better proliferation rate and a significantly lower apoptosis rate (P＜0.05). Conclusion NGF-PBCA-NPs obtained by emulsion polymerization method are uniform in morphology and possess a high drug entrapment efficiency and a high loading efficiency.They can restrain significantly the in vitro growth of Aβ1-40-PC12.

Amyloid beta-protein induces apoptosis of neurons in rat hippocampus and expression changes of cyclophilin A in these neurons / 中华神经医学杂志

Objective To explore the neuron injury in rat hippocampus induced by Aβ25-35 and the cyclophilin A (CyPA) expression changes in these neurons. Methods Sixty healthy Wister rats were equally randomized into experimental group and control group (n=30); AD rat models in the experimental group were established by injection of Aβ25-35 into the bilateral hippocampus of rats,and rats of the control group were received NS injection. The morphological features of neurons in the CA1 area of hippocampus were observed by HE staining; the neuron apoptosis was determined with TUNEL staining; the mRNA and protein expressions of CyPA were detected by PT-PCR and Western blotting,respectively. Results Aβ25-35 caused damage and apoptosis of neurons in the CA1 area of hippocampus; with time being prolonged,the cell injury aggravated and apoptosis increased in the CA1 area ofhippocampus; significant differences were noted as compared with those in control group 1,7 and 14 d after the inducement (P＜0.05).After injection of Aβ25-35 into the hippocampus of rat,the mRNA and protein expressions of CyPA were obviously changed:in early stage,the expressions increased,and then,the expressions decreased gradually; significant differences were noted as compared with those in control group 1 and 7 d after the inducement (P＜0.05); the protein expression of CyPA in the experimental group 14 d after the inducement was significantly decreased as compared with that in the control group (P＜0.05). Conclusion Aβ25-35 plays a neurotoxicity role through aggravating the apoptosis of neurons; and the increment of CyPA expressions maybe play an endogenously protective role in these damage.

Effects of cyclophilin A on apoptosis of PC12 cells induced by Aβ25-35 and its mechanism / 中华神经医学杂志

Objective To explore the effect of cyclophilin A (CyPA) on apoptosis of PC 12 cells induced by Aβ25-35 and its potential mechanism. Methods PC 12 cells were divided into normal control group (0 μmol/L Aβ25-35), Aβ25-35 inducement group (10 μmol/L Aβ25-35) and drug protection groups (0.1, 1,10 and 100 nmol/L CyPA+10 μmol/L Aβ25-35). Cells in the drug protection groups were pretreated by CyPA of different concentrations for 30 min, and then co-cultured with Aβ25-35 We evaluated the survival rate of PC12 cells with MTT assay, analyzed the apoptosis of PC12 cells with Hoechst33258 staining, and detected the mRNA expressions of Bcl-2 and Bax with PT-PCR and the protein levels of Bcl-2 and Bax with Western blotting. Results Cells pretreated wth CyPA of 1, 10 and 100 nmol/L enjoyed an obvious elevation of survival rate of PC 12 cells, a significant reduction of apoptosis induced by Aβ25-35,an obvious increase of mRNA expression of Bcl-2 and protein level of Bcl-2, and a statistical decrease of mRNA expression of Bax and protein level of Bax as compared with those cells of the Aβ25-35 inducement group (P<0.05);and these effects were dose-dependent. Conclusion CyPA could resist the toxic role of Aβ25-35 on PC 12 cells and reduce the apoptosis in a dose-dependent manner by up-regulation of anti-apoptosis gene Bcl-2 and down-regulation of apoptosis gene Bax.

Effect of protein kinase B on the learning and memory functions of mice with chronic lead exposure / 中华神经医学杂志

Objective To assess the effect of protein kinase B(PKB)on the learning and memory functions of mice with chronic lead exposure. Methotis Since the first day after birth,50 Kunming mice were randomized into 5 groups(n=10)and exposed to a 6-week acetic lead exposure at the concentrations of 1.2,2.4,4.8,7.2,and 9.6 mmol/L administered in the drinking water,with another 10 having normal water as the control group.After the 6-week exposure.water maze test Was performed to assess the spatial learning and memory abilities of the mice.The total PKB(t-PKB) and phosphorylated PKB(p-PKB)in the cortical brain neurons were determined with Western bloRing.Results The mean latency of finding the platform in the lead exposure groups was significantly higher than that in the control group(P＜0.05).Chronic acetic lead exposure resulted in obviously impaired spatial learning and memory ability in the mice,and the severity ofthe damage was positively correlated to lead concentrations in the blood and brain tissue(r=678,P=0.000;r=0.643,P=0.000).Lead exposure caused a significant dose-dependent reduction in P-PKB level in the cortical neurons,and p-PKB level in the cortical neurons was inversely correlated to blood lead concentrations(r=0.820,P=0.028) and to the spatial learning and memory ability(r=-0.671,p=000).The level of t-PKB in the cortical brain neurons showed no significant changes in response to lead exposure. Conclusion Chronic acetic lead exposure can impair the learning and memory functions of mice possibly in association with reduced content of p-PKB in the cortical brain neurons.

Effect of conditioned medium from coculture of astrocytes and β-amyloid-induced apoptotic PC12 cells on the differentiation of neural stem cells and the possible mechanism / 中华神经医学杂志

Objective To observe the effect of the conditioned medium obtained from the coculture of astrocytes and β-amyloid (Aβ1-40)-induced apoptotic PC12 cells on the neuronal differentiation efficiency of neural stem cells (NSCs), and investigate the possible involvement of the neurotrophins proteins including brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF)and neurotrophin-3 (NT-3) in the induced differentiation of the NSCs. Methods PCI2 cells wereinduced by Aβ1-40 for different time lengths (0, 4, 6, 12, and 24 h), and after detection of the apoptotic rates using flow cytometry, the cells were cocultured with astrocytes for 2 days. The astrocyte-conditioned medium (ACM), after analysis of the neurotrophins proteins using erLzyme-linked immunosorbent assay (ELISA), was mixed with DMEM/F12 medium at the proportion of 1:3 to induce the differentiation of the NSCs. The cell differentiation was identified by laser confocal microscopy, NSE immunofluorescent labeling, and neuronal counting. Results Flow cytometry showed that induction by Aβ1-40 for 6 h resulted in the highest apoptosis rate of PC12 cells (P<0.05). BDNF content in the ACM derived from the coculture of astrocytes and the PC12 cells induced for 6 h was significantly increased, and the NSCs induced in this ACM showed the highest neuronal differentiation rates, showing significant difference from those induced by other ACMs (P<0.05). Conclusion The ACM derived from the coculture of Aβ1-40-induced PC12 cells and astrocytes can increase the neuronal differentiation rates of NSCs in vitro, and BDNF in the ACM may play a role in this process.

Protective effect of cyclophilin A against Alzheimer's amyloid beta-peptide (25-35)-induced oxidative stress in PC12 cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>beta-amyloid peptide (Abeta) is considered responsible for the pathogenesis of Alzheimer's disease (AD). Possible mechanisms underlying Abeta-induced neuronal cytotoxicity include excessive production of reactive oxidative species (ROS) and apoptosis. Cyclophilin A (CypA), exhibits antioxidant properties and protects neurons against oxidative stress induced injury. This study was conducted to demonstrate whether CyPA added to cultured PC12 cells could alleviate Abeta-induced oxidative stress and protect them from apoptosis.</p><p><b>METHODS</b>PC12 cells were pre-incubated for 30 minutes with recombinant human cyclophilin A (rhCyPA) in 0.1 nmol/L, 1.0 nmol/L, 10 nmol/L and 100 nmol/L and then incubated with 10 micromol/L Abeta(25-35). In every group, cell viability, apoptotic morphology, apoptotic rate, intracellular ROS accumulation, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of PC12 cells and mitochondrial transmembrane potential were detected. Subsequently, the expression of the active form of caspase-3 was determined by Western blotting.</p><p><b>RESULTS</b>It was shown that cultures treated with 1.0 nmol/L, 10 nmol/L or 100 nmol/L rhCyPA + Abeta(25-35) had significantly higher cell viability and a lower rate of apoptosis compared with the cultures exposed only to Abeta(25-35). In addition, rhCyPA attenuated Abeta(25-35)-induced overproduction of intracellular ROS and Abeta(25-35)-induced a decrease in activity of the key antioxidant enzymes SOD and GSH-Px. Furthermore, rhCyPA also attenuated Abeta(25-35)-induced mitochondrial dysfunction and the activation of caspase-3.</p><p><b>CONCLUSION</b>CyPA may act as an ROS scavenger, and prevent Abeta(25-35)-induced neurotoxicity through attenuating oxidative stress induced by Abeta(25-35).</p>

Exercise test on the patients with normokalaemic periodic paralysis from a Chinese family with a mutation in the SCN4A gene / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Normokalaemic periodic paralysis (normoKPP) is characterized by transient and recurrent myoasthenia, and some patients also show muscle stiffness induced by cold exposure (paramyotonia congenita, PMC). It is caused by a mutation in the muscle voltage gated sodium channel alpha subunit (SCN4A) gene. Due to the diversity of the clinical manifestations of patients, it is difficult for clinicians to differentiate some of patients with atypical normoKPP from those who suffer from other periodic paralysis and nondystrophic myotonia. So far, for normoKPP there are almost no ways to assist definite diagnosis besides genetic screening. This research was designed to evaluate an exercise test (ET) in confirming the diagnosis of normoKPP and in assessing the therapeutic effectiveness of some drugs on this disease.</p><p><b>METHODS</b>ET, described by McMains, was performed on six subjects from a Chinese family, including four patients with overlapping disease of normoKPP and PMC caused by a mutation of SCN4A Met1592Val that is identified by genetic analysis and two normal control members. The change of compound muscle action potential (CMAP) was recorded. Besides the family, two patients were also tested during treatments with acetazolamide.</p><p><b>RESULTS</b>All patients showed a slight increase in CMAP immediately after exercise, followed by an abnormal gradual decline, which reached its nadir 25-30 minutes after exercise. CMAP amplitude dropped by more than 40% in patients but less than 23% in controls. In the patients who received treatment with acetazolamide, the change of CMAP amplitude was less than 28% and, at any fixed times, less than pretreatment values.</p><p><b>CONCLUSIONS</b>The ET may be used as a predictive, easy and reliable method of diagnosing normoKPP under conditions without genetic screening help, and is an objective way to evaluate the therapeutic effectiveness. According to different response patterns, the ET may also be helpful in reducing the scope of genetic screening.</p>

Beneficial effects of BV2 cell on proliferation and neuron-differentiating of mesenchymal stem cells in the circumstance of injured PC12 cell supernatant / 神经科学通报·英文版

Objective The microglias is the representative of immune cells in the brain. It plays dual roles of both repairing and damaging in injured nervous system, and works as an inevitable component of the circumstance of injured neurons. This study was aiming at the effects of the microglias on the biological activities of mesenchymal stem cells (MSCs) in the circumstance of injured neurons. Methods MSCs were obtained by primary culture. We adopted PC12 cells (PC12) and BV2 cells (BV2) to substitute for neurons and microglias, respectively. PC12 were injured by aged Abeta(1-40) and the supernatant of the injured PC12 was used to set up the circumstance of injured neurons. Transwells were used for co-culture of BV2 and MSCs, which allowed the independent detection of cells after co-culture. Immunofluorescence was used to identify MSCs and neuron-differentiating cells with CD44 and neuron specific enolase (NSE) staining, respectively. MTT assay was adopted to measure the proliferation. Results In the circumstance of both BV2 presence and injured PC12 supernatant incubation, either the proliferation or the differentiation of MSCs reached the highest, which seemed to be contradictory, but we gave our explanations. With the BV2 co-culture, the proliferation of MSCs tend to be higher, but the neuron-differentiating MSCs were similar to those incubated without BV2 co-culture either in normal or injured in PC12 supernatant. With the incubation of injured PC12 supernatant, the neuron-differentiating cells were significantly higher than that of control (P < 0.05). Conclusion In the circumstance of injured neurons, microlgias tend to promote the MSCs proliferation. Although not helpful in neuron-differentiating, microglias did not exert any negative effect either.

Clinical research of curative effect and safety of deproteinised calf serum enteric capsules on nerve function defect caused by cerebral ischemia damage / 中华神经科杂志

Objective To observe the clinical curative effect and security of the deproteinised calf serum enteric capsules on nerve functions defect caused by cerebral isehemia damage.Methods A randomized,double-blind,multicentric trial design was conducted.Treatment group:based on routine treatment,the patients were given deproteinised calf serum enteric capsules for 90 days.Control group: based on routine treatment,the patients were given placebo for 90 days.The NIHSS,BI,mRS measuring scale were used for evaluation on the 7th,14th and 28th day before and after treatment.BI and mRS measuring scale were used for evaluation on the 90th day.The change of numerical value was inspected before and after treatment.Results Before treatment,there was no significant difference in the NIHSS,BI and mRS measuring scale evaluation between the groups of treatment and control.After treatment,there were significant differences in the NIHSS,BI and mRS measuring scale evaluation between the groups of treatment and control on the 14th and 28th day (P

Effect of Batroxobin on focal cerebral ischemia and reperfusion injury in streptozotocin-induced diabetic rats / 中国康复理论与实践

@#ObjectiveTo observe the effect of Batroxobin and Urokinase on brain of diabetic rats following focal cerebral ischemia and reperfusion injury. To investigate the preventive mechanism of Batroxobin following focal cerebral ischemia and reperfusion injury in diabetic rats after thrombolysis therapy.MethodsDiabetic rat was induced by administrating streptozotocin intraperitoneally. Rat model of focal cerebral ischemia and reperfusion injury induced by intraluminal filament occlusion of middle cerebral artery(MCA) that removed 2h later was used. Batroxobin and Urokinase were administrated intravenously in different groups. Infarct volume,cerebral hemorrhage and matrix metalloproteinase (MMP)-2,MMP-9 were detected at 2h,24h,48h after ischemia and reperfusion injury.ResultsThe significant decrease of infarct volume were observed in Batroxobin and Urokinase groups. There were 5 rats observed cerebral hemorrhage in Urokinase group and no cerebral hemorrhage in Batroxobin group. The number of MMP-2 and MMP-9 positive cells in Batroxobin and Batroxobin Urokinase groups decreased compared with saline and Urokinase groups. ConclusionBatroxobin can decrease the infarct volume significantly without the complication of cerebral hemorrhage after ischemia and reperfusion injury in diabetic rats, which maybe relate to down regulation of the expression of MMP-2 and MMP-9.

Effect of hyperbric oxygen on infarct volume and matrix metalloproteinase after permanent focal cerebral ischemia in adult rats / 中国康复理论与实践

@#ObjectiveTo investigate the effect of hyperbric oxygen (HBO) on infarct volume and relevant mechanism after permanent focal cerebral ischemia in adult rats.MethodsRat model of focal cerebral ischemia induced by intraluminal filament occlusion of middle cerebral artery (MCA) was used. HBO(2.0 ATA) was applied to HBO group. Infarct volume, matrix metalloproteinase 2 (MMP-2) and MMP-9 were detected at 6h, 24h, 48h, 72h,120h and 10d after ischemia.ResultsThe infarct volume obviously decreased at 120h and 10d and expression of MMP-9 lowered at 48—120h in HBO groups. There was no significant change in MMP-2.Conclusion HBO can reduce infarct volume after cerebral ischemia, which may be related to downregulation of MMP-9 levels.