Flunarizine and lamotrigine prophylaxis effects on neuron-specific enolase, S-100, and brain-specific creatine kinase in a fetal rat model of hypoxic-ischemic brain damage

BACKGROUND： Calcium antagonists may act as neuroprotectants, diminishing the influx of calcium ions through voltage-sensitive calcium channels. When administered prophylactically, they display neuroprotective effects against hypoxic-ischemic brain damage in newborn rats. OBJECTIVE： To investigate the neuroprotective effects of flunarizine （FNZ）, lamotrigine （LTG） and the combination of both drugs, on hypoxic-ischemic brain damage in fetal rats. DESIGN AND SETTING： This randomized, complete block design was performed at the Department of Pediatrics, Shenzhen Fourth People＇s Hospital, Guangdong Medical College. MATERIALS： Forty pregnant Wistar rats, at gestational day 20, were selected for the experiment and were randomly divided into FNZ, LTG, FNZ ＋ LTG, and model groups, with 10 rats in each group. METHODS： Rats in the FNZ, LTG, and FNZ ＋ LTG groups received intragastric injections of FNZ （0.5 mg/kg/d）, LTG （10 mg/kg/d）, and FNZ （0.5 mg/kg/d） ＋ LTG （10 mg/kg/d）, respectively. Drugs were administered once a day for 3 days prior to induction of hypoxia-ischemia. Rats in the model group were not administered any drugs. Three hours after the final administration, eight pregnant rats from each group underwent model establishment hypoxia-ischemia brain damage to the fetal rats. Cesareans were performed at 6, 12, 24, and 48 hours later; and 5 fetal rats were removed from each mother and kept warm. Two fetuses without model establishment were removed by planned cesarean at the same time and served as controls. A total of 0.3 mL serum was collected from fetal rats at 6, 12, 24, and 48 hours, respectively, following birth. MAIN OUTCOME MEASURES： Serum protein concentrations of neuron-specific enolase and S-100 were measured by ELISA. Serum concentrations of brain-specific creatine kinase were measured using an electrogenerated chemiluminescence method. RESULTS： Serum concentrations of neuron-specific enolase, S-100, and brain-specific creatine kinase were significantly higher in the hypoxic-ischemic fetal rats, compared with the non-hypoxic-ischemic group. Serum concentrations of neuron-specific enolase, S-100, and brain-specific creatine kinase were significantly less in the FNZ, LTG, and FNZ ＋ LTG groups following ischemia, compared with the model group （P 〈 0.01）. However, these values were significantly greater in the FNZ and LTG groups, compared with the FNZ ＋ LTG group, following ischemia （P 〈 0.01）. CONCLUSION： Preventive antenatal use of oral FNZ and LTG has positive neuroprotective effects on intrauterine hypoxic-ischemic brain damage. The combined effect of these two drugs is superior.

Development and validation of a rapid chromatographic method for the analysis of flunarizine and its main production impurities

A rapid selective method for the analysis of flunarizine and its associated impurities was developed and validated according to ICH guidelines. The separation was carried out using a Thermo Scientific Hypersil Gold C18 column （50 mm × 4.6 mm i.d., 1.9 μm particle size） with a gradient mobile phase of acetonitrile-ammonium acetate-tetrabutylammoniumhydrogen sulfate buffer, at a flow rate of 1,8 mL/min and UV detection at 230 nm. Naturally aged samples were also tested to determine sample stability. A profile of sample and impurity breakdown was also presented.

Flunarizine inhibits sensory neuron excitability by blocking voltage-gated Na＋ and Ca2＋ currents in trigeminal ganglion neurons

Background Although flunarizine has been widely used for migraine prophylaxis with clear success, the mechanisms of its actions in migraine prophylaxis are not completely understood. The aim of this study was to investigate the effects of flunarizine on tetrodotoxin-resistant Na＋ channels and high-voltage activated Ca2＋ channels of acutely isolated mouse trigeminal ganglion neurons. Methods Sodium currents and calcium currents in trigeminal ganglion neurons were monitored using whole-cell patch-clamp recordings. Paired Student＇s t test was used as appropriate to evaluate the statistical significance of differences between two group means. Results Both tetrodotoxin-resistant sodium currents and high-voltage activated calcium currents were blocked by flunarizine in a concentration-dependent manner with the concentration producing half-maximal current block values of 2.89 μmol/L and 2.73 μmol/L, respectively. The steady-state inactivation curves of tetrodotoxin-resistant sodium currents and high-voltage activated calcium currents were shifted towards more hyperpolarizing potentials after exposure to flunarizine. Furthermore, the actions of flunarizine in blocking tetrodotoxin-resistant sodium currents and high-voltage activated calcium currents were use-dependent, with effects enhanced at higher rates of channel activation. Conclusion Blockades of these currents might help explain the peripheral mechanism underlying the preventive effect of flunarizine on migraine attacks.

儿童难治性癫外周血P糖蛋白的表达及药物干预

目的测定儿童难治性癫及初诊癫患儿外周血白细胞多药耐药基因产物P糖蛋白(P-gp),建立儿童难治性癫耐药性客观指标及预测儿童难治性癫客观指标;同时观察氟桂利嗪的疗效。方法应用流式细胞仪检测41例难治性癫儿童及45例初诊癫儿童外周血多药耐药基因产物P-gp的表达,44例健康儿为对照组。难治性癫组患儿加用氟桂利嗪(弗瑞林)2.5～5 mg,qn口服治疗,对其疗效进行临床验证。结果难治性癫组P-gp表达阳性23例(56.1%),初诊癫组P-gp表达阳性10例(22.2%),对照组P-gp表达阳性3例(6.8%),三组比较差异有统计学意义(χ2=26.77,P<0.01)。难治性癫组与对照组比较χ2=24.27,P<0.01;难治性癫组与初诊癫组比较χ2=10.41,P<0.01;初诊癫组与对照组比较χ2=4.23,P<0.05。难治性癫组P-gp表达阳性23例中17例(73.9%)无效,6例(26.1%)有效,P-gp表达阴性18例中3例(16.7%)无效,15例(83.3%)有效,两者比较差异有统计学意义(χ2=10.10,P<0.01)。初诊癫组P-gp表达阳性10例中7例(70.0%)转为难治性癫,P-gp表达阴性35例中3例(18.6%)转为难治性癫。两者比较差异有统计学意义(χ2=16.98,P<0.01)。难治性癫组抗癫药物治疗无效者20例加用氟桂利嗪治疗3个月,P-gp表达阳性17例中有效11例(64.7%),11例有效者复查P-gp有6例(54.5%)转为阴性;无效6例(35.3%),复查P-gp表达仍阳性。P-gp表达阴性3例中1例有效,2例无效,复查P-gp表达阴性。结论外周血多药耐药基因产物P-gp在儿童难治性癫中表达增强,可作为儿童难治性癫耐药性的客观指标;P-gp表达阳性的初诊癫患儿多转为难治性癫,P-gp可作为预测儿童难治性癫的客观指标;氟桂利嗪有一定抗癫及逆转P-gp的表达作用。

Alternating hemiplegia of childhood misdiagnosed as hysteria: a case report

Background Alternating hemiplegia of childhood(AHC)is a rare pediatric syndrome characterized by recurring episodes of hemiplegia or quadriplegia,and frequently accompanied by dystonic posturing,choreoathetosis movements,anomalous ocular motions,and a gradual deterioration in cognitive function.The principal etiology of this disorder is traced back to mutations in the ATP1A3 gene.Case presentation Here,we report a 16-year-old girl with recurrent hemiplegia since her infancy.This patient has experienced paroxysmal limb weakness and aphasia for over 15 years,and has kept seeking medical attention but without receiving effective treatment.A misdiagnosis of hysteria persisted for over 4 years until the patient’s admission to our hospital.Whole-exome sequencing identified a known pathogenic heterozygous c.2270T>C(p.Leu757Pro)mutation in her ATP1A3 gene.Notably,her clinical manifestations,including pathological emotional responses and autonomic dysfunction,differed from the established profile associated with the same ATP1A3 mutation,which typically present with intellectual disability,a rostrocaudal symptom gradient,choreoathetosis,and dysarthria.The patient was finally diagnosed with AHC and treated with flunarizine thus significantly ameliorated hemiplegic episodes.Conclusions This case enhances our understanding of the intricate clinical manifestations of AHC,which require careful differentiation from various diseases such as epilepsy,hysteria,and paroxysmal dyskinesias.In the diagnosis of patients presenting with suspected symptoms,adhering to a systematic approach for localizing and diagnosing neurological disorders is crucial to prevent misdiagnosis and inappropriate treatments.Additionally,when AHC is suspected in a patient,genetic testing should be considered as part of the diagnostic approach.

Clinical observation on cervical chiropractic for cervical spondylosis of vertebral artery type

Objective：To observe the curative effect of cervical chiropractic for cervical spondylosis of vertebral artery type （CSA） and to explore its possible mechanism.Methods：Sixty CSA patients were randomly divided into two groups.Thirty patients in the treatment group were treated with chiropractic manipulation for cervical vertebrae,once every other day,7 treatments in total.The other 30 patients in the control group received oral administration of flunarizine hydrochloride,10 mg per night,for 14 d.Cervical vertigo and functional assessment scale and transcranial cerebral Doppler （TCD） were measured before treatment and 7 days after treatment.Results：The total effective rate was 96.7% in the treatment group and 83.3% in the control group.The curative rate was 66.7% in the treatment group and 20% in the control group.The differences between the two groups in the total effective rate and the curative rate were statistically significant （both P〈0.05）.The scores of cervical vertigo symptom and functional assessment after treatment in both groups were significantly higher than those before treatment （both P〈0.01）;the difference between the two groups was statistically significant （P〈0.05）.After treatment,the maximum systolic velocity （Vs）,the maximum diastolic velocity （Vd）,the mean velocity （Vm）,the pulsatility index （PI） and the vascular resistance index （RI） in both groups were significantly improved compared with those before the treatment （all P〈0.01）;there were significant differences between the two groups （all P〈0.05）.Conclusion：Cervical chiropractic is an effective method for CSA,and its curative effect is better than that of flunarizine hydrochloride alone.Its mechanism may relate to correcting cervical instability.

The observation on different effectiveness between the embedding needle therapy and medication in the preventative treatment of chronic migraine

Objective: To evaluate the therapeutic effects of the embedding needle therapy in the preventative treatment of chronic migraine.Methods: A total of 36 patients with chronic migraine were divided into an embedding needle therapy group and a flunarizine group according to random number table, 18 cases in each group. In the embedding needle therapy group, the thumb-tack needles were used at Sizhúkōng(丝竹空TE 23), Hànyàn(颔厌GB 4), Wángǔ(完骨GB 12) and Cuánzhú(攒竹BL 2) and the needles were embedded for 24 h. The embedding needle therapy was given three times a week. In the flunarizine group, flunarizine was prescribed, 10 mg a day. Separately, at the end of the 4-week treatment and in 4 weeks of the follow-up, the number of days of headache attack per month(days/month) and the headache duration(hours/month)in the cases with moderate to severe headache were observed in the two groups.Results: The number of days of headache attack in the cases with moderate to severe headache was reduced after treatment as compared with the number before treatment in the two groups. The headache duration was reduced obviously after treatment in the cases of moderate to severe headache in the two groups. In the 4-week follow-up, all the indexes were still reduced as compared with the indexes before treatment, indicating the significant differences in the two groups(both P < 0.05). In the embedding needle therapy group, at the end of the 4-week treatment and in 4 weeks of the follow-up, the number of days of headache attack per month was(8.1 ± 2.4) days and(7.2 ± 1.6) days separately, which were significant difference as compared with(9.7 ± 2.3) days and(8.9 ± 1.4) days in the flunarizine group(both P < 0.05). Regarding the headache duration, at the end of the 4-week treatment and in 4 weeks of the follow-up, it was(105.6 ± 10.5) h/month and(92.1 ± 7.2) h/month respectively in the embedding needle therapy group, indicating the significant differences as compared with(135.4 ± 8.9) h and(116.1 ± 8.2) h in the flunarizine group respectively(both P < 0.05).Conclusion: The 4-week treatment with the embedding needle therapy obviously reduces the number of days of headache attack and the headache duration, presents the preventative effects in the patients with chronic migraine.

Clinical research of atlantoaxial instability treated with the combination of ulna-tibia needling therapy and“point-to-surface”tuina manipulation

Objective:To observe the clinical therapeutic effect on atlantoaxial instability treated with the combination of ulna-tibia needling therapy and"point-to-surface"tuina manipulation.Methods:A total of 64 outpatients diagnosed as atlantoaxial instability were collected and randomized into a control group and an observation group,32 cases in each one.In the control group,flunarizine hydrochloride capsules were used for oral administration.In the observation group,the combined treatment of ulna-tibia needling therapy and"point-to-surface"tuina manipulation was adopted.The ulnatibia needling therapy was exerted at the cutaneous region of hand taiyang meridian on the ulnar region(from carpal joint to elbow joint)on both sides.The"point-to-surface"tuina manipulation included"onepoint and two-surface"technique,"upper-to-lower pressing"method and"lifting-trembling"method.The treatment was provided once daily,consecutively for 5 times.The scores of neck symptoms and physical signs,atlantoaxial axle separation degree and clinical therapeutic results were taken as the observation indicators to evaluate the treatment effect.Results:After treatment,the curative rate and the total effective rate in the observation group were all higher significantly than the control group(all P<0.01).The scores of clinical symptoms and physical signs were statistically different in the self-comparison of each group before and after treatment(all P<0.05).After treatment,the scores of clinical symptoms and physical signs,the excellence rate of atlantoaxial axle separation and the effective rate in the observation group were all significantly better than those of the control group(all P<0.01).Conclusion:The combination of ulna-tibia needling therapy and"point-to-surface"tuina manipulation achieves the better clinical therapeutic effect as compared with flunarizine hydrochloride.