Comparison of vitamin B levels in febrile children with and without febrile seizures: A prospective single-center study

Abstract

Febrile seizures (FS) are among the most common neurological disorders in childhood, and their pathogenesis may be influenced by multiple factors. As scientific research progresses, the significant role of B vitamins in the nervous system has become progressively clearer. In this study, children clinically diagnosed with FS were selected as the experimental group, and febrile children without a history of convulsions or a family history of seizures were selected as the control group to investigate the potential correlation between FS and whole-blood B vitamin levels. The concentrations of B vitamins in whole blood were measured using liquid chromatography-mass spectrometry, and statistical analyses were conducted using SPSS 25.0, followed by significance testing. The levels of vitamins B1, B2, B5, B6, B7, and B12 in the experimental group were significantly higher than those in the control group, particularly vitamins B5 and B7 (P < .05). Additionally, statistically significant differences were observed between sodium and calcium ion levels. These results suggest that febrile convulsions are the complex outcome of multiple factors, including known ion channel defects. Vitamin B may play a role in febrile convulsions and warrants further investigation.

1. Introduction

Febrile seizures (FS) are characterized by convulsions that occur between the ages of 6 months and 5 years, often coinciding with the onset of fever or a rapid increase in body temperature. This seizure episode occurred in the absence of: central nervous system infections, febrile infection-related epilepsy syndrome,^[1] and acute provoking factors, including metabolic imbalances, neurotoxins, or structural brain abnormalities.

Febrile seizures are among the most prevalent neurological disorders in childhood, with an incidence ranging from 2% to 5%.^[2,3] Unfortunately, many parents have limited understanding of febrile convulsions and often feel overwhelmed and helpless when their children experience seizures. This lack of understanding not only heightens parental anxiety but may also delay the child’s access to timely and effective treatment. Therefore, it is crucial to enhance awareness of febrile seizures and their prevention.

Based on clinical manifestations, febrile seizures are classified into 2 types: simple febrile seizures (SFS) and complex febrile seizures (CFS). SFS is characterized by solitary, generalized tonic-clonic seizures occurring within a 24-hour period and lasting <15 minutes. In contrast, CFS exhibits focal characteristics, lasts longer than 15 minutes, or involves multiple recurrent episodes within the same 24-hour period. Approximately 20% to 35% of febrile seizures are classified as CFS.^[4]

Emerging evidence underscores the essential role of B vitamins as enzymatic cofactors in cellular metabolism and neurophysiological regulation, particularly in maintaining neuronal energy homeostasis, neurotransmitter synthesis, and myelin integrity.^[5] Notably, their neuroprotective properties have been increasingly recognized for mitigating oxidative stress and neuronal excitability. A critical hypothesis-driven question arises: Do pediatric patients with febrile seizures exhibit significantly lower concentrations of B vitamins compared to febrile controls without convulsions? Few clinical studies have explored the correlation between vitamin B levels and febrile seizures. Therefore, this study aimed to measure B vitamin levels in the whole blood of children experiencing febrile seizures and compare these levels with those of children who had fever but did not experience seizures.

2. Materials and methods

2.1. Clinical data

We selected 32 children with febrile seizures admitted to our hospital between January 2023 and August 2024 as the case group, and 32 febrile children without seizures during the same period as the control group. The target sample size was initially determined to be 140 participants through a priori power analysis, based on the methodology described by Ozkale et al.^[6] Due to time and budgetary constraints, the final sample consisted of 64 cases. We conducted a post hoc power analysis to assess the statistical power of the current sample size. Using G*Power software (version 3.1.9.7), the statistical power was calculated to be 0.72 (72%), based on the observed effect size (Cohen’s d = 0.64) and significance level (α = 0.05). The sample size in our study has a 72% probability of detecting a moderately high effect (d = 0.64). However, the significant results observed (P = .019) suggest that the effect size is substantial and clinically meaningful.

The case group consisted of 23 males and 15 females, aged 6 to 58 months, with an average age of 27.87 ± 7.13 months. The control group consisted of 21 males and 11 females, aged 6 to 59 months, with an average age of 28.13 ± 7.22 months. There was no statistically significant difference in the general data between the 2 groups (P > .05), indicating their comparability (Table 1).

Table 1.

Demographic characteristics of children in the study and control groups.

Groups	Male	Female	Age (mo)
Cases (n = 32)	59.4%	40.6%	25.0 (18.25, 36.75)
Controls (n = 32)	68.8%	31.3%	22.75 (22.75, 53.5)
P value	.434		.077

2.2. Inclusion criteria

The case group included children who met the 2011 diagnostic criteria for febrile seizures set by the American Academy of Pediatrics. These criteria specified that seizures must occur during a fever (axillary temperature ≥ 38°C), exclude central nervous system infections and other acute illnesses that could cause seizures, and not involve a previous diagnosis of epilepsy. Children were divided into 2 subgroups: SFS and CFS. The control group consisted of children with fever who did not experience seizures.

2.3. Exclusion criteria

Children with a history of afebrile seizures, diagnosed central nervous system infections, or congenital metabolic diseases were excluded.

2.4. Data sources

All blood analyses were performed using the first blood sample collected after informed consent was obtained upon admission. Liquid chromatography-mass spectrometry was used to detect the levels of group B vitamins in the whole blood of the 2 groups of children (water-soluble vitamin determination kit, Yingsheng 9900MD, Yingsheng Company, China).

2.5. Statistical analysis

All collected data were analyzed statistically using SPSS software (version 25.0). Normality tests were conducted on each data point, and metric data following a normal distribution were expressed as mean ± SD. An independent sample t test was used for comparisons between 2 groups; non-normally distributed quantitative data were expressed as M (P25, P75) and analyzed using the Mann–Whitney U test, a 2-independent sample non-parametric test. Count data were expressed as frequency and percentage, and the chi-square test was used for comparisons between the 2 groups. Statistical significance was determined at P ≤ .05.

3. Results

A total of 64 children were enrolled in the study, with 32 in the case group and 32 in the control group. No significant differences were observed between the demographic characteristics of the children in the case group and control group (Table 1).

The levels of vitamins B1, B2, B5, B6, B7, and B12 were significantly higher in the case group than in the control group, with significant differences observed for vitamins B5 and B7 (P < .05). Conversely, the vitamin B3 level in the case group was lower than that in the control group; however, this difference was not statistically significant (P > .05; Table 2).

Table 2.

Comparison of vitamin B levels between study group and control group.

Variables	Cases	Controls	Z value	P value
Vitamin B1	4.25 (2.80, 5.72)	3.38 (1.70, 4.93)	−1.558	.119
Vitamin B2	13.56 (10.73, 18.60)	13.04 (9.15, 16.98)	−1.074	.283
Vitamin B3	41.80 (29.64, 85.22)	58.99 (43.67, 88.73)	−1.719	.086
Vitamin B5	58.22 (40.66, 91.36)	38.06 (32.58, 65.02)	−2.336	.019
Vitamin B6	2.24 (0.85, 3.69)	1.68 (1.01, 2.95)	−0.638	.524
Vitamin B7	0.19 (0.13, 0.24)	0.15 (0.10, 0.20)	−2.189	.029
Vitamin B9	13.52 (4.30, 27.85)	13.29 (7.48, 21.57)	−0.309	.757

Table 3 presents the serum biochemical characteristics of the case group and control group. Regarding inflammatory factors, statistically significant differences (P < .05) were observed in the levels of white blood cells, neutrophil percentage, lymphocyte percentage, and procalcitonin between the case and control groups. However, no statistically significant difference was found in the levels of high-sensitivity C-reactive protein and lactate dehydrogenase between the groups. Statistically significant differences (P < .05) were observed in sodium, chloride, and calcium levels between the case group and control group, whereas no statistically significant differences were found in potassium and magnesium levels.

Table 3.

Comparison of biochemical and complete blood count parameters of children in the study and control groups.

Variables	Cases	Controls	Z value	P value
WBC	11.25 (8.83, 1618)	6.90 (5.86, 10.0)	−4.062	<.01
NEUT%	75.6 ± 9.61	59.56 ± 13.14	5.58	<.01
LYMPH%	77.65 (70.15, 83.45)	29.80 (24.80, 36.0)	−4.895	<.01
NLR	5.29 (3.41, 12.87)	2.09 (1.58, 5.94)	−4.820	<.01
PLT	254.24 ± 83.88	267.86 ± 59.741	0.329	.74
Plateletcrit	0.75 ± 0.06	0.22 ± 0.05	2.23	.034
hs-CRP	9.77 (4.16, 66.4)	12.25 (4.9, 20.27)	−0.725	.468
PCT	0.33 (0.14, 1.02)	0.14 (0.09, 0.23)	−2.300	.021
LDH	318.0 (270.25, 359.0)	326.5 (284.75, 357.5)	−0.690	.490
Potassium	4.12 (3.92, 4.43)	3.98 (3.87, 4.44)	1.806	.071
Sodium	134.94 ± 2.21	136.75 ± 2.06	−3.388	<.01
Chlorine	102.19 ± 2.60	104.0 ± 2.67	−2.744	<.01
Calcium	2.29 ± 0.08	2.16 ± 0.09	5.553	<.01
Magnesium	0.84 (0.79, 0.92)	0.87 (0.82, 0.94)	−0.733	.464

hs-CRP = high-sensitivity C-reactive protein, LDH = lactate dehydrogenase, LYMPH = lymphocyte, NEUT = neutrophil, NLR = neutrophil–lymphocyte ratio, PCT = procalcitonin, PLT = platelet, WBC = white blood cell.

We analyzed the data from the SFS group and CFS group and found that the differences in the levels of all types of vitamin B between these 2 groups were not significant (P > .05). The differences between the SFS group and CFS group were only statistically significant for PLT levels and platelet stress (P < .05). Differences in the levels of the remaining serological indices between the groups were not statistically significant. For further details, please refer to Tables 4 and 5.

Table 4.

Comparison of vitamin B levels between SFS and CFS.

Variables	SFS	CFS	Z value	P value
Vitamin B1	3.23 (2.84, 6.44)	4.45 (2.62, 5.61)	−0.170	.88
Vitamin B2	12.95 (11.21, 16.88)	15.14 (9.64, 19.95)	−0.208	.85
Vitamin B3	52.26 (34.55, 101.14)	38.68 (22.50, 80.0)	−0.206	.21
Vitamin B5	61.1 (36.5, 101.24)	57.99 (43.96, 91.68)	−0.359	.73
Vitamin B6	1.77 (0.69, 4.27)	2.57 (0.69, 4.27)	−0.170	.88
Vitamin B7	0.20 (0.12, 0.27)	0.18 (0.15, 0.23)	−0.170	.88
Vitamin B9	9.59 (3.73, 20.7)	23.23 (5.34, 30.58)	−1.057	.29

CFS = complex febrile seizures, SFS = simple febrile seizures.

Table 5.

Comparison of biochemical and complete blood count parameters of children with SFS and CFS.

Variables	SFS	CFS	Z value	P value
WBC	16.25 ± 8.96	10.4 ± 2.74	1.192	.243
NEUT%	79.4 ± 10.63	81.2 ± 2.72	1.346	.188
LYMPH%	13.11 ± 8.91	8.72 ± 2.75	−1.390	.175
NLR	9.75 ± 7.80	9.89 ± 2.54	1.534	.136
PLT	277.5 (227.0, 323.0)	196.0 (178.0, 278)	−2.512	.012
Plateletcrit	0.75 ± 0.06	0.22 ± 0.05	2.23	.034
hs-CRP	9.19 (2.86, 14.83)	7.09 (4.44, 13.75)	−0.04	.968
PCT	0.36 (0.19, 1.05)	0.14 (0.11, 0.70)	−1.659	.097
LDH	326.33 ± 44.25	284.50 ± 50.47	0.847	.408
Potassium	4.18 ± 0.30	4.05 ± 0.17	−1.049	.33
Sodium	133.78 ± 1.64	136.00 ± 2.16	−2.001	.055
Chlorine	102.0 ± 1.73	101.75 ± 3.77	0.647	.533
Calcium	2.28 ± 0.07	2.24 ± 0.05	0.08	.936
Magnesium	0.80 ± 0.06	0.99 ± 0.11	−0.744	.463

CFS = complex febrile seizures, hs-CRP = high-sensitivity C-reactive protein, LDH = lactate dehydrogenase, LYMPH = lymphocyte, NEUT = neutrophil, NLR = neutrophil–lymphocyte ratio, PCT = procalcitonin, PLT = platelet, SFS = simple febrile seizures, WBC = white blood cell.

4. Discussion

Although scholars both domestically and internationally have conducted extensive research on febrile convulsions, the underlying mechanisms remain unclear. Most researchers suggest that genetic susceptibility, viral infections, family history, and incomplete development of the nervous system may contribute to the condition.^[7] Studies have identified sodium-ion channel genes associated with the pathogenesis of febrile convulsions including SCN1A,^[8] SCN2A,^[9] SCN9A,^[10] SCN1B,^[11] and HCN2.^[12] These genes can enhance neuronal excitability by modifying the activity, opening duration, and current magnitude of sodium channels, as well as regulating the transmission of the inhibitory neurotransmitter gamma-aminobutyric acid, which may further trigger convulsions. It has been found that the inward flow of calcium in neurons primarily depends on voltage-gated calcium channels and glutamate receptor channels. Fu et al^[13] found that a significant amount of excitatory amino acids is released during convulsions, which activate NMDA receptors in hippocampal pyramidal cells. This activation opens NMDA receptor-regulated calcium channels, leading to a substantial influx of calcium and excitotoxic neuronal damage. Over the past few years, Postnikova et al^[14] have utilized hyperthermia-induced FS in 10-day-old rats, demonstrating that FS rapidly reduces synaptic calcium-permeable-AMPA receptors in both the hippocampus and the entorhinal cortex. This process is associated with a significant decrease in the calcium permeability of the membranes of principal neurons. The reduction in calcium permeability may protect neurons and potentially serve a compensatory role in preventing hyperexcitability, toxicity, and neuronal death. Consequently, both sodium and calcium ions contribute to this process during convulsions.

With increasing research on micronutrients in recent years, neurotrophic B vitamins have been shown to play a crucial role as coenzymes in the nervous system. Vitamin B5, also known as pantothenic acid, is a water-soluble vitamin that belongs to the vitamin B family.^[15] It serves as a substrate for the synthesis of coenzyme A and acyl carrier protein. Coenzyme A serves as an acyl carrier and functions as a cofactor in various enzymatic reactions involved in the synthesis and degradation of fatty acids, carbohydrates, cholesterol, acetylcholine, and other essential substances.^[16] Pantothenic acid influences epithelial tissues and organs, including the digestive tract, adrenal glands, neural tubes, and skin in both animals and humans. It enhances immune function and promotes the biosynthesis of glutathione, which helps mitigate cell apoptosis and damage.^[17]

Although research on the association between vitamin B5 and neurological disorders is limited, several potential associations have been identified. Patassini et al^[18] discovered that pantothenic acid salts were widely distributed in the putamen and cerebellum of healthy individuals. They also observed that patients with Huntington’s disease, including those in the pre-symptomatic stage, generally exhibited lower levels of pantothenic acid salts in their brains. Consequently, they proposed that adequate storage of pantothenic acid in the brain may be essential for sustaining the high rate of myelin synthesis necessary for the integrity of myelinated neurons. Mari-Bauset et al^[19] also found that children with autism spectrum disorder have insufficient levels of vitamin B5. Furthermore, researchers have suggested that vitamin B5 deficiency in the brain may contribute to neurodegenerative changes and dementia associated with Alzheimer’s disease.^[20]

Overall, vitamin B5 plays a crucial role in nervous system function by protecting the myelin sheath. However, when febrile children experience seizures, neurotransmitters such as acetylcholine significantly increase, and the potential for enhanced pantothenic reactivity should not be overlooked. These findings align with the results of this experiment, suggesting that children with febrile seizures have higher levels of vitamin B5 than those with a normal fever. However, further research is required to confirm the specific mechanisms of action and their impact on neurological diseases. Ensuring an adequate intake of vitamin B5 is essential for maintaining overall health, particularly that of the nervous system.

Vitamin B7, commonly known as biotin, cannot be synthesized by the body and must be obtained from various sources, including eggs, liver, pork, leafy greens, and gut microbiota. Previous studies have shown that the active form of biotin, biotin-AMP, plays a crucial role in intracellular signal transduction. Biotin-AMP activates soluble guanylate cyclase, which in turn promotes the production of the second messenger cyclic guanosine monophosphate and activates downstream protein kinase G. This process leads to the expression of various carboxylases (PC, PCC, ACC-1) and is involved in the metabolism of carbohydrates, fats, and proteins.^[21,22] Ebru Canda found that biotin deficiency leads to insufficient biotin recovery, impaired function of biotin-dependent carboxylases, and may result in the production of neurotoxic and epileptogenic metabolites.^[23] Overall, both pantothenic acid and biotin exhibit antioxidant properties that can neutralize free radicals and diminish cellular oxidative damage.^[24] Although the observed increase in vitamins B5 and B7 in this study is not entirely understood, it may be interpreted as an adaptive mechanism by the brain to counteract oxidative stress and excitability.^[25] However, the relationship between these vitamins and neurological disease requires further investigation.

Although this study found that children with febrile seizures had higher blood concentrations of vitamins B5 and B7 compared to those with normal fever, several limitations must be acknowledged. The modest sample size (N = 64) and single-center design may limit statistical power and generalizability, especially for subgroup analyses. Furthermore, the absence of longitudinal data limits our understanding of long-term neurological implications. Future multi-center studies with larger and more diverse cohorts are needed to validate these findings. Mechanistic investigations using experimental models should explore how B vitamins influence neuronal excitability or oxidative stress during hyperthermia. Additionally, interventional trials assessing dietary supplementation or genetic profiling of vitamin B metabolism pathways could elucidate their therapeutic potential in high-risk populations. These efforts may ultimately inform personalized strategies for preventing febrile seizures and neuroprotection.

Acknowledgments

The authors are grateful to all the participants involved in the present study for their enthusiasm and commitment.

Author contributions

Data curation: Ying Huang, Xinxin Wen, Hongxuan Guo, Jianwei Cao.

Formal analysis: Ying Huang, Xinxin Wen, Hongxuan Guo, Jianwei Cao.

Funding acquisition: Jianwei Cao.

Investigation: Xinxin Wen, Jianwei Cao.

Methodology: Jianwei Cao.

Project administration: Jianwei Cao.

Resources: Jianwei Cao.

Software: Ying Huang, Xinxin Wen, Hongxuan Guo.

Supervision: Jianwei Cao.

Validation: Ying Huang, Hongxuan Guo, Jianwei Cao.

Visualization: Ying Huang.

Writing – original draft: Ying Huang, Xinxin Wen.

Writing – review & editing: Jianwei Cao.