Participants
24 CTN patients (9 males, 15 females; average age 55 ± 13 years; average duration 3.4 ± 4.2 years; average VAS 6.1 ± 1.4) and 22 HC (13 males, 9 females; average age 55 ± 11 years) were included in this study. Inclusion criteria: reference to (ICDH-3)[13]. Exclusion criteria: I) other types of chronic pain conditions, II) history of other central nervous system diseases or mental illness, III) other somatic or psychiatric conditions, IV) unsuitable for magnetic resonance scanning. Demographics and behavioral results of CTN and HC groups were listed in Table 1. And there is no significant difference between CTN and HC groups in age, gender, and handedness. The present study was approved by the Medical Research Ethics Committee of The First Affiliated Hospital of Nanchang University. All individuals include healthy controls provided signed informed consent to participate in the study.
Table 1
Demographics and behavioral results of CTN and HC groups
|
CTN
|
HC
|
P
|
Gender(Male/Female)
|
9/15
|
13/9
|
0.143
|
Age(years)
|
55 ± 13
|
55 ± 11
|
0.96
|
Handedness(R/L)
|
24/0
|
22/0
|
༞0.99
|
Duration(years)
|
3.4 ± 4.2
|
-
|
-
|
Side(R/L)
|
16/8
|
-
|
-
|
VAS
|
6.1 ± 1.4
|
-
|
-
|
Independent t-tests comparing the age of the two groups. Male/female and Handedness were analyzed using a chi-squared test. CTN, classical trigeminal neuralgia; HC, healthy control; R, right; L, left; VAS, visual analogue scale (P < 0.05 represented statistically significant differences)
MRI parameters
All magnetic resonance imaging (MRI) data are acquired with a 3.0T MRI scanner (Siemens AG). The total scan time was approximately 15 min. All participants were instructed to close their eyes, keep relax, and breathe smoothly, and do not do any task or systematic thinking until the scans were completed. T1-weighted MRI and resting-state functional MRI data were acquired. Parameters were as follows: 176 structural images ([TR] = 1900 msec; [TE] = 2.26 msec; thickness = 1 mm; gap = 0.5 mm; matrix size = 256x256; FOV = 250x250 mm; turning angle = 90°;voxel size = 1 mm × 1 mm × 1 mm) and 240 functional images ([TR] = 2000 msec; [TE] = 30 msec; thickness = 4 mm; gap = 1.2 mm; matrix size = 64x64; turning angle = 90°; FOV = 220x220 mm, 29 axial, voxel size = 3 mm × 3 mm × 3 mm). At the same time, conventional axial T1WI, T2WI, and fluid-attenuated inversion recovery sequences were acquired for excluding intracranial lesions before the BOLD and T1 sequence. Besides, before further analysis, we carefully examined the images of each subject to ensure the quality of the data.
fMRI data processing
We applied voxel-based morphometry (VBM) to quantify GMV for individual participants, unlike traditional methods of regions of interest (ROI), VBM is not susceptible to selection bias, it can provide an unbiased method to identify brain structural abnormalities through MRI scans. I) Convert the data format of initial T1 images into an analytical image format by SPM8(http://www.fil.ion.ucl.ac.uk/spm); II)Segment the converted images into gray matter (GM), white matter(WM), and cerebrospinal fluid (CSF) with a default 0.0001 bias regularization; III) Registration of nonlinear deformations of GM images of all subjects by using the DARTEL (Diffeomorphic Anatomical Registration Through Exponentiated Lie Algebra) package (Ashburner, 2007) provided by SPM8; IV) Create a customized template by using the GM segmented images from all subjects; V) Register the individual GM images to the template and normalize to the Montreal Neurological Institute (MNI) space; vi) Be spatially smoothed with an 8mm full-width at half maximum (FWHM) Gaussian kernel to improve the signal-to-noise ratio of the image; VII) Extract the signal of the resulting normalized and smoothed GM images that were used for training.
Statistical analysis
Demographic and clinical variables of CTN and HC groups were statistically analyzed using SPSS 26.0 software. Independent t-tests comparing the age of the two groups. Gender and Handedness were analyzed using the chi-squared test, P < 0.05 was considered to indicate a statistically significant difference.
SVM Classification
Machine learning in neuroimage data research can be divided into five steps[14]: I)Feature extraction; II)Feature selection or feature dimension reduction; III)Model training and model testing; IV)Evaluation the prediction ability of the model; V)Feature localization that contributes to the prediction.
To discriminate CTN from HC based on GMV, we applied an SVM-based algorithm from the Libsvm software library (https://www.csie.ntu.edu.tw/~cjlin/libsvm/) on the Matlab platform. The flow chart for SVM classification was shown in Fig. 1, which followed the same classification procedure published previously[15]. First of all, extract the signal of the 46 patients’ GMV as a features dataset and normalized it to (0,1). Due to the small sample size, the performance of the classifier was tested by leaving-one-out cross validation(LOOCV). The advantage of LOOCV over other cross-validation is that it can train the data as much as possible so that to obtain a more accurate classifier. Thus, within each iteration, we considered one participant as the testing dataset and the remaining participants as the training dataset. Next, Spearman, T-test, F-score, and PCA was respectively used for feature selection or feature dimension reduction to avoid overfitting and discard non-informative features. A) Spearman: Spearman correlation coefficient measures the strength of the relationship between the two variables, for each feature, it was calculated from the two groups based on GMV. B) T-test: In the process of feature selection, by calculating the test statistics, comparing the size of the statistics between features. C) F-score: Generally speaking, the larger F-score of a feature, the greater the value of this feature in classification. In the practical application of machine learning, calculating the F-score of all features. Based on the above three methods, arrange the statistical values in descending order, and then select the top 5% statistical values as features for training. D) PCA: All the features were decomposed into a series of principal components(PC), and only with a cumulative contribution of more than 90% were retained.
The misclassification parameter C of the SVM was optimized using nested cross-validation on the current training dataset. The process for each iteration includes finding the optimal model used to classify the test dataset. During model training, linear SVM assigns a specific weight to each feature to reflect its importance in the classification[16]. And the features that survive in each iteration were retained, this makes it possible to derive the spatial pattern underlying the classification from the mean weight across iterations for the surviving features. A positive weight indicates TN patients having higher GMV than HC in that particular region, while a negative weight indicates the opposite. After 46 iterations, the posterior balanced accuracy was calculated to evaluate the classification performance. By the way, the PCA method can not return a weight map.
Besides, to examine the degree to which the classification was driven by CTN symptoms rather than other confounds unrelated to CTN, we make a correlation between the deci_value for each subject and the VAS scores and pain duration, respectively. The method was similar to previous researches[17, 18].
Finally, the reliability of the model was evaluated by permutation test. In the permutation test, the label of the sample is randomly replaced and then repeat the above steps 1000 times. If the classifier does not acquire the corresponding relationship between the sample data and the label, then the frequency distribution of the average classification accuracy after generalization in the permutation test should obey the normal distribution with an average of 50%. If the average classification accuracy after generalization based on real tags falls outside the 95% confidence interval based on random tags, it is considered that the SVM classifier does get reliable learning from training data.