Association of sLR11 gene polymorphism with T2DM and carotid atherosclerosis

Abstract

OBJECTIVE:

To investigate the association of single nucleotide polymorphisms (SNPs) of soluble low-density lipoprotein receptor 11 (sLR11) genes with type-2 diabetes mellitus (T2DM) and carotid atherosclerosis (CAS) in Korean and Han nationalities in the Yanbian area.

METHODS:

530 T2DM patients were divided into two groups according to the intima-media thickness (IMT) of the carotid artery: CAS group ( $n=$ 256, T2DM patients with carotid artery IMT $\geqslant$ 1.0 mm and plaque) and non-CAS group (NCAS group, $n=$ 274, T2DM patients with carotid IMT $<$ 1.0 mm). IMT and plaque were measured by color Doppler ultrasound. SNP typing and sequencing were performed by PCR-LDR.

RESULTS:

1. Allele frequency and genotype frequency distribution results: Differences in genotype and allele frequency distribution between the CAS and NGT groups, the NCAS and NGT groups, and the CAS and NCAS groups were not statistically significant ( $P>$ 0.05). The dominant and recessive modes were analyzed, but the difference in genotype frequency among these three groups was not statistically significant ( $P>$ 0.05). Differences in genotype frequency distribution between Korean and Han populations in all three groups were not statistically significant ( $P>$ 0.05). 2. Correlation analysis with clinical indicators: LDL-C levels in TT and AT patients in the CAS group were significantly higher than those in AA patients ( $P>$ 0.05), representing the dominant mode of inheritance..

CONCLUSION:

This study is the first to determine that the sLR11 gene rs3824968 polymorphic of factor T may increase the risk of CAS in T2DM patients by regulating the concentration of LDL-C, showing the dominant mode of inheritance.

Keywords

sLR11 gene polymorphism type 2 diabetes mellitus carotid intima media thickness

1. Introduction

The main pathological change of diabetic macrovascular disease is atherosclerosis (AS), and the mechanism involves the induction of smooth muscle cell proliferation, thickening of the arterial wall, atherosclerotic plaque formation and vascular calcification [1]. In the formation of AS, the migration of vascular smooth muscle cells (SMCs) from the media to the intima is a critical step [2]. In 1996, Yamazaki et al. [3] first discovered that low density lipoprotein receptor (LDLR) relative with 11 ligand-binding repeats (LR11) detected by immunological methods in serum is a member of the LDLR family. Furthermore, it was also observed that LR11 was highly expressed on the intima of SMCs, which was followed by medial SMCs near the intimal boundary of the AS lesion [4]. Subsequently, they found that the serum concentration of soluble LDLR relative with soluble LR11 (sLR11) was positively correlated carotid artery intima-media thickness (IMT) in patients with dyslipidemia. Furthermore, multivariate regression analysis revealed that sLR11 levels served as a new quantitative circulatory marker of IMT, and was independent of the classic risk factors for AS [5]. A recent study revealed that sLR11 levels are positively correlated with IMT in middle-aged and non-obese type-2 diabetes mellitus (T2DM) patients. At present, the association of sLR11 gene polymorphism with T2DM and carotid atherosclerosis (CAS) has not been reported in any study worldwide. However, the analysis of single nucleotide polymorphism (SNPs) of the sLR11 gene has been used in studies on the pathogenesis of Alzheimer’s disease (AD). Studies have revealed that the gene polymorphism of apolipoprotein E (APOE) was closely related to T2DM and AD [7, 8, 9, 10]. We inferred that T2DM and AD may have a similar etiology. Therefore, in this study, the positive motif rs3824968 related to AD [11] was selected to investigate the association of the rs3824968 polymorphism of the sLR11 gene with T2DM and CAS in Korean and Han nationalities in the Yanbian area, and provide a genetics theoretical basis for the effective prevention and treatment of AS in diabetes patients.

2. Subjects and methods

2.1 Subjects

The T2DM group: A total of 530 patients with T2DM were enrolled into this study and divided into two groups: CAS group ( $n=$ 256) and non-CAS group (NCAS group, $n=$ 274). The normal glucose tolerance control (NGT) group: This group comprised of 246 subjects, who had no family history of diabetes mellitus, were diagnosed without diabetes, coronary heart disease, cerebral infarction, disorder of lipid metabolism, inflammation, or infections, and had normal liver and kidney function and normal white blood cell count. Differences in age and gender between these two groups were not statistically significant ( $P>$ 0.05); hence, these two groups were comparable. All subjects were informed of the study contents, and provided an informed consent. The study was also approved by the Ethics Committee of the affiliated Hospital of Yanbian University, Yanji, China. Diagnostic criteria were based on the 1999 Diagnostic Criteria of Diabetes.

2.2 Methods

2.2.1 General information of subjects

Gender, age, course of disease, blood pressure, body mass index (BMI) were recorded from all subjects; and fasting blood glucose (FBG), 2-hour postprandial blood glucose, glycated hemoglobin (HbA1c), total cholesterol (CHO), triglyceride (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL) were detected. The ACUSON-128 probe (frequency: 7.5 MHz) was used to determine IMT at the site 1 cm under the enlargement of the common carotid artery, where the thickness of the thickest site and two sites 1 cm from the thickest site in the lengths near and beyond the heart were measured. A total of six sites on the left and right sides were measured, and the average value was defined as the IMT value. In order to ensure the stability of the IMT values, all operations were conducted by the same personnel.

Fasting peripheral venous blood was withdrawn in the early morning for detection of the following biochemical indexes: high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), CHO, TG and HbA1c. Then, 0.5 mL of whole blood was placed in an anticoagulant tube containing ethylenediaminetetraacetic acid disodium salt (EDTA-Na ${}_{2}$ ), and the indexes were determined using the microcolumn method. The remaining blood samples were stored in a low temperature freezer ( $-$ 80 ${}^{\circ}$ C) for extraction of genomic DNA. Whole blood DNA was extracted using a kit provided by Promega.

2.2.2 Primers and probes: PCR primers and LDR probe sequences are shown in Table 1

Table 1
Primer probe sequence

Primers and probe	Sequence (5’-3’)	Size of the products (bp)
PCR primers
Upstream primer	TGGACAGCTGACTTCTCTGG	151
Downstream primers	ACATCCTGGGGCAATGAAGA
LDR probe
rs3824968_modify	P-GCAGAGGGCATTTTTTTGGGCCTCA
	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
	TTTTTTTTTTTTTTTTTTTTTTTT-FAM
rs3824968_A	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTT	170
	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
	TTTTGTAGTAGACATTATATACACAAGAT
rs3824968_T	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT	172
	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
	TTTTGTAGTAGACATTATATACACAAGAA

2.2.3 PCR-LDR genotyping procedure

Step 1: Multiple cycles of polymerase chain reaction (PCR) amplification was performed in a 20- $\mu$ l system. The formula was as follows: PCR buffer with a final concentration of 1 $\times$ , Mg ${}^{2+}$ with a final concentration of 3 mM, dNTP with a final concentration of 2 mM, primer with a final concentration of 0.05 pmol/ $\mu$ l, and 1 U of Taq enzyme. PCR reaction conditions: denaturation at 94 ${}^{\circ}$ C for 30 seconds, annealing at 59 ${}^{\circ}$ C for 90 seconds, and extension at 65 ${}^{\circ}$ C for 30 seconds; a total of 40 cycles were performed. Step 2: Multiple cycles of ligase detection reaction (LDR) amplification was performed in a 10- $\mu$ l system. The formula was as follows: ligase buffer with a final concentration of 1 $\times$ , 4 $\mu$ l of PCR products, and 2 U of Taq ligase. LDR reaction conditions were as follows: denaturation at 94 ${}^{\circ}$ C for 15 seconds, and ligation at 50 ${}^{\circ}$ C for 25 seconds; a total of 40 cycles were performed. Step 3: One $\mu$ L of LDR products and 0.5 uL of ROX internal control (size standard) was used, added with 8 $\mu$ L of Hi-Di Formamide loading buffer, denatured for two minutes, placed under an ice bath for two minutes, the product was detected on an 3730 sequencer, and fragment analysis was performed using Genemapper 4.0.

2.2.4 Statistical analysis

Allele frequency, genotype frequency and Hardy-Weinberg genetic equilibrium tests were performed using the SHEsis online software (http://202.120.7.14/analysis/) to determine the population representation of the sample. Data were statistically analyzed using statistical software SPSS 22.0. The comparison of measurement data between groups was performed using one-way analysis of variance (ANOVA), in which pairwise comparison was conducted using the least significant difference test. Risk factors for CAS were screened by logistic regression analysis. $P<$ 0.05 was considered statistically significant.

3. Results

3.1 Detection by 3% agarose gel electrophoresis

Detection by 3% agarose gel electrophoresis was performed. The PCR products were observed to determine the amount that would be added to LDR as a template.

Figure 1.

Electrophoretogram of PCR products. The markers were 100, 200, 300, 400 and 500, respectively. The size of the destination fragment is: 151 bp.

Figure 2.

Genemapper analysis result.

Results are shown in Fig. 1. The main purpose of this experiment was to determine whether the PCR was successful. It cannot be observed by electrophoresis whether each site is normal.

151 bp PCR product of sLR11 gene polymorphism (rs3824968). $M=$ Marker (100,200,300,400,500), Lanes 1, 2, 3 (the destination fragment is: 151 bp).

3.2 Typing and sequencing results

Genemapper 4.0 analysis results are shown in Fig. 2, that the rs3824968 was divided into AA, AT and TT genotypes. And the results of the whole blood genome sequencing are shown in Fig. 3, peaks demonstrated SNP homozygous AA (Fig. 3A), heterozygous AT (Fig. 3B) and homozygous TT (Fig. 3C) genotypes. These figures refer to the reverse complementary sequences. In Fig. 3A, the arrow points to a map of the homozygous AA genotype sequence with the base T as a single peak. In Fig. 3B, the arrow points to two A/T peaks, and represents the map of the heterozygous AT genotype sequence. In Fig. 3C, the arrow points to a single A peak, and represent the map of the homozygous TT genotype sequence. Genemapper analysis result and randomly selected sample sequencing results completely match,accuracy rate is 100%.

Figure 3.

A: Sequencing graph of homozygote AA; B Sequencing graph of heterozygote AT; C: Sequencing graph of homozygote TT. The Genemapper analysis result and randomly selected sample sequencing results completely match. The accuracy rate is 100%.

3.3 Statistical results

3.3.1 Comparison of the rs3824968 polymorphism genotype of the sLR11 gene and allele frequency distribution

In the Yanbian area, the differences in genotype and allele frequency distribution between the CAS and NGT groups, the NCAS and NGT groups, and the CAS and NCAS groups were not statistically significant ( $P>$ 0.05). The difference in genotype frequency in the dominant and recessive modes among the three groups was not statistically significant ( $P>$ 0.05). The differences in genotype and allele frequency distribution between Korean and Han populations in the CAS, NCAS and NGT groups were not statistically significant ( $P>$ 0.05). The above data was in accordance with the Hardy-Weinberg equilibrium law test, and conforms to the population representation of the sample in this study (Tables 2–4).

Table 2
sLR11 gene rs3824968 allele and genotype frequency distribution in NGT, NCAS, and CAS groups [n(%)]

Group	n	Genotype			Allele
		AA	AT	TT	A	T
NGT group	246	77(31.3)	131(53.3)	38(15.4)	258(57.9)	207(42.1)
NCAS group	274	87(31.8)	137(50.0)	50(18.2)	311(56.8)	237(43.2)
$\chi^{2}$		0.875			0.146
$P$		0.656			0.702
NGT group	246	77(31.3)	131(53.3)	38(15.4)	258(57.9)	207(42.1)
CAS group	256	84(32.8)	128(50.0)	44(17.2)	296(57.8)	216(42.2)
$\chi^{2}$		0.579			0.001
$P$		0.749			0.971
OR (95% CI)		–			0.995(0.775–1.279)
NCAS group	274	87(31.8)	137(50.0)	50(18.2)	311(56.8)	237(43.2)
CAS group	256	84(32.8)	128(50.0)	44(17.2)	296(57.8)	216(42.2)
$\chi^{2}$		0.130			0.122
$P$		0.937			0.727
OR (95% CI)		–			1.044(0.819–1.332)

Genotype and allele frequency distribution were not statistically different between the CAS group and NGT group, NCAS group and NGT group, and CAS group and NCAS group, $P>$ 0.05.

Table 3

Distribution under different models of sLR11 gene rs3824968 genotype frequency[n(%)]

Group	Dominance pattern		Recessive pattern
	AA	AT $+$ TT	TT	AT $+$ AA
NGT group	77(31.3)	169(68.7)	38(15.4)	208(84.6)
NCAS group	87(31.8)	187(68.2)	50(18.2)	224(81.8)
$\chi^{2}$	0.012		0.723
$P$	0.912		0.395
OR (95% CI)	1.021(0.705–1.479)		0.818(0.516–1.299)
NGT group	77(31.3)	169(68.7)	38(15.4)	208(84.6)
CAS group	84(32.8)	172(67.2)	449(17.2)	212(82.8)
$\chi^{2}$	0.132		0.728
$P$	0.717		0.598
OR (95% CI)	1.072(0.737–1.560)		0.880(0.548–1.415)
NCAS group	87(31.8)	187(68.2)	50(18.2)	224(81.8)
CAS group	84(32.8)	172(67.2)	449(17.2)	212(82.8)
$\chi^{2}$	0.068		0.102
$P$	0.794		0.749
OR (95% CI)	1.050(0.729–1.511)		1.075(0.688–1.681)

There was no significant difference in genotype frequency distribution among the three groups, $P>$ 0.05.

Table 4

Allele and genotype distribution in Korean and Han of sLR11 gene rs3824968 allele and genotype frequency in the CAS group, NCAS group and NGT group

Group	Allele/Genotype	Korean	Han	OR [95% CI]	$P$
NGT group	A	93(56.0)	192(58.9)
				1.125[0.771 $\sim$ 1.641]	0.542
	T	73(44.0)	134(41.1)
	A A	24(28.9)	53(32.5)
	A T	45(54.2)	86(52.8)		0.813
	T T	14(16.9)	24(14.7)
NCAS group	A	91(51.7)	220(59.1)
				1.352[0.943 $\sim$ 1.939]	0.101
	T	85(48.3)	152(40.9)
	A A	20(22.7)	67(36.0)
	A T	51(58.0)	86(46.2)		0.080
	T T	17(19.3)	33(17.7)
CAS group	A	91(52.3)	205(60.7)
				1.406[0.972 $\sim$ 2.033]	0.070
	T	83(47.7)	133(39.3)
	A A	24(27.6)	60(35.5)
	A T	43(49.4)	85(50.3)		0.158
	T T	20(23.0)	24(14.2)

Allele and genotype frequency distribution in Korean and Han of three groups have no significant different, $P>$ 0.05.

3.3.2 Analysis of risk factors for CAS

The regression coefficients of LDL-C and TG were 0.451 and 0.135, respectively, regression coefficient was positively correlated with CAS. From the foregoing, LDL-C and TG are risk factors for CAS (Table 5).

Table 5
Logistic stepwise regression analysis of risk factors for carotid atherosclerosis

	Regression coefficients	Standard error	Wald value	$P$	OR (95% CI)
LDL-C (mmol/L)	0.451	0.178	6.447	0.011	1.570(1.108–2.225)
TG (mmol/L)	0.135	0.067	4.043	0.044	1.145(1.003-1.306)

$P<$ 0.05 between two groups.

3.3.3 Comparison of clinical characteristics between different genotype groups in the CAS group

The CHO level order was AT group $>$ TT group $>$ AA group, and the difference between the AT and AA groups was statistically significant ( $P<$ 0.05). The LDL-C level order was TT group $>$ AT group $>$ AA group, and differences between the AT and AA groups, and between the TT and AA groups were statistically significant ( $P<$ 0.05, Table 6). AA group and AT $+$ TT group was significantly different ( $P1<$ 0.05, Table 7).

Table 6
Comparison of clinical data of different genotypes in the CAS group

Groups	AA group		AT group		TT grosup		$P_{1}$	$P_{2}$	$P_{3}$
Age (years)	61.38	$\pm$ 8.75	61.34	$\pm$ 10.19	62.48	$\pm$ 9.89	0.978	0.544	0.504
BMI (kg/m2)	29.92	$\pm$ 5.01	25.17	$\pm$ 2.90	24.80	$\pm$ 3.72	0.168	0.119	0.579
SBP (mmHg)	138.99	$\pm$ 20.26	135.20	$\pm$ 21.54	133.73	$\pm$ 21.61	0.230	0.182	0.690
DBP (mmHg)	84.24	$\pm$ 11.53	83.23	$\pm$ 10.89	81.80	$\pm$ 11.07	0.521	0.239	0.460
CHO (mmol/L)	5.18	$\pm$ 1.27	5.58	$\pm$ 1.40	5.26	$\pm$ 1.34	0.037	0.760	0.174
TG (mmol/L)	2.43	$\pm$ 2.04	2.79	$\pm$ 2.44	2.10	$\pm$ 1.43	0.241	0.407	0.069
HDL-C (mmol/L)	1.27	$\pm$ 0.43	1.30	$\pm$ 0.57	1.21	$\pm$ 0.42	0.718	0.507	0.319
LDL-C (mmol/L)	2.75	$\pm$ 0.83	3.09	$\pm$ 0.87	3.12	$\pm$ 0.82	0.005	0.019	0.087
HbAlc (%)	8.28	$\pm$ 1.85	8.88	$\pm$ 5.78	8.23	$\pm$ 1.87	0.317	0.949	0.383

TG, HDL-C, HDL-C taking logarithm; $P_{1}$ : AA group and AT group, $P_{2}$ : AA group and TT group, $P_{3}$ : AT group and TT group, $P<$ 0.05 was different, $P<$ 0.01 was significantly different.

Table 7

Comparison of LDL-C of different models in the CAS group

Group	Dominance pattern			Recessive pattern
	AA	AT $+$ TT	$P1$	TT	AT $+$ AA	$P2$
LDL-C	2.75 $\pm$ 0.83	3.10 $\pm$ 0.85	0.002	3.12 $\pm$ 0.82	2.95 $\pm$ 0.87	0.234

AA group and AT $+$ TT group, $P1<$ 0.05 was significantly different.

4. Discussion

Diabetes mellitus is a common metabolic disease characterized by hyperglycemia. The risk of cardiovascular disease in patients with diabetes mellitus is 2–4 times of that in normal subjects. The main pathological change of macrovascular disease in patients with T2DM is AS. IMT thickening occurs earlier than AS. IMT is an early quantitative clinical marker of AS in patients with diabetes mellitus. A study revealed that the LR11 (SORL1) gene is located on chromosome 11 (11q24.1), and is the same gene as the sorLA gene in brain, which is consists of 177,491 base containing 51 exons. Studies have confirmed that as a novel circulatory marker for IMT risk of carotid artery, the level of serum sLR11 was correlated with carotid artery ITM thickening and plaque formation, acute coronary syndrome and diabetic retinopathy in T2DM patients [12, 13, 14, 15]. Most experiments have revealed that sLR11 gene polymorphism is associated with the pathogenesis of AD. According to statistical data, approximately 65% of patients with diabetes mellitus have mild-moderate cognitive impairment [16, 17]. The risk of AD in patients with diabetes mellitus is higher than in the normal population. Studies have revealed that in patients with diabetes mellitus and AD, atrophy of the hippocampus and amygdala could be observed by MRI [18, 19]. Therefore, T2DM may be related to the occurrence and development of AD. A study in China revealed that the level of plasma cholesterol was significantly higher in individuals carrying the ApoE $\varepsilon$ 4 allele than in individuals carrying the ApoE $\varepsilon$ 2 and ApoE $\varepsilon$ 3 allele, which increased the risk of AD in T2DM patients. SLR11 is a member of the LDL receptor family, which can bind with inherent LDL receptor-related proteins and ApoE on the endoplasmic reticulum. Therefore, sLR11 may be a susceptible gene following ApoE. At present, studies on the association of the sLR11 gene polymorphism with T2DM and CAS have not been reported. In this study, the AD-related positive motif rs3824968 in an Asian population was selected [11] to investigate the association of sLR11 gene polymorphism with T2DM and CAS in a Korean and Han population in the Yanbian area.

This experiment revealed that a single-base mutation A $\to$ T on the rs3824968 locus on the sLR11 gene was detected in the T2DM population and normal population in the Yanbian area, and that the sLR11 gene rs3824968 polymorphism was not associated with T2DM. Recent studies conducted by Chinese scholars revealed that in the Chinese Han population, the sLR11 gene rs2070045 polymorphism was not significantly related to T2DM. Therefore, we considered that the sites associated with AD are not necessarily associated with T2DM. To date, no studies have reported the correlation between the polymorphism of other sites and T2DM. Hence, this is exactly what we are going to do next.

The sLR11 gene polymorphism is affected by genetic heterogeneity. A large number of experiments revealed that sLR11 gene variation is associated with the risk of AD in different ethnic populations. At present, the study subjects are mainly European, Hispanic, African American, Japanese, Israeli Arab and Chinese Han nationality populations. At present, there is still a lack of reports on ethnic differences in sLR11 gene polymorphism among the ethnic minorities in china. In the present study, the genotype and allele frequency distribution of the Korean and Han nationality populations in the Yanbian area were selected and compared. Results revealed that the difference in sLR11 gene rs3824968 polymorphism in the NGT, NCAS and CAS groups between Korean and Han nationalities was not statistically significant. The reason may be related to the small sample size.

At present, it is considered that diabetic hyperglycemia may directly induce the expression of sLR11, and the phenotype of smooth muscle cells in the vascular wall is transformed into the so-called synthetic type. These possibilities are currently being studied [20]. This study revealed that LDL-C and TG are risk factors for CAS. This is consistent with the results of other studies, in which serum sLR11 level was positively correlated with LDL-C and HbA1c [21], and that serum sLR11 level was correlated to the formation of AS [6]. Recent studies have revealed that the deposition of cholesterol and other lipids in the blood vessel wall plays a decisive role in the early formation of AS. These lipid settlings are mainly derived from lipoproteins in the blood, especially from LDL [22]. The concentration of LDL on the surface of the inner wall of blood vessels is proportional to the lipoprotein entering the vessel wall. With the increase in TG and LDL levels, arterial intima thickening and elasticity decrease has become more significant. The present study revealed that in the CAS group, the level order of LDL-C closely correlated to CAS was TT group $>$ AT group $>$ AA group, showing an increasing trend. This suggests that rs3824968 polymorphic T factor may increase the concentration of LDL, allowing more LDL to enter into the arterial intima through the LR. As a result, the peroxidized LDL is engulfed by macrophages, which form foam cells and deposit beneath the intima. This finally makes SMCs to migrate into the intima and proliferate, leading to hyperplasia and hypertrophy of the intima. Then, atherosclerotic plaques are finally formed. Therefore, the rs3824968 polymorphic T factor may increase the risk of CAS in T2DM patients.

In this experiment, no direct risk factors for T2DM were found. However, this study is the first to determine that the sLR11 gene rs3824968 polymorphic of factor T may increase the risk of CAS in T2DM patients by regulating the concentration of LDL-C, showing the dominant mode of inheritance. This would provide some direction and basis for research work in the future.

Footnotes

Acknowledgments

The study was supported by grants from the National Natural Science Foundation of China (no. 81060067 and no. 81460158 (Study on relationship of genetic susceptibility and machanism of ARL15 associated with type 2 diabetes mellitus with macroangiophy). The author is Kang-Juan Yang).

Conflict of interest

None to report.

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Association of sLR11 gene polymorphism with T2DM and carotid atherosclerosis

Abstract

OBJECTIVE:

METHODS:

RESULTS:

CONCLUSION:

Keywords

1. Introduction

2. Subjects and methods

2.1 Subjects

2.2 Methods

2.2.1 General information of subjects

2.2.2 Primers and probes: PCR primers and LDR probe sequences are shown in Table 1

Table 1 Primer probe sequence

2.2.4 Statistical analysis

3. Results

3.1 Detection by 3% agarose gel electrophoresis

3.3.1 Comparison of the rs3824968 polymorphism genotype of the sLR11 gene and allele frequency distribution

Table 2 sLR11 gene rs3824968 allele and genotype frequency distribution in NGT, NCAS, and CAS groups [n(%)]

Table 5 Logistic stepwise regression analysis of risk factors for carotid atherosclerosis

Table 6 Comparison of clinical data of different genotypes in the CAS group

Footnotes

Acknowledgments

Conflict of interest

References

Table 1
Primer probe sequence

Table 2
sLR11 gene rs3824968 allele and genotype frequency distribution in NGT, NCAS, and CAS groups [n(%)]

Table 5
Logistic stepwise regression analysis of risk factors for carotid atherosclerosis

Table 6
Comparison of clinical data of different genotypes in the CAS group