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Table 1 shows the baseline characteristics of the participants according to NAD+ quartile. The mean age of participants was 43.0 ± 11.5 years, and the mean level of NAD+ in the whole population was 31.3 ± 5.3 μmol/L. The mean levels of NAD+ were 25.0 ± 2.6, 29.3 ± 0.9, 32.5 ± 1.0, and 38.3 ± 3.5 μmol/L for the Q1, Q2, Q3, and Q4 groups, respectively. Participants with high NAD+ levels were more likely to have many more RBC, higher SBP, higher TG levels, and a history of dyslipidemia than those with low NAD+ levels. There were no significant differences in the other variables of interest, such as age and BMI, across the NAD+ quartile groups.
Table 1. Baseline characteristics of the study population within quartiles of NAD+ level
Characteristics Overall (n = 665) NAD+ groups P-value Q1 (< 27.6)
(n = 166)Q2 (27.6–30.9)
(n = 165)Q3 (30.9–34.4)
(n = 167)Q4 (≥ 34.4)
(n = 167)NAD+ (μmol/L) 31.3 ± 5.3 25.0 ± 2.6 29.3 ± 0.9 32.5 ± 1.0 38.3 ± 3.5 < 0.01 Age (y) 43.0 ± 11.5 41.0 ± 9.4 43.9 ± 12.8 43.9 ± 11.8 43.4 ± 11.6 0.06 Age at menarche (y) 13.9 ± 1.5 13.8 ± 1.3 13.9 ± 1.6 14.1 ± 1.7 13.8 ± 1.2 0.11 Age at menopausal (y) 50.0 ± 3.7 50.2 ± 3.7 50.6 ± 3.3 49.9 ± 3.3 49.5 ± 4.5 0.54 Body mass index (N, %) 0.39 < 24 kg/m2 414 (62.3) 109 (65.7) 104 (63.0) 106 (63.5) 95 (56.9) ≥ 24 kg/m2 251 (37.7) 57 (34.3) 61 (37.0) 61 (36.5) 73 (43.1) Education level (N, %) 0.39 Middle school or below 206 (31.0) 46 (27.7) 59 (35.8) 48 (28.7) 53 (31.7) College or above 459 (69.0) 120 (72.3) 106 (64.2) 119 (71.3) 114 (68.3) Physical activity (N, %) 0.17 Inactive 188 (30.1) 52 (33.1) 38 (24.5) 41 (26.5) 57 (36.7) Moderate active 116 (18.6) 32 (20.4) 31 (19.6) 27 (17.4) 26 (16.8) Active 321 (51.4) 73 (46.5) 89 (56.3) 87 (56.1) 72 (46.5) Hypertension (N, %) 106 (17.2) 18 (11.6) 28 (17.7) 28 (18.1) 33 (21.4) 0.14 Dyslipidemia (N, %) 254 (38.2) 47 (28.3) 64 (38.8) 66 (39.5) 77 (46.1) 0.01 SBP (mmHg) 121.1 ± 17.3 117.5 ± 14.8 121.9 ± 19.4 122.5 ± 16.2 122.3 ± 18.3 0.04 DBP (mmHg) 75.3 ± 12.0 74.0 ± 11.1 75.2 ± 11.7 75.7 ± 11.2 76.6 ± 13.6 0.26 RBC (1012/L) 4.4 ± 0.3 4.3 ± 0.4 4.4 ± 0.3 4.4 ± 0.3 4.5 ± 0.3 < 0.01 LDL-C (mmol/L) 2.2 ± 0.7 2.1 ± 0.7 2.2 ± 0.6 2.3 ± 0.8 2.2 ± 0.8 0.22 HDL-C (mmol/L) 1.3 ± 0.3 1.4 ± 0.3 1.3 ± 0.3 1.3 ± 0.3 1.3 ± 0.3 0.48 TG (mmol/L) 1.4 ± 0.9 1.3 ± 0.8 1.5 ± 1.0 1.3 ± 0.7 1.6 ± 1.0 < 0.01 TC (mmol/L) 5.1 ± 1.0 4.9 ± 0.9 5.1 ± 0.8 5.2 ± 1.0 5.2 ± 1.0 0.08 ALT (U/L) 26.4 ± 21.8 24.2 ± 17.6 27.0 ± 27.2 28.2 ± 24.8 26.2 ± 16.0 0.38 AST (U/L) 24.3 ± 14.1 22.8 ± 8.6 24.6 ± 15.2 25.9 ± 20.9 23.7 ± 7.8 0.24 Note. ALT, alanine aminotransferase; AST, aspartate aminotransferase; DBP, diastolic blood pressure; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; NAD+, nicotinamide adenine dinucleotide; RBC, red blood cell; SBP, systolic blood pressure; TC, total cholesterol; TG, triglycerides. Data are shown as frequency and percentage for categorical variables and mean ± standard deviation for continuous variables. -
Among the 665 eligible participants, nabothian cysts occurred in 184 (27.7%) participants with, single cysts in 100 (15.0%), and multiple cysts in 84 (12.6%) (Supplementary Table S1, available in www.besjournal.com). The prevalence of nabothian cysts showed a decreasing trend across NAD+ quartiles (P for trend = 0.09). Figure 2A does not show a significant correlation between age and whole-blood NAD+ levels in participants with nabothian cysts (r = 0.04) and in those without nabothian cysts (r = 0.07). Figure 2B shows the prevalence of nabothian cysts according to NAD+ quartiles. The prevalence of total nabothian cysts among participants in the NAD+ quartiles Q1, Q2, Q3, and Q4 were 37.4%, 24.9%, 27.0%, and 21.6%, respectively. The lowest NAD+ level group (Q1) had the highest percentage of total nabothian cysts as well as single and multiple nabothian cysts (P < 0.05).
Figure 2. Association between whole-blood NAD+ levels and nabothian cysts in the study population. (A) Correlation between age and NAD+ levels in non-nabothian or nabothian cysts. (B) Comparison of the prevalence of nabothian cysts among the NAD+ quartile groups. NAD+, nicotinamide adenine dinucleotide; Non-NC, non-nabothian cyst; NC, nabothian cyst; Q1, first quartile; Q2, second quartile; Q3, third quartile; Q4, fourth quartile. *indicates statistical difference between the quartile of interest and the first quartile of NAD+ levels. *P < 0.05, **P < 0.01.
Table S1. Descriptive statistics of the characteristics of eligible participants with or without nabothian cysts
Characteristic Non-nabothian cysts
(n = 481)Nabothian cysts P value Total
(n = 184)Single-cystic
(n = 100)Multiple-cystic
(n = 84)NAD+ (μmol/L) 31.7 ± 5.4 30.3 ± 5.1 30.1 ± 5.1 30.5 ± 5.0 0.002 Age (y) 43.9 ± 12.4 40.8 ± 8.2 40.2 ± 8.5 41.4 ± 8.0 < 0.001 Age at menarche (y) 13.9 ± 1.5 13.7 ± 1.3 13.6 ± 1.3 13.7 ± 1.2 0.01 Age at menopausal (y) 49.9 ± 3.9 50.8 ± 3.0 50.7 ± 3.3 50.7 ± 2.9 0.26 Body mass index (N, %) 0.02 < 24 kg/m2 286 (59.5) 128 (69.6) 75 (75.0) 53 (63.1) ≥ 24 kg/m2 195 (40.5) 56 (30.4) 25 (25.0) 31 (36.9) Education level (N, %) 0.01 Middle school or below 161 (33.9) 43 (23.4) 14 (13.2) 23 (30.3) College or above 318 (66.1) 141 (76.6) 63 (81.8) 53 (69.7) Physical activity (N, %) 0.15 Inactive 128 (28.7) 60 (33.5) 24 (32.0) 27 (37.0) Moderate active 78 (17.5) 38 (21.2) 20 (26.7) 13 (17.8) Active 240 (53.8) 81 (45.3) 31 (41.3) 33 (45.2) Hypertension (N, %) 89 (18.9) 25 (13.7) 5 (6.5) 14 (18.7) 0.111 Dyslipidemia (N, %) 206 (42.8) 48 (26.1) 18 (23.4) 22 (29.0) < 0.001 SBP (mmHg) 122.3 ± 18.0 117.7 ± 15.2 115.3 ± 13.5 120.5 ± 16.7 0.003 DBP (mmHg) 75.6 ± 11.6 74.7 ± 12.6 73.1 ± 12.5 76.6 ± 12.7 0.38 RBC (1012/L) 4.4 ± 0.3 4.4 ± 0.4 4.4 ± 0.3 4.5 ± 0.4 0.31 LDL-C (mmol/L) 2.2 ± 0.7 2.1 ± 0.8 2.0 ± 0.8 2.2 ± 0.7 0.03 HDL-C (mmol/L) 1.3 ± 0.3 1.4 ± 0.3 1.3 ± 0.3 1.4 ± 0.2 0.12 TG (mmol/L) 1.5 ± 1.0 1.2 ± 0.7 1.1 ± 0.5 1.3 ± 0.8 < 0.001 TC (mmol/L) 5.2 ± 1.0 4.9 ± 0.9 4.8 ± 0.9 5.0 ± 0.8 0.002 ALT (U/L) 27.1 ± 23.9 24.3 ± 14.8 22.7 ± 13.5 26.3 ± 16.1 0.07 AST (U/L) 24.6 ± 15.9 23.3 ± 8.0 22.4 ± 7.1 24.3 ± 9.0 0.16 Note. ALT, alanine aminotransferase; AST, asparate aminotransferase; DBP, diastolic blood pressure; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; NAD+, nicotinamide adenine dinucleotide; RBC, red blood cell; SBP, systolic blood pressure; TC, total cholesterol; TG, triglycerides. Data are shown as frequency and percentage for categorical variables and mean ± standard deviation for continuous variables. -
Analysis of the association between NAD+ and nabothian cysts is shown in Table 2. In the unadjusted model, as compared with the NAD+ Q4, ORs with 95% CI of NAD+ Q1 was 2.17 (1.34–3.52) for total nabothian cysts, 1.89 (1.04–3.44) for the single cyst, and 2.61 (1.31–5.19) for multiple cysts. In the multivariate model, as compared with the NAD+ Q4, ORs with 95% CI of NAD+ Q1 was 2.02 (1.21–3.40) for total nabothian cysts, 1.56 (0.82–2.97) for the single cyst, and 2.98 (1.44–6.18) for multiple cysts.
Table 2. Association of whole-blood NAD+ levels with nabothian cysts in all participants
Nabothian cysts Events, N (%) Odds ratios (95% CI) Unadjusted Adjusted Total 184 (27.7) Q1 62 (37.4) 2.17 (1.34, 3.52) 2.02 (1.21, 3.40) Q2 41 (24.9) 1.20 (0.72, 2.00) 1.13 (0.66, 1.95) Q3 45 (27.0) 1.34 (0.81, 2.22) 1.26 (0.74, 2.14) Q4 36 (21.6) Ref Ref Single-cystic 100 (17.2) Q1 33 (24.1) 1.89 (1.04, 3.44) 1.56 (0.82, 2.97) Q2 25 (16.8) 1.20 (0.64, 2.24) 1.04 (0.54, 2.02) Q3 20 (14.1) 0.98 (0.51, 1.88) 0.88 (0.44, 1.75) Q4 22 (14.4) Ref Ref Multiple-cystic 84 (14.9) Q1 29 (21.8) 2.61 (1.31, 5.19) 2.98 (1.44, 6.18) Q2 16 (11.4) 1.21 (0.57, 2.58) 1.26 (0.57, 2.80) Q3 25 (17.0) 1.92 (0.95, 3.86) 2.02 (0.97, 4.22) Q4 14 (9.7) Ref Ref Note. CI, confidence interval; Ref, reference; Q1, first quartile; Q2, second quartile; Q3, third quartile; Q4, fourth quartile. Adjustments were made for age, body mass index, history of dyslipidemia, systolic blood pressure, triglyceride level, and number of red blood cells. Subgroup analysis showed associations of NAD+ levels with nabothian cyst in participants stratified by age (< 40 vs. ≥ 40 years), BMI (< 24 vs. ≥ 24 kg/m2), SBP (< 120 vs. ≥ 120 mmHg), and dyslipidemia as see Supplementary Table S2 (available in www.besjournal.com). Age, BMI, SBP, and history of dyslipidemia had no impact on the association between whole-blood NAD+ levels and nabothian cysts (P > 0.05 for interaction).
Table S2. Association of whole blood NAD+ contents with nabothian cysts in each subgroup
Nabothian cysts Age Body Mass Index Systolic Blood Pressure Dyslipidemia Adjusted OR
(95% CI)P for inter-action Adjusted OR
(95% CI)P for inter-action Adjusted OR
(95% CI)P for inter-action Adjusted OR
(95% CI)P for inter-action < 40 y
(n = 302)≥ 40 y
(n = 363)< 24 kg/m2
(n = 414)≥ 24 kg/m2
(n = 251)<120 mmHg
(n = 377)≥ 120 mmHg
(n = 288)Yes
(n = 254)No
(n = 411)Total Q1 1.95
(0.93, 4.11)1.99
(0.96, 4.11)0.93 1.53
(0.81, 2.89)3.69
(1.51, 9.00)0.81 2.03
(1.05, 3.92)2.52
(1.12, 5.69)0.58 2.04
(1.10, 3.82)2.05
(0.81, 5.18)0.53 Q2 0.98
(0.45, 2.13)1.21
(0.57, 2.59)1.19
(0.62, 2.26)0.88
(0.30, 2.54)1.50
(0.77, 2.92)0.82
(0.33, 2.01)1.26
(0.65, 2.43)0.88
(0.32, 2.38)Q3 1.41
(0.65, 3.05)1.09
(0.52, 2.28)1.11
(0.58, 2.11)1.52
(0.58, 3.98)1.80
(0.91, 3.56)1.02
(0.45, 2.32)1.14
(0.59, 2.20)1.40
(0.57, 3.44)Q4 Ref Ref Ref Ref Ref Ref Ref Ref Single-cystic Q1 1.55
(0.65, 3.70)1.47
(0.55, 3.95)0.76 1.17
(0.55, 2.47)4.04
(1.10, 14.84)0.37 1.58
(0.74, 3.38)2.61
(0.84, 8.14)0.82 1.43
(0.67, 3.03)2.20
(0.63, 7.73)0.74 Q2 0.97
(0.40, 2.33)1.08
(0.38, 3.02)1.02
(0.48, 2.15)0.97
(0.20, 4.68)1.42
(0.66, 3.03)0.70
(0.18, 2.73)1.08
(0.50, 2.33)0.87
(0.22, 3.47)Q3 0.74
(0.27, 2.02)1.02
(0.38, 2.72)0.63
(0.28, 1.42)1.98
(0.50, 7.88)1.05
(0.45, 2.46)1.08
(0.33, 3.54)0.70
(0.31, 1.61)1.43
(0.41, 5.06)Q4 Ref Ref Ref Ref Ref Ref Ref Ref Multiple-cystic Q1 3.23
(1.02, 10.34)2.62
(1.02, 6.76)0.83 2.62
(0.98, 7.02)3.68
(1.24, 10.89)0.67 3.66
(1.21, 11.05)2.48
(0.90, 6.80)0.52 3.77
(1.46, 9.69)1.82
(0.53, 6.25)0.36 Q2 0.96
(0.26, 3.57)1.41
(0.51, 3.88)1.51
(0.64, 4.21)0.83
(0.22, 3.15)1.96
(0.62, 6.24)0.91
(0.30, 2.72)1.66
(0.59, 4.63)0.85
(0.22, 3.27)Q3 3.41
(1.10, 10.57)1.24
(0.46, 3.37)2.39
(0.92, 6.24)1.27
(0.37, 4.38)4.56
(1.52, 13.67)1.06
(0.37, 2.99)2.32
(0.88, 6.13)1.44
(0.44, 4.77)Q4 Ref Ref Ref Ref Ref Ref Ref Ref Note. OR, odds ratio; CI, confidence interval; Ref, reference. Adjustments made for age, body mass index, history of dyslipidemia, systolic blood pressure, triglyceride level, and number of red blood cells. -
Figure 3 shows the adjusted dose-response association of whole-blood NAD+ levels with nabothian cysts. The risk of total and single nabothian cysts decreased linearly with increasing NAD+ levels (Figure 3A and B). The occurrence of multiple cysts showed non-linear decline along with the NAD+ level (Figure 3C). The risk of multiple nabothian cysts was stable before NAD+ levels of < 28 μmol/L, after which it linearly declined at NAD+ levels of 28 to 35 μmol/L, and then became stable after NAD+ levels of > 35 μmol/L.
Figure 3. Dose-response association of whole-blood NAD+ levels with nabothian cysts. (A) Restricted cubic spline curves for the association of NAD+ levels with total nabothian cysts; (B) single nabothian cyst; and (C) multiple nabothian cysts. The model was adjusted for age, body mass index, history of dyslipidemia, systolic blood pressure, triglyceride level, and number of red blood cells. The black line represents the odds ratios and the gray shaded area represents the 95% CI. NAD+, nicotinamide adenine dinucleotide; CI, confidence interval.
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Characteristics of Participants at Baseline
Nabothian Cyst Prevalence among NAD+ Quartiles
Association of Whole-blood NAD+ Levels with Nabothian Cysts
Restricted Cubic Spline Curves for the Associations of NAD+ with Nabothian Cyst
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