| Literature DB >> 28837106 |
Xiaojun Zhu1, Panjun Gao2, Yishuo Gu3, Pei Xiao4, Mengxuan Liu5, Juan Chen6, Yacai Cen7, Wenjun Ma8, Tao Li9.
Abstract
Since the number of greenhouse workers are increasing in China, this observational cross-sectional study was designed to evaluate lung function and discuss the potential risk factors, to provide evidence in the surveillance of greenhouse workers' health. 678 greenhouse workers in Gansu Province, China were enrolled. A questionnaire which included demographic and occupational information was used. Vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in 1 s (FEV₁), and FEV₁:FVC ratios (FEV₁/FVC), maximal expiratory flow after 50% of the FVC has not been exhaled (MEF50), maximal expiratory flow after 25% of the FVC has not been exhaled (MEF25) and maximal mid-expiratory flow curve (MMEF) were measured as lung function indicators. The mean values and standard deviations (SDs) of VC% predicted, FVC% predicted, FEV₁% predicted and FEV₁/FVC ratio were 106.07 ± 13.36, 107.60 ± 13.95, 97.19 ± 14.80 and 89.76 ± 10.78 respectively. The positive rates of above four and abnormal lung ventilation function were 2.9%, 2.8%, 11.2%, 4.6% and 6.5% respectively. Gender, age, BMI and number of greenhouses owned were influence factors of lung ventilation function (p < 0.05). The mean values and SDs of MEF50% predicted, MEF25% predicted and MMEF% predicted were 69.63 ± 24.95, 54.04 ± 24.94 and 66.81 ± 24.53. The positive rates of above three and abnormal small airway function were 45.0%, 72.1%, 47.2% and 49.4% respectively. Age, education and number of greenhouses owned were influence factors for small airway function (p < 0.05). Working in a greenhouse might influence lung function of the workers. Small airway function indicators could be used as priority indicators for the surveillance of greenhouse workers' health.Entities:
Keywords: greenhouse working; lung function; small airway function
Mesh:
Year: 2017 PMID: 28837106 PMCID: PMC5615493 DOI: 10.3390/ijerph14090956
Source DB: PubMed Journal: Int J Environ Res Public Health ISSN: 1660-4601 Impact factor: 3.390
Equation of calculating individual predicted value of lung function indicators developed by European Respiratory Society (ERS) in 1993.
| Indicators | Equations | |
|---|---|---|
| Male | Female | |
| VC | 0.0466 × (Height × 100) − 0.024 × Age − 3.28 | 0.061 × (Height × 100) − 0.028 × Age − 4.65 |
| FVC | 0.0576 × (Height × 100) − 0.026 × Age − 4.34 | 0.0443 × (Height × 100) − 0.026 × Age − 2.89 |
| FEV1 | 0.0395 × (Height × 100) − 0.025 × Age − 2.60 | 0.0430 × (Height × 100) − 0.029 × Age − 2.49 |
| MEF50 | 0.0379 × (Height × 100) − 0.031 × Age − 0.35 | 0.0245 × (Height × 100) − 0.025 × Age + 1.16 |
| MEF25 | 0.0261 × (Height × 100) − 0.026 × Age − 1.34 | 0.0105 × (Height × 100) − 0.025 × Age + 1.11 |
| MMEF | 0.0194 × (Height × 100) − 0.043 × Age + 2.70 | 0.0125 × (Height × 100) − 0.034 × Age + 2.92 |
Variable assignment of independent variables of logistic regression analysis.
| Variable | Assignment |
|---|---|
| Gender | 0 = male, 1 = female |
| Age, years | 1 = ≤47, 2 = >47 |
| BMI | 1 = <18.5, 2 = 18.5 − 23.9, 3 = <23.9 |
| Education | 1 = Primary school and below, 2 = Middle school, 3 = High school and above |
| Smoking | 0 = never−smokers, 1 = ever−smokers |
| Working lifetime, years | 1 = 0−5, 2 = 6−10, 3 = 11−20, 4 = >20 |
| Number of greenhouses owned | 1 = 1, 2 = 2, 3 = 3, 4 = >3 |
| Mask using | 0 = No, 1 = Yes |
Dummy variable contrast coding for logistic regression.
| Variable | N | Coding | |||
|---|---|---|---|---|---|
| (1) | (2) | (3) | |||
| BMI | <18.5 | 31 | 0 | 0 | - |
| 18.5–23.9 | 336 | 1 | 0 | - | |
| >23.9 | 311 | 0 | 1 | - | |
| Education | Primary school and below | 268 | 1 | 0 | - |
| Middle school | 319 | 0 | 1 | - | |
| High school and above | 91 | 0 | 0 | - | |
| Working lifetime, years | 0–5 | 104 | 0 | 0 | 0 |
| 6–10 | 218 | 1 | 0 | 0 | |
| 11–20 | 317 | 0 | 1 | 0 | |
| >20 | 39 | 0 | 0 | 1 | |
| Numbers of greenhouses owned | 1 | 184 | 0 | 0 | 0 |
| 2 | 281 | 1 | 0 | 0 | |
| 3 | 143 | 0 | 1 | 0 | |
| >3 | 70 | 0 | 0 | 1 | |
The mean values and standard deviations (SDs) of lung ventilation function indicators among greenhouse workers.
| N (%) | VC% Predicted | FVC% Predicted | FEV1% Predicted | FEV1/FVC | |
|---|---|---|---|---|---|
| Total | 678 (100) | 106.07 ± 13.36 | 107.60 ± 13.95 | 97.19 ± 14.80 | 89.76 ± 10.78 |
| Gender | |||||
| Male | 291 (42.9) | 102.04 ± 12.83 a | 104.66 ± 13.89 a | 93.86 ± 15.27 a | 89.67 ± 10.92 |
| Female | 387 (57.1) | 109.10 ± 12.95 | 109.81 ± 13.60 | 99.69 ± 13.94 | 89.83 ± 10.69 |
| Age (years) | |||||
| ≤47 | 365 (53.8) | 105.31 ± 11.96 | 106.78 ± 12.17 | 97.94 ± 12.49 | 90.10 ± 10.76 |
| >47 | 313 (46.2) | 106.95 ± 14.79 | 108.55 ± 15.75 | 96.30 ± 17.09 | 89.36 ± 10.80 |
| BMI | |||||
| <18.5 | 31 (4.6) | 98.85 ± 11.87 a,b | 99.95 ± 13.68 a,b | 89.96 ± 16.37 a,b | 86.80 ± 90.17 |
| 18.5–23.9 | 336 (49.6) | 106.35 ± 13.27 | 108.07 ± 14.01 | 98.07 ± 14.66 | 90.17 ± 10.57 |
| >23.9 | 311 (45.9) | 106.49 ± 13.42 | 107.85 ± 13.74 | 96.95 ± 14.63 | 89.62 ± 10.90 |
| Education | |||||
| Primary school and below | 268 (39.5) | 107.45 ± 14.84 | 108.72 ± 15.38 | 97.74 ± 16.41 | 90.40 ± 10.04 |
| Middle school | 319 (47.1) | 105.34 ± 12.24 | 106.90 ± 13.05 | 97.45 ± 13.04 | 89.59 ± 10.99 |
| High school and above | 91 (13.4) | 104.57 ± 12.21 | 106.74 ± 12.42 | 94.61 ± 15.49 | 88.48 ± 12.05 |
| Smoking | |||||
| Never-smokers | 454 (67) | 107.71 ± 13.16 a | 108.75 ± 13.70 a | 98.90 ± 14.15 a | 89.64 ± 11.24 |
| Ever-smokers | 224 (33) | 102.75 ± 13.15 | 105.26 ± 14.19 | 93.71 ± 15.50 | 89.82 ± 10.56 |
| Number of greenhouses owned | |||||
| 1 | 184 (27.1) | 106.07 ± 14.72 | 107.46 ± 15.27 | 97.50 ± 16.39 | 90.55 ± 1050 |
| 2 | 281 (41.4) | 106.06 ± 13.02 | 107.54 ± 13.77 | 96.62 ± 14.64 | 89.52 ± 10.49 |
| 3 | 143 (21.1) | 106.96 ± 12.31 | 108.72 ± 12.51 | 97.92 ± 14.11 | 89.90 ± 11.84 |
| >3 | 70 (10.3) | 104.32 ± 13.09 | 105.90 ± 13.94 | 97.02 ± 12.42 | 88.40 ± 10.39 |
| Working lifetime (years) | |||||
| 0–5 | 104 (15.3) | 102.82 ± 12.29 a,b | 103.68 ± 13.37 a,b | 95.10 ± 14.86 a,b | 89.12 ± 11.23 |
| 6–10 | 218 (32.2) | 106.56 ± 13.45 | 108.05 ± 14.19 | 98.65 ± 14.60 | 89.21 ± 11.90 |
| 11–20 | 317 (46.8) | 106.46 ± 13.21 | 108.15 ± 13.55 | 96.98 ± 14.66 | 90.01 ± 9.97 |
| >20 | 39 (5.8) | 108.88 ± 15.61 | 111.02 ± 15.74 | 96.20 ± 16.55 | 92.50 ± 9.03 |
| Mask using | |||||
| No | 497 (73.3) | 105.41 ± 13.69 a | 107.14 ± 14.33 | 96.58 ± 14.75 | 89.59 ± 11.26 |
| Yes | 181 (26.7) | 107.89 ± 12.24 | 108.87 ± 12.80 | 98.84 ± 14.85 | ± 9.33 |
a: p < 0.05; b: ptrend < 0.05.
Positive rates of abnormal lung ventilation function among greenhouse workers.
| N (%) | VC% Predicted <80% | FVC% Predicted <80% | FEV1% Predicted <80% | FEV1/FVC <70% | Abnormal Lung Ventilation Function | |
|---|---|---|---|---|---|---|
| Total | 678 (100) | 20 (2.9) | 19 (2.8) | 76 (11.2) | 31 (4.6) | 44 (6.5) |
| Gender | ||||||
| Male | 291 (42.9) | 11 (3.8) | 10 (3.4) | 43 (14.8) a | 17 (5.8) | 23 (7.9) |
| Female | 387 (57.1) | 9 (2.3) | 9 (2.3) | 33 (8.5) | 14 (3.6) | 21 (5.4) |
| Age (years) | ||||||
| ≤47 | 365 (53.8) | 8 (2.2) | 6 (1.6) a | 26 (7.1) a | 15 (4.1) | 21 (5.8) |
| >47 | 313 (46.2) | 12 (3.8) | 13 (4.2) | 50 (16.0) | 16 (5.1) | 23 (7.3) |
| BMI | s | |||||
| <18.5 | 31 (4.6) | 1 (3.2) | 3 (9.7) | 8 (25.8) a | 3 (9.7) | 3 (9.7) |
| 18.5–23.9 | 336 (49.6) | 10 (3.0) | 8 (2.4) | 37 (11.0) | 11 (3.3) | 17 (5.1) |
| >23.9 | 311 (45.9) | 9 (2.9) | 8 (2.6) | 31 (10.0) | 17 (5.5) | 24 (7.7) |
| Education | ||||||
| Primary school and below | 268 (39.5) | 10 (3.7) | 10 (3.7) | 33 (12.3) | 10 (3.7) | 18 (6.7) |
| Middle school | 319 (47.1) | 9 (2.8) | 8 (2.5) | 29 (9.1) | 15 (4.7) | 19 (6.0) |
| High school and above | 91 (13.4) | 1 (1.1) | 1 (1.1) | 14 (15.4) | 6 (6.6) | 7 (7.7) |
| Smoking | ||||||
| Never-smokers | 454 (67) | 12 (2.6) | 13 (2.9) | 43 (9.5)a | 18 (4.0) | 28 (6.2) |
| Ever-smokers | 224 (33) | 8 (3.6) | 6 (2.7) | 33 (14.7) | 13 (5.8) | 16 (7.1) |
| Number of greenhouses owned | ||||||
| 1 | 184 (27.1) | 11 (6.0) a,b | 10 (5.4) a,b | 21 (11.4) | 8 (4.3) | 17 (9.2) |
| 2 | 281 (41.4) | 5 (1.8) | 5 (1.8) | 37 (13.2) | 13 (4.6) | 15 (5.3) |
| 3 | 143 (21.1) | 2 (1.4) | 1 (0.7) | 12 (8.4) | 5 (3.5) | 6 (4.2) |
| >3 | 70 (10.3) | 2 (2.9) | 3 (4.3) | 6 (8.6) | 5 (7.1) | 6 (8.6) |
| Working lifetime (years) | ||||||
| 0–5 | 104 (15.3) | 3 (2.9) | 5 (4.8) | 12 (11.5) | 7 (6.7) | 9 (8.7) |
| 6–10 | 218 (32.2) | 8 (3.7) | 8 (3.7) | 23 (10.6) | 9 (4.1) | 16 (7.3) |
| 11–20 | 317 (46.8) | 7 (2.2) | 6 (1.9) | 34 (10.7) | 14 (4.4) | 18 (5.7) |
| >20 | 39 (5.8) | 2 (5.1) | 0 (0.0) | 7 (17.9) | 1 (2.6) | 1 (2.6) |
| Mask using | ||||||
| No | 497 (73.3) | 18 (3.6) | 16 (3.2) | 62 (12.5) | 26 (5.2) | 37 (7.4) |
| Yes | 181 (26.7) | 2 (2.9) | 3 (1.7) | 14 (7.7) | 5 (2.8) | 7 (3.9) |
a: p < 0.05; b: ptrend < 0.05.
Influence factors of lung ventilation function among greenhouse workers.
| Variables in the Equation | B | Wald | OR (95% CI) | ||
|---|---|---|---|---|---|
| VC% predicted | Number of greenhouses owned | 7.383 | 0.061 | ||
| Number of greenhouses owned (1) | −1.244 | 5.130 | 0.024 | 0.288 (0.098, 0.846) | |
| Number of greenhouses owned (2) | −1.509 | 3.759 | 0.053 | 0.221 (0.048, 1.017) | |
| Number of greenhouses owned (3) | −0.791 | 1.019 | 0.313 | 0.453 (0.098, 2.106) | |
| Mask using | −1.208 | 2.576 | 0.108 | 0.299 (0.068, 1.306) | |
| FVC% predicted | Age | 0.868 | 2.872 | 0.090 | 2.383 (0.873, 6.505) |
| Number of greenhouses owned | 6.727 | 0.081 | |||
| Number of greenhouses owned (1) | −1.142 | 4.199 | 0.040 | 0.319 (0.107, 0.951) | |
| Number of greenhouses owned (2) | −1.882 | 3.138 | 0.076 | 0.152 (0.019, 1.222) | |
| Number of greenhouses owned (3) | −0.046 | 0.004 | 0.947 | 0.955 (0.249, 3.668) | |
| FEV1% predicted | Gender | −0.596 | 5.672 | 0.017 | 0.551 (0.338, 0.900) |
| Age | 0.862 | 11.163 | 0.001 | 2.368 (1.428, 3.927) | |
| BMI | 5.385 | 0.068 | |||
| BMI (1) | −0.948 | 4.295 | 0.038 | 0.388 (0.158, 0.950) | |
| BMI (2) | −1.066 | 5.319 | 0.021 | 0.345 (0.139, 0.852) | |
| FEV1/FVC | - | ||||
| Lung ventilation function | Mask using | −0.693 | 2.700 | 0.100 | 0.500 (0.219, 1.143) |
(1), (2) and (3) were the dummy variable contrast coding for logistic regression which were showed as Table A3.
The mean values and SDs of small airway function indicators among greenhouse workers.
| Variables | N (%) | MEF50% Predicted | MEF25% Predicted | MMEF% Predicted |
|---|---|---|---|---|
| Total | 678 (100) | 69.63 ± 24.95 | 54.04 ± 24.94 | 66.81 ± 24.53 |
| Gender | ||||
| Male | 291 (42.9) | 69.44 ± 25.42 | 54.74 ± 25.77 | 67.05 ± 24.84 |
| Female | 387 (57.1) | 69.77 ± 24.62 | 53.51 ± 24.31 | 66.63 ± 24.33 |
| Age (years) | ||||
| ≤47 | 365 (53.8) | 71.33 ± 25.13 | 55.63 ± 26.04 | 68.84 ± 24.59 a |
| >47 | 313 (46.2) | 67.66 ± 24.63 | 52.17 ± 23.49 | 64.45 ± 24.28 |
| BMI | ||||
| <18.5 | 31 (4.6) | 64.38 ± 25.74 | 53.07 ± 26.86 | 60.42 ± 2750 |
| 18.5–23.9 | 336 (49.6) | 70.54 ± 25.57 | 53.75 ± 24.84 | 67.54 ± 24.31 |
| >23.9 | 311 (45.9) | 69.18 ± 24.19 | 54.43 ± 24.92 | 66.67 ± 24.45 |
| Education | ||||
| Primary school and below | 268 (39.5) | 71.42 ± 24.91 a,b | 53.43 ± 23.36 | 68.01 ± 24.10 b |
| Middle school | 319 (47.1) | 69.88 ± 25.26 | 55.05 ± 26.02 | 67.33 ± 25.08 |
| High school and above | 91 (13.4) | 63.52 ± 23.25 | 52.26 ± 25.65 | 61.47 ± 23.39 |
| Smoking | ||||
| Never-smokers | 454 (67) | 69.61 ± 25.20 | 53.30 ± 24.28 | 66.44 ± 24.58 |
| Ever-smokers | 224 (33) | 69.68 ± 24.50 | 55.53 ± 26.22 | 67.57 ± 24.48 |
| Number of greenhouses owned | ||||
| 1 | 184 (27.1) | 69.92 ± 24.03 | 55.67 ± 24.43 | 67.60 ± 24.54 |
| 2 | 281 (41.4) | 69.28 ± 24.80 | 51.98 ± 24.16 | 65.69 ± 24.25 |
| 3 | 143 (21.1) | 71.49 ± 26.52 | 57.49 ± 28.11 | 69.54 ± 26.01 |
| >3 | 70 (10.3) | 66.50 ± 24.81 | 50.94 ± 21.56 | 63.67 ± 22.26 |
| Working lifetime (years) | ||||
| 0–5 | 104 (15.3) | 68.96 ± 23.64 | 53.62 ± 23.98 | 66.17 ± 25.31 |
| 6–10 | 218 (32.2) | 68.19 ± 24.66 | 53.24 ± 25.14 | 65.72 ± 24.68 |
| 11–20 | 317 (46.8) | 69.93 ± 25.40 | 53.93 ± 25.09 | 66.96 ± 24.24 |
| >20 | 39 (5.8) | 77.03 ± 25.87 | 60.45 ± 25.06 | 73.45 ± 23.83 |
| Mask using | ||||
| No | 497 (73.3) | 69.18 ± 24.26 | 53.79 ± 24.92 | 66.44 ± 24.35 |
| Yes | 181 (26.7) | 70.89 ± 26.52 | 54.69 ± 25.04 | 67.83 ± 25.05 |
a: p < 0.05; b: ptrend < 0.05.
Positive rates of abnormal small airway function among greenhouse workers.
| Variables | N (%) | MEF50% Predicted <65% | MEF25% Predicted <65% | MMEF% Predicted <65% | Abnormal Small Airway Function |
|---|---|---|---|---|---|
| Total | 678 (100) | 305 (45.0) | 489 (72.1) | 320 (47.2) | 335 (49.4) |
| Gender | |||||
| Male | 291 (42.9) | 133 (45.7) | 206 (70.8) | 139 (47.8) | 145 (49.8) |
| Female | 387 (57.1) | 172 (44.4) | 283 (73.1) | 181 (46.8) | 190 (49.1) |
| Age (years) | |||||
| ≤47 | 365 (53.8) | 154 (42.2) | 258 (70.7) | 157 (43.0)a | 164 (44.9) a |
| >47 | 313 (46.2) | 151 (48.2) | 231 (73.8) | 163 (52.1) | 171 (54.6) |
| BMI | |||||
| <18.5 | 31 (4.6) | 16 (51.6) | 22 (71.0) | 18 (58.1) | 18 (58.1) |
| 18.5–23.9 | 336 (49.6) | 150 (44.6) | 243 (72.3) | 152 (45.2) | 161 (47.9) |
| >23.9 | 311 (45.9) | 139 (44.7) | 224 (72.0) | 150 (48.2) | 156 (50.2) |
| Education | |||||
| Primary school and below | 268 (39.5) | 113 (42.2) | 187 (69.8) | 116 (43.3) b | 124 (46.3) |
| Middle school | 319 (47.1) | 143 (44.8) | 236 (74.0) | 154 (48.3) | 159 (49.8) |
| High school and above | 91 (13.4) | 49 (53.8) | 66 (72.5) | 50 (54.9) | 52 (57.1) |
| Smoking | |||||
| Never-smokers | 454 (67) | 201 (44.3) | 333 (73.3) | 211 (46.5) | 224 (49.3) |
| Ever-smokers | 224 (33) | 104 (46.4) | 156 (69.6) | 109 (48.7) | 111 (49.6) |
| Number of greenhouses owned | |||||
| 1 | 184 (27.1) | 75 (40.8) | 120 (65.2) | 76 (41.3) | 81 (44.0) |
| 2 | 281 (41.4) | 135 (48.0) | 213 (75.8) | 143 (50.9) | 148 (52.7) |
| 3 | 143 (21.1) | 59 (41.3) | 102 (71.3) | 64 (44.8) | 67 (46.9) |
| >3 | 70 (10.3) | 36 (51.4) | 54 (77.1) | 37 (52.9) | 39 (55.7) |
| Working lifetime (years) | |||||
| 0–5 | 104 (15.3) | 49 (47.1) | 73 (70.2) | 45 (43.3) | 51 (49.0) |
| 6–10 | 218 (32.2) | 96 (44.0) | 162 (74.3) | 102 (46.8) | 105 (48.2) |
| 11–20 | 317 (46.8) | 148 (46.7) | 229 (72.2) | 159 (50.2) | 166 (52.4) |
| >20 | 39 (5.8) | 12 (30.8) | 25 (64.1) | 14 (35.9) | 13 (33.3) |
| Mask using | |||||
| No | 497 (73.3) | 222 (44.7) | 361 (72.6) | 232 (46.7) | 241 (48.5) |
| Yes | 181 (26.7) | 83 (45.9) | 128 (70.7) | 88 (48.6) | 94 (51.9) |
a: p < 0.05; b: ptrend < 0.05.
Influence factors of small airway function among greenhouse workers.
| Variables in The Equation | B | Wald | OR (95%CI) | ||
|---|---|---|---|---|---|
| MEF50% predicted | - | ||||
| MEF25% predicted | Number of greenhouses owned | 7.113 | 0.068 | ||
| Number of greenhouses owned (1) | 0.513 | 6.074 | 0.014 | 1.671 (1.111, 2.513) | |
| Number of greenhouses owned (2) | 0.283 | 1.375 | 0.241 | 1.327 (0.827, 2.129) | |
| Number of greenhouses owned (3) | 0.588 | 3.291 | 0.070 | 1.800 (0.954, 3.397) | |
| MMEF% predicted | Age | 0.506 | 8.417 | 0.004 | 1.659 (1.178, 2.335) |
| Education | 6.870 | 0.032 | |||
| Education (1) | −0.487 | 3.923 | 0.048 | 0.614 (0.379, 0.995) | |
| Education (2) | −0.059 | 0.055 | 0.815 | 0.943 (0.577, 1.542) | |
| Small airway function | Age | 0.562 | 9.959 | 0.002 | 1.754 (1.237, 2.485) |
| Education | 5.424 | 0.066 | |||
| Education (1) | −0.446 | 3.202 | 0.074 | 0.640 (0.393, 1.043) | |
| Education (2) | −0.068 | 0.072 | 0.788 | 0.934 (0.568, 1.535) | |
| Working lifetime | 6.334 | 0.096 | |||
| Working lifetime (1) | 0.015 | 0.004 | 0.952 | 1.015 (0.632, 1.629) | |
| Working lifetime (2) | 0.137 | 0.353 | 0.552 | 1.146 (0.731, 1.798) | |
| Working lifetime (3) | −0.775 | 3.724 | 0.054 | 0.461 (0.210, 1.012) |
(1), (2) and (3) were the dummy variable contrast coding for logistic regression which were showed as Table A3.
Relationship between mask using and possible influence factors.
| Mask Using | |||
|---|---|---|---|
| No | Yes | ||
| BMI | 0.172 | ||
| <18.5 | 25 | 6 | |
| 18.5–23.9 | 236 | 100 | |
| >23.9 | 236 | 75 | |
| Education | 0.967 | ||
| Primary school and below | 195 | 73 | |
| Middle school | 235 | 84 | |
| High school and above | 67 | 24 | |
| Number of greenhouses owned | 0.901 | ||
| 1 | 136 | 48 | |
| 2 | 202 | 79 | |
| 3 | 106 | 37 | |
| >3 | 53 | 17 | |
a p value was calculated with Chi-square test.