Invest Clin 64(4): 460 - 470, 2023 https://doi.org/10.54817/IC.v64n4a3
Corresponding author: Yuan Dong. First Ward, Department of Kidney Transplantation, The Second People’s Hospi-
tal of Shanxi Province, Taiyuan, Shanxi Province, China. Email: dongyuansphsp@sdsch.cn
Influencing factors of post-transplantation
diabetes mellitus in kidney transplant
recipients and establishment of a risk
prediction model.
Yuan Dong
First Ward, Department of Kidney Transplantation, The Second People’s Hospital
of Shanxi Province, Taiyuan, Shanxi Province, China.
Keywords: kidney transplant; diabetes mellitus; influencing factor; prediction model.
Abstract. The aim was to explore the influencing factors of post-trans-
plantation diabetes mellitus (PTDM) in kidney transplant recipients and to es-
tablish a risk prediction model. A retrospective analysis was performed on the
clinical data of 408 patients subjected to kidney transplantation from May 2015
to March 2022. With the simple random sampling method, they were divided
into a training set (n=306) and a test set (n=102) at a ratio of 3:1. According
to the occurrence of PTDM, the training set was further classified into PTDM
and non-PTDM groups. The influencing factors of PTDM were identified by least
absolute shrinkage and selection operator and multivariate logistic regression
analysis. A nomogram prediction model was constructed and validated. Non-PT-
DM and PTDM groups had significantly different preoperative body mass index
(BMI), family history of diabetes mellitus, 2-h preoperative and postprandial
blood glucose, 2-h preoperative and postprandial peptide index, postoperative
hypomagnesemia, whole blood concentration of tacrolimus, triacylglycerol,
glycated albumin and fasting blood glucose (P<0.05). BMI, family history of
diabetes mellitus, 2-h preoperative and postprandial blood glucose, and post-
operative whole blood tacrolimus concentration were independent risk factors
for PTDM. In contrast, the 2-h preoperative and postprandial peptide index was
an independent protective factor (P<0.05). The incidence of PTDM in patients
receiving kidney transplantation correlates with the family history of diabetes
mellitus, preoperative BMI, 2-h postprandial blood glucose, 2-h postprandial
peptide index, and postoperative whole blood tacrolimus concentration.
Prediction of post-transplantation diabetes mellitus 461
Vol. 64(4): 460 - 470, 2023
Factores que influyen en la diabetes mellitus post-trasplante
en receptores de trasplante renal y el establecimiento
de un modelo de predicción de riesgo.
Invest Clin 2023; 64 (4): 460 – 470
Palabras clave: trasplante renal; diabetes mellitus; factores de influencia; modelo de
predicción.
Resumen. El propósito del trabajo fue explorar los factores que influyen
en la diabetes mellitus post-trasplante (PTDM) en receptores de trasplante re-
nal y establecer un modelo de predicción. Se realizó un análisis retrospectivo
de los datos clínicos de 408 pacientes sometidos A trasplante renal de mayo
de 2015 a marzo de 2022. La muestra se obtuvo con el método de generar
números aleatorios en una computadora, y fueron divididos en un conjunto de
entrenamiento (n=306) y un conjunto de prueba (n=102) en una proporción
de 3:1. De acuerdo con la ocurrencia de PTDM, el conjunto de entrenamiento
fue clasificado en grupos PTDM y no PTDM. Los factores de influencia de PTDM
se identificaron mediante el operador de menor contracción y selección abso-
luta y el análisis de regresión logística multivari. Se construyó y validó un mo-
delo de predicción de nomograma. Los grupos no PTDM y PTDM presentaron
diferencias significativas en el índice de masa corporal (IMC) preoperatorio,
antecedentes familiares de diabetes mellitus, glucosa sanguínea preoperatoria
y postprandial 2-h, índice de péptido preoperatorio y postprandial 2-h, hipo-
magnesemia posoperatoria, concentración sanguínea total de tacrolimus, tria-
cilglicerol, albúmina glicosilada sanguínea en ayunos (p<0,05). Entre ellos,
el IMC, los antecedentes familiares de diabetes mellitus, la glucemia preope-
ratoria y postprandial de 2-h y la concentración de tacrolimus en sangre total
postoperatoria fueron factores de riesgo independientes para PTDM, mientras
que el índice de péptido preoperatorio y postprandial de 2-h fue un factor de
protección independiente (p<0,05). La incidencia de PTDM en pacientes que
reciben trasplante renal tiene correlaciones con los antecedentes familiares de
diabetes mellitus, IMC preoperatorio, glucosa sanguínea postprandial 2-h, ín-
dice de péptido postprandial 2-h y concentración de tacrolimus en sangre total
posoperatoria.
Received: 01-12-2022 Accepted: 03-06-2023
INTRODUCTION
Kidney transplantation is currently con-
sidered effective in treating end-stage renal
disease. The five-year survival rate reaches
over 80% among kidney transplant recipients
1, but some still experience different postop-
erative complications. A common metabolic
complication after kidney transplantation
is post-transplantation diabetes mellitus
(PTDM), increasing the risk of cardiovas-
cular and cerebrovascular diseases and re-
sulting in deaths and seriously affecting the
prognosis of patients 2,3. PTDM, with an inci-
dence of about 4-25%, usually occurs within
one year after surgery 4. It may be triggered
462 Dong
Investigación Clínica 64(4): 2023
by such factors as the patient’s age, family
history of diabetes mellitus, high-fat diet,
and donor type 5,6. Thus, exploring the risk
factors of PTDM in kidney transplant recipi-
ents and constructing a risk prediction mod-
el is of great significance in reducing the
incidence rate of PTDM and improving the
prognosis of patients. This study conducted
a retrospective analysis of the clinical data
of 312 patients experiencing living-donor
kidney transplantation in our hospital from
May 2015 to August 2021. On this basis, the
influencing factors in the development of
PTDM in patients were identified, and a no-
mogram prediction model was built to pro-
vide a clinical reference.
PATIENTS AND METHODS
General data
A retrospective analysis was performed
on the clinical data of 408 patients who re-
ceived kidney transplantation in our hospi-
tal (the Second People’s Hospital of Shanxi)
from May 2015 to March 2022. These patients
were assigned to a training set (n=306) and
a test set (n=102) at a ratio of 3:1 by gener-
ating random numbers on a computer. These
two sets were used to construct a risk predic-
tion model and validate the model’s predic-
tion performance, respectively. The training
set [160 males and 146 females, (34.02 ±
7.71 years old)] and the test set [53 males
and 49 females, (34.15±7.32 years old)] did
not have significant differences in the gen-
eral data (P>0.05). The inclusion criteria
were: (1) Patients who received allogeneic
kidney transplantation for the first time,
(2) had a follow-up time ≥ one year, and (3)
whose age ≥ 18 years old. Exclusion criteria
were patients who (1) had no family history
of diabetes before surgery, (2) experienced
more than one kidney transplantation, (3)
experienced the preoperative use of gluco-
corticoids for > three months, (4) had in-
complete clinical data, or (5) died within
one year after transplantation. This study
was reviewed and approved by the ethics
committee of our hospital, and all enrolled
patients were informed and signed the in-
formed consent.
Postoperative immunosuppressive regimen
The postoperative immunosuppressive
regimen for patients was orally taking cy-
closporine A (3-5 mg·kg–1·d–1) or tacrolimus
(0.05-0.10 mg·kg-1·d–1) + mycophenolate
mofetil (1.0-1.5 g/d) or sodium mycophe-
nolate (720-1080 mg/d) or mizoribine (3-4
mg·kg–1·d–1). The dose was adjusted based on
the plasma concentration of cyclosporine A
or tacrolimus. Then methylprednisolone (30
mg/d) was taken orally from the fourth day
after surgery, and the dose was reduced to
5 mg on the seventh day after surgery and
continually taken.
Clinical data collection
The clinical data of patients collected in-
cluded (1) preoperative clinical data: age, gen-
der, family history of diabetes mellitus, body
mass index (BMI), causes of end-stage renal
disease, type of dialysis, dialysis time, type of
donor’s kidney, warm ischemia time, cold isch-
emia time, glycated albumin, 2-h postprandial
blood glucose, and 2-h postprandial peptide
index, and (2) postoperative data: delayed
functional recovery of the transplanted kidney,
rejection, cytomegalovirus, hypomagnesemia,
postoperative immune induction drugs, whole
blood concentration of tacrolimus, whole
blood concentration of cyclosporine, triglyc-
eride, glycated albumin, total cholesterol, cre-
atinine, urea nitrogen, uric acid and estimated
glomerular filtration rate.
Diagnostic criteria
Patients were diagnosed six weeks af-
ter kidney transplantation according to the
diagnostic criteria issued by the American
Diabetes Association (ADA) in 2019 7 if they
had stable immunosuppression, stable renal
function, and no acute infection. Those sat-
isfying the diagnostic criteria were included
in the PTDM group, while the rest of the pa-
tients were included in the non-PTDM group.
Prediction of post-transplantation diabetes mellitus 463
Vol. 64(4): 460 - 470, 2023
Statistical analysis
The statistical analysis of data was per-
formed with the IBM SPSS® 23.0 software.
Measurement data were expressed as mean
± standard deviation (`x ± s), and the t-
test was applied to compare the two groups.
Count data were expressed as a percent-
age (%), and the c2 test was used to com-
pare groups. The independent risk factors of
PTDM were analyzed with the least absolute
shrinkage and selection operator (LASSO)
and multivariate logistic regression. The
nomogram prediction model was built by R
software, and its predictive value, accuracy,
and clinical practicability were evaluated
using the receiver operating characteristic
(ROC) curve, calibration curve, and deci-
sion curve, respectively. A significance level
of α=0.05 was utilized.
RESULTS
Univariate analysis results of PTDM in
patients
Among the 306 patients, the incidence
rate of PTDM within one year after surgery
was 24.84% (76/306). The non-PTDM group
and the PTDM group had statistically signifi-
cant differences in preoperative BMI, family
history of diabetes mellitus, 2-h preoperative
and postprandial blood glucose, 2-h preop-
erative and postprandial peptide, postopera-
tive hypomagnesemia, whole blood concen-
tration of tacrolimus, triglyceride, glycated
albumin and fasting blood glucose (P<0.05)
and no statistically significant differences in
other clinical data (P>0.05) (Table 1).
Multivariate analysis results of PTDM
in patients
The occurrence of PTDM was taken as
the dependent variable, and a total of 29 in-
dependent variables were included. LASSO
reduced the dimensionality of independent
variables to avoid model overfitting. The
optimal penalty coefficient λ of the model
was identified by the 10-fold cross-validation
method. When λ kept increasing to one stan-
dard error, it was the optimal value of the
model. Nine predictors were screened out,
including BMI, family history of diabetes
mellitus, 2-h preoperative and postprandial
blood glucose, 2-h preoperative and post-
prandial peptide index, postoperative hypo-
magnesemia, whole blood concentration of
tacrolimus, triacylglycerol, glycated albumin
and fasting blood glucose (Fig. 1).
With the occurrence of PTDM as the
dependent variable (yes =1, no =0), the
above nine predictors were included in the
multivariate logistic regression model. It
was found that BMI, family history of diabe-
tes mellitus, 2-h preoperative and postpran-
dial blood glucose, and postoperative whole
blood concentration of tacrolimus were
independent risk factors for PTDM. In con-
trast, the 2-h preoperative and postprandial
peptide index was an independent protective
factor for PTDM (P<0.05) (Table 2).
Model establishment
By means of the “rms” program pack-
age, the nomogram prediction model was
built based on the five independent influ-
encing factors for predicting the occurrence
of PTDM in patients. The results showed
that the five independent influencing fac-
tors obtained 263 points (56.75 points for
the family history of diabetes mellitus, 82.5
points for 2-h preoperative and postprandi-
al blood glucose PG >6.65 mmol/L, 66.25
points for 2-h preoperative and postprandial
CPI<5.26, 26 points for BMI >23.85 kg/m2,
and 31.50 points for whole blood concentra-
tion of tacrolimus >8.62 C0) in total and
the corresponding risk value of PTDM was
0.875, meaning that the probability of PTDM
predicted by the model was 87.50% (Fig. 2).
Discrimination evaluation
of the nomogram model
Here, the discrimination of the mod-
el was evaluated by the ROC curve. The
training set obtained the area under the
curve (AUC) of 0.758 (95%CI: 0.682-0.834,
p<0.001) and the C-index of 0.882. The test
464 Dong
Investigación Clínica 64(4): 2023
Table 1
Clinical data of the two groups of patients.
Preoperative data
Item
Non-PTDM group
(n=230)
PTDM group
(n=76)
t/c2
value
p
Age (years old)** 34.02±7.71 34.15±7.32 0.892 0.215
Male* 160 (69.57) 53 (69.74) 0.112 0.902
BMI (kg/m2)** 22.45±1.32 24.61±1.45 5.943 <0.001
Family history of diabetes mellitus* 23 (10.00) 26 (34.21) 6.934 <0.001
Smoking* 57 (24.78) 24 (31.58) 1.082 0.345
Type of dialysis before transplantation 1.023 0.093
Hemodialysis* 187 (81.30) 57 (75.00)
Peritoneal dialysis* 43 (18.70) 19 (25.00)
Dialysis time (month)** 25.92±8.12 24.81±7.96 1.009 0.116
Causes of end-stage renal disease 0.863 0.345
Glomerulus nephritis* 171 (74.35) 57 (75.00)
IgA nephropathy* 29 (12.61) 8 (10.53)
Polycystic kidney* 18 (7.83) 6 (7.89)
Others* 12 (5.21) 5 (3.13)
Type of donor kidney 0.834 0.226
Living body* 34 (14.78) 17 (22.37)
Corpse* 196 (85.22) 59 (77.63)
Warm ischemia time (min)** 7.56±5.43 7.67±4.76 0.782 0.324
Cold ischemia time (h)** 5.71±1.24 5.85±1.13 0.343 0.872
Glycated albumin (%)** 13.52±1.12 14.15±1.34 0.345 0.668
2-h postprandial blood glucose (mmol/L)** 5.33±1.32 7.29±1.45 4.012 0.012
2-h postprandial peptide index** 5.42±1.31 4.61±1.10 3.024 0.015
Delayed functional recovery of transplanted
kidney*
22 (9.57)
7 (10.34)
0.283
0.692
Rejection* 6 (2.84) 4 (5.17) 0.091 0.804
Cytomegalovirus* 16 (7.95) 4 (5.17) 0.224 0.782
Hypomagnesemia* 41 (21.59) 27 (41.38) 5.9723 <0.001
Postoperative immune induction drugs
Basiliximab* 46 (20.00) 20 (26.32) 0.852 0.203
Rabbit anti-human thymocyte immunoglobulin* 128 (55.65) 35 (46.05) 0.773 0.334
Antithymocyte immunoglobulin* 132 (57.39) 38 (50.00) 0.765 0.204
Whole blood trough concentration
of tacrolimus (C0)**
7.19±2.21
9.34±3.12
6.245
<0.001
Whole blood trough concentration
of cyclosporine (C0)**
158.23±21.32
161.14±20.34
2.034
0.098
Triacylglycerol (mmol/L)** 1.96±0.21 2.38±0.32 7.304 <0.001
Prediction of post-transplantation diabetes mellitus 465
Vol. 64(4): 460 - 470, 2023
set had an AUC of 0.732 (95% CI: 0.682-
0.782, P<0.001) and a C-index of 0.878.
The prediction model had a C-index >0.75
in both sets, showing high discrimination
(Fig. 3).
Calibration evaluation of the nomogram
model
According to the calibration curve of
the prediction model plotted, the predic-
tion probability curve of the model well fit
the reference probability, and no significant
difference was revealed by the Hosmer-Lem-
eshow test results (P>0.05), indicating high
accuracy of the model (Fig. 4).
Efficiency evaluation of the nomogram
model
According to the plotted clinical deci-
sion curve, the model was far away from the
extreme curve in both the training and test
sets and obtained high a net benefit, indicat-
ing high reliability and practicability of the
constructed nomogram model (Fig. 5).
Preoperative data
Item
Non-PTDM group
(n=230)
PTDM group
(n=76)
t/c2
value
p
Glycated albumin (%)** 12.78±1.23 15.11±1.23 5.492 <0.001
Fasting blood glucose (mmol/L)** 4.32±0.34 5.18±0.34 7.472 <0.001
Albumin (g/L)** 42.45±1.34 42.45±1.26 0.603 0.402
Total cholesterol (mmol/L)** 3.09±0.34 3.17±0.32 0.282 0.828
Urea nitrogen (mmol/L)** 13.83±1.23 10.45±1.25 0.447 0.548
Creatinine (μmol/L)** 151.31±24.34 151.72±20.23 0.682 0.392
Uric acid (μmol/L)** 309.124±24.23 295.19±20.83 0.332 0.672
Estimated glomerular filtration rate
[mL (min·1.73 m2)]**
60.45±5.72
72.98±5.15
1.114
0.092
Measurement data were expressed as mean ± standard deviation (`x ± s) and the t-test was applied to the compa-
rison between the two groups. Count data were expressed as a percentage (%), and the c2 test was applied to the
comparison between groups. C0: Whole blood trough concentration. *: n (%); **: (`x ± s).
Table 1
CONTINUATION
Fig. 1. LASSO regression analysis results for 27 predictors. A: Coefficient curve of 27 variables, B: Optimal
clinical features selected by 10-fold cross-validation.
Coefficient paths
Cross-validation plot
Coefficients
Binomial deviance
466 Dong
Investigación Clínica 64(4): 2023
DISCUSSION
PTDM is a common complication after
kidney transplantation, the pathogenesis of
which remains unclear. Its correlation with
insulin resistance and insufficient insulin se-
cretion is accepted in most literature 8,9, while
hyperglycemia is closely associated with insulin
production and target tissue demand. In addi-
tion, PTDM is also a high-risk factor inducing
cardiovascular and cerebrovascular diseases in
kidney transplantation, possibly resulting in the
reduction or loss of transplanted kidney func-
tion and increased risk of postoperative death
in patients 10. For this reason, exploring the in-
fluencing factors of PTDM in kidney transplant
recipients is significant in improving patients’
prognosis and postoperative survival rate.
Table 2
Multivariate logistic regression analysis results of related factors affecting PTDM in patients.
Factor βSE Wald p OR 95%CI
BMI 1.825 1.538 2.417 0.009 3.474 2.045~4.856
Family history of diabetes mellitus 2.672 2.358 3.983 0.006 4.728 3.049~5.861
2-h preoperative and postprandial blood glucose 0.501 0.146 11.775 0.012 1.156 1.024~1.572
2-h preoperative and postprandial peptide index -0.342 0.172 0.835 0.003 0.710 0.518~0.849
Postoperative hypomagnesemia 0.794 0.519 7.68 0.066 2.213 0.986~4.733
Whole blood concentration of tacrolimus 2.583 2.067 4.075 0.004 4.369 2.358~5.592
Postoperative triglycerides 0.507 0.179 0.750 0.038 1.661 0.731~2.439
Postoperative glycated albumin 0.502 0.492 0.757 0.024 1.652 0.915~2.903
Postoperative fasting blood glucose 1.578 1.326 2.805 0.091 2.152 0.937~3.498
BMI, family history of diabetes mellitus, 2-h preoperative and postprandial blood glucose, and postoperative whole
blood concentration of tacrolimus were independent risk factors for PTDM, while 2-h preoperative and postprandial
peptide index was an independent protective factor for PTDM. BMI: Body mass index; CI: confidence interval; OR:
odds ratio; PTDM: post-transplantation diabetes mellitus; SE: standard error.
Fig. 2. Nomogram prediction model for predicting PTDM in patients.
Prediction of post-transplantation diabetes mellitus 467
Vol. 64(4): 460 - 470, 2023
Fig. 3. ROC curves of prediction model in training and test sets. A: Training set, B: test set.
Fig. 4. Calibration curves of nomogram prediction model. A: Training set, B: test set.
Fig. 5. Clinical decision curve analysis results of prediction model in training and validation sets. A: Training
set, B: test set.
468 Dong
Investigación Clínica 64(4): 2023
A total of 58 patients (24.78%) in this
study’s test set (n=234) had PTDM within
one year after surgery. PTDM is a major
cause of postoperative serious infection and
even death in patients. Herein, preoperative
BMI, family history of diabetes mellitus, 2-h
preoperative and postprandial blood glu-
cose, 2-h preoperative and postprandial pep-
tide index, postoperative hypomagnesemia,
the whole blood concentration of tacroli-
mus, triacylglycerol, glycated albumin, and
fasting blood glucose were all determined in
the univariate analysis to be influencing fac-
tors of PTDM in patients. BMI, family history
of diabetes mellitus, 2-h preoperative and
postprandial blood glucose, and postopera-
tive whole blood concentration of tacrolimus
were independent risk factors for PTDM. In
contrast, the 2-h preoperative and postpran-
dial peptide index was an independent pro-
tective factor for PTDM, as revealed by the
multivariate logistic regression analysis re-
sult. The close correlation of BMI with the
occurrence of PTDM in kidney transplant
recipients has been reported in previous lit-
erature 11.
According to a study on the Korean
population 12, kidney transplant recipients
with BMI ≥25 kg/m2 suffered a 3.64 times
higher risk of PTDM than those with BMI<
25 kg/m2. A possible mechanism is that
obesity triggers chronic inflammation and
stimulates pancreatic beta cells, thus caus-
ing insulin resistance and reduced glucose
clearance rate, eventually increasing the risk
of PTDM. It was found in a study 13 that a
family history of diabetes presented a sig-
nificant correlation with the risk of PTDM.
People with a family history of diabetes may
be subjected to abnormal glucose metabo-
lism, which in turn influences the function
of pancreatic β-cells and thus causes abnor-
mal changes in postoperative blood glucose
levels and even the occurrence of PTDM.
Hence, for patients with a family history of
diabetes mellitus, measures should be taken
to closely monitor their blood glucose and
carry out timely interventions to reduce the
incidence rate of PTDM. A related study 14
published by the ADA showed that the ma-
jority of patients experience an abnormal
glucose tolerance stage before diabetes de-
velopment, and those showing abnormal
glucose tolerance possibly become potential
diabetic patients. The study of Sato et al. 15
unveiled that preoperative glucose tolerance
was a risk factor for postoperative diabetes
in transplant recipients. The 2-h postpran-
dial peptide index, which reflects the func-
tion of pancreatic islet B cells and reduces
with the increasing duration of type 2 diabe-
tes mellitus, is related to insulin sensitivity
and is considered a protective factor against
PTDM in transplant recipients 16. Moreover,
tacrolimus is a typical drug for treating anti-
rejection reactions. Its significantly positive
correlation with the occurrence of PTDM and
stronger sugar-causing effect than cyclospo-
rine A 17 has been revealed. Additionally, for
patients receiving kidney transplantation,
the administration of tacrolimus can reduce
the synthesis and secretion of insulin in the
body, increasing the body’s blood glucose
level and thus resulting in diabetes 18. Fur-
ther, some believe that other important in-
fluencing factors on the occurrence of PTDM
in transplant recipients include hypomagne-
semia and rejection 19. However, no statisti-
cally significant difference in rejection was
found between the two groups of patients in
this study. In addition, postoperative hypo-
magnesemia was found in the multivariate
analysis not to be an independent influence
factor of PTDM, possibly related to the small
sample size of this study, which failed to
present statistical differences.
Based on the influencing factors on the
occurrence of PTDM in kidney transplant
recipients, the nomogram model was built
in this study, whose predictive performance
was evaluated with the ROC curve, calibra-
tion curve, and clinical decision curve. The
results showed that the predicted value ap-
proximated the actual observed value, sig-
nifying high discrimination and clinical va-
lidity of the model. Compared with a single
Prediction of post-transplantation diabetes mellitus 469
Vol. 64(4): 460 - 470, 2023
influencing factor, the prediction model can
better identify patients subjected to high-
risk liver metastasis, which boosts the clini-
cal application of the research results.
This study still has some limitations. First,
the subjects were collected from a single center,
and the types of potential predictive variables
collected were limited by clinical practice. Sec-
ond, the prediction model was built through
retrospective analysis, while limited clinical
data were collected, and further validation in a
prospective cohort was not carried out. Hence,
the results may have bias. The research should
be further improved by prolonging the follow-
up time and increasing the collected data on
influencing factors.
In conclusion, the model established in
this study showed that BMI, family history
of diabetes mellitus, 2-h postprandial blood
glucose, postoperative whole blood tacroli-
mus concentration, and 2-h postprandial
peptide index were independent influenc-
ing factors for predicting the occurrence of
PTDM. Based on this model, attention can
be paid to these factors, and early interven-
tion can be taken to reduce the incidence
rate of PTDM. Thus, this model is potentially
applicable to clinical practice.
Funding
This study was financially supported by
the Project No. 2020014.
Conflicts of interest
The author reported no potential con-
flict of interest.
Author ORCID number
• Yuan Dong: 0000-0001-9630-3281
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