After obtaining the necessary permission from Kerman School of Nursing and Seyyed Al-Shohada Hospital, the researcher referred to the neonatal intensive care unit of Seyyed Al-Shohada Hospital in Kerman. After providing a complete explanation of the research objectives to nurses and mothers of infants hospitalized in the neonatal ward, he obtained a complete list of hospitalized infants. Based on the inclusion criteria, 120 infants were randomly selected and then randomly assigned to three groups using a table of random numbers by researcher. The sample size was determined by the formula.
Inclusion criteria included: no barrier for oral feeding and no sedatives and painkillers for at least 3 hours before venipuncture (18)), the infant should be awake before venipuncture, the mother has not received any drugs, the infant should not have any cleft lip and palate abnormalities, mother should not be diabetic, and infant should be breastfed. The research exclusion criteria also included all infants who have congenital anomalies such as congenital heart disease, gastrointestinal abnormalities, hypoxic lesions, respiratory failure, current or past history of NEC, suspected or confirmed sepsis, recent abdominal surgery, diagnosis of sepsis (positive blood culture) during the study, infant undergoing NPO at least two consecutive times, and discharging before the intervention completion.
In the first group, the infant’s mother was asked to be present in the ward during venipuncture and hug her infant. The duration of hugging by mother was five minutes before the intervention. Then, the mother was asked to breastfeed her infant. During breastfeeding, venipuncture was done from the veins behind the infant’s hand. During venipuncture, the neonatal infant pain scale (NIPS) and the infant’s physiologic checklist were completed by the researcher. In the second group (glucose 10% feeding), the infant’s mother was asked to be present during the venipuncture. Two minutes before the venipuncture via syringe, two ccs of oral glucose10% was given to the infant, and two minutes later, venipuncture was done while the mother has hugged her infant. The pain questionnaire and physiological criteria checklist were completed by the researcher before and five minutes after venipuncture. In the third group (control), venipuncture was done without any other intervention. The instrument of determining the severity of pain and the checklist of physiological criteria were completed five minutes before, during venipuncture, and five minutes after venipuncture.
All infants were fed half to one hour before the test. To do venipuncture from the veins on back of the hand, the desired area was first cleaned with an alcohol-soaked swab, and the venipuncture was done using a needle No. 22 in all infants by a nurse. If venipuncture failed in the first time of entering the needle inside the skin, that infant would be excluded. Behavioral changes and physiological criteria were recorded by the researcher.
It should be noted that during the pilot study, the crying and restlessness of infants lasted two minutes on average, so the researcher allocated a 2-minute time for observing before, during, and after the intervention.
Study instruments: The instruments used in this study include 3 questionnaires. The first instrument was demographic information questionnaire that included maternal characteristics (age, number of children, number of deliveries, type of delivery, history of underlying disease and neonatal characteristics (sex, gestational age, birth age, birth rank, weight, height, head circumference, first-minute and fifth-minute Apgar). Also, this part of the questionnaire included the sex of infant, birth age, the weight of infant’s birth, the history of multiple pregnancies, the first-minute and fifth-minute Apgar, and birth rank. The second instrument included neonatal infant pain scale. This instrument can be used in infants up to 6 weeks after birth. The validity and reliability of this scale have been investigated in studies (18)Calculation of validity and reliability: Reliability and validity of NIPS have been confirmed by Mollahadi and Moradi (95% (r = 95%) (14) and other studies (19)The validity of Purltan Nellcor 295-NB Bennett pulse oximetry and chronometer devices was confirmed based on the manufacturer company and performing calibration. To evaluate the reliability of the instrument during the research, single pulse oximetry was used, and it was done in a similar situation (on supine position and connecting probe to the right leg of the infant). The infants’ physiological data checklist in this questionnaire included physiological information of infants, including respiration rate, heart rate, arterial blood oxygen levels, and temperature. Its content validity was examined and its reliability was obtained at 0.87 using Cronbach's alpha.
Data analysis
Descriptive and analytical statistics were used to analyze the data. According to the results of the Kolmogorov-Smirnov test, the study data did not have a normal distribution, so non-parametric tests were used. Central and distribution indices (frequency, percentage, mean, and standard deviation) were used to report demographic characteristics, Kruskal-Wallis test was used to compare means of pain severity and physiological indicators, and Mann-Whitney U test was used to compare variables before and after intervention in each group.