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Objective: To investigate the effects of enriched environments on behavioral development at toddler period of preterm who had experienced early repeated operative pain. Methods: A cross-sectional study was conducted. A total of 80 high-risk preterm children of 2 years of age, who had experienced repeated pain stimuli in the neonatal intensive care unit (NICU), were enrolled as preterm group from the High-risk Children Clinic of Children's Hospital of Nanjing Medical University from October 2016 to March 2021. Furthermore, 39 full-term healthy children, aged 2 years, who were undergoing routine check-ups during the same period, were selected as the full-term group. The preterm group was further divided into preterm intervention group and preterm non-intervention group based on the implementation of enriched environment interventions. Data of neonatal characteristics from 3 groups were collected. Growth and development indicators at the age of 2 years were measured. Neuropsychological development evaluated by Gesell developmental scale. Behavioral development evaluated by child behavior check list. The salivary cortisol levels in response to novelty (baseline, task, end) were collected. The family environment, including maternal parenting pressure, were evaluated through a survey questionnaire. One-way ANOVA and least significant difference (LSD) tests were used to compare physical development, maternal parenting stress, Gesell neuropsychological development, and behavioral problems among the 3 groups. A repeated-ANOVA and LSD tests were employed to compare the patterns of salivary cortisol secretion. Pearson correlation analysis was used to explore the influencing factors related to neuropsychological and behavioral development and cortisol level. Results: There were 44 cases in the preterm intervention group (17 males, gestational age of (31.3±2.8) weeks), and 36 in the preterm non-intervention group (29 males, gestational age of (32.5±2.6) weeks). The full-term group consisted of 39 children (23 males, gestational age of (39.3±2.1) weeks). At 2 years of age, the height, weight, and head circumference of the preterm intervention group and non-intervention group were all lower than those of the full-term group (all P <0.05).The Gesell developmental schedule showed that the preterm non-intervention group scored all lower in gross motor, fine motor, adaptive, language and personal-social domains compared to the full-term group (91±7 vs. 97±6, 88±9 vs. 94±6, 89±8 vs. 99±8, 84±10 vs. 100±15, 89±7 vs. 95±6), with statistical significance (all P <0.01). The preterm intervention group scored all higher than the preterm non-intervention group in gross motor, fine motor, adaptive and language domains (all P <0.05), with no significant difference compared to the full-term group (all P >0.05). The number of needle painful procedures during hospitalization in NICU of the non-intervention group was negatively correlated to the adaptive development quotient ( r =-0.48, P <0.05). Furthermore, the preterm non-intervention group exhibited higher scores in social withdrawal, depression, somatic complaints, aggression, and destructive behaviors compare to the full-term group and preterm intervention group ( F =8.07, 5.67, 7.72, 7.90, 7.06; all P <0.05); while the preterm intervention group showed no significant difference compared to full-term group (all P >0.05). Behavioral problems (social withdrawal and depression) in the preterm non-intervention group were positively correlated with maternal parenting stress ( r =0.66, 0.50; both P <0.05). In response to novel visual stimuli and cognitive challenges, the preterm non-intervention group had significantly higher salivary cortisol levels compared to the full-term group ( P =0.006), which were negatively correlated with the frequency of early painful procedures ( r =-0.83, -0.80; both P <0.01). There was no significant difference in cortisol secretion pattern between the intervention group and the full-term group ( P =0.772). Conclusion: Enriched environmental interventions can improve neuropsychological development, decrease behavioral problems, and down-regulate consistent high cortisol response to task in preterm infants who have experienced repeated procedural pain in the NICU by the age of 2 years.

Objective: To establish a liquid chromatography tandem mass spectrometry method to determine the cortisol in saliva. Methods: Take 0.5 ml saliva sample into a 2 ml centrifuge tube, add excess sodium chloride and 1ml acetonitrile to it, then vortex for 3 min, centrifuge for 10 min at 15 000 r/min, and take 800 μl of the upper layer to another centrifuge tube. Finally, the sample was concentrated by a vacuum concentrator and brought to 200 μl with the initial mobile phase. Then, the sample was analyzed by liquid chromatography tandem mass spectrometry. The target compound was quantified by external standard curve method. Results: The linear range of the method was 0.02 - 5.00 ng/ml, r =0.999 9, the method limit of the detection was 0.002 ng/ml, the method limit of quantitative was 0.02 ng/ml, and the spiked recoveries were 89.60% - 98.60%. The intra - assay precision was 1.90% - 3.30%, and the inter - assay precision was 4.20% - 9.00%; samples could be stored at -20 °C for at least 14 days. The determination of cortisol could not be interfered by other endogenous substances in the sample. Conclusion: The method is simple in pretreatment, high sensitivity, good reproducibility and good recovery, and it is suitable for the quantitative analysis of cortisol in saliva for normal and occupationally stressed populations.

Objective: To determine the possible relations between hair cortisol concentration (HCC) as a biomarker of chronic stress and childhood overweight and obesity. Methods: Children from grade 1 to 3 from 2 primary schools in Bengbu were invited to participate in the study; and those who suffer from mental disorders, endocrine diseases and those who took hormone drugs were excluded. Parental questionnaire was implemented to collect information on physical activity, screen time, sleep duration, sugar-beverage consumption, as well as socio-economic status. Height and weight were examined. Eligible hair samples from 1 263 children (598 boys and 665 girls) were obtained, and the cortisol content was determined. Multivariate logistic regression model analysis was recruited to examine the association between HCC quartiles and overweight and obesity among boys and girls, respectively. Results: HCC in P (50) ( P (25)- P (75)) among girls and boys was separately 11.86 (10.57-13.15) and 11.71 (10.54-13.09) µg/kg ( Z=- 0.886, P= 0.376); HCC in P (50) ( P (25)- P (75)) among non-overweight, overweight and obesity boys was separately 11.62 (10.45-12.82), 12.24 (10.88-13.55) and 12.30 (10.99-13.53) µg/kg (χ(2)=8.24, P= 0.016); HCC in P (50) ( P (25)- P (75)) among non-overweight, overweight and obesity girls was separately 11.35 (10.06-12.62), 12.07 (11.04-13.21) and 12.59 (11.12-13.63) µg/kg (χ(2)=36.16, P< 0.001). After age, physical activity, screen time, sleep duration, sugared beverages and socio-economic status adjusted, compared with HCC in Q 1 level, girls whose HCC in Q 2, Q 3 Q 4 level had higher rates in overweight, with OR (95 %CI ) at 1.10 (1.04-3.32), 2.59 (1.51-4.45) and 3.16 (1.79-5.57), respectively. Girls with HCC in Q 3 and Q 4 level had higher rates in obesity, with OR (95 %CI ) at 2.22 (1.16-4.26) and 5.62 (3.04-10.41). Among boys, highest quartiles in HCC was risk factor for overweight, OR (95 %CI ) was 2.14 (1.18-3.90). Conclusion: Hair cortisol had a significant relation with overweight and obesity in 6-9 years old childhood especially among girls.

Objective: To evaluate the key indicators in the pituitary-target gland axes in the animal model of Shen-yang deficiency syndrome (SYDS)., Methods: The 8 biological indicators [thyroid stimulating hormone (TSH), 3, 3', 5-triiodothyronine (T3), thyroxine (T4), luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), adrenocorticotropic hormone (ACTH), and cortisol (CORT)] in the pituitary-target gland axes were grouped using factor analysis. Then the sensitivity of every indicator was calculated according to the sensitivity function defined in this paper, so as to find all the most sensitive indicators in every group as key indicators of SYDS., Results: The key indicators in the early period of SYDS were T, LH, T4, and CORT. The key indicators in the middle period were LH,T, CORT, and ACTH. The key indicators in the late period were LH, T, CORT, and FSH., Conclusions: T, LH, and CORT were the common key indicators of the three periods, and other different key indicator of SYDS in the early, middle and late period were T4, ACTH, and FSH respectively, which changed from the thyroid axis to the adrenal axis and then to the gonadal axis as the period changed. The key indicators in the late period were mainly in the gonadal axis, showing gonadal dysfunction in the late period.

Objective: To evaluate the stress level during parachuting training by salivary biomarker and to study the dynamic characteristics., Methods: Twenty recruits of military parachuting training completed 8 trainings in a month. The saliva samples were collected at 2 h and 1h before boarding and at 0.5 h after landing on the 1st, 4th and 7th trainings. The levels of cortisol, chromogranin A and α-amylase in saliva samples were detected., Results: The concentrations of cortisol, chromogranin A and activity of α-amylase increased significantly from pre-boarding to landing during 3 trainings. The concentrations of cortisol, chromogranin A and activity of α-amylase at 2 h before boarding and at 0.5 h after landing decreased significantly with the training times. However, the changes of 3 biomarkers at 1 h before boarding among 3 trainings were not significant., Conclusion: The levels of stress increased significantly for 20 recruits from pre-boarding to landing during parachuting trainings. The stress levels of 20 recruits before boarding and after landing significantly decreased with parachuting training times.

Objective: To evaluate the usefulness of the measurement of midnight salivary cortisol (SC) in the initial diagnosis of Cushing's syndrome (CS)., Methods: A total of 18 patients with pathologically confirmed CS were recruited in the study. Thirteen patients with pure obesity and 36 healthy people served as controls. Salivary samples at 8:00, 16:00, 24:00 and 8:00 the next day after the 1 mg dexamethasone suppression test(DST) were collected by a commercially-available saliva collection device, and assayed by Electro Chemiluminescence Immuno Assay (ECLIA)., Results: There were highly significant correlations between salivary cortisol and plasma cortisol (PTC) at the 4 time points. The correlation coefficient was 0.73 (0.71, 0.74, 0.78 and 0.61 at the four time points, respectively) (P < 0.05). Midnight salivary cortisol level in Cushing's Syndrome [(10.58 +/- 5.17) nmol/L] was significantly higher than healthy people [(1.55 +/- 0.60) nmol/L]. The receiver operating characteristics curve (ROC) analysis showed a sensitivity of 100.00% and specificity of 84.00% at 8:00 with the cut-off point of 6.21 nmol/L., Conclusion: The measurement of midnight salivary cortisol is a convenience and noninvasive screening test for the initial diagnosis of Cushing's Syndrome.

An analytical method based on ultra performance liquid chromatography has been developed for the simultaneous determination of 15 hormones (triamcinolone, hydrocortisone, prednisone, cortisone, 6a-methylprednisolone, betamethasone, dexamethasone, prednisolone acetate, hydrocortisone acetate, estriol, beta-estradiol, estrone, diethylstilbestrol, testosterone and progesterone) in cosmetics. Various cosmetic samples were extracted with methanol under ultrasonication, and then cleaned up by a Oasis HLB solid phase extraction cartridge. The 15 hormones were separated and detected within 6 min on a Waters ACQUITY UPLC BEH C18 column (1.7 microm, 2.1 mm x 50 mm) by gradient elution with acetonitrile and water system. In the range of 1-25 ng, the calibration curves of 15 hormones showed good linearity with the correlation coefficients larger than 0.9995. The mean recoveries at the three spiked levels (2, 10, 20 mg/kg) were 88.2%-102.4% with the relative standard deviations of 1.6%-7.4%.

Objective: The aim of the study was to elucidate the interaction and clinical significance between melatonin and cortisol and the status of disease in patients with bronchial asthma., Methods: Ten mild persistent and 10 moderate-to-severe persistent asthma patients were recruited to participate in the study. Fifteen normal subjects served as contrds Salivary free melatonin and cortisol were measured simultaneously by radioimmunoassay in all subjects, and 8 salivary samples were collected in a series during a 24-hour period in each subject. The intensity of light was restricted to natural light at room during the daytime and less than 50 lux at night., Results: The results showed that salivary free melatonin levels were significantly lower in mild (15.5 +/- 5.3) micro g/L and moderate-to-severe (7.1 +/- 2.5) micro g/L persistent asthma patients as compared to control subjects (28.9 +/- 8.7) micro g/L (F = 4.47, P < 0.05; F = 7.61, P < 0.01, respectively). The results also showed that salivary free cortisol levels were significantly lower in mild (3.1 +/- 0.5) micro g/L and moderate-to-severe (4.2 +/- 0.5) micro g/L persistent asthma patients as compared to control subjects (5.9 +/- 0.7) micro g/L (F = 10.45, P < 0.01; F = 5.21, P < 0.05, respectively). The amplitude of salivary free cortisol level was reduced in mild and moderate-to-severe persistent asthma patients, and salivary cortisol peak level was significantly delayed in mild and moderate-to-severe persistent asthma patients (P < 0.05, P < 0.01, respectively). There were no significant correlations between the salivary free melatonin and cortisol levels in control and mild persistent asthma patients (r = 0.174, P = 0.057; r = -0.138, P = 0.221, respectively). However, a significant negative correlation was found between the salivary free melatonin and cortisol levels in moderate-to-severe persistent asthma patients (r = -0.275, P = 0.013)., Conclusions: There were lower salivary free melatonin and cortisol levels in asthmatic patients. A significant negative correlation was found between melatonin and cortisol levels in moderate-to-severe persistent asthma patients. Furthermore, the reciprocal inhibition of melatonin and cortisol was closely associated with the status of disease in asthmatic patients.

The method of reverse phase evaporation is used to coat phosphatidylcholine(PC) directly on the surface of silica, which is used as biomembrane chromatographic solid phase to study the interactions between drugs and biomembrane. It was observed that the solid phase coated phosphatidylcholine had a good stability at 20 degrees C-30 degrees C, and the stability would be improved by the presence of appropriate amount of cholesterol in phosphatidylcholine. The content of cholesterol in phosphatidylcholine, the nature of buffer, the concentration of salt in buffer and the pH of mobile phase could all affect chromatographic retention of drugs on the prepared biomembrane column. Six compounds, polyethylene glycol, mannitol, salicylic acid, warfarin, hydrocortisone, and cortisone have been tested. The biomembrane chromatographic stationary phase coated PC with silica as matrices can be simply prepared and it is possible to simulate the human's physiological environment by the biomembrane chromatographic system, so it is a useful method to study drug absorption and distribution in human body.

Chloramphenicol and hydrocortisone ear drops are possessed of anti-inflammatory and antiallergic functions. It can be used effectively in the treatment of tympanitis. It is composed of chloramphenicol and hydrocortisone. In this paper, we report a reversed phase HPLC method for the determination of chloramphenicol and hydrocortisone in the drops. The operatings conditions were YWG-C18 column (5 microns, 5 mm x 150 mm) at 30 degrees C, mobile phase of methanol-water (60:40, V/V) and UV detector at 244 nm. Prednisone acetate was selected as the internal standard. The linear relationship of calibration curves was good in the range of 40-200 and 8-40 mg/L for chloramphenicol and hydrocortisone, respectively (r = 0.9999). The average recovery and RSD of chloramphenicol and hydrocortisone were 99.91%, 0.86% and 99.58%, 1.34% (n = 6) respectively. The total time for separation and determination was within 9 min. The method was simple, sensitive, rapid and accurate.

The authors have observed the aging change of the adrenocortical function and the effect of the Kidney-tonifying herbs on such a change. The result showed that there was no statistical difference in the plasma cortisol levels between the aged and the young persons, and so was in the plasma corticosterone levels between the aged and the young rats. As the circulating steroids level depends on the balance of the secretion ability of the adrenal cortex and the peripheral degradation rate of the hormone, the authors measured the corticosterone concentration of the culture solution of the adrenocortical cells in vitro of the rats with different age to observe the secretion ability of the adrenocortical cell exclusively. The result showed that the corticosteróne concentration was significantly lower in the culture solution of the adrenocortical cell of the aged rats than that of the young rats, which suggests that the secretion ability of the adrenal cortex declines with aging. In addition, the authors determined the reserve power of the adrenal cortex with different age and found that the increased multiple of the salivary cortisol concentration 60 minutes after the stimulation of ACTH analogue was markedly lower in the aged persons than that in the young persons. The response of the cultured adrenocortical cells to secret corticosterone to the stimulation of ACTH analogue was also significantly lower in the aged rats than that in the young rats, which suggests that the reserve power of the adrenal cortex was also decreased with aging.(ABSTRACT TRUNCATED AT 250 WORDS)