Objective To compare broth microdilution and agar dilution methods for in vitro testing of activities of fluconazole,ketoconazole and itraconazole against clinical Malassezia isolates.Methods Broth microdilution and agar dilution methods were used to determine the minimal inhibitory concentration(MIC)of fluconazole,ketoconazole and itraconazole for 27 clinical strains(5 species)of Malassezia.Results The minimal inhibitory concentration(MIC)ranges of fluconazole,ketoconazole and itraconazole were 0.25-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.125 mg/L respectively as shown by broth microdilution method,2-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.25 mg/L respectively as revealed by agar dilution method.Both methods demonstrated that itraconazole possessed the strongest activity against Malassezia species,followed by ketoconazole and fluconazole.The agreement rate in MICs between the two methods was 78.8％,85.2％ and 88.9％,respectively for fluconazole,ketoconazole and itraconazole,with the intraclass correlation coefficients (ICCs)being 0.88,0.80 and 0.76 respectively.Conclusions Fluconazole,ketoconazole and itraconazole are highly active against Malassezia species in vitro,and itraconazole is the most active.Broth microdilution and agar dilution method coincide well in,and are applicable for,the antifungal susceptibility testing of Malassezia species in vitro.

Objective: To analyze the clinical features, etiology and outcome of neonatal sepsis, and to clarify the characteristics of early-onset sepsis and late-onset sepsis, which may provide clinical evidence for early diagnosis and treatment of neonatal sepsis. Methods: Two hundred and twenty-nine cases of neonatal sepsis who were hospita-lized in the First Neonatal Wards, Shengjing Hospital Affiliated to China Medical University First Neonatal Wards from June 2013 to June 2016, and according to the onset time, they were divided into the early-onset sepsis (EOS) group and the late-onset sepsis(LOS) group, and SPSS 20.0 statistical software was used for comparative analysis of the data concerning the general situation, clinical manifestation, laboratory examination, pathogen distribution, complications and outcome between 2 groups, and Logistic stepwise regression was used to detect multiple factors. Results: Sixty-eight patients (29.69%) were in the EOS group, and among them the full-term infants with normal weight accounted for 51.5%; 161 cases (70.31%) were in the LOS group with extremely low birth weight premature infants occupying 52.2%.The mothers with gestational hypertension, after the birth of mechanical ventilation, parenteral nutrition(>3 d), indwelling peripherally inserted central catheters (>5 d) had significant correlations with sepsis(all P<0.05). They had fever and jaundice, complicated with purulent meningitis in the EOS group.The apnea and abdominal distension seemed more easily complicated with necrotizing enterocolitis in the LOS group.There as no difference in laboratory test indexes such as platelet, C-reactive protein (CRP) (all P>0.05). Procalcitonin and CRP had higher sensitivity in the diagnosis of sepsis.EOS was given priority to Gram-positive bacteria, which was far more common coagulase negative staphylococcus [26.5%(18/68 cases)], and LOS was due to mainly Gram-negative bacteria and the main pathogens were klebsiella pneumoniae in 86 cases (53.4%). The mortality in the EOS group was significantly higher than that in the LOS group[7.4%(5/68 cases) vs.2.5%(4/161 cases)], but the difference was not statistically significant (χ²=4.72, P>0.05). Conclusions Because time of neonatal sepsis onset time is different, the clinical characteristics, the distribution of etiology and outcome also show some differences, and full-terms often have EOS which is more common in the premature babies with LOS.There are various high risk factors for sepsis in perinatal and postnatal period.Therefore, early and timely diagnosing the neonatal sepsis has great significance in reducing the mortality of neonatal sepsis.

Objective: To analysis the transcutaneous carbon dioxide pressure(TcpCO₂)in very low birth weight infant (VLBW) and extremely low birth weight infant (ELBW) after the time of tube drawing, and further guidance the management of premature ventilator breathing. Methods: Premature infants with gestational age less than 32 weeks, birth weight below 1 500 g, hospitalized from March 2016 to December 2018 in our hospital, who needed early mechanical ventilation due to respiratory distress within 24 hours after birth and intended to be withdrawn were enrolled.Fifty-five infants in the observation group were monitored by TcpCO₂ and blood gas was collected at 1 h, 6 h, 12 h, 24 h, 48 h, and 72 h after weaning; 55 infants in the control group were collected arterial blood gas only.When TcpCO₂ monitoring and PaCO₂ indicated hypercapnia, clinical intervention was actively given.The value of blood gas PaCO₂ in the two groups and the TcpCO₂ value of the observation group were recorded at different time points (1 h, 6 h, 12 h, 24 h, 48 h, 72 h) after weaning, and the failure of weaning was recorded in both group. Results: There was no significant difference in the general information (gender, gestational age, birth weight, and whether prenatal maternal glucocorticoids) between the two groups (P>0.05). There were no significant differences in mechanical ventilation time, noninvasive ventilation mode and time between the two groups(P>0.05). The failure rate of withdrawal within 24 h and 72 h was lower in the observation group than the control group[3.6% (2/55) vs.14.0% (8/55), 7.3% (4/55) vs. 21.0% (12/55)], the difference was statistically significant (P<0.05). There was a significant difference in blood gas PaCO₂ between the two groups at 6 h, 12 h, and 24 h after weaning (P<0.05), but had no significant difference between the two groups at 1 h, 48 h, and 72 h (P>0.05). The values of TcpCO₂ and PaCO₂ had no significant difference in the observation group at the same time point (P>0.05), and there was a correlation between them(r=0.761, P<0.05). Conclusion VLBW and ELBW undergoing mechanical ventilation are given continuous TcpCO₂ monitorings after weaning, which not only has the characteristics of continuous and non-invasive monitoring, but also help to identify hypercapnia early and give symptomatic treatment, and reduce the incidence of weaning failure.And it proves a good correlation between TcpCO₂ and PaCO₂.

Bronchopulmonary dysplasia(BPD)is one of the most serious lung diseases in premature infants and an important cause of death in preterm infants.The pathophysiological mechanism of BPD is still unclear.The preventive and targeted treatment strategies for this disease are also very limited.However, various biomarkers have been found in blood, alveolar lavage fluid, exhaled gas condensate, and urine.It is used to identify high-risk children with BPD early and to predict the severity and prognosis of the disease.

Caffeine citrate, as one type of methylxanthine, is often used to treat apnea in premature infants.Recent studies have found that it is not only used for extubation before weaning, shortening the time of mechanical ventilation, preventing apnea and reducing the occurance of intermittent hypoxic episodes, but also exerting lung protection by inhibiting inflammatory reactions, mitigating oxidative stress, improving pulmonary function and pulmonary angiogenesis.Thus, it could effectively reduce the risk of bronchopulmonary dysplasia in premature infants.

Very low birth weight infants (VLBW) and extremely low birth weight infants (ELBW) are prone to complications such as feeding intolerance and infectious diseases such as necrotizing enterocolitis due to the prematurity of the immunological and gastrointestinal system.In recent years, probiotics are being widely used in preterm infants to improve gut microbiota and enhance the resistance to foreign substance.In this way, probiotics plays a role in lessening the incidence of feeding intolerance, reducing the mortality and morbidity of severe NEC and shortening of hospital stay.

Objective: Previous studies have found abnormal proliferation and transdifferentiation of alveolar epithelial cells(AECs)in hyperoxic lung injury of neonatal rats.The purpose of this study was to clarify the expression of zonula occludens 1(ZO-1) and ZO-1 related nucleic acid binding protein(ZONAB)in AECs in hyperoxic lung injury model, in order to investigate its effect on the proliferation and transdifferentiation of AECs in the injured lung tissue. Methods: Full-term neonatal Wistar rats were randomly divided into two groups within 12 h after birth, model group(inhaled oxygen concentration 85%)and control group(inhaled air). Lung specimens were collected at 7, 14 and 21 days after exposure.The expression of ZONAB in typeⅡalveolar epithelial cells(AECⅡ)was observed by double immunofluorescence staining.At the same time, AEC Ⅱ was isolated from lung tissues of animal models at these time points, and the expression levels of ZO-1, ZONAB protein and mRNA in lung tissues and AECⅡof the two groups were detected by Western blot and Real-Time PCR.In addition, AEC Ⅱ was isolated from lung tissue of normal newborn rats and then divided into model group(85% oxygen concentration)and control group(21% oxygen concentration). After 48 hours of culture in vitro, the expression levels of ZO-1, ZONAB protein and mRNA were detected, and the expression level and location of ZONAB were observed by immunofluorescence staining. Results: Double immunofluorescence staining showed that the expression of ZONAB in AECⅡin model group was significantly lower than that in control group.The protein and mRNA expression levels of ZO-1 and ZONAB in AECⅡisolated from lung tissue of model group were both significantly lower than those from control group, starting from 7 d after hyperoxia exposure.AECⅡisolated from lung tissue of normal newborn rats, were then incubated for 48 hours under hyperoxia or normoxia in vitro, the protein and mRNA expression levels of ZO-1 and ZONAB significantly decreased in model group compared with those in control group.The results of immunofluorescence staining showed that the expression of ZONAB was higher in AECⅡof the control group, and ZONAB was mostly located in the junction and nucleus of cells, while the expression of ZONAB in the model group significantly decreased than that in the control group, and the expression sites were clustered in the cytoplasm, with little expression in the junction and nucleus. Conclusion ZO-1, as a tight junction-related protein, is down-regulated in hyperoxic lung injury model.In addition to destroying pulmonary epithelial barrier to mediate pulmonary edema, it also participated in the regulation of proliferation and differentiation of AECs by regulating transcription factor ZONAB, suggesting that this may be another pathway leading to hyperoxic lung injury.

Objective: To explore the effect of family need-based health education mode on family members under stress caused by transferring their children out of pediatric intensive care unit(PICU). Methods: A total of 40 family members of children transferred from PICU of Shengjing Hospital of China Medical University from July to December 2017 were selected as the control group, and 40 family members of children transferred from April to September 2018 were selected as the observation group. In the control group, daily routine health education mode were implemented by nurses on the relatives, while in the observation group, nurses provide the family members of children with corresponding graphic and text materials of health education according to the 2 copies of Critical Care Family Needs Inventory completed by their relatives within 24 hours and 72-96 hours of admission, and implement the daily health education combined with the Flipped Classroom method. The level of uncertainty in illness, stress in transfer and the satisfaction for family-centered nursing were compared between the groups. Results: In the observation group, the total score of the family members′ level of uncertainty in illness and stress in transfer were (80.05 ± 3.96) and (39.65 ± 2.41) points respectively, which were lower than those in the control group (99.70 ± 3.78) and (67.15 ± 2.38) points, and all the differences were statistically significant (t=-22.60, -50.21, P<0.01). The satisfaction for family-centered nursing in the observation group was higher than the control group, and the difference was statistically significant (P < 0.05). Conclusions The implementation of health education mode based on family needs is beneficial to reduce the level of uncertainty in illness of the family members of critically ill children transferred out of PICU, relieve the stress in transfer, and thus improving the family-centered high-quality nursing services in PICU.

Female adolescent is in the beginning of sexual maturity period and get into stage of give birth. The key points of management for the inguinal hernia of female adolescent is to cure the hernia, decrease the incidence of recurrence and the complications and protect the round ligament uterus, avoid damage to reproductive function. Tension free hernia repair is the mainly method for inguinal hernia management.Laparoscopic hernia repair is good in decrease the incidence of post operation recurrence and the chronic pain in female adolescent inguinal hernia.

The 2019 European Guideline on Respiratory Distress Syndrome(RDS)updated and supplemented the 2016 version according to newly-published evidence and literature,in order to optimize the management of RDS. The updates includes risk factors of preterm births,the standardized use of prenatal glucocorticoids protocol,the evidence-based delivery room management and the early application of protective pulmonary ventilation after birth;pulmonary surfactant administration as the crucial treatment for RDS;the incidence chronic lung disease can be reduced by non-invasive respiratory support. Prenatal glucocorticoids use and caffeine administration can significantly shorten the duration of mechanical ventilation. In addition,good temperature control,precise fluid and nutrition management,perfusion maintenance and cautious antibiotic use can help improve the outcome of preterm births.