Development and validation of a diagnostic prediction model for severe periventricular-intraventricular hemorrhage in newborns: insights from a retrospective analysis utilizing the MIMIC-III database

Abstract Objective: Periventricular-intraventricular hemorrhage is the most common type of intracranial bleeding in newborns, especially in the first 3 days after birth. Severe periventricular-intraventricular hemorrhage is considered a progression from mild periventricular-intraventricular hemorrhage and is often closely associated with severe neurological sequelae. However, no specific indicators are available to predict the progression from mild to severe periventricular-intraventricular in early admission. This study aims to establish an early diagnostic prediction model for severe PIVH. Method: This study was a retrospective cohort study with data collected from the MIMIC-III (v1.4) database. Laboratory and clinical data collected within the first 24 h of NICU admission have been used as variables for both univariate and multivariate logistic regression analyses to construct a nomogram-based early prediction model for severe periventricular-intraventricular hemorrhage and subsequently validated. Results: A predictive model was established and represented by a nomogram, it comprised three variables: output, lowest platelet count and use of vasoactive drugs within 24 h of NICU admission. The model's predictive performance showed by the calculated area under the curve was 0.792, indicating good discriminatory power. The calibration plot demonstrated good calibration between observed and predicted outcomes, and the Hosmer-Lemeshow test showed high consistency (p = 0.990). Internal validation showed the calculated area under a curve of 0.788. Conclusions: This severe PIVH predictive model, established by three easily obtainable indicators within the NICU, demonstrated good predictive ability. It offered a more user-friendly and convenient option for neonatologists.

MicroRNA-124 negatively regulates STAT3 to alleviate hypoxic-ischemic brain damage by inhibiting oxidative stress.

MicroRNA-124 (miR-124) is implicated in various neurological diseases; however, its significance in hypoxic-ischaemic brain damage (HIBD) remains unclear. This study aimed to elucidate the underlying pathophysiological mechanisms of miR-124 in HIBD. In our study performed on oxygen-glucose deprivation followed by reperfusion (OGD)/R-induced primary cortical neurons, a substantial reduction in miR-124 was observed. Furthermore, the upregulation of miR-124 significantly mitigated oxidative stress, apoptosis, and mitochondrial impairment. We demonstrated that miR-124 interacts with the signal transducer and activator of transcription 3 (STAT3) to exert its biological function using the dual-luciferase reporter gene assay. As the duration of OGD increased, miR-124 exhibited a negative correlation with STAT3. STAT3 overexpression notably attenuated the protective effects of miR-124 mimics, while knockdown of STAT3 reversed the adverse effects of the miR-124 inhibitor. Subsequently, we conducted an HIBD model in rats. In vivo experiments, miR-124 overexpression attenuated cerebral infarction volume, cerebral edema, apoptosis, oxidative stress, and improved neurological function recovery in HIBD rats. In summary, the neuroprotective effects of the miR-124/STAT3 axis were confirmed in the HIBD model. MiR-124 may serve as a potential biomarker with significant therapeutic implications for HIBD.

Development and validation of a diagnostic prediction model for severe periventricular-intraventricular hemorrhage in newborns: insights from a retrospective analysis utilizing the MIMIC-III database.

OBJECTIVE: Periventricular-intraventricular hemorrhage is the most common type of intracranial bleeding in newborns, especially in the first 3 days after birth. Severe periventricular-intraventricular hemorrhage is considered a progression from mild periventricular-intraventricular hemorrhage and is often closely associated with severe neurological sequelae. However, no specific indicators are available to predict the progression from mild to severe periventricular-intraventricular in early admission. This study aims to establish an early diagnostic prediction model for severe PIVH. METHOD: This study was a retrospective cohort study with data collected from the MIMIC-III (v1.4) database. Laboratory and clinical data collected within the first 24 h of NICU admission have been used as variables for both univariate and multivariate logistic regression analyses to construct a nomogram-based early prediction model for severe periventricular-intraventricular hemorrhage and subsequently validated. RESULTS: A predictive model was established and represented by a nomogram, it comprised three variables: output, lowest platelet count and use of vasoactive drugs within 24 h of NICU admission. The model's predictive performance showed by the calculated area under the curve was 0.792, indicating good discriminatory power. The calibration plot demonstrated good calibration between observed and predicted outcomes, and the Hosmer-Lemeshow test showed high consistency (p = 0.990). Internal validation showed the calculated area under a curve of 0.788. CONCLUSIONS: This severe PIVH predictive model, established by three easily obtainable indicators within the NICU, demonstrated good predictive ability. It offered a more user-friendly and convenient option for neonatologists.

MiR-155 promotes inflammation and apoptosis via targeting SIRT1 in hypoxic-ischemic brain damage.

Hypoxic-ischemic brain damage (HIBD) is one of the major causes of infant death and long-term neurological disturbances, which puts great pressure on families and society. Previous studies have reported that neuroinflammation regulates the pathogenesis of HIBD. MiR-155 has been reported to participate in many brain injuries; however, its direct implication and related mechanisms are not illuminated in HIBD. Herein, we identified that miR-155 plays a vital role in HIBD both in in vitro and in vivo models. We found that miR-155 promoted inflammation and apoptosis via targeting SIRT1 and negatively regulated its expression levels in oxygen-glucose deprivation/reoxygenation (OGD/R) in an in vitro model. Silencing of SIRT1 reversed the effects of miR-155 inhibitor on apoptosis and the NF-κB pathway in OGD/R-treated PC12 cells and microglia (BV2) cells. Moreover, in a neonatal rat HIBD model, miR-155 enhanced apoptosis and inflammation in the brains of rats with HIBD in vivo. Together, our results demonstrated that miR-155 exerted a negative effect in HIBD by targeting SIRT1, which could contribute to the treatment of neonatal patients with hypoxic-ischemic brain damage.

Denoising method of machine tool vibration signal based on variational mode decomposition and Whale-Tabu optimization algorithm.

The noise from other sources is inevitably mixed in the vibration information of CNC machine tools obtained using the sensors. In this work, a de-noising method based on joint analysis is proposed. The variational mode decomposition (VMD), correlation analysis (CA), and wavelet threshold (WT) denoising are used to denoise the original signal. First, VMD decomposes noisy signals into multiple intrinsic mode functions (IMFs). The penalty factor and decomposition level of VMD parameters are selected by the optimization algorithm by combining the whale optimization algorithm (WOA) and tabu search (TS). The minimum permutation entropy of IMF is used as the fitness function of the proposed fusion algorithm. Then, the IMF is divided into three categories by using the cross-correlation number. They include the pure components, signals containing noise, and complete noise components. Then, the WT method is used to further denoise the signals, and signal reconstruction is performed with the pure component to obtain the denoised signal. This joint analysis denoising method is named TS-WOA-VMD-CA-WT. The simulation results show that the fusion optimization algorithm proposed in this work has better performance as compared to the single optimization algorithm. It performs effectively when applied to the actual machine tool vibration signal denoising. Therefore, the proposed TS-WOA-VMD-CA-WT method is superior to other existing denoising techniques and has good generality, which is expected to be popularized and applied more widely.

Transcranial ultrasound diagnostic value of hemodynamic cerebral changes in preterm infants for early-onset sepsis.

Background: Due to the limitation of blood culture diagnosis, this study sought to evaluate the cerebral hemodynamic changes by Doppler ultrasound for timely and objective diagnosis techniques in preterm infants with early onset-neonatal sepsis. Methods: In this retrospective study, 86 preterm infants treated at the Department of Neonatology, Renmin Hospital of Wuhan University from January 1, 2019 to March 31, 2021, were divided into the following 2 groups: (I) the early onset neonatal sepsis (EONS) group (G1, n=41); (II) the normal control group (G2, n=45). The cerebral hemodynamic changes were examined by transcranial ultrasound. Stata15.0 and SPSS26.0 software were used for the data analysis. The pair-wise comparisons of the receiver operating characteristic (ROC) curves were on the MedCalc18.2.1 software. For all the statistical analyses, P value <0.05 was considered significant. Results: Sex, birth weight, and gestational age did not differ significantly between the groups (P>0.05); the peak systolic velocity (PSV), mean velocity (MV), end diastolic velocity (EDV) (cm/s), resistivity index (RI), pulsatility index (PI) of the anterior cerebral artery (ACA), middle cerebral artery (MCA), and posterior cerebral artery (PCA) differed significantly between 2 groups (P<0.05). In relation to the diagnostic sensitivity, the area under the receiver operating characteristic (AUROC) analyses showed that compared to IL-6 (0.95, 1.00), EDV of the ACA, and PSV, EDV and MV of the MCA and PCA had a higher sensitivity than the others (AUROC: 1, all 95% CI: 1.00, 1.00). The diagnostic points of the EDV and MV of the ACA were 9.8 and 17.3 cm/s, respectively, the PSV, EDV, and MV of the MCA were 55.9, 10.9, and 20.4 cm/s, respectively, and the PSV, EDV, and MV of the PCA were 27.5, 7.5, and 9.8 cm/s, respectively. Conclusions: The study showed that PI increases and RI decreases, MV increases, and cerebral blood flow increases in EONS. Further, the EDV and MV of the ACA and the PSV, EDV, and MV of the MCA and PCA showed higher sensitivity than IL-6.

Inhibition of ADAM10 ameliorates doxorubicin-induced cardiac remodeling by suppressing N-cadherin cleavage.

The present research was designed to examine the effects of disintegrin metalloproteinases 10 (ADAM10) on the doxorubicin (DOX)-induced dilated cardiomyopathy (DCM) and the mechanisms involved, with a focus on ADAM10-dependent cleavage of N-cadherin. The present study constructed recombinant lentiviral vectors expressing short hairpin RNA (shRNA) targeting the ADAM10 gene. H9C2 cells were treated with the recombinant lentivirus or GI254023 (an ADAM10 inhibitor). The expression level of N-cadherin and its C-terminal fragment1 (CTF1) was tested by western blotting and flow cytometry. The adhesion ability was analyzed using a plate adhesion model. Cardiac function and morphology were assessed in control and lentivirus-transfected rats with or without DOX treatment. The inhibition of ADAM10 activity significantly increased the expression of full-length N-cadherin on the cellular surface and reduced CTF1 generation in vivo and in vitro. The adhesion ability was also increased in ADAM10-knockdown H9C2 cells. Furthermore, DOX-induced myocardial dysfunction was ameliorated in rats transfected with ADAM10-shRNA lentivirus. These findings demonstrated that ADAM10 specifically cleaves N-cadherin in cardiomyocytes. ADAM10-induced N-cadherin cleavage results in changes in the adhesive behavior of cells. Therefore, ADAM10 may serve as a therapeutic target to reverse cardiac remodeling in DCM.

N-acetylserotonin Derivative Exerts a Neuroprotective Effect by Inhibiting the NLRP3 Inflammasome and Activating the PI3K/Akt/Nrf2 Pathway in the Model of Hypoxic-Ischemic Brain Damage.

Neonatal hypoxic-ischemic encephalopathy (HIE) is one of the main causes of neonatal disability and death. As a derivative of N-acetylserotonin, N-[2-(5-hydroxy-1H-indol-3-yl) ethyl]-2-oxopiperidine-3-carboxamide (HIOC) can easily cross the blood-brain barrier and have a long half-life in the brain. In this study, the hypothesis was verified that HIOC plays a neuroprotective role in the HIE model and its potential mechanism was evaluated. Firstly, an HIE rat model was established to deliver HIOC, revealing that it can reduce cerebral infarction volume, cerebral edema, and neuronal apoptosis. The results of immunofluorescence staining, Western blots and RT-PCR further showed that HIOC could inhibit the activation of the NLRP3 inflammasome and the expression of related proteins. Finally, the activation of the phosphatidylinositol-3-kinase (PI3K)/Akt/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway by HIOC was verified in vitro and in vivo. It was discovered that HIOC could increase the nuclear translocation of Nrf2, and that this induction can be reversed by the PI3K/Akt pathway inhibitor LY294002. In general terms, the neuroprotective effect of HIOC was confirmed in the HIE model, which is related to the activation of the Pi3k/Akt/Nrf2 signal pathway and the inhibition of the NLRP3 inflammasome.

The value of interleukin-6 (IL-6) within 6 hours after birth in the prompt diagnosis of early-onset neonatal sepsis.

BACKGROUND: To investigate the value of interleukin-6 (IL-6) for neonates within 6 hours after birth in the prompt diagnosis of early-onset neonatal sepsis (EONS). METHODS: The clinical and laboratory data of 129 neonates in the neonatal intensive care unit (NICU) of our center from March 31, 2017, to February 29, 2020, were retrospectively analyzed. These patients were divided into two groups on their disease conditions: the EONS group (n=66) and the healthy control group (n=63). All enrolled patients were born in our hospital's Obstetrics Department and were admitted to the NICU within 2 hours after birth. The first session of the blood test was conducted within 4-6 hours after birth for the measurements of IL-6, C-reactive protein (CRP1), serum amyloid A1 (SAA1), and serum immunoglobulin M (IgM). The second session of the blood test was performed 12-24 hours after birth for procalcitonin (PCT), CRP2, and SAA2. All the tests were completed in our clinical laboratory. The non-parametric test (Mann-Whitney U test) was used to compare all parameters between these two groups. The receiver operating characteristic (ROC) curves were drawn to compare the diagnostic sensitivities and specificities. The pairwise comparisons of the ROC curves were on the MedCalc18.2.1 software. A P value of <0.05 was considered statistically significant. RESULTS: Gender, birth weight, and gestational age were matched between the EONS group and the control group (all P>0.05). The differences of IL-6, CRP2, PCT, and SAA2 were statistically significant between these two groups (all P<0.05), while there was no significant difference in CRP1, SAA1, and IgM (all P>0.05). The area under the ROC curve (AUC) is 1 (95% CI: 0.918-1.000), 1 (95% CI: 0.918-1.000), 1 (95% CI: 0.918-1.000), and 0.977 (95% CI: 0.878-0.999), respectively, for IL-6, CRP2, PCT, and SAA2. Pairwise comparisons among four biomarkers showed the diagnostic specificity and sensitivity of IL-6 were not significantly different from those of CRP2, PCT, and SAA2 (all P>0.05). CONCLUSIONS: IL-6 is a quick and independent diagnostic biomarker for EONS, and its sensitivity and specificity are inferior to the conventional inflammation markers, including CRP, PCT, and SAA.

Clinical values of common biomarkers for efficacy monitoring of antibiotics in early-onset neonatal sepsis.

BACKGROUND: To investigate the clinical values of the common biomarkers including blood routine (B-Rt), C-reactive protein (CRP), serum amyloid A (SAA) and procalcitonin (PCT) for efficacy monitoring of antibiotics in early-onset neonatal sepsis (EONS). METHODS: The clinical and laboratory data of 78 neonates with confirmed EONS in the neonatal intensive care unit (NICU) of our center from July 1, 2019 to June 30, 2020 were retrospectively analyzed. All the subjects were treated with cefotiam (50 mg/kg q12h) and augmentin (30 mg/kg q12h) within 12 hours after birth. Blood samples were collected 0-12 hours after birth for blood culture, measurements of B-Rt, CRP and SAA. Subsequently, blood sampling was performed at intervals of 12-24, 24-48, 48-96, and 96-144 hours for measurements of B-Rt, CRP, SAA and PCT. Statistical analyses were performed in the SPSS 20.0 software package. P value of <0.05 was considered statistically significant. RESULTS: WBC count showed no significant change among different intervals (12-24, 24-48, 48-96, and 96-144 hours); in contrast, NEU%, CRP, SAA and PCT significantly differed across all intervals. SAA had sensitivities of 75.86%, 93.1%, 44.83%, and 3.45%, respectively; specificities of 100% across all intervals; and AUCs of 0.879 (P<0.0001), 0.966 (P<0.0001), 0.724 (P<0.0001), and 0.500, respectively (P=1). PCT had sensitivities of 100%, 100%, 79.31%, and 51.72%, respectively; specificities of 100% across all intervals; and AUCs of 1 (P<0.0001), 1 (P<0.0001), 0.793 (P<0.0001), and 0.517 (P>0.8551), respectively. CONCLUSIONS: WBC count, NEU% and CRP showed no clinical significance for any intervals for efficacy monitoring of antibiotic treatment. SAA and PCT had similar monitoring values at 12-24 and 24-48 hours. SAA is thus more valuable than PCT for efficacy monitoring of antibiotics at the 48-96 and even at the 96-144 hours intervals in EONS.

Diagnostic utility of procalcitonin as a biomarker for late-onset neonatal sepsis.

BACKGROUND: To explore the diagnostic utility of procalcitonin (PCT) as a biomarker for late-onset neonatal sepsis (LONS). METHODS: The clinical and laboratory data of 131 neonatal patients in the neonatal intensive cares unit (NICU) of our center (Department of Neonatology, Renmin Hospital of Wuhan University) from June 1, 2015, to May 31, 2018, were retrospectively analyzed. These patients were divided into 3 groups based on their disease conditions: the bacterial sepsis (BS) group (n=47), the fungal sepsis (FS) group (n=39), and the normal control group (n=45, without sepsis). Blood cultures, routine blood tests, and testing for PCT and C-reactive protein (CRP) were performed in all 3 groups. Both PCT and CRP were measured by using enzyme-linked immunosorbent assay (ELISA). Blood culture was performed in an automated blood culture system. Routine blood tests were performed by using a fully automatic hematology analyzer. RESULTS: Serum PCT level was significantly different between the BS group and control group (P<0.01) but showed no significant difference between the FS group and control group (P>0.05); the difference in CRP was statistically significant between the FS group and control group (P<0.01) but was not statistically significant between the BS group and control group (P>0.05). The areas under the receiver operating characteristics (ROC) curve were 0.979 and 0.826 for PCT/CRP in the BS group and FS group, with a best cutoff value of 0.93 and 33.27, respectively; the sensitivities and specificities of PCT/CRP in these 2 groups were 0.962/0.679 and 0.964/0.964, respectively. CONCLUSIONS: Compared with CRP, PCT is more sensitive in diagnosing BS but is not sensitive for diagnosing FS. Therefore, PCR is a useful biomarker in distinguishing BS from FS in neonates with late-onset sepsis.

Silencing of long non-coding RNA CRNDE promotes autophagy and alleviates neonatal hypoxic-ischemic brain damage in rats.

Hypoxic-ischemic (HI) brain damage (HIBD) leads to high neonatal mortality and severe neurologic morbidity. Autophagy is involved in the pathogenesis of HIBD. This study aims to investigate the effect of long non-coding RNA colorectal neoplasia differentially expressed (CRNDE) on HIBD and to validate whether autophagy is involved in this process. A HIBD model in rat pups and a HI model in rat primary cerebrocortical neurons were established. Autophagy was evaluated by western blot. The HIBD in rats was evaluated by hematoxylin and eosin staining, TUNEL staining, triphenyl tetrazolium chloride staining, and morris water maze test. The HI injury in vitro was evaluated by determining cell viability and apoptosis. The results showed that CRNDE expression was time-dependently increased in the brain after HIBD. Administration with CRNDE shRNA-expressing lentiviruses alleviated pathological injury and apoptosis in rat hippocampus, decreased infarct volume, and improved behavior performance of rats subjected to HIBD. Furthermore, CRNDE silencing promoted cell viability and inhibited cell apoptosis in neurons exposed to HI. Moreover, CRNDE silencing promoted autophagy and the autophagy inhibitor 3-methyladenine counteracted the neuroprotective effect of CRNDE silencing on HI-induced neuronal injury both in vivo and in vitro. Collectively, CRNDE silencing alleviates HIBD, at least partially, through promoting autophagy.

Long non-coding RNA CRNDE deteriorates intrauterine infection-induced neonatal brain injury.

BACKGROUND: This study aimed to test the hypothesis that long non-coding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) could exacerbate brain injury caused by intrauterine infection in neonatal rats. METHODS: Intrauterine infection was induced in pregnant rats by lipopolysaccharide (LPS). After delivery, newborn rats with brain injury caused by intrauterine infection were randomly divided into control, control shRNA, and CRNDE shRNA groups. CRNDE expression in serum and amniotic fluid of pregnant rats and neonatal brain tissues were determined by quantitative real-time PCR (qRT-PCR). Morris water maze (MWM) task was used to test the spatial learning and memory ability. Histological examination and apoptosis detection were performed by hematoxylin and eosin (H&E) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, respectively. Immunohistochemistry was conducted to evaluate the activation of astrocytes and microglia. RESULTS: LncRNA CRNDE was highly expressed in serum and amniotic fluid of maternal rats and in brain tissues of offspring rats. Furthermore, shRNA-mediated CRNDE downregulation could rescue the spatial learning and memory ability, improve brain histopathological changes and cell death, and inhibit the activation of astrocytes and microglia caused by LPS. CONCLUSION: CRNDE silencing possessed a cerebral protective effect in neonatal rats with brain injury caused by interauterine infection.

Clinical analysis of 10 neonates born to mothers with 2019-nCoV pneumonia.

BACKGROUND: The newly identified 2019-nCoV, which appears to have originated in Wuhan, the capital city of Hubei province in central China, is spreading rapidly nationwide. A number of cases of neonates born to mothers with 2019-nCoV pneumonia have been recorded. However, the clinical features of these cases have not been reported, and there is no sufficient evidence for the proper prevention and control of 2019-nCoV infections in neonates. METHODS: The clinical features and outcomes of 10 neonates (including 2 twins) born to 9 mothers with confirmed 2019-nCoV infection in 5 hospitals from January 20 to February 5, 2020 were retrospectively analyzed. RESULTS: Among these 9 pregnant women with confirmed 2019-nCoV infection, onset of clinical symptoms occurred before delivery in 4 cases, on the day of delivery in 2 cases, and after delivery in 3 cases. In most cases, fever and a cough were the first symptoms experienced, and 1 patient also had diarrhea. Of the newborns born to these mothers, 8 were male and 2 were female; 4 were full-term infants and 6 were born premature; 2 were small-for-gestational-age (SGA) infants and 1 was a large-for-gestational-age (LGA) infant; there were 8 singletons and 2 twins. Of the neonates, 6 had a Pediatric Critical Illness Score (PCIS) score of less than 90. Clinically, the first symptom in the neonates was shortness of breath (n=6), but other initial symptoms such as fever (n=2), thrombocytopenia accompanied by abnormal liver function (n=2), rapid heart rate (n=1), vomiting (n=1), and pneumothorax (n=1) were observed. Up to now, 5 neonates have been cured and discharged, 1 has died, and 4 neonates remain in hospital in a stable condition. Pharyngeal swab specimens were collected from 9 of the 10 neonates 1 to 9 days after birth for nucleic acid amplification tests for 2019-nCoV, all of which showed negative results. CONCLUSIONS: Perinatal 2019-nCoV infection may have adverse effects on newborns, causing problems such as fetal distress, premature labor, respiratory distress, thrombocytopenia accompanied by abnormal liver function, and even death. However, vertical transmission of 2019-nCoV is yet to be confirmed.

The influence factors of neonatal respiratory distress syndrome in Southern China: a case-control study.

Objective: To explore the relevant influencing factors of neonatal respiratory distress syndrome (NRDS) in southern China and provide scientific basis for improving the quality of life for neonates.Methods: A retrospective analysis of 320 cases with NRDS neonates admitted from January 2015 to December 2017 in a neonatal department of a Maternal and Child Health Hospital in South China was conducted. Three hundred twenty non-NRDS patients admitted to the same hospital during the same period were also included as control. The basic characteristics were compared and the risk and protective factors for NRDS were evaluated by logistic regression analysis.Results: Univariate analysis showed that the difference in age, gestational age, fetal sex, mode of delivery, asphyxia, intrauterine distress, and gestational diabetes in the case group and the control group were significantly different (p < .05). The multivariate logistic regression analysis revealed that the age of pregnant women (OR ± 1.539, 95% CI ± 1.427-1.660), intrauterine distress (OR ± 2.427, 95% CI ± 1.079-5.458), and gestational diabetes (OR ± 2.881, 95% CI ± 1.271-6.532) were independent risk factors for NRDS. Meanwhile, gestational age (OR ± 0.588, 95% CI ± 0.508-0.681) was an independent protective factor for NRDS.Conclusions: The age of pregnant women, intrauterine distress, and gestational diabetes can increase the risk of NRDS, while long gestational age can reduce the risk of NRDS. Early detection, early diagnosis and early treatment of children with NRDS have achieved the purpose of improving the quality of life of children.

Tanshinone IIA improves hypoxic ischemic encephalopathy through TLR­4­mediated NF­κB signal pathway.

Hypoxic ischemic encephalopathy (HIE) is the most common brain injury following hypoxia and/or ischemia caused by various factors during the perinatal period, resulting in detrimental neurological deficits in the nervous system. Tanshinone IIA (Tan­IIA) is a potential agent for the treatment of cardiovascular and cerebrovascular diseases. In this study, the efficacy of Tan­IIA was investigated in a newborn mouse model of HIE. The dynamic mechanism of Tan­IIA was also investigated in the central nervous system of neonate mice. Intravenous injection of Tan­IIA (5 mg/kg) was administered and changes in oxidative stress, inflammation and apoptosis­associated proteins in neurons. Histology and immunohistochemistry was used to determine infarct volume and the number of damaged neurons by Fluoro­Jade C staining. The effects of Tan­IIA on mice with HIE were evaluated by body weight, brain water content, neurobehavioral tests and blood­brain barrier permeability. The results demonstrated that the apoptosis rate was decreased following Tan­IIA administration. Expression levels of pro­apoptotic proteins, caspase­3 and caspase­9 and P53 were downregulated. Expression of Bcl­2 anti­apoptotic proteins was upregulated by Tan­IIA treatment in neuro. Results also found that Tan­IIA treatment decreased production of inflammatory cytokines such as interleukin­1, tumor necrosis factor­α, C­X­C motif chemokine 10, and chemokine (C­C motif) ligand 12. Oxidative stress was also reduced by Tan­IIA in neurons, as determined by the expression levels of superoxide dismutase, glutathione and catalase, and the production of reactive oxygen species. The results demonstrated that Tan­IIA treatment reduced the infarct volume and the number of damaged neurons. Furthermore, body weight, brain water content and blood­brain barrier permeability were markedly improved by Tan­IIA treatment of newborn mice following HIE. Furthermore, the results indicated that Tan­IIA decreased Toll­like receptor­4 (TLR­4) and nuclear factor­κB (NF­κB) expression in neurons. TLR­4 treatment of neuronal cell in vitro addition stimulated NF­κB activity, and further enhanced the production of inflammatory cytokines and oxidative stress levels in neurons. In conclusion, these results suggest that Tan­IIA treatment is beneficial for improvement of HIE through TLR­4­mediated NF­κB signaling.

Berberine ameliorates neonatal necrotizing enterocolitis by activating the phosphoinositide 3-kinase/protein kinase B signaling pathway.

Neonatal necrotizing enterocolitis (NEC) is a severe acquired disease that predominantly affects the small intestine of neonates. NEC is caused by a combination of metabolic products, dysfunctions of the blood vessels, mucus and other unknown factors. Berberine may induce beneficial effects on necrotic and cardiovascular diseases due to its anti-inflammatory and anti-apoptotic effects on epithelial cells. In the present study, the therapeutic effects of berberine were investigated and the potential mechanisms by which it functions within a neonatal NEC mouse model were analyzed. Inflammation and levels of associated factors were measured in the serum of mice with NEC prior to and following treatment with berberine. Apoptotic rates in epithelial cells were analyzed following treatment with berberine. The expression of genes associated with apoptosis and apoptosis signaling were determined in epithelial cells in the small intestines of mice with NEC following treatment with berberine. The phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway was investigated in epithelial cells isolated from mice following treatment with either berberine or PBS. Histology and immunohistochemistry were used to determine the area of infarction and apoptosis. Body weight and food intake were measured to evaluate the physical effects of berberine on mice with NEC. The results indicated that berberine attenuated the inflammation caused by NEC in mice after 10 days of treatment. The apoptosis rate of epithelial cells isolated from experimental mice was decreased following berberine treatment. Western blot analysis indicated that the expression of the anti-apoptotic genes c-Myc and p53 were upregulated by berberine, whereas caspase-3 and -9 levels were downregulated in epithelial cells following treatment with berberine. In addition, the expression and phosphorylation levels of PI3K and AKT were downregulated in epithelial cells following treatment with berberine. An in vitro assay indicated that treatment with PI3K alone increased the expression of AKT and promoted the apoptosis of epithelial cells. Treatment with berberine markedly increased epidermal growth factor (EGF) and Bcl-2 expression levels, the activity of epithelial cells and decreased the infarction area of the small intestine. Accordingly, the body weight and food intake of mice with NEC were increased following berberine treatment. Therefore, the results of the present study demonstrate that berberine inhibits inflammation and apoptosis via the PI3K/AKT signaling pathway and may therefore attenuate the progression of NEC. These results suggest that berberine may be a potential therapeutic agent for the treatment of patients with NEC.

The chemiluminescence immunoassay for aflatoxin B1 based on functionalized magnetic nanoparticles with two strategies of antigen probe immobilization.

A rapid and sensitive chemiluminescence immunoassay (CLIA) based on magnetic nanoparticles (MNPs) was developed to detect aflatoxin B1 (AFB1), which is a potent carcinogen in nature. We prepared monodisperse MNPs (300 nm in diameter) according to the solvothermal synthesis reaction and the MNPs were coated with silica by the Stöber method. Triethox was used as a one-step carboxylation reagent, and 3-aminopropyltriethoxysilane (APTES) an amination reagent, to modify the MNPs. We prepared two types of solid phase antigens using the above synthesized functionalized MNPs coupled with the later prepared AFB1-oxime active ester and the purchased BSA-AFB1 respectively. 2',6'-dimethylcarbonylphenyl-10-sulfopropylacridinium-9-carboxylate 4'-N-hydroxysuccinimide (4'-NHS) ester (NSP-DMAE-NHS), as a novel luminescent reagent, was used to label anti-AFB1 antibodies. The two CLIA calibration curves based on the two types of solid phase antigens were obtained and compared. The acquired limit of detection (LOD) was about 0.001 ng/mL for the two functionalized MNPs-based immunoassays, and the half maximal inhibitory concentration (IC50 ) was 0.51 ng/mL for the MNPs-AFB1-based method and 0.72 ng/mL for the MNPs-BSA-AFB1-based method.

Intraventricular injection of human dental pulp stem cells improves hypoxic-ischemic brain damage in neonatal rats.

OBJECTIVE: To investigate the effect of intraventricular injection of human dental pulp stem cells (DPSCs) on hypoxic-ischemic brain damage (HIBD) in neonatal rats. METHODS: Thirty-six neonatal rats (postnatal day 7) were assigned to control, HIBD, or HIBD+DPSC groups (n = 12 each group). For induction of HIBD, rats underwent left carotid artery ligation and were exposed to 8% to 10% oxygen for 2 h. Hoechst 33324-labeled human DPSCs were injected into the left lateral ventricle 3 days after HIBD. Behavioral assays were performed to assess hypoxic-ischemic encephalopathy (HIE), and on postnatal day 45, DPSC survival was assessed and expression of neural and glial markers was evaluated by immunohistochemistry and Western blot. RESULTS: The HIBD group showed significant deficiencies compared to control on T-maze, radial water maze, and postural reflex tests, and the HIBD+DPSC group showed significant improvement on all behavioral tests. On postnatal day 45, Hoechst 33324-labeled DPSC nuclei were visible in the injected region and left cortex. Subsets of DPSCs showed immunostaining for neuronal (neuron-specific enolase [NSE], Nestin) and glial markers (glial fibrillary acidic protein [GFAP], O4). Significantly decreased staining/expression for NSE, GFAP, and O4 was found in the HBID group compared to control, and this was significantly increased in the HBID+DPSC group. CONCLUSION: Intraventricular injection of human DPSCs improves HIBD in neonatal rats.