In hepatocytes,the endoplasmic reticulum and mitochondria work together in a reciprocal manner to regulate the physiological functions of the cells.Nevertheless,its precise mechanism remains unknown.Endoplasmic reticulum-associated degradation (ERAD) is a key process for maintaining pro-tein quality in the endoplasmic reticulum.Recent studies have found that ERAD can regulate the adipo-cyte mitochondrial function through mitochondria associated membranes (MAMs).However,it is still unknown how exactly ERAD and mitochondria interact with one another in hepatocytes.In this study,chemical induction and gene deletion were used to induce ERAD deficiency in HepG2 cells.To investi-gate the effect of ERAD on mitochondrial function in hepatocytes,we employed fluorescent labelling,flow analysis,immunoblotting and other methods.The results demonstrated that ERAD dysfunction resulted in a reduction in the mitochondrial membrane potential and ATP levels in HepG2 cells (P<0.01) .Fur-thermore,ERAD dysfunction also disrupted the endoplasmic reticulum and mitochondrial Ca2+homeosta-sis in HepG2 cells,and improved mitochondrial energy metabolism function when endoplasmic reticulum Ca2+release was reduced (P<0.05) .These findings were further validated in ERAD dysfunctional pri-mary hepatocytes,which had been induced by pharmacological agents.The data above indicate that ERAD plays a key regulatory role in endoplasmic reticulum-mitochondrial interactions in hepatocytes.En-hanced ERAD function may boost mitochondrial activity in hepatocytes.This study offers fresh perspec-tives on disorders linked to mitochondrial malfunction.

In hepatocytes,the endoplasmic reticulum and mitochondria work together in a reciprocal manner to regulate the physiological functions of the cells.Nevertheless,its precise mechanism remains unknown.Endoplasmic reticulum-associated degradation (ERAD) is a key process for maintaining pro-tein quality in the endoplasmic reticulum.Recent studies have found that ERAD can regulate the adipo-cyte mitochondrial function through mitochondria associated membranes (MAMs).However,it is still unknown how exactly ERAD and mitochondria interact with one another in hepatocytes.In this study,chemical induction and gene deletion were used to induce ERAD deficiency in HepG2 cells.To investi-gate the effect of ERAD on mitochondrial function in hepatocytes,we employed fluorescent labelling,flow analysis,immunoblotting and other methods.The results demonstrated that ERAD dysfunction resulted in a reduction in the mitochondrial membrane potential and ATP levels in HepG2 cells (P<0.01) .Fur-thermore,ERAD dysfunction also disrupted the endoplasmic reticulum and mitochondrial Ca2+homeosta-sis in HepG2 cells,and improved mitochondrial energy metabolism function when endoplasmic reticulum Ca2+release was reduced (P<0.05) .These findings were further validated in ERAD dysfunctional pri-mary hepatocytes,which had been induced by pharmacological agents.The data above indicate that ERAD plays a key regulatory role in endoplasmic reticulum-mitochondrial interactions in hepatocytes.En-hanced ERAD function may boost mitochondrial activity in hepatocytes.This study offers fresh perspec-tives on disorders linked to mitochondrial malfunction.

OBJECTIVES: To investigate the risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture and establish a nomogram model for predicting the risk of neonatal asphyxia. METHODS: A retrospective study was conducted with 613 cases of neonatal asphyxia treated in 20 cooperative hospitals in Enshi Tujia and Miao Autonomous Prefecture from January to December 2019 as the asphyxia group, and 988 randomly selected non-asphyxia neonates born and admitted to the neonatology department of these hospitals during the same period as the control group. Univariate and multivariate analyses were used to identify risk factors for neonatal asphyxia. R software (4.2.2) was used to establish a nomogram model. Receiver operator characteristic curve, calibration curve, and decision curve analysis were used to assess the discrimination, calibration, and clinical usefulness of the model for predicting the risk of neonatal asphyxia, respectively. RESULTS: Multivariate logistic regression analysis showed that minority (Tujia), male sex, premature birth, congenital malformations, abnormal fetal position, intrauterine distress, maternal occupation as a farmer, education level below high school, fewer than 9 prenatal check-ups, threatened abortion, abnormal umbilical cord, abnormal amniotic fluid, placenta previa, abruptio placentae, emergency caesarean section, and assisted delivery were independent risk factors for neonatal asphyxia (P<0.05). The area under the curve of the model for predicting the risk of neonatal asphyxia based on these risk factors was 0.748 (95%CI: 0.723-0.772). The calibration curve indicated high accuracy of the model for predicting the risk of neonatal asphyxia. The decision curve analysis showed that the model could provide a higher net benefit for neonates at risk of asphyxia. CONCLUSIONS The risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture are multifactorial, and the nomogram model based on these factors has good value in predicting the risk of neonatal asphyxia, which can help clinicians identify neonates at high risk of asphyxia early, and reduce the incidence of neonatal asphyxia.
Infant, Newborn ; Humans ; Male ; Pregnancy ; Female ; Nomograms ; Retrospective Studies ; Cesarean Section ; Risk Factors ; Asphyxia Neonatorum/etiology*

Purpose: To identify more accurate predictors of upper urinary tract dilatation (UUTD) in neurogenic bladder (NB) children, we studied the relationship among urodynamic parameters at different bladder filling stages, detrusor leak point pressure (DLPP) and UUTD. Methods: A total of 158 children (3–16 years) with NB were included and then divided into 2 groups according to whether their NB diagnosis was complicated with UUTD: the UUTD group (39 patients) and those without UUTD group (control group, 119 patients). The bladder filling phase was divided into 3 equal parts: the early, middle, and end filling stages. The bladder compliance (BC) and detrusor pressure (△Pdet) at each phase and DLPP at the end filling stage were recorded. Results: A BC<8 mL/cm H2O both in the middle and end stages is more specific than a BC<9 mL/cm H2O in the end stage (72%, 73%, vs. 66%), and △Pdet >8 cm H2O in the early stage, 20 cm H2O in the middle stage and 25 cm H2O in the end stage are more sensitive than △Pdet >40 cm H2O in the end stage (82%, 85%, 85%, vs. 49%). A DLPP cutoff value of 20 cm H2O showed higher sensitivity for predicting UUTD than 40 cm H2O. Conclusions Low BC and a high △Pdet in the middle and end filling stages are more accurate factors than classic indicators for predicting UUTD. In addition, a DLPP value of >20 cm H2O in the end bladder filling stage shows high sensitivity.

OBJECTIVES: To investigate the incidence of extrauterine growth retardation (EUGR) and its risk factors in very preterm infants (VPIs) during hospitalization in China. METHODS: A prospective multicenter study was performed on the medical data of 2 514 VPIs who were hospitalized in the department of neonatology in 28 hospitals from 7 areas of China between September 2019 and December 2020. According to the presence or absence of EUGR based on the evaluation of body weight at the corrected gestational age of 36 weeks or at discharge, the VPIs were classified to two groups: EUGR group (n=1 189) and non-EUGR (n=1 325). The clinical features were compared between the two groups, and the incidence of EUGR and risk factors for EUGR were examined. RESULTS: The incidence of EUGR was 47.30% (1 189/2 514) evaluated by weight. The multivariate logistic regression analysis showed that higher weight growth velocity after regaining birth weight and higher cumulative calorie intake during the first week of hospitalization were protective factors against EUGR (P<0.05), while small-for-gestational-age birth, prolonged time to the initiation of total enteral feeding, prolonged cumulative fasting time, lower breast milk intake before starting human milk fortifiers, prolonged time to the initiation of full fortified feeding, and moderate-to-severe bronchopulmonary dysplasia were risk factors for EUGR (P<0.05). CONCLUSIONS It is crucial to reduce the incidence of EUGR by achieving total enteral feeding as early as possible, strengthening breastfeeding, increasing calorie intake in the first week after birth, improving the velocity of weight gain, and preventing moderate-severe bronchopulmonary dysplasia in VPIs.
Female ; Fetal Growth Retardation ; Gestational Age ; Hospitalization ; Humans ; Incidence ; Infant ; Infant, Newborn ; Infant, Premature ; Infant, Very Low Birth Weight ; Prospective Studies ; Risk Factors

OBJECTIVETo observe the effects of Naoyikang (NYK) on expression of choline acetyltransferase (ChAT) in brain of rats with Alzheimer' s disease (AD).

METHODBilateral infusions of Ibotenic acid (IBO) into nucleus basalis of Meynert (NBM) using hamilton syringe and stereotaxic apparatus were adopted to establish the rat model of AD. After intragastrically administrated with different solution for 28 days, immunohistochemistry and Western-blot were adopted to study the expression of ChAT in frontal cortex of AD rats.

RESULTNYK could improve the morphology and increase the number of ChAT immunoreactive neurons, and significantly promote ChAT protein expression.

CONCLUSIONNYK may be able to increase the synthesis of acetylcholine (ACh) through elevating the expression of ChAT protein, thus improving the level of brain ACh so as to protect central cholinergic neurons.

Alzheimer Disease ; enzymology ; Animals ; Blotting, Western ; Brain ; drug effects ; enzymology ; Choline O-Acetyltransferase ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation ; drug effects ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effect of catalpol from Radix Rehmanniae on A?25-35-induced apoptosis of PC12 cells.Methods PC12 cells were routinely cultivated and treated by A?25-35(final concentration,20 ?mol/L) 24 hours after the addition of catalpol or saline.Forty-eight hours later,cells were examined for viability and apoptosis by MTT method and TUNEL method,respectively,while Bax and Bcl-2 mRNA expression were analyzed by semi-quantitive RT-PCR. Results Catalpol could significantly elevate the viability at 1?10-5 mol/L and 1?10-4 mol/L(P

Objective To establish mouse model of dementia by intracranial injection of A?25-35 and small amount of ibotenic acid(IBO) and to explore whether the effects of catalpol can affect the brain M receptor density and the short term memory. Methods The mice were randomly divided into three groups: control group,model group,treat group which were given orally for 2 months with 50 mg?kg-1?d-1 of catalpol.Dementia model was developed by single unilateral injection of 0.3 ?L of a solution of A?(1?L normal saline containing 4 ?g of A?25-35 and 1 ?g of ibotenic acid) into right basal ganglion region according the atlas of mouse brain with the aid of a stereotaxic equipment.The track of injection was observed by HE staining.The learning/memory ability was measured by Y-maze perfor-mance.The brain muscarinic receptor density was analyzed with single-site binding assay using 3H-quinuclidinyl benzilae(QNB).Results Two months after model development,the learning ability as well as the density of muscarinic receptor in brain were significantly decreased in model mice compared with those in control mice.Parallel models treated with daily oral administration of Catalpol for two months improved the learning ability and increased the brain muscarinic receptor density when compared with model mice.The correlation coefficient between total M receptor densities and the learning/memory ability was significant when examined with linear regresion.Conclusion A dementia model was established in mice.Dementia model was developed by single unilateral injection of 0.3 ?L of a solution of A?(1 ?L normal saline containing 4 ?g of A?25-35 and 1 ?g of ibotenic acid) into right basal ganglion region was established in mice.Catalpol can significantly improve the learning and increase the brain muscarinic receptor density of the model.

Objective To investigate whether catalpol is the active component responsible for the Yin tonic effect of Radix Rehmanniae.Methods Young NH mice were injected with triiodothyronine to produce the hyperthyroidism model,while old mice were used as the model of natural aging.The single point radioligand binding assay was carried out to determine the ?-adrenergic receptor density and M-cholinergic receptor density.The learning ability(short term memory) was determined by the Y-maze avoidance test. Results In the ?-adrenergic receptor experiment,the densities were(15.7?5.2) and(20.9?7.2) fmol/mg protein in normal control group and in T3 control group(P

Objective To investigate the effects of ZMS,the active component of Zhimu,on amyloid precursor protein(APP) and ?-site APP cleaving enzyme 1(BACE1) in HEK293sw cells. Methods HEK293sw cells were pretreated with different concentrations of ZMS in routine culture medium for 24 h,and then serum-free medium with ZMS was used for further culture for 48 h.Cells were examined for expression of APP and ?-secretase BACE1 with Western blotting and RT-PCR,and were then examined for BACE1 activity with fluorescence method. Results 1 ?mol/L and 10 ?mol/L ZMS significantly decreased the expression of BACE1 mRNA and protein,while had no effect on the expression of APP mRNA and protein.It was indicated by fluorescence analysis that 10 ?mol/L ZMS significantly decreased the ?-secretase activity.Conclusion ZMS significantly decreases the activity and expression of ?-secretase BACE1,while has no effect on the expression of APP in HEK293sw cells.