Extracellular matrix (ECM) stiffness is closely related to the physiological and pathological states of breast tissue. The current study was aimed to investigate the effect of silk fibroin/collagen composite hydrogels with adjustable matrix stiffness on the growth and phenotype of normal breast epithelial cells. In this study, the enzymatic reaction of horseradish peroxidase (HRP) with hydrogen peroxide (H₂O₂) was used to change the degree of cross-linking of the silk fibroin solution. The rotational rheometer was used to characterize the composite hydrogel's biomechanical properties. Human normal mammary epithelial cell line MCF-10A were inoculated into composite hydrogels with various stiffness (19.10-4 932.36 Pa) to construct a three dimensional (3D) culture system of mammary epithelial cells. The CCK-8 assay was applied to detect the cell proliferation rate and active states in each group. Hematoxylin-Eosin (HE) staining and whole-mount magenta staining were used for histological evaluation of cell morphology and distribution. The results showed that with the increase of matrix stiffness, MCF-10A cells exhibited inhibited proliferation rate, decreased formation of acinus structures and increased branching structures. Meanwhile, with the increase of matrix stiffness, the polarity of MCF-10A cells was impeded. And the increase of matrix stiffness up-regulated the expression levels of mmp-2, mmp-3, and mmp-9 in MCF-10A cells. Among the genes related to epithelial-mesenchymal transition (EMT), the expression level of the epithelial marker gene E-cadherin was significantly down-regulated, while the interstitial cell marker gene Vimentin was up-regulated, and the expression levels of Snail, Wnt5b and Integrin β1 in the Wnt pathway were up-regulated. These results suggest that the silk fibroin/collagen composite hydrogels with adjustable matrix stiffness regulates the proliferation and the phenotype of MCF-10A cells. The effects of increased matrix stiffness may be closely related to the changes of the polar structures and function of MCF-10A cells, as well as the occurrence of ECM-remodeling and EMT.
Collagen/metabolism* ; Epithelial Cells/metabolism* ; Fibroins/pharmacology* ; Humans ; Hydrogels/metabolism* ; Hydrogen Peroxide ; Phenotype

Birth Weight ; Female ; Gestational Age ; Humans ; Incidence ; Infant ; Infant, Newborn ; Infant, Very Low Birth Weight ; Pregnancy ; Risk Factors ; Sepsis/etiology*

OBJECTIVE: To study the influence of twin pregnancy by assisted reproductive technology (ART) versus twin pregnancy by spontaneous conception (SC) on neonatal outcomes. METHODS: A retrospective analysis was performed for the clinical data of 3 356 live twins with a gestational age of ≥24 weeks who were born in Nanjing Maternal and Child Health Hospital from 2017 to 2019, with 2 006 twins (1 003 pairs) in the ART group and 1 350 (675 pairs) in the SC group. The two groups were compared in terms of the mother's general information and pregnancy comorbidities and the general information, diseases, and outcomes of neonates. RESULTS: Compared with the SC group, the ART group had a significantly higher maternal age ( CONCLUSIONS Compared with twin pregnancy by SC, twin pregnancy by ART does not increase the neonatal mortality rate and risk of adverse outcomes.
Cesarean Section ; Female ; Humans ; Infant, Newborn ; Pregnancy ; Pregnancy Outcome ; Pregnancy, Twin ; Premature Birth ; Reproductive Techniques, Assisted ; Retrospective Studies

OBJECTIVE: To explore the genetic basis for a child suspected for hypokalemic periodic paralysis. METHODS: Clinical data of the patient was collected, and venous blood samples were taken from the patient and his parents for the extraction of genomic DNA. Next generation sequencing (NGS) with target capture was carried out to detect potential variants. Suspected variants were validated by Sanger sequencing. RESULTS: The child developed fatigue without obvious reason at the age of 15. Laboratory test revealed hypokalemia but normal serum magnesium. Genetic testing discovered that he has carried two variants in the SLC12A3 gene, namely c.179C>T and c.539C>A. The patient was diagnosed with Gitelman syndrome. CONCLUSION For children with hypokalemia, genetic testing should be considered for the differential diagnosis of Gitelman syndrome from hypokalemia due to other causes.

Colla Corii Asini(Ejiao)is an important Chinese medicine used in China for thousands of years, and is well known for its famous tonic properties. The herbalogical study was detailed carried out based on the naming, habitat, harvesting, processing, medicinal properties and clinical efficacy. The results showed that the name of Ejiao could be traced back to Shennong's Materia Medica, and various names of Lvpi Jiao, Penfu Jiao and Fuzhi Jiao were recorded in other ancient books. In the many intervening centuries, the main materials of Ejiao had been replaced from cow leather before Tang Dynasty to donkey skin in the middle to late Tang Dynasty. This phenomenon could be probably caused by complicated social factors of various periods and different efficacy of Ejiao made by all kinds of raw materials. Ejiao was merely processed with the simple methods before Tang Dynasty, which subsequently improved avariety of methods to enhance the supplementation action. Most importantly, Ejiao has a wide clinic application along with the development of processing theories and methods, which can be found in various Classics, especially in imperial medical case record in Qing Dynasty.
Animals ; Cattle ; China ; Drugs, Chinese Herbal ; Female ; Gelatin ; Materia Medica ; Medicine, Chinese Traditional

Objective To explore the genetic etiology of a girl featuring epilepsy,speech delay and mild mental retardation.Methods Peripheral blood samples of the child and her parents were collected.Genomic DNA was extracted and subjected to next generation sequencing.Suspected variant was confirmed by Sanger sequencing.Results The child was found to carry a de novo heterozygous c.3592G＞ A (p.V1198M) variant of the SMARCA2 gene,which was predicted to be pathogenic by bioinformatic analysis.Conclusion The child was diagnosed with Nicolaides-Baraitser syndrome due to heterozygous variant of the SMARCA2 gene.

OBJECTIVE: To explore the genetic cause for a child with growth retardation by next generation sequencing (NGS). METHODS: Clinical data of the patient was collected. Peripheral venous blood samples were taken from the neonate and his parents. Targeted capturing and NGS were carried out to detect mutations of genes associated with inborn errors of metabolism. Suspected mutations were validated by Sanger sequencing. RESULTS: The 15-month-old female patient was admitted to hospital for growth retardation for 4 months. Hypomyelination was found upon cranium MRI. Genetic testing revealed two novel insertional mutations in the GLB1 gene in the patient, namely c.2006-2007insT and c.475-476 insGGTCC. CONCLUSION The c.2006-2007insT and c.475-476 insGGTCC mutations of the GLB1 gene probably underlie the GM1 gangliosidosis resulting in the growth retardation in the child.
Female ; Gangliosidosis, GM1 ; genetics ; Humans ; Infant ; Infant, Newborn ; Mutation ; Pedigree ; beta-Galactosidase ; genetics

OBJECTIVE: To detect pathogenic mutation of DOCK6 gene in a patient with convulsive seizure and refractory epilepsy. METHODS: CytoScan HD-Array and next generation sequencing were used to detect the potential mutation in the patient. RESULTS: The proband has carried compound heterozygous mutations of c.188C>T (p.Arg63Gln) and c.5374C>T (p.Glu1792Lys) of the DOCK6 gene, which were respectively inherited from his mother and father. Neither mutation was reported previously. Bioinformatic analysis indicated that the two amino acids are highly conserved. Based on the ACMG guidelines, the c.188C>T mutation was predicted to be likely pathogenic, while the c.5374C>T mutation was of uncertain significance. CONCLUSION The compound heterozygous mutations of c.188C>T (p.Arg63Gln) and c.5374C>T (p.Glu1792Lys) of the DOCK6 gene probably underlie the disease in this patient.
Child ; Diabetes Mellitus, Type 2 ; Ectodermal Dysplasia ; genetics ; Guanine Nucleotide Exchange Factors ; genetics ; Humans ; Limb Deformities, Congenital ; genetics ; Mutation ; Pedigree ; Scalp Dermatoses ; congenital ; genetics

Objective To observe the dynamic change of serum cholinesterase (ChE) activity in severely burned patients,analyze the correlation between serum cholinesterase activity and both the severity and prognosis of burn injury.Methods The clinical data of 203 patients with large area burn (TBSA ＞30％) were collected from the Department of Burns and Plastic Surgery,First Affiliated Hospital of PLA General Hospital from January 2014 to December 2016 and the data of 30 healthy subjects as control group were retrospectively analyzed.According to the total burn area,these patients were divided into 3 groups:30％-49％ TBSA group (n=77),50％-79％ TBSA group (n=70),more than 80％ TBSA group (n=56).The serum was extracted from the two groups at 1,3,5,7,9,11,14,21,28,35 and 42 days after injury.Serum ChE activity was compared between the patients and the controls,and between the patient groups.The comparisons were also conducted between the 3 patient groups on the altitude of the decrease in serum ChE activity 21 days after injury,when the serum ChE activity reached the lowest level after injury,and the correlation between the burn area and the serum ChE activity was analyzed at each time point.In addition,according to the prognosis,the patients were divided into the survival group (n=172) and the death group (n=31).The serum ChE activity at each time point and the range of serum ChE activity decrease on day 21 and day 28 after injurywere compared between the two groups.Results 1)The serum ChE activity was lower in the patient groups than in the control group (8.18 ± 1.15kU/L) at all the time points observed except the first day after injury (P＜0.01).2) On the 21st and 28th days after injury,the serum AhE activity was higher in 30％-49％TBSA group than in the 50％-79％ TBSA group and in ≥ 80％ TBSA group,and the activity was higher in 50％-79％ TBSA group than in ≥ 80％ TBSA group (P＜0.05).Pearson correlation analysis on the serum ChE activity and burn area showed a significant positive correlation on day 3,5 and 7 after injury (P＜0.01) and a significant negative correlation on day 14,21 and 28 day after the injury (P＜0.05,P＜0.01),and no significant correlation at the remaining time points (P＞0.05).Up to the 21st day after injury,serum ChE decreased less significantly in 30％-49％ TBSA group (1.18 ± 1.70kU/L) than in 50％-79％ TBSA group (2.20 ± 2.01kU/L)and in ≥80％ TBSA group (3.35 ± 1.89kU/L),and less significantly in 50％-79％ TBSA group than in ≥80％ TBSA group,with statistically significant differences (P＜0.05).3) The serum ChE activity decreased to the lowest value (4.89 ± 1.48kU/L) on day 21 after burn in the survival group and to the lowest value (4.21 ± 1.37kU/L) on day 28 in the death group,with statistically significant difference (P＜0.05).On day 28 after injury,the serum ChE activity was significantly higher (4.92 ± 1.46kU/L) in the survival patients than in the patients died (4.21 ± 1.37kU/L) at the 21st day after injury.On day 21 and 28 after injury,the magnitude of decrease in serum ChE activity was less significant in the survival group (2.26 ± 1.93kU/L and 2.43 ± 1.87kU/L respectively) than in the death group (3.61 ± 2.20kU/L and 4.22 ± 1.94kU/L) (P＜0.05).Conclusions After burn,the activity of serum ChE decreased significantly,and there was a tendency to decrease firstly and increase aftetwards.The activity reached the lowest value on day 21 after injury.The area of burn was negatively correlated with the activity of serum ChE,and this relation was more remarkable on day 14,21 and 28 after injury.The greater the decrease of serum ChE activity,the worse the prognosis of the patients.As such,serum ChE activity has a certain value in reflecting the severity of burn injury and predicting the prognosis of patients.

Objective To investigate the time distribution of pre-hospital emergency stroke patients,so as to provide recommendations for stroke prevention and control.Methods The information of 2844 stroke patients from 2013 to 2016 was collected from emergency medical center of Wenzhou,and circular distribution method was used to analyze the seasonal stroke incidence.Results Both seasonal and daily occurrence of cerebralvascular accidents showed weak(all r<0.3) trends of centralization(all P<0.05).The average angle for each year was not equal,and the average angle of 2014 was significantly different from the other years(P<0.05). The peak time point of every year(except 2014)was on the 4th of January and the peak time period was from the 2nd of September to the 9th of May of next year. The average angle for every day was(230.965±108.999)°(P<0.05),which suggested that the peak time point was 14:35,and the peak time period was from 7:19 to 21:51.Conclusion Stroke morbidity has obvious seasonality in Wenzhou,and the peak time was in winter and spring. Besides,the cerebralvascular accidents occures more often in the day time.The time distribution characteristics of stroke patients should be considered when studying the causes of stroke.