OBJECTIVE To investigate the protective effects of total flavonoids from bamboo (Phyllostachys pubescens) leaves (BLE) on UVB-irradiated oxidative damage in HaCaT keratinocytes.METHODS The HaCaT cells were pretreated with BLE for 24 h and then exposed to UVB(20 mJ/cm2) for 2 h.Cell viability was determined by MTT assay.The levels of reactive oxygen species (ROS),lipid peroxidation and antioxidant enzymes including catalase (CAT),superoxide dismutase (SOD) and glutathione peroxidase(GSH-Px) were measured.mRNA transcription levels of these antioxidant enzymes were determined by RT-PCR assay.In addition,the production of TNF-αand IL-6 were detected by ELISA assay.RESULTS BLE significantly reduced the UVB-induced HaCaT cell damage and improved the cell survival rates (P<0.05).Compared to the control group,BLE decreased the levels of ROS and lipid peroxidation,and increased the activities and mRNA transcription levels of endogenous antioxidant enzymes(SOD,CAT and GSH-Px)in UVB-irradiated HaCaT cells.In addition,BLE also in-hibited the secretion of TNF-αand IL-6 in UVB-irradiated HaCaT cells.CONCLUSION BLE can exhibit cytoprotective activi-ties against UVB-induced oxidative damage in HaCaT cells through the reduction of ROS,inhibition of lipid peroxidation and regulation of antioxidant enzymes activities.In addition,BLE can also decrease the levels of TNF-αand IL-6 in UVB-irradiated HaCaT cells.

Objective The neuronal ceroid lipofuscinosis (CLN are a group of severe lysosomal storage diseases.The patients present with clinically and genetically heterogeneous neurodegenerative disorders.This study aims to investigate the clinical characteristics and the gene mutations of a rare Chinese family with 3 siblings affected by CLN7.Methods The proband,a 5-year-old girl,visited us because of intermittently seizures and mental retardation for 2 years and a half in December,2015.Clinical investigation,brain magnetic resonance imaging(MRI),biochemical and the gene analysis were performed for the etiological study.Results The proband had seizures at the age of 2 and a half years,with the progressive motor deterioration,speech disturbance,mental regression and vision loss.Her brain MRI showed diffusive cerebral atrophy.The blood aminoacids,acylcarnitine and urine organic acid profiles were normal.Lysosomal palmitoyl protein thioesterase and tripeptidyl peptidase activities of peripheral leukocytes were normal.A compound heterozygous mutation of c.1351-1G ＞ A and c.300T ＞ G was detected on her MFSD8 gene,supporting the diagnosis of CLN7.Both of the 2 mutations were novel.Each of her parents carried one of the mutations.Two brothers of the proband had similar clinical process.Her elder brother died at the age of 7 due to severe encephalopathy of unknown etiology.The younger brother showed dyskinesia from the age of 2 years and seizures from the age of 4 years.A compound heterozygous mutation on MFSD8 gene,c.1351-1G ＞ A and c.300T ＞ G,was found from the younger brother,as same as the proband.Conclusions CLN7 is a rare disorder of CLN.In this study,the diagnosis of the 3 siblings with similar clinical process were much delayed.Gene analysis was key for the diagnosis.Two novel mutations were found on MFSD8 of the family.There is still no effective treatment for neurol ceroid lipofuscinosis.The prognosis is poor.Based on the mutation diagnosis,prenatal diagnosis for the next sibling is possible to the prevention of the disease.

AIM:To observe the effect of thyroxine on the expression of T-type calcium channels Cav3. 1, Cav3. 2 and Cav3. 3 in rat myocardium, and to explore the possible biological mechanism between the changes of the ex-pression of T-type calcium channels and the arrhythmia in hyperthyroid heart disease. METHODS:Healthy SD rats (n=20) were randomly divided into normal control group (n=10) and hyperthyroid heart disease group (n=10). The animal model was established by intraperitoneal injection of levothyroxine for 35 d. The contents of T3 and T4 in serum, the heart-to-body weight ratio, the diameter of cardiac myocytes and electrocardiograph were measured to evaluate hyperthyroid heart disease. Moreover, the mRNA and protein expression levels of T-type calcium channels in the myocardium were measured by RT-PCR, immunohistochemistry and Western blot. RESULTS:After intraperitoneal injection of levothyroxine for 35 d, compared with the normal control group, the serum contents of T3 and T4, the heart-to-body weight ratio and the diameter of cardiac myocytes were significantly increased in hyperthyroid heart disease group (P<0.05), and arrhythmia occurred in hyperthyroid heart disease group. By immunohistochemistry and Western blot, the protein expression of Cav3. 1 in-creased significantly (P<0.05), while the protein expression of Cav3.2 decreased significantly (P<0.01). However, no change of the Cav3. 3 protein was observed. The results of RT-PCR were the same as immunohistochemistry and Western blot. CONCLUSION:Thyroxine promotes the expression of Cav3. 1 in the myocardium but inhibits the expression of Cav3. 2 at mRNA and protein levels, which might be involved in arrhythmia in hyperthyroid heart disease.

OBJECTIVETo investigate the association between birth defects and dietary nutrient intake in a high risk area of China.

METHODSA dietary survey was performed and serum folic acid was measured in women whose pregnancy was affected by neural tube defects (NTDs) or unaffected by any birth defects (BDs) in Zhongyang and Jiaokou Counties in Shanxi Province of China.

RESULTSThe local average consumption of foods including dark green vegetables, fruits, fat and meat, and nutrient intake (e.g., energy, protein, retinol, riboflavin, vitamin E, and selenium) were lower than the national average level. In women of childbearing age, these regions, the intake of nutrients was much lower than the recommended nutrient intake (9%-77%). The case-control dietary nutrition study of women whose pregnancy was affected by BDs (including NTDs and congenital heart defects) demonstrated that, in early pregnancy, adequate nutrition (i.e., eating meat, fresh vegetables, fruit more than once a week) was a protective factor, while eating germinated potatoes was a risk factor. The geometrical mean (p5-p95) of serum folic acid in women with NTD birth defects was 9.6 nmol/L (3.6, 23.03), which was significantly lower than that in normal women (14.03 nmol/L).

CONCLUSIONWomen of childbearing age in the two counties of Shanxi Province, China, have a marked insufficient intake of some nutrients, especially folic acid, zinc, vitamins A and B12. This nutrient deficiency may be an important risk factor for the high prevalence of birth defects in these regions. Therefore, adequate dietary nutrition in early pregnancy can prevent BDs.

Adolescent ; Adult ; Aged ; Case-Control Studies ; China ; epidemiology ; Female ; Folic Acid ; blood ; Humans ; Incidence ; Middle Aged ; Neural Tube Defects ; epidemiology ; Nutritional Status ; Surveys and Questionnaires

Intrauterine infection is an important cause of some birth defects worldwide. The most common pathogens include rubella virus, cytomegaloviurs, ureaplasma urealyticum, toxoplasma, etc. General information about these pathogens in epidemiology, consequence of birth defects, and the possible mechanisms in the progress of birth defects, and the interventions to prevent or treat these pathogens' infections are described. The infections caused by rubella virus, cytomegaloviurs, ureaplasma urealyticum, toxoplasma, etc. are common, yet they are proved to be fatal during the pregnant period, especially during the first trimester. These infections may cause sterility, abortion, stillbirth, low birth weight, and affect multiple organs that may induce loss of hearing and vision, even fetal deformity and the long-term effects. These pathogens' infections may influence the microenvironment of placenta, including levels of enzymes and cytokines, and affect chondriosome that may induce the progress of birth defect. Early diagnosis of infections during pregnancy should be strengthened. There are still many things to be settled, such as the molecular mechanisms of birth defects, the effective vaccines to certain pathogens. Birth defect researches in terms of etiology and the development of applicable and sensitive pathogen detection technology and methods are imperative.
Animals ; Congenital Abnormalities ; etiology ; Female ; Humans ; Infant, Newborn ; Placenta Diseases ; complications ; Pregnancy ; Pregnancy Complications, Infectious ; Pregnancy Outcome ; Pregnancy Trimester, First ; Rubella ; complications ; Toxoplasma ; pathogenicity ; Ureaplasma urealyticum ; pathogenicity

Objective: To study the protective effects of Lycium ruthenicum Murr. juice on alcoholic liver injury in rats and explore the regulatory mechanism of toll-like receptors 4 (TLR4)/p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in this process. Methods: Sixty male SD rats were randomly divided into control group (C), model group (M), low-dose Lycium ruthenicum Murr. juice group (LLM), medium-dose Lycium ruthenicum Murr. juice group (MLM) and high-dose Lycium ruthenicum Murr. juice group (HLM), 12 rats in each group. The group M, LLM, MLM and HLM were treated with 20 ml/kg (8 g/(kg·d)) ethanol (400 g/L) intragastrically and the gavage was divided into two sessions, group C was treated with an equal volume of distilled water at the same time point. Four hours before the first alcohol gavage session, rats in each dose group of Lycium ruthenicum Murr. juice were administered with 2.4, 4.8, 9.6 ml/(kg·d) Lycium ruthenicum Murr. juice respectively, and the other groups were given equal volume of distilled water at the corresponding time points. Four weeks later, the rats were sacrificed 24 hours after the end of the last experiment, blood and liver were collected. The liver index was calculated. The morphology of the liver was observed by HE staining. The expressions of hepatic TLR4, p38 MAPK and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) were detected by immunohistochemistry. The activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by colorimetry. The levels of hepatic tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-10 (IL-10) and interleukin-18 (IL-18) were detected by enzyme linked immunosorbent assay. Results: Compared with group C, the alcoholic liver injury model was established successfully in Group M. Compared with group M, related indicators in each dose group of Lycium ruthenicum Murr. juice were improved, the improvement of hepatic morphology in group HLM was the most significant, the liver index, the levels of serum ALT, AST and hepatic TLR4, p38 MAPK/p-p38 MAPK ratio, TNF-α, IL-1β, IL-18 were decreased (P＜ 0.05 or P＜0.01), while the level of hepatic IL-10 was increased (P＜0.01). Comparison among the dose groups of Lycium ruthenicum Murr. juice, the levels of liver index, serum AST and hepatic TLR4, p38 MAPK/p-p38 MAPK ratio, TNF-α, IL-18 in HLM were lower than those in LLM (P＜0.05 or P＜0.01); the level of hepatic IL-10 in HLM was higher than that in LLM and MLM (P＜0.05 or P＜0.01); the other indicators in each dose group had no statistical difference (P＞0.05). Conclusion: Lycium ruthenicum Murr. juice can improve the inflammatory stress by regulating TLR4/p38 MAPK signaling pathway, relieve alcoholic liver injury in rats, and the effect of high-dose group is better than the others.
Animals ; Fruit and Vegetable Juices ; Interleukin-10 ; Interleukin-18 ; Liver/metabolism* ; Liver Diseases, Alcoholic/therapy* ; Lycium/chemistry* ; Male ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; Tumor Necrosis Factor-alpha ; p38 Mitogen-Activated Protein Kinases/metabolism*

Objective:To analyze the clinical features, genetic characteristics, treatment and follow-up results of patients with hydrocephalus caused by methylmalonic acidemia combined with homocysteinuria, and to discuss the optimal strategies for assessing and treating such patients.Methods:From January 1998 to December 2020, 76 patients with hydrocephalus due to methylmalonic acidemia combined with homocysteinuria in the Department of Pediatrics in 11 hospitals including Peking University First Hospital were diagnosed by biochemical, genetic analysis and brain imaging examination. The patients were divided into operation-group and non-operation-group according to whether they underwent ventriculoperitoneal shunt. The clinical features, laboratory examinations, genotype, and follow-up data were retrospectively analyzed. Data were compared between the two groups using rank sum test, and categorical data were compared using χ 2 test. Results:Among the 76 patients (51 male, 25 female), 5 were detected by newborn screening, while 71 were diagnosed after clinical onset, 68 cases (96%) had early-onset, 3 cases (4%) had late-onset. The most common clinical manifestations of 74 cases with complete data were psychomotor retardation in 74 cases (100%), visual impairment in 74 cases (100%), epilepsy in 44 cases (59%), anemia in 31 cases (42%), hypotonia or hypertonia in 21 cases (28%), feeding difficulties in 19 cases (26%) and disturbance of consciousness in 17 cases (23%). Genetic analysis was performed in 76 cases, all of whom had MMACHC gene variations, including 30 homozygous variations of MMACHC c.609G>A. The most common variations were c.609G>A (94, 62.7%), followed by c.658_660del (18, 12.0%), c.567dupT (9, 6.0%) and c.217C>T (8, 5.3%). Therapy including cobalamin intramuscular injection, L-carnitine and betaine were initiated immediately after diagnosis. A ventriculoperitoneal shunt operation was performed in 41 cases (operation group), and 31 patients improved after metabolic intervention (non-operation group). There was no significant difference in the age of onset, the age of diagnosis, the blood total homocysteine, methionine, and urinary methylmalonic acid concentration between the two groups (all P>0.05). The symptoms of psychomotor development, epilepsy, and visual impairments improved gradually after a long-term follow-up in the operation group. Conclusions:Hydrocephalus is a severe complication of methylmalonic acidemia combined with homocysteinuria. The most common clinical manifestations are psychomotor retardation, visual impairment, and epilepsy. It usually occurs in early-onset patients. Early diagnosis and etiological treatment are very important. Hydrocephalus may improve after metabolic intervention in some patients. For patients with severe ventricular dilatation, prompt surgical intervention can improve the prognosis.

Objective:To observe the effect of isopimpinellin on primary hippocampal neuron cells γ-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT) and receptor genes expressions, in order to explore its hypnotic mechanism. Method:The primary hippocampal neurons of neonatal Sprague-Dawley rats were cultured in vitro. And subsequent experiments were conducted in the optimal state of cell growth, and the purity was identified by immunohistochemistry of neuron-specific enolase. Hippocampal neurons were randomly divided into five groups, namely blank control group, diazepam group (25 mg·L^-1), and low-dose (5 mg·L^-1), moderate-dose (10 mg·L^-1) and high-dose (20 mg·L^-1) isopimpinellin groups. Early apoptosis of hippocampus neuron cells were detected using flow cytometry technique after 24 h administration, and the changes in the levels of GABA and 5-HT were detected using enzyme-linked immunosorbent. The changes in mRNA expressions of receptor genes relating to gamma-aminobutyric acid type A receptor(GABAA) genes GABRA₁,GABRA₅,GABBR₁, gamma-aminobutyric acid type B receptor genes (GABAB) GABRB₂, 5-hydroxytryptamine 1A receptor (5-HT1_A)5-HT1_A(A),5-HT1_A(B),5-HT1_A(C) were detected by real-time quantitative PCR(Real-time PCR). Result:On the 7^th day, the hippocampal neurons grew in a good condition, and the purity was above 90%. Apoptosis rates of hippocampal neurons in the low-dose and moderate-dose groups were significantly lower than that in the blank control group (P<0.01). The level of GABA secreted by hippocampal neurons in the high-dose isopimpinellin group were significantly higher than that in the blank control group (P<0.01). The mRNA expression levels of GABRA₁,GABRA₅,5-HT1_A(A),5-HT1_A(C) in the moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (P<0.05, P<0.01). The mRNA expression levels of GABBR₁,5-HT1_A(B) in the low-dose, moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (P<0.05, P<0.01). Conclusion:The hypnotic mechanism of isopimpinellin may be related to the inhibition of hippocampal neuron apoptosis, the increase of the content of inhibitory neurotransmitter GABA, and the up-regulation of GABA and 5-HT-related receptor genes.

Objective:This study explores the key core targets of Guishenwan in the treatment of thin endometrium and related signaling pathways through the method of network pharmacology，and further uses animal experiments to verify the obtained targets and verify that Guishenwan are effective for thin endometrium. Method:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） was used to retrieve the effective chemical components，active component targets and target abbreviations of the eight Chinese medicines in Guishenwan，the GeneCards database and Online Mendelian Inheritance in Man（OMIM）database were used to retrieve thin endometrial related targets gene.Use Wayne software to take the intersection of the drug target of Guishenwan and the disease target of the thin endometrium，and import the intersection target into the STRING database and Cytoscape 3.7.2 software for visual analysis to obtain the "drug-disease" protein protein interaction（PPI） network， then input the intersection target into Enrichr database and DAVID database for gene ontology（GO） enrichment analysis and Kyoto encyclopedia of genes and genomes（KEGG） enrichment analysis. Using the obtained possible core and key targets as the theoretical basis，a thin endometrial model in rats was established. After Guishenwan and estrogen intervention for 21 days，the endometrial thickness of rats was observed by hematoxylin-eosin staining（HE） staining. Western blot and quantitative real time polymerase chain reaction（Real-time PCR） detect the protein and mRNA expression levels of the four core key targets of epidermal growth factor receptor（EGFR），matrix metalloproteinase 9（MMP9），interleukin-1beita（IL-1β） and mitogen activated protein kinase 14（MAPK14）. Result:The Venn software obtained 130 intersection targets in total， imported 130 intersection targets into the STRING database and Cytoscape database，and obtained a protein interaction network diagram including 33 nodes and 107 edges. DAVID 6.8 database for GO analysis. The function annotation analysis involving 167 biological processes（BP），22 cell components（CC），39 molecular functions（MF）. DAVID 6.8 database for KEGG enrichment analysis， and thin endometrium related A total of 34 pathways. Western blot and Real-time PCR were used to analyze the expression results of EGFR，MMP9，IL-1β，MAPK14 protein and genes in the endometrial tissues of the 6 groups of rats. Guishenwan can enhance the expression of EGFR，MMP9，IL-1β，MAPK14 protein and mRNA on the thin endometrium. Conclusion:According to the theoretical analysis of network pharmacology and the results of animal experiments，it is found that Guishenwan can effectively improve the related indicators of thin endometrium，and promote the expression of EGFR，MMP9，IL-1β，MAPK14 protein and genes. The intimal tissue proliferates and improves the symptoms of thin intima.

Objective:To investigate the factors affecting phenotypes in the patients of methylmalonic acidemia combined with homocysteinemia cblC type with MMACHC c. 609G>A homologous variant. Methods:A retrospective study on the clinical manifestations, complications, treatment, and outcome in 164patients of cblC type with MMACHC c. 609G>A homologous variant was conducted.The patients were diagnosed by biochemical and genetic analysisfrom January 1998 to December 2020. Results:Among the 164 patients, 2 cases were prenatally diagnosed and began treatment after birth. They are 3 and 12 years old with normal physical and mental development. Twenty-one cases were diagnosed by newborn screening. Among them, 15 cases had with normal development. They were treated fromthe age of two weeks at the asymptomatic period. Six cases began treatment aged 1 to 3 months after onset. Their development was delayed. One hundred and forty-one cases were clinically diagnosed. Their onset age ranges from a few minutes after birth to 6 years old. 110 cases had early-onset (78.0%). 31 cases had late-onset (22.0%). Five of them died. 24 patients lost to follow-up. Of the 141 clinically diagnosed patients, 130 (92.2%) with psychomotor retardation, 69 (48.9%) with epilepsy, 39 (27.7%) with anemia, 30 (21.3%) had visual impairment, 27 (19.1%) had hydrocephalus, 26 (18.4%) had feeding difficulties, 7 (5.0%) with liver damage, and 5 (3.5%) with metabolic syndrome. The frequency of hydrocephalus and seizures was significantly higher in the early-onset group. The urinary methylmalonic acid increased significantly in the patients with epilepsy. During the long-term follow-up, the level of plasma total homocysteine in the seizure-uncontrolled group was significantly higher than that in the seizure-controlled group, the difference had a statistical significance ( P<0.05). Conclusion:Most of the patients with MMACHC c. 609G>A homozygous variant had early-onset disease, with a high mortality and disability rate. If not treated in time, it will lead to neurological damage, resulting in epilepsy, mental retardation, hydrocephalus, and multiple organ damage. Pre-symptomatic diagnosis and treatment are crucial to prevent irreversible neurological damage. Neonatal screening and prenatal diagnosis are important to improve the outcome of the patients.