BACKGROUND: Intrauterine growth restriction (IUGR) is associated with adverse metabolic outcomes during adulthood. Histone modifications and changes in DNA methylation-affected genes are important for fetal development. This study aimed to confirm the epigenetic mechanisms in IUGR. METHODS: IUGR models were established in Sprague-Dawley rats using a maternal nutritional restriction approach during pregnancy. The abundance of insulin-like growth factor 2 (IGF2), phosphoinositide 3-kinase (PI3K), AKT serine/threonine kinase 2 (AKT2), and PPAR gamma coactivator 1 alpha (PGC-1α) was examined by real-time polymerase chain reaction (RT-PCR) and Western blotting analysis. Chromatin immunoprecipitation RT-PCR was employed to analyze histone modification in CCCTC-binding factor (CTCF)1-4 binding sites of the IGF2/H19 imprinting control region (ICR). The methylation states of CTCF1-4 binding sites were studied by pyrosequencing. RESULTS: The IUGR models were constructed successfully. IGF2 mRNA abundance in the placenta, fetal liver, and newborn liver was decreased in the IUGR group (P <0.01). Meanwhile, as compared with the control group, the expression levels of AKT2, PI3K, and PGC-1α were lower in newborn and 8-week-old livers in the IUGR group (P <0.05). In addition, knocking down IGF2 reduced the protein expression levels of AKT2-P and PGC-1α (P <0.05). In CTCF binding sites 1-4 of the IGF2/H19 ICR, AcH3 enrichment was significantly lower in CTCF1-3 in newborn and 8-week-old IUGR rats. H3K4me3 enrichment was significantly lower in the CTCF1-4 of newborn and 8-week-old IUGR groups (P <0.01). H3K9me2 enrichment was significantly higher in the IUGR group (P <0.01). The CpG dinucleotide methylation levels of CTCF1 and CTCF3, but not those of CTCF2 and CTCF4 binding sites in IUGR rat fetal, 4-week old, and 8-week-old livers decreased significantly (P <0.05). CONCLUSION The methylation status and histone modification in the IGF2/H19 ICR are related to growth and lipid metabolism via the PGC-1α/PI3K/AKT2 pathway in IUGR rats.

Objective : To explore the protective effect and mechanism of salvianolic acid salt on acute lung injury induced by one lung ventilation(OLV) in rabbits. Methods : 24 rabbits were randomly divided into three groups : control group , model group and treatment group , with 8 rabbits in each group. The W/D value of lung was measured , and the morphological changes of lung tissue were observed by HE staining and electron microscopy; The contents of arachidonic acid(AA) , leukotriene B4( LTB4) , thromboxaneA2( TXA2) and prostacyclin 2( PGI2) in rabbit lung tissue were detected by ELISA ; The expressions of cyclooxygenase⁃2(COX⁃2) ,5 ⁃lipoxygenase(5 ⁃LOX) in lung tissue were detected by RT⁃qPCR; The expressions of COX⁃2 , 5 ⁃LOX , clara cell secretory protein(CCSP) and cytoplasmic phospholipase A2(C⁃PLA2) in lung tissue were detected by Western blot. Results : The W/D ratio of model group was higher than that of control group , but decreased after treatment ( P < 0. 05) ; HE staining showed that in the model group , the capillaries were dilated and congested , there were red exudates in the alveoli , the walls of alveoli were thickened , and inflammatory cells were infiltrated , but the treatment of salvianolic acid salt was relieved ; The mitochondria swelling , crista breaking and even disappearing were observed by transmission electron microscope in the model group , and alveolar epithelial cell Ⅰ (AT⁃ Ⅰ ) ultrastructural damage in the treatment group significantly improved compared with that in the model group ; The AA , LTB4 , TXA2 and PGI2 contents in the treatment group were lower than those in the model group (P < 0. 05) . RT⁃qPCR showed that the expression of 2 and 5 ⁃LOX decreased after Salvianolic acid salt treatment (P < 0. 05) ; Western blot showed that the expression of COX⁃2 , 5 ⁃LOX and C ⁃PLA2 in treatment group was lower than that in model group , while the expression of CCSP increased (P < 0. 05 ) . Conclusion Salvianolic acid salt has protective effect on lung injury induced by one⁃lung ventilation. Its mechanism may be related to down⁃regulation of COX⁃2 , 5 ⁃LOX and C ⁃PLA2 expression , up⁃regulation of CCSP expression.

OBJECTIVE: To explore the correlation between the genotypes and metabolic markers and microstructure of bones in children with Gitelman syndrome (GS). METHODS: For 15 children with GS and 10 healthy individuals, baseline data and bone metabolic markers including parathyroid hormone, alkaline phosphatase, osteocalcin, N-terminal propeptide of type I procollagen, beta isomer of the C-terminal telopeptide of type I collagen and 25-hydroxyvitamin D, high-resolution peripheral quantitative computed tomography indicators (volumetric bone mineral density, bone microstructure indicators) were collected. Genetic testing was carried out to determine their genotypes. RESULTS: The volumetric bone mineral density, bone geometry and bone microstructure parameters of the GS group were better than those of the healthy controls (P<0.05). Variants of the SLC12A3 gene were identified in 9 of the 15 patients but none of the 10 healthy controls. CONCLUSION The phenotype of GS children is influenced by the interaction of genetic variants, though the phenotype associated with high frequency mutations showed no specificity. There is also a correlation between their genotype and the bone microstructure.
Biomarkers ; Bone and Bones ; Child ; Collagen Type I/genetics* ; Genotype ; Gitelman Syndrome ; Humans ; Osteocalcin/genetics* ; Peptide Fragments ; Solute Carrier Family 12, Member 3

Objective:To investigate whether miRNA-296-5p can inhibit enterovirus 71 (EV71) virus replication in neural cells SK-N-SH by targeting the phosphatase and tensin homology deleted on chromosome 10 (PTEN)/phosphatidylinositol 3-kinase (PI3K)/serine/threonine kinases (AKT) signaling pathway.Methods:Serum samples were collected from patients with EV71 virus-infected hand-foot-mouth disease and normal physical examination, and the expressions of serum inflammatory factors procalcitonin (PCT), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) , interleukin-6 (IL-6), IL-1β, and IL-13 were detected by enzymelinked immunosorbent assay (ELISA). Human neuroblastoma SK-N-SH cells infected by EV71 virus in logarithmic growth phase were set up as control group, miRNA-296-5p mimic group and miRNA-296-5p inhibitor group. The transfection was carried out according to the Lipofectamine 2000tm cell transfection reagent. The expression of EV71-VP1 gene mRNA and protein and PTEN/PI3K/Akt signal pathway related molecules in three groups of cells was observed by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blot.Results:ELISA test results showed that the levels of serum inflammatory factors PCT, CRP, TNF-α, IL-1β, IL-6 and IL-13 in patients with EV71 virus-infected hand, foot and mouth disease were significantly higher than those in normal physical examination ( P<0.05). The results of qRT-PCR and Western blot showed that the mRNA and protein levels of mirna-296-5p and PTEN in SK-N-SH were significantly decreased after EV71 virus infection, while the mRNA and protein levels of EV71-VP1 and molecules related to PI3K/AKT signaling pathway were significantly increased ( P<0.05). The expression of PTEN was significantly increased in the miRNA-296-5p mimic group, and the expression of EV71-VP1 and the activation of the PI3K/AKT signaling pathway were inhibited, while the effect was reversed in the miRNA-296-5p inhibitor group ( P<0.05). Conclusions:MiRNA-296-5p inhibits the replication of EV71 virus in neural cells SK-N-SH by targeting the PTEN/PI3K/AKT signaling pathway, while reducing the cellular inflammatory response, targeting miRNA-296-5p and downstream PTEN/PI3K/AKT The signal pathway is expected to provide therapeutic targets and theoretical basis for the treatment of hand-foot-mouth disease caused by clinical EV71 virus.

Objective:To explore the effect of gradual dural decompression(GDD)in preventing intraoperative encephalocele in severe traumatic brain injury (TBI).Methods:A retrospective case-control study was conducted to analyze the clinical data of 78 patients with severe TBI admitted to Affiliated Hospital of North Sichuan Medical College from January 2015 to October 2018, including 55 males and 23 females, aged 15-68 years with an average age of 38.2 years. Thirty-eight patients were treated by GDD (GDD group). During the operation, the hematoma was cleared by opening a window in the area with more hematoma, then gradually cleared in the area without opening a window. Finally, the dural "junction" was cut and the skull was closed quickly after covering the dural patch. The other 40 patients were operated by fast open dural (FOD) operation (FOD group). The incidence of intraoperative encephalocele, intracranial pressure data at the time, 24 hours, 72 hours and 7 days after operation, incidence of delayed intracranial hematoma and glasgow outcome score (GOS) at 3 months after operation were compared between the two groups.Results:All patients (except for 26 deaths) were followed up for 3-14 months, with an average of 8 months. None had intraoperative encephalocele in GDD group, compared to 8 patients in FOD group ( P<0.05). Intracranial pressure in GDD group was respective (18.4±3.6)mmHg, (20.4±4.0)mmHg, (27.7±4.6)mmHg and (28.3±4.5)mmHg at the time, 24 hours, 72 hours and 7 days after operation, showing no significant differences compared to FOD group [(19.1±3.4)mmHg, (20.9±3.9)mmHg, (27.0±3.5)mmHg, (27.6±3.4)mmHg, respectively] ( P>0.05). There was no significant difference in the incidence of delayed intracranial hematoma between the two groups [16% (6/38) in GDD group and 16% (5/32) in FOD group (except 8 cases of intraoperative encephalocele)] ( P>0.05). Three months after operation, the good rate of GOS score of GDD group was 61% (23/28) higher than that of FOD group [28% (11/40)], and the mortality rate of GDD group was 21% (8/38) lower than that of FOD group [45% (18/40)] ( P<0.05). There was no significant difference between the two groups in medium severe disability and plant survival rate ( P>0.05). Conclusions:For patients with severe TBI, compared with FOD, GDD is more effective in reducing the incidence of intraoperative encephalocele, improving the prognosis and decreasing mortality.

Objective To explore the clinical features,diagnoses,differential diagnoses and treatments of superior cerebellar artery aneurysms.Methods The clinical data of 16 patients with superior cerebellar artery aneurysms,admitted to our hospital from January 2013 to March 2018,were retrospectively collected.Their clinical manifestations,imaging features,surgical effects and related problems in the process of diagnoses and treatments were analyzed.Results Among the 16 patients,11 were caused by aneurysm rupture;8 had subarachnoid hemorrhage alone,and three had subarachnoid hemorrhage accompanied by ventricular hemorrhage;CT and CTA confirmed that 8 were superior cerebellar artery aneurysms,two were posterior cerebral artery aneurysms,and one was with unclear diagnosis.In the other 5 patients,three had eyelid ptosis and two had abducent nerve palsy;CT,CTA or MR imaging showed that two were considered as ventral brainstem occupying lesions,and three did not have clear diagnosis.Finally,all patients were diagnosed as having superior cerebellar artery aneurysms by three-dimensional DSA.Five patients were treated with interventional embolization first,and one was treated with surgical clipping because of vertebral artery stenosis and difficulty of catheter access;two patients were transferred to our department for surgical clipping due to aneurysm rupture after embolization treatment in other hospitals;and 9 patients were treated by surgical clipping directly.After treatments,one patient was in bed for a long time due to cerebellar infarction and systemic complications,and the other 15 patients recovered well;two of them underwent ventricular peritoneal shunt due to hydrocephalus.Conclusions Superior cerebellar artery aneurysm has onset of subarachnoid hemorrhage mostly,and oculomotor and abductor nerve paralysis,and space occupying manifestation around the brainstem sometimes.For patients with suspicious posterior circulation aneurysms whose diagnosis or location are unclear,three-dimensional DSA examination should be performed early to confirm the diagnosis.Treatment should be taken as soon as possible once the superior cerebellar artery aneurysm is defined.Interventional embolization may be the first choice,but it is necessary to master the methods of surgical clipping in order to treat the disease timely.

Objective:To evaluate the clinical value of surfactant protein-A (SP-A) in exudate pleural effusion (EPE).Methods:This clinical study was prospective,observational and cross-sectional.Two hundred and fifteen patients with pleural effusion were divided into the transudate pleural effusions (TPE) group and the EPE group.TPE patients served as the control group.The concentrations of pleural effusions SP-A (SP-Apl) and serum SP-A (SP-Ase) were measured by ELISA,and receiver operator characteristic (ROC) curve and multivarate Cox analysis of SP-A was analysed for its clinical value.Results:SP-Apl concentrations in the EPE group were significantly higher than that in the TPE group [(189.8±43.4) ng/mLvs (22.3±5.1) ng/mL,P＜0.01];SP-Ase concentrations in the EPE group were higher than that in the TPE group [(78.9±11.3) ng/mL vs (25.8±12.4) ng/mL,P＜0.05];SP-Apl concentrations were significantly higher than the concentrations of SP-Ase in the EPE group (P＜0.01).In EPE group,SP-Apl and SP-Ase concentration in the patients with primary lung adenocarcinomas were the highest.The cut off value of SP-Apl concentrations was more than 484.5 ng/mL,yielding a 85.4％ sensitivity and 95.2％ specificity for diagnosing primary lung adenocarcinomas,with an area under the curve (AUC) of 0.943 (95％ CI 0.852 to 0.934,P＜0.01);when SP-Ase concentration was more than 84.2 ng/mL,it yielded a 76.4％ sensitivity and 94.3％ specificity for diagnosing primary lung adenocarcinomas,with an AUC of 0.910 (95％ CI 0.921 to 0.953,P＜0.01).Conclusion:While SP-Apl concentration is more than 484.5 ng/mL and/or SP-Ase concentration is more than 84.2 ng/mL,it may be helpful for the diagnosis of primary lung adenocarcinomas with the usage ofpleural effusion.

The relationship between vitamin D deficiency and idiopathic central precocious puberty (ICPP) has been recently documented. In this study, 280 girls diagnosed with ICPP and 188 normal puberty control girls of similar ages were enrolled and retrospectively studied. The ICPP group had significantly lower serum 25-hydroxyvitamin D (25[OH]D) levels than the control group. Furthermore, a nonlinear relationship was found between serum 25[OH]D and ICPP, and a cut-off point for serum 25[OH]D was found at 31.8 ng/ml for ICPP with and without adjusting the different confounding factors. Girls with serum 25[OH]D ≥ 31.8 ng/ml had a lower odds ratio (unadjusted: OR 0.36, 95% CI 0.15 to 0.83, P < 0.05; height and weight adjusted: OR 0.44, 95% CI 0.18 to 1.08, P = 0.072; BMI adjusted: OR 0.36, 95% CI 0.16 to 0.84, P < 0.05). The ICPP subjects with 25[OH]D deficiency had a higher body mass index (BMI) than the subjects from the two other subgroups. Correlation analysis showed that vitamin D level is correlated with BMI and some metabolic parameters in the ICPP group. Our study suggested that vitamin D status may be associated with ICPP risk and may have a threshold effect on ICPP.
Body Mass Index ; Child ; China ; Female ; Humans ; Linear Models ; Logistic Models ; Multivariate Analysis ; Puberty, Precocious ; blood ; complications ; Retrospective Studies ; Vitamin D ; analogs & derivatives ; blood ; Vitamin D Deficiency ; epidemiology

Objective To conduct mutation screening of SCN1A 3′ untranslated region (UTR) on Dravet syndrome (DS) patients without mutations in the SCN1A coding region and promoter region, and functional analysis of the variant from DS patients.Methods Twenty-eight DS patients without mutations in the SCN1A coding region and promoter region were screened for SCN1A 3′ UTR mutations using PCR and direct sequencing.Functional analysis of the detected mutation was done via luciferase assay, mRNA stability analysis and RNA electrophoretic mobility shift assay (RNA-EMSA).Results A novo variant (c.*20A>G) in SCN1A 3′ UTR was found in one DS patient.The variant (c.*20A>G) reduced the luciferase gene xpression by 30% through increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing luciferase gene mRNA stability (t=8.5,P<0.01).Conclusions A functional variant was detected from one patient with DS.This variant negatively regulated the gene expression by increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing mRNA stability.

With the accelerated pace of the world population aging,it has provided great opportunities for the rapid development of traditional Chinese medicine (TCM) with natural medicine as its main component.However,the lack of legal status of TCM has long been a great influence on Chinese medicine.There is a big difference in the degree of recognition of TCM among people along the way.If people are still able to accept herbal medicine,they are very exclusive of zoic medication.Although zoic medication is an important part of TCM,zoic medication and animal rights appeal have a sharp contradiction.The question is how to solve the dilemma between the protection and utilization of zoic medication,utility and lofty.This article analyzed the understanding difference and consensus of zoic medication from the ethics perspective under both Chinese and western culture.It clarified the understanding and practice of zoic medication in TCM.It also illustrated the rational protection and sustainable using of zoic medication in TCM.