Objective By detecting the DNA methylation and gene expression of long-chain 3-hydroxyacyl CoA dehydrogenase (LCHAD) in trophoblast cells, analyze the correlation of DNA methylation and gene expression in early-onset preeclampsia (EPE), hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome and antiphospholipid syndrome (APS), to investigate the molecular basis of long-chain fatty acid oxidation changes in different preeclampsia and pathological pregnancy. Methods Primary human cytotrophoblast cells and HTR8/Svneo cells were treated with serum from patients with EPE (14 cases), HELLP (12 cases), APS (14 cases), and normal pregnant women (NP, 14 cases). The methylation level of LCHAD gene promoter region through the MassARRAY platform and mRNA expression level by real-time fluorescent quantitative PCR technique were conducted. Results (1) Cytosine-phosphate-guanine (CpG)sites in human LCHAD DNA promoter region:CpG sites were detected in the range of 558 bp before LCHAD gene transcription start site, the detected CpG sites were 11 sites including 8 single sites and 3 complex sites. The position of these sites were at-984,-960,-899,-853,-811,-796,-774,-727,-615,-595,-579 respectively. (2) The sites of-899,-853,-615 and-595 showed increased methylation level in EPE and HELLP groups. The methylation level at-899,-853 and-615 sites in EPE and HELLP groups were significantly higher than those in NP group(P<0.01). The methylation level at-853 site was higher in EPE group than that in HELLP group(P<0.05). The-595 site showed the unmethylated in EPE, HELLP and APS groups. There were significantly difference between the 3 groups and EPE group(P<0.01). (3) The gene expression of LCHAD mRNA in EPE(0.048±0.005), HELLP(0.045±0.006)and APS(0.044±0.004) groups were significantly lower than NP group(0.076 ± 0.009;P<0.01). (4) The correlation of methylation level and gene expression in all groups: the methylation level at-899,-853,-727,-615 and-579 sites were negatively correlated with gene mRNA expression in EPE group (P<0.05). The methylation level at-899,-853 and-615 sites were negatively correlated with gene mRNA expression in HELLP group(P<0.05). Conclusions The variation of LCHAD DNA methylation of trophoblast cells are found among EPE, HELLP syndrome and APS. The different correlation of LCHAD DNA methylation and gene expression are different in pathological groups. LCHAD DNA methylation of EPE and HELLP syndrome were significantly increased and negatively correlated with LCHAD gene mRNA expression. These results further revealed the molecular basis of long-chain fatty acid oxidation in different preeclampsia and pathological pregnancy.

The contents of adenosine, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, parishin and sulfur dioxide residue were compared in differently-processed Gastrodiae Rhizoma to provide the basis for a reasonable processing method of Gastrodiae Rhizoma. The analysis was performed on a Merck Purospher STAR column (4.6 mm x 250 mm, 5 μm) with a mobile phase consisting of methanol and water (containing 0.1% formic acid) under gradient elution at a flow rate of 1.0 mL x min(-1). The eluates were detected at 270 nm, and the column temperature was 35°C. The content of adenosin, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxy-benzaldehyde and parishin in processing of boiling or sulfur-fumigated were lower than that of in processing of steaming. Furthermore, the sulfur dioxide residue of sulphur-fumigated groups exceed 400 mg x kg(-1). This stable and reliable method will contribute to the quality control of different processed Gastrodiae Rhizoma.
Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Gastrodia ; chemistry ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods

Objective To investigate the effect of arsenic trioxide(As_2 O_3)nanoparticles on rabbit vascular smooth muscle cells in vitro in comparison with normal form As_2 03.Methods The rabbit vascular smooth muscle cells were cultured in vitro.Nano and normal forms of As_2O_3 with drug concentrations of 3?mol/L were added into the cells.Cell proliferation curve was drawn according to the light absorption values of MTT test.Flow cytometry was applied to observe the apoptosis.DNA was extracted and underwent electrophoresis.Results Cell proliferation treated with the 3?mol/L concentration of As_2O_3 was inhibited. Cell growth was inhibited markedly with increased treatment time,and the inhibition effect of nano drug form seemed stronger than that of normal form.MTT light absorption values of cells treated at 24,48 and 72 h showed statistically significant difference(H=10.934,15.039,15.539,P

OBJECTIVETo explore the diagnostic value of CT in midgut malrotation.

METHODSThe CT appearances of 16 patients with midgut malrotation were analyzed retrospectively.

RESULTSThe features of CT manifestation in 16 cases were as follows: (1) Horizontal part of duodenum could not reach medioventral line or could reach it but encircled right-down behind the superior mesenteric artery(SMA). (2) Ectopic ileocecal junction. (3) Jejunum located in right-middle abdomen while ileum in left abdomen. (4) A clockwise or counterclockwise rotation of the superior mesenteric vein (SMV) around the SMA. (5) Mid-gut volvulus.(6)Accompanied by other malformations.

CONCLUSIONAmbulation of duodenum, location of the small intestine and colon as well as anatomical position of mesenteric vessels should be intensively observed in order to exclude midgut malrotation.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Digestive System Abnormalities ; diagnostic imaging ; Female ; Humans ; Infant ; Infant, Newborn ; Intestine, Small ; diagnostic imaging ; Jejunum ; diagnostic imaging ; Male ; Mesentery ; diagnostic imaging ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; Torsion Abnormality ; diagnostic imaging ; Young Adult

OBJECTIVE: To evaluate the value of capsule endoscopy in children with small intestinal diseases with hematochezia as the chief complaint. METHODS: A retrospective analysis was performed on the clinical data and capsule endoscopy findings of 93 children with hematochezia who were admitted to Children's Hospital of Fudan University from May 2015 to January 2019 and underwent capsule endoscopy. According to the capsule endoscopy findings of the jejunum and the ileum, they were divided into a positive lesion group with 39 patients and a negative lesion group with 54 patients. Related clinical data and the features of lesion on capsule endoscopy were analyzed for the two groups. RESULTS: There were no significant differences in age, sex, duration of capsule endoscopy, gastric transit time, and small intestinal transit time between the positive lesion and negative lesion groups (P>0.05). The positive lesion group had a significantly lower level of hemoglobin than the negative lesion group (P<0.05). Hemoglobin level was negatively correlated with the rate of positive lesions on capsule endoscopy (r=-0.342, P=0.001). Among the 39 patients with positive lesions on capsule endoscopy, the detection of Meckel's diverticulum was the highest (41%), followed by inflammatory bowel disease (21%). CONCLUSIONS Capsule endoscopy has a certain value in detecting small intestinal diseases, especially diseases in the jejunum and the ileum, in children with lower gastrointestinal hemorrhage.
Capsule Endoscopy ; Child ; Gastrointestinal Hemorrhage ; Humans ; Intestinal Diseases ; Jejunum ; Meckel Diverticulum ; Retrospective Studies

OBJECTIVEA reliable method for genotyping blood group antigens Dib, k, Jsb1910 and Jsb2019 was developed. Through screening for rare blood types, the National Rare Blood Bank of China may be enriched.

METHODSThe controls for allele detection of blood groups Dib, k, Jsb1910 and Jsb2019 were prepared via polymerase chain reaction (PCR)-mediated gene site-directed mutagenesis (SDM) technique. Sequence-specific primers were designed according to known single nucleotide polymorphism (SNP) sites of alleles of blood groups antigens Dib, k, Jsb1910 and Jsb2019, a multiplex PCR system was developed by optimizing PCR reaction system. And 4190 random healthy donors samples were screened for the blood group antigens.

RESULTSUsing SDM technique, controls for alleles in blood group Dib, k, Jsb1910 and Jsb2019 were successfully generated. And a multiplex PCR system for genotyping above blood groups was developed. After verification, the system has performed with good stability and reproducibility. Two Di (b-) samples have been discovered from 4190 samples, no k- and Js(b-) sample was found.

CONCLUSIONMultiplex PCR features rapid detection, high throughput and low cost, and can be used for screening for donors of rare blood types. Information of donors may be registered in a database, which in turn can help those with rare blood types or require long-term blood transfusion to obtain matched blood, thereby reduce the adverse reactions of blood transfusion.

Blood Group Antigens ; genetics ; Erythrocytes ; immunology ; Genotyping Techniques ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Mutagenesis, Site-Directed ; Polymorphism, Single Nucleotide

Objective To investigate the effect of emodin on spinal cord edema induced by acute spinal cord injury (SCI) and its mechanism. Methods A total of 180 healthy female Sprague-Dawley rats were randomly divided into sham group(group A),model group(group B),and low-dose group(group C),middle-dose group(group D)and high-dose group(group E)of emodin,with 36 cases in each group.The SCI model was established with the modified Allen's method.Function-al recovery was evaluated with Basso-Beattie-Bresnahan(BBB)score and inclined plate test three days,seven days,14 days and 28 days after modeling.Three days after modeling,the pathological changes of the spinal cord were observed by HE staining; the water content of spinal cord was detected by dry-wet weight method, the blood-spinal cord barrier(BSCB)permeability was detected by Evans blue(EB)staining,and the expression of aquaporin-4 (AQP-4) and matrix metalloproteinase-2 (MMP-2) mRNA and protein were detected by RT-PCR and Western blotting respectively.Results The BBB score and inclined plate scores were better in groups C,D and E than in group B(P<0.05)seven days, 14 days and 28 days after modeling, especially in group E (P<0.05). Three days after modeling, HE staining showed that there was a large hemorrhage in the section of group B,the nerve cells were swollen and damaged, and a large number of inflammatory cells were infiltrated,the tissue gap was widened and the edema was severe. The above pathological changes were better in groups C,D and E than in group B,especially in group E.The spi-nal cord water content was higher in group B than in group A(P<0.05),and was lower in groups D and E than in groups B and C(P<0.05).EB content was higher in group B than in group A(P<0.05),and was lower in groups C, D and E than in group B (P<0.05). The expression of AQP-4, MMP-2 mRNA and protein were lower in groups C,D and E than in group B(P<0.05),especially in group E(P<0.05). Conclusion Emodin can alleviate spinal cord edema,and improve hind limb movement function after SCI,which could be related with the down-regulation of AQP-4 and MMP-2 expression,and the reduction of the permeability of BSCB.

Objective To investigate the effect of cold-inducible RNA binding protein(CIRP)on the viability of cryopreserved sciat-ic nerve and nerve regeneration after allograft. Methods Sciatic nerve segments of 15 mm from male Sprague-Dawley rats were placed in DMEM solution and pretreat-ed with 4 ℃, 15 ℃ and 32 ℃ for 24 hours (group A, group B and group C, respectively). Fresh nerve group (group D)without pretreatment was set up.The mRNA and protein level of CIRP was detected by RT-PCR and Western blotting,respectively.The above nerves were cryopreserved in liquid nitrogen for four weeks.The via-ble cells of the nerve segments after cryopreservation were observed by calcein-AM/propidium iodide staining. The expression of Bax and Bcl-2 was detected by Western blotting.After cryopreservation,the nerve segments were cultured in vitro for one week, the protein level of nerve growth factor (NGF) and glial cell line-derived neurotrophic factor(GDNF)was detected by Western blotting.In addition,the above four cryopreserved groups were transplanted to the Wistar rats by allografting(groups A',B',C'and D').Fresh nerve allograft group(E')and isograft group(F')were set up.At four-week post operation,the expression of CD4of the nerve and plasma level of interleukin(IL)-6 and interferon(IFN)-γ were detected by immunohistochemistry and ELISA,respectively.At 20-week postoperation, the muscle compound action potential (CMAP) and motor nerve conduction velocity (MNCV) were examined by electrophysiological examination. The number and the thickness of myelinated nerve fibers were analyzed by toluidine blue staining. The ultrastructure of the sciatic nerve was observed by electron microscopy. Results The mRNA and protein of CIRP were significantly higher in group C than in groups A and B(P<0.05).After 4 weeks of cryopreservation,compared with groups A,B and D,the viable cells increased,the expression of Bax decreased and the expression of Bcl-2 increased in group C (P<0.05). The expression of NGF and GDNF in-creased in group C than in groups A and B(P<0.05).At four-week postoperation,the expression of CD4and plas-ma concentration of IL-6 and IFN-γ significantly decreased in group C'than in group E'(P<0.05),however,no significant difference was found in group C'compared with groups D'and F'(P>0.05).At 20-week postopera-tion, CMAP, MNCV, the number of axon, and thickness of myelin sheath were significantly better in group C' than in groups A',B',D'and E'(P<0.05).Compared with groups A',B',D'and E',the myelinated nerve fibers were more, the fiber thickness was more uniform, the fiber distribution was wider, and the myelin sheath was thicker in groups C'and F'. Conclusion CIRP was induced at 32℃in the sciatic nerve,which exerted a significant protective effect on the viability of the nerves during cryopreservation,and promoted nerve regeneration and functional recovery after transplanta-tion.

OBJECTIVETo construct the recombinant fowlpox virus (rFPV) coexpressing HIV-1 gag-gp120 and hIL-6.

METHODSThe recombinant expressing plasmid pUTA-GE-IL6 was successfully constructed by inserting gag-gp120 gene and hIL-6 gene into the downstream of the combined promoter ATI-p7.5 and p7.5 tandem promoter respectively. After transfecting the plasmid into chicken embryonic fibroblast (CEF) cells preinfected with FPV 282E4 strain and selecting the recombinant virus under the pressure of BUdR. The recombinant virus was analyzed by nucleic acid probe hybridization and immunoblotting. In addition, the formation of virus-like particle and the expression of interested proteins in the recombinant virus-infected p815 cells were observed, and the immunogenicity of the recombinant virus was also analyzed.

RESULTSThere was colorable dot for the positive recombinant virus, immunoblotting analysis showed that the recombinant virus could expressed both gag-gp120 and IL-6. Virus-like particles (VLP) were formed in virus-infected cells, and the interested proteins could be expressed in mammalian cells infected by the recombinant virus. The immunity index from the immunized mice showed that the recombinant virus had good immunogenicity.

CONCLUSIONThe recombinant fowlpox virus coexpressing gag-gp120 and IL-6 was successfully constructed, which may provide basis for the preparation of live vector genetic engineering vaccine and macromolecule particle vaccine against HIV-1.

Animals ; Antibodies, Viral ; blood ; Blotting, Western ; Cells, Cultured ; Chick Embryo ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Fibroblasts ; cytology ; metabolism ; ultrastructure ; Fowlpox ; blood ; immunology ; virology ; Fowlpox virus ; genetics ; immunology ; Gene Products, gag ; genetics ; metabolism ; Genetic Vectors ; genetics ; HIV Envelope Protein gp120 ; genetics ; metabolism ; HIV-1 ; genetics ; metabolism ; Immunization ; methods ; Interleukin-6 ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Transfection ; Viral Vaccines ; genetics ; immunology ; metabolism

OBJECTIVETo study the association between the single nucleotide polymorphisms (SNPs) in FXYD6 gene and schizophrenia in a family-trios population.

METHODSSix SNPs (rs10790212, rs11544201, rs555577, rs1815774, rs4938446 and rs497768) in the FXYD6 gene were genotyped by allele-specific PCR method in 101 nuclear families, and transmission disequilibrium test (TDT) was performed.

RESULTSSNPs rs10790212 and rs11544201 showed significant association with schizophrenia (P<0.05). Furthermore, significant association of schizophrenia with the haplotype rs10790212-rs11544201 was found (P<0.05).

CONCLUSIONFXYD6 gene might play an important role in schizophrenia susceptibility and functional analysis of FXYD6 are needed.

Adolescent ; Adult ; Alleles ; Female ; Genetic Predisposition to Disease ; Haplotypes ; Humans ; Ion Channels ; genetics ; Linkage Disequilibrium ; genetics ; Male ; Polymorphism, Single Nucleotide ; genetics ; Schizophrenia ; genetics ; Young Adult