BACKGROUND/OBJECTIVES: Previous research has shown maternal betaine supplementation alleviates fetal-derived hepatic steatosis. Therefore, this study examined the anti-inflammatory effect of maternal betaine intake in offspring mice and its mechanism.MATERIALS/METHODS: Female C57BL/6J mice and their offspring were randomly divided into 3 groups according to the treatment received during gestation and lactation: control diet (CD), fatty liver disease (FLD), and fatty liver disease + 1% betaine (FLD-BET). The FLD group was given a high-fat diet and streptozotocin (HFD + STZ), and the FLD-BET group was treated with HFD + STZ + 1% betaine. After weaning, the offspring mice were given a normal diet for 5 weeks and then dissected to measure the relevant indexes. RESULTS: Compared to the CD group, the offspring mice in the FLD group revealed obvious hepatic steatosis and increased serum levels of alanine aminotransferase, interleukin (IL)-6, and tumor necrosis factor (TNF)-α; maternal betaine supplementation reversed these changes. The hepatic mRNA expression levels of IL-6, IL-18, and Caspase-1 were significantly higher in the FLD group than in the CD group. Maternal betaine supplementation reduced the expression of IL-1β, IL-6, IL-18, and apoptosis-associated speck-like protein containing C-terminal caspase recruitment domain (ASC). Maternal betaine supplementation also reversed the increasing protein expressions of nitric oxide dioxygenase-like receptor family pyrin domain containing 3 (NLRP3), ASC, Caspase-1, IL-1β, and IL-18 in offspring mice exposed to HFD + STZ. Maternal betaine supplementation decreased the homocysteine (Hcy) and s-adenosine homocysteine (SAH) levels significantly in the livers. Furthermore, the hepatic Hcy concentrations showed significant inverse relationships with the mRNA expression of TNF-α, NLRP3, ASC, and IL-18. The hepatic SAH concentration was inversely associated with the IL-1β mRNA expression. CONCLUSIONS The lipotropic and anti-inflammatory effect of maternal betaine supplementation may be associated with the inhibition of NLRP3 inflammasome in the livers of the offspring mice.

Objective To compare the efficacy,safety and cost between ultrasound-guided percutaneous microwave ablation and surgical resection in patients with papillary thyroid microcarcinoma.Methods A total of 89 patients highly suspected papillary thyroid microcarcinoma by cervical ultrasonography were proved by ultrasound guided fine-needle aspiration biopsy in The Second Affiliated Hospital of Zhejiang University School of Medicine from January 2014 to February 2017.Totally 49 patients underwent microwave ablation(microwave group)while 40 patients underwent surgical resection(surgical group).T test was used to compare operation time,hospitalization expenses and the hospitalization time between the microwave group and the surgical group.Chi-squared test was applied to compare complications rate between the two groups.T test was used to compare the level of thyroid-related hormone before and after operation in the two groups.Results In the microwave group,the operation time,the hospitalization expenses and the hospitalization time were less [(55.85±5.05)min vs(25.73±9.46)min,(25435.91±5763.35)CNY vs(11307.48±3884.62)CNY and(6.78±3.03)d vs(2.92±0.78)d].These differences were statistically significant(t=-18.985,-13.084 and-7.747,P＜0.001).No severe complications occurred in the two groups.The difference of complications rate between the two groups was not statistically significant [6.1%(3/49)vs 10.0%(4/40),χ 2=-0.452,P=0.779].The level of 3'-triiodothyronine(FT3)and 4'-triiodothyronine(FT4)were higher after the operation in the microwave group,but these differences were not statistically significant.The level of FT3 and FT4 were lower [(4.5±0.50)pmol/L vs(3.90±0.72)pmol/L,(13.94±2.41)pmol/L vs(12.69±2.88)pmol/L],while the level of TSH was higher [(3.66±6.29)mIU/L vs(10.12±15.61)mIU/L] after operation in the surgical group.These differences were statistically significant(t=6.214,P＜0.001; t=2.808,P=0.008; t=-3.035,P=0.004).Conclusions Ultrasound-guided percutaneous microwave ablation is characterized by minimal invasion,good cosmetic effect,low cost and definite curative effect.It offered a new choice for the patients who refuse to undergo surgical resection.

OBJECTIVETo explore the function of a novel nonsense mutation c.1476G>A of ITGB3 gene using an in vitro expression system.

METHODSAn eukaryotic expression vector containing ITGB3 c.1476G>A cDNA was generated by site-directed mutagenesis and transformed into E.coli. Plasmid DNA was extracted and sequenced to confirm the target mutations. Wild-type and mutant recombination plasmids were transfected into Chinese hamster ovarian cancer (CHO) cells by nonliposome method, and the stable expression cells were harvested by G418 screening. The ITGB3 gene mRNA transcription and GPIIIa expression level in CHO cells were detected with real-time quantitative PCR, Western blotting and flow cytometry, respectively.

RESULTSThe eukaryotic expression vectors of wild ITGB3 cDNA and c.1476G>A mutant were successfully constructed. CHO cells with stable expression were obtained after transfection and screening. Compared with the wild-type transfected cells, the amount of CD61 antigen expression was 37% and mRNA transcription level was only 6% in the mutant-transfected cells. Full length GPIIIa protein was found only in the stably wild-type-transfected cells, but not in mutant-transfected cells by Western blotting analysis.

CONCLUSIONThe ITGB3 c.1476G>A mutation can decrease the transcription level and further affect GPIIIa synthesis and CD61 antigen expression.

Animals ; Base Sequence ; Blood Platelets ; cytology ; metabolism ; CHO Cells ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Cricetinae ; Cricetulus ; Humans ; Integrin beta3 ; genetics ; metabolism ; Molecular Sequence Data ; Plasmids ; genetics ; metabolism ; Point Mutation

OBJECTIVETo study the serological characteristics and molecular mechanism for a rare Pk phenotype of the P1Pk blood group system.

METHODSThe blood group of the proband was identified by serological techniques. The coding region and flanking intronic sequences of the β-1,3-N-acetylgalactosyltransferase gene (B3GALANT1) associated with the Pk phenotype were analyzed using polymerase chain reaction sequence-based typing.

RESULTSThe proband was identified as having a rare Pk phenotype including anti-P in her serum. The blood group of her daughter and husband showed a P2 phenotype. The nucleotide sequences of the B3GALANT1 gene of her husband and two randomly-chosen individuals were the same as the reference sequence (GenBank AB050855). Nucleotide position 433 C>T homozygous mutation in the B3GALANT1 was found in the proband, which has resulted in a stop codon at amino acid position 145, which may produce a premature protein capable of decreasing or inhibiting the activity of the β -1,3-N-acetylgalactosyltransferase. The nucleotide position 433 C/T heterozygous in the B3GALANT1 was found in her daughter.

CONCLUSIONThe Pk phenotype resulted from 433 C>T mutation in the B3GALANT1 gene has been identified.

ABO Blood-Group System ; genetics ; Adult ; Base Sequence ; Blood Grouping and Crossmatching ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; N-Acetylgalactosaminyltransferases ; genetics ; Pedigree ; Phenotype ; Point Mutation

OBJECTIVETo explore the molecular basis for an individual with rare p phenotype in the P1Pk blood group system.

METHODSErythrocyte blood group antigens and antibodies in serum were identified in the proband and five family members with a serological method. Coding regions and flanking untranslated regions of the α1,4-galactosyltransferase gene (A4GALT) encoding P1Pk antigens were amplified with polymerase chain reaction and directly sequenced. The haplotypes of A4GALT in the parents of the proband were also analyzed by cloning sequencing.

RESULTSThe proband was found with a rare p phenotype with anti-Tja antibody in his serum by serological method. The other family members all had a common P2 phenotype. The results of DNA sequencing showed that a cytosine was inserted at nucleotide position 1026 to 1029 (1026_1029insC) of both alleles of the A4GALT gene in the proband. The mutation has caused a reading frame shift and formed a mutant protein by extending 92 amino acid residues. The other family members were either heterozygous for the insertion or of the wild type at above position.

CONCLUSIONThe 1026_1029insC mutation of the A4GALT gene is probably responsible for the p phenotype identified for the first time in Chinese population. The individual with the p phenotype possesses anti-Tja antibody.

ABO Blood-Group System ; genetics ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Female ; Frameshift Mutation ; Galactosyltransferases ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutagenesis, Insertional ; Pedigree ; Phenotype ; Young Adult

OBJECTIVETo analyze specific expression of blood group genes using nucleated erythroid cells cultured from un-mobilized peripheral stem cells in vitro.

METHODSHematopoietic stem cells(HSC) bearing the CD34 antigen were isolated from peripheral blood by centrifugation and magnetic beads sorting, followed by suspension culture in vitro. Cells were collected from medium on various stages and analyzed by immunofluorescence. The RNA transcription of RH and ABO blood group genes was analyzed using culture cells on day 12.

RESULTSA total of(3.19±0.13) ×10 (4) CD34+cells were isolated from about 50 mL peripheral blood with a recovery rate of 67.3%±2.7%. The cells amount in erythroid-lineage culture system on day 9 reached a plateau of a 237.1±15.5-fold amplification of the initial cell input. The stem cell-specific CD34 antigen was dropped off, while the erythroid-specific CD235a and CD240D antigens were increased in culture period. RHD/CE and ABO genes can be amplified using RNA extracted from culture cells on day 12, and genotypes of Rh and ABO systems by DNA sequencing were consistent with their serologic phenotypes.

CONCLUSIONA method was established to analyze the gene expression of erythroid blood group derived from un-mobilized peripheral stem cells cultured in vitro. It can be used to study the expression of various erythroid-specific genes.

Antigens, CD34 ; analysis ; genetics ; Base Sequence ; Blood Group Antigens ; analysis ; genetics ; Cells, Cultured ; Erythrocytes ; cytology ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Humans ; Molecular Sequence Data

OBJECTIVETo delineate serological features and genetic basis for a rare p phenotype of P1Pk blood group system found in a Chinese individual.

METHODSSerological assaying was carried out for a proband with unexpected antibody found in his serum using specific antibodies and panel cells. Coding regions and flanking introns of α 1,4-galactosyltransferase gene (A4GALT) associated with the p phenotype were screened with polymerase chain reaction and DNA sequencing.

RESULTSA rare p phenotype of the P1Pk blood group system has been identified with red blood cells from the proband, whose serum contained anti-Tja antibody which can agglutinate and hemolyze with other common red blood cells. Other members of the proband's family were all normal with P1 or P2 phenotype. DNA sequencing has identified in the proband a homozygous 26 bp deletion at position 972 to 997 of the A4GALT gene. The deletion has caused a shift of the reading frame, resulting in a variant polypeptide chain with additional 83 amino acid residues compared with the wild-type protein. Other family members were either heterozygous for above deletion or non-deleted.

CONCLUSIONA 26 bp deletion at position 972 to 997 of the A4GALT gene has been identified in a Chinese individual with p phenotype.

ABO Blood-Group System ; genetics ; Alleles ; Base Sequence ; Galactosyltransferases ; genetics ; Genetic Association Studies ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Phenotype ; Sequence Deletion

Objective To investigate the effect of local mild hypothermia on the cerebral hemodynamic parameters,plasma Endothelin-1 (ET-1s) and calcitonin gene-related peptide (CGRPs) of the subarachnoid hemorrhage patients (SAH).Methods Sixty patients were divided randomly into local mild hypothermia group and control group (n =30 patients each group).The middle cerebral artery average blood flow rates (VMCAs) and pulse index (PIs) were detected with transcranial Doppler (TCD),plasma ET-1 s and CGRPs were tested on the D1,D7,D10,and D14,respectively.Results The VMCAs in the mild hypothermia group were lower on the D7,D10,and D14 [7 d:(95.46 ±22.48)cm/s vs (110.35 ±32.38) cm/s,t =2.07,P ＜ 0.05 ; 10 d:(85.57 ± 17.47) cm/s vs (97.64 ± 20.55) cm/s,t =2.45,P＜0.05 ;14 d:(57.16 ± 14.36)cm/s vs (70.56 ± 19.42) cm/s,t =3.04,P ＜ 0.01],PIs and plasma ET-1s were lower on the D10 and D14 compared with the control group [PIs:10 d:0.76 ±0.21 vs 0.88±0.25,t =2.01,P ＜0.05;14 d:0.72±0.18 vs 0.84 ±0.19,t =2.51,P ＜0.05] [ET-1s:10 d:(71.37 ± 16.63) pg/ml vs (81.46 ±21.38)pg/ml,t =2.04,P ＜0.05 ;14 d:(55.73 ± 15.18) pg/ml vs (68.28 ± 20.57) pg/ml,t =2.69,P ＜ 0.01].Plasma CGRPs were higher compared with the control group on the D7,D10,and D14 [7 d:(26.55 ±6.45)pg/ml vs (23.64 ±4.56)pg/ml,t =2.02,P ＜0.05;10 d:(24.15 ±7.35)pg/ml vs (20.52 ±6.18) pg/ml,t =2.07,P ＜0.05;14 d:(30.37 ±6.28)pg/ml vs (26.88 ± 4.39) pg/ml,t =2.49,P ＜ 0.05].Conclusions The mild hypothermia treatment could reduce the plasma ET-1s,improve plasma CGRPs,and improve the prognosis of the patients.

Objective To study the gene expression profiles of megakaryocytes(MKs) from human cord blood CD34+ cells in vitro expanded and to understand megakaryopoiesis at the molecular level. Methods CD34+ cells were isolated using density gradient centrifugation and magnetic activated cell sorting. The cells were cultured and stimulated with recombinant human TPO ( 100 ng/ml). After 12 days, the MKs fraction was separated using an anti-CD41 monoclonal antibody by immunomagnetic sorting. The gene expression profiles of MKs, non-MKs as well as meg-01 cells were studied by gene chip assay. THBSI, HOX A9,β-actin, lL-8,Annexin A6, FGF-8 were selected to validate the gene chip results by RT-PCR. Results A total of 116 genes between MKs and non-MKs cells were significantly different, 52 genes were up-regulated and 64 genes were down-regulated. In addition, 158 genes between MKs and meg-01 cells were significantly different, 71 genes were up-regulated and 87 genes were down-regulated. THBSI showed higher expression in MKs than in non-MKs. HOXA9 showed lower expression in MKs than in non-MKs. The expression of β-actin did not show any significant difference in MKs and non-MKs. IL-8 showed higher expression in MKs than in meg-01 cells, while ANXA6 showed lower expression in MKs than in meg-01 cells. The expression of FGF-8 did not show any significant difference between MKs and meg-01 cells. Conclusions MKs, non-MKs and meg-01 cells show different gene expression profiles. The regulatory genes include stress response genes,immune related genes, DNA synthesis and repair genes, metabolism genes, pro-onco genes and tumor suppressor genes.

Objective To observe the curative effect of simvastatin to treat carotid atheroma. Methods 106 patients with carotid atheroma were randomly divided into two groups. The treatment group( n=56) was given orally 20 micro-gram of simvastatin every night, while the matched control group( n = 50) was given 100 microgram of aspirin enteric-coated everyday for 4 months. Results After four months of follow-up, the carotid artery inner-intermediate thickness and the speckle's size,examined by the color Doppler, were obviously decreased ( P 0.05). Conclusion Simvastatin can obviously ameliorate the carotid atheroma. It can be served as one of the normal selections for treating the carotid atheroma.