Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective:To evaluate the feasibility of delineating subvolume target in radiotherapy for brain tumors using Gd-based contrast clearance difference.Methods:Twenty-six patients with malignant brain tumors were scanned with MRI. The first and second acquisitions of standard T 2-weighted images (T 2WI) and T 1-weighted images (T 1WI) were performed at 5 min and 60 min after injection of contrast agent. Delayed contrast extravasation (DCEM) MRI computed by Brainlab comprised regions of contrast agent clearance representing active tumors and regions of contrast accumulation representing non-tumor tissues. Based on T 2WI images, 14 patients with liquefaction necrosis were divided into group A, and 12 patients without liquefaction necrosis into group B, respectively. Then, gross target volume (GTV) was delineated on T 1WI images. Based on the GTV, active tumor (GTV tumor) and non-tumor regions (GTV non-tumor) were delineated on T 1WI-DCEM fusion images, while liquefaction necrosis (GTV liquefaction) and non-liquefaction (GTV non-liquefaction) were delineated on T 1-T 2WI fusion images. Finally, the differences between different subvolumes were compared by paired t-test. Results:In group A, the GTV non-liquefaction and GTV liquefaction were (13.65±18.15) cm 3 and (6.30±7.57) cm 3. The GTV tumor was (10.40±13.52) cm 3 and the GTV non-tumor was (9.55±14.57) cm 3. The GTV non-liquefaction was significantly increased by 16.3% on average compared with the GTV tumor ( P<0.05). The GTV non-tumor was significantly increased by 16.3% on average compared with the GTV liquefaction ( P<0.05). In group B, The GTV non-tumor was significantly reduced by 68.8% on average compared with the GTV tumor ( P<0.05). Conclusions:Compared with T 2WI, DCEM has advantages in identifying the liquefaction area and can clearly differentiate the subvolume of active tumors from non-liquefaction necrosis. DCEM provides evidence for guiding the delineation of subvolume in primary and metastatic brain tumors.

Objective: To explore the structural domains of the CENP-E protein that interact with Mps1 protein.Methods: Two recombinant vectors named pEGFP-CENPE2(containing 674-1085 amino acids of CENP-E protein) and pEGFP-CENPE 3(containing 1200~2134 amino acids of CENP-E protein) were transfected into human embryo kidney 293(HEK293) cells respectively.The respective energy transfer efficiency(Ef) between either EGFP-CENPE2 and Mps1,or EGFP-CENPE3 and Mps1 were detected by FRET through selective photobleaching of the acceptors.Results: Both recombinant proteins expressed in HEK293 cells transfected by the recombinant plasmids were found to co-localize with the Mps1 protein as confirmed by confocal microscopy.The Ef between EGFP-CENPE3 and Mps1 protein was [(12.63?0.48)%,n=30] and that between EGFP-CENPE3 and Mps1 protein was [(3.17?0.21)%,n=30] as revealed by the results from FRET,the result of FRET was confirmed by co-Immunoprecipitate(CO-IP) method.When compared with that between the control and Mps1,the Ef between EGFP-CENPE3 and Mps1 was significantly higher(p

OBJECTIVETo investigate the therapeutic effects of vitamine B(6) (Vit B(6)) and Xuefu Zhuyu Decoction (XFZY, for activating blood circulation to remove stasis) in patients with localized scleroderma(LSD).

METHODSThirty-three patients were treated with XFZY and Vit B(6), with 15 cases taking orally prednisone acetate and 20 healthy volunteers as the control. Their level of soluble interleukin-2 receptor (sIL-2R) and tumor necrosis factor-alpha (TNF-alpha) in the patients with LSD before and after treatment were observed.

RESULTSThe level of sIL-2R and TNF-alpha in the serum from the patients with LSD were higher than those of healthy volunteers (P < 0.01). After treatment with Vit B(6) and XFZY, the level of sIL-2R and TNF-alpha from the patients with LSD decreased significantly (P < 0.01), but there were no difference between the group taking Vit B(6) plus XFZY and the group given prednisone.

CONCLUSIONThe activating blood circulation to remove stasis approach in treating LSD with integrative Chinese and Western drugs got better results, and metabolic disorder of tryptophan might be correlated with the etiology of LSD.

Adolescent ; Adult ; Child ; Controlled Clinical Trials as Topic ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Receptors, Interleukin-2 ; blood ; Scleroderma, Localized ; blood ; drug therapy ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; metabolism ; Vitamin B 6 ; therapeutic use

A series of tacrine-methoxybenzene hybrids (5a-5i) were designed, synthesized and evaluated as inhibitors of cholinesterases (ChEs). All the compounds had better ChEs inhibitory activities than tacrine with IC50 values at the nanomolar range. Compound 5h exhibited the strongest inhibition on acetylcholinesterase (AChE) with an IC50 value of 6.74 nmol x L(-1) and compound 5f showed the most potent inhibition on butyrylcholinesterase with IC50 value of 3.83 nmol x L(-1). Kinetic and molecular modeling studies showed that these hybrids targeted both the catalytic active site and the peripheral anionic site of AChE.
Acetylcholinesterase ; metabolism ; Anisoles ; chemical synthesis ; chemistry ; pharmacology ; Binding Sites ; Butyrylcholinesterase ; metabolism ; Catalytic Domain ; Cholinesterase Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Drug Design ; Inhibitory Concentration 50 ; Tacrine ; chemical synthesis ; chemistry ; pharmacology

The aim of this study was to explore the characteristics of Toll-like receptor expression in mesenchymal stem cells derived from bone marrow of healthy donor (BM-MSCs). BM-MSCs were isolated from bone marrow of healthy donor by Ficoll method. Expressions of CD34, CD45, HLA-DR, CD44 and CD71 in BM-MSCs were detected by flow cytometry. CD71 in BM-MSCs was assayed by immunocytochemistry. The adipocyte and osteoblast induction of BM-MSCs were detected by alizarin red stain and oil red stain respectively. TLR 1 - 10 mRNA levels in BM-MSCs were evaluated by semiquantitative RT-PCR. The results showed that expressions of CD34, CD45 and HLA-DR in BM-MSC were negative while the expressions of CD44 and CD71 were positive. CD71 in BM-MSCs was positive. After induced by osteoblast and adipocyte inductor, BM-MSCs were positive for alizarin red staining and oil red staining respectively. All of TLR 1 - 10 mRNA were found in BM-MSCs with high expression levels of TLR2, TLR3, TLR4, TLR7, TLR8, TLR9 and low expression levels of TLR1, TLR5, TLR6, TLR10. In conclusion, different levels of TLR 1 - 10 mRNA were expressed in BM-MSCs of healthy donor.
Bone Marrow Cells ; metabolism ; Cell Differentiation ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; metabolism ; RNA, Messenger ; genetics ; Toll-Like Receptors ; metabolism

OBJECTIVETo investigate the changes in the proliferation and migration of human ovarian cancer cells (CAOV-3) after knocking down COX-2 gene by RNA interference.

METHODSThe recombinant plasmid of pGenesil-1-siRNA-COX-2 was constructed and transfected into CAOV-3 cells. The transcription of COX-2 gene was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), and the protein expression of COX-2 was determined by Western blotting . MTT assay was used to investigate the proliferation of the transfected CAOV-3 cells, and the cell migration was evaluated using a transwell migration assay.

RESULTSCOX-2 mRNA and protein levels were significantly reduced after pGenesil-1-siRNA-COX-2 transfection into CAOV-3 cells, which showed obvious reduction in the cell proliferation and migration.

CONCLUSIONRNA interference allows obvious COX-2 gene knocking down in human ovarian cancer cells to result in lowered cell growth rate and migration ability. COX-2 gene may become a new therapeutic target for ovarian cancer.

Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclooxygenase 2 ; genetics ; Female ; Humans ; Ovarian Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics

OBJECTIVE: To investigate the frequency of transfusion transmitted virus (TTV) infection in healthy blood donors in Hangzhou area and the mutation of TTV genomic fragment. METHODS DNA in serum samples of 203 healthy donors was extracted by phenol-chloroform method to detect TTV by semi-nested polymerase chain reaction and nucleotide sequences of partial amplification products were determined after T-A cloning. RESULTS TTV infection rate in 203 cases of blood donors in Hangzhou area was 15.3%. The homology of the amplified products of partial TTV positive samples compared with thereported nucleotide and putative amino acid sequences of TTV TA278 were 63.51% approximate, equals 67.12% and 59.46% approximate, equals 66.22% respectively. CONCLUSIONS TTV infection rate in the blood donors in Hangzhou is relatively high. The TTV infecting blood donors in the area may be a kind of novel genotype.

Objective To investigate the effect of high power microwave(HPM) irradiation on neuropeptide Y (NPY) and neural nitric oxide synthase (nNOS) expression in the cerebral cortex and hippoeampus of Wistar rats. Methods A total of 110 Wistar rats were used for this study.Three groups of 30 Wistar rats were exposed to HPM irradiation at intensities of 3,10,30 and 100 mW/cm~2,respectively.Twenty rats served as controls and were ex- posed to sham HPM irradiation.At 6 h,and at 1,3,7,14 and 28 d after irradiation,five rats from each group were sacrificed,and their cerebral cortices and hippocampi were harvested.HE staining was used to highlight any change in the structure of the cerebral cortex or hippocampus.Immunohistochemistry techniques and image analysis were used to study the changes in NPY and nNOS expression.Results 10 to 100 mW/cm~2 HPM irradiation caused pyc- nosis and deep staining of some neurons in the cerebral cortex and hippocampus.The increase in nNOS expression and decrease in NPY expression observed were significant at 3 days after irradiation.Conclusion HPM irradiation can induce injury in neurons of the cerebral cortex and hippoeampus,and abnormal NPY and nNOS expression.