BACKGROUND: Results from clinical trials suggested that clopidogrel and ticlopidine had side effects of granulopenia, and aspirin could inhibit endothelial progenitor cell proliferation. There is no report of effects of these drugs on human bone marrow mesenchymal stem cells (hBMSCs) in stem cell transplantation. OBJECTIVE: To investigate the effects of antiplatelet drugs including clopidogrel, ticlopidine and aspirin on hBMSC proliferation and secretion. DESIGN, TIME AND SETTING: The cytology in vitro observation was performed at the Laboratory of Toxicology, Shanghai Municipal Center for Disease Control and Prevention from March to December 2006.MATERIALS: The second passage of hBMSCs was kindly donated from Shanghai Tissue Engineering Research & Development Center, Shanghai Ninth People's Hospital. Clopidogrel (Lot number J20040006) and ticlopidine (Lot number H19980186) were obtained from Hangzhou Sanofi-Synthelabo Minsheng Pharmaceutical CO., Ltd. Aspirin (Lot number 20050059) was obtained from Bayer Vital GmbH. METHODS: The standard culture medium consisted of DMEM-LG, 10% heat-inactivated FBS, 100 U/mL penicillin and 100 μg/mL streptomycin. After being cultured in vitro expanded out to passage 6, hBMSCs were treated with antiplatelet drugs of different concentrations and compared with control group. MAIN OUTCOME MEASURES: Cell proliferation was assessed by 3- (4, 5-dimethylthiazol -2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay, level of vascular endothelial growth factor (VEGF) of culture medium was detected by enzyme-linked immunoadsordent assay (ELISA), and surface antigens of hBMSCs were analyzed by the flow cytometry. RESULTS: A570 values of hBMSCs treated by clopidogrel or ticlopidine of 0.02,0.1,0.4,2,10,40 μmol/L were higher than control group (P < 0.01), while A570 values of aspirin group of 60, 600, 2 000 μmol/L were lower than control group(P < 0.05). Antiplatelet drugs had no obvious effect on cell surface antigens(CD34, CD105, CD106)expressed by hBMSCs. Treated by high dose clopidogrel or ticlopidine (40 μmol/L), VEGF level from hMSCs was lower than that of control group(P < 0.01), but VEGF level of low dose (0.02 μmol/L) ticlopidine group was higher than control group(P < 0.01), and there was no significantly difference of VEGF level among low dose clopidogrel group (0.02 μmol/L), aspirin group (5, 2 000 μmol/L), and control group(P > 0.05). CONCLUSION: Clopidogrel and ticlopidine improve proliferation of hBMSCs, but aspirin inhibits proliferation of hBMSCs. High dose of clopidogrel and ticlopidine suppress VEGF secretion of hBMSCs, while low dose of ticlopidine promote it. Antiplatelet drugs have no obvious effect on hBMSCs differentiation.

Fatty acid synthase (FAS) catalyses the reaction between acetyl-CoA and malonyl-CoA to produce fatty acids. It is one of the most important enzyme in lipid biosynthesis. FAS of the oleaginous yeast Rhodosporidium toruloides has two acyl carrier protein (ACP) domains and a distinct subunit composition compared with FASs of other species. As ACP is a protein cofactor crucial for fatty acid chain elongation, more ACPs in the FAS may facilitate the reaction. To study the biochemical and structural properties of this novel FAS from R. toruloides, plasmids were constructed and transformed into Escherichia coli BL21 (DE3). The strain ZWE06 harboring plasmids pET22b-FAS1 and pET24b-FAS2 could co-overexpress the two subunits. The recombinant FAS was purified by sequentially using ammonium sulphate precipitation, sucrose density gradient centrifugation and anion exchange chromatography. The specific activity of the recombinant FAS was 548 mU/mg. The purified complex would be used to study enzyme kinetics and protein structure of FAS, and heterogeneous expression and purification will facilitate revealing the mechanism of this novel FAS with double ACPs.
Acyl Carrier Protein ; Basidiomycota ; enzymology ; Chromatography ; Escherichia coli ; metabolism ; Fatty Acid Synthases ; biosynthesis ; genetics ; Fatty Acids ; biosynthesis ; Plasmids ; Recombinant Proteins ; biosynthesis ; genetics

Objective To test whether multiplex ligation-dependent probe amplification(MLPA)could be used for the prenatal detection of the most common aneuploidies of chromosomes 13,18,21,X,and Y.Methods 34 cases including 22 blood samples(12 with trisomy 21,1 with monosomy X,one male witll extra Y and 8 healthy persons),4 cord blood samples with Down syndrome and 8 amniotic fluid samples ( 1 with trisomy 21 and 7 normal fetuses)were recruited into this study.All samples were confirmed by karvotype analysis. DNA was extracted from blood and amniotic lysate was incubated with proteinase K.MLPA was used to determine the relative copy numbers.Results The resuhs were available within 48 h and were concordant with karyotype analysis in all but one case of amniotic fluid that was suggested to be triploid sample 69,XXY by MLPA or contaminated by maternal blood.This sample actually was found containing a number of red blood cells after centfifugation in test. In total,the concordance rate with clinical characteristics was 97.1%.The Ratio values of 13,18,21,X in normal samples were approaching 1.0 except chromosome Y having slightly higher variation in relative copy number.The difference of ratio means between the normal and trisomy 21 samples was statistically significant by one-way ANOVA(F=298.906.P=0.000).Conclusion Computer assisted MLPA with high sensitivity is a rapid,simple,automatic and reliable method for detection of common chromosomal aneuploidies.

Objective To study the effect of blood glucose level and duration of type 2 diabetes mellitus on carotid intima-media thickness (CIMT) and quantitative carotid stiffness (QCS) in the patients with type 2 diabetes mellitus. Methods Two hundred and seventy-four type 2 diabetic patients (diagnosed according to the World Health Organization criteria) were involved in this study. The subjects were divided into three groups according to the HbAlclevels and duration of diabetes mellitus: 60 with duration ≤2 years (as newly diagnosed) , 96 with duration≥5 years and HbAlc <6.5%, and 118 with duration≥5 years and HbAlc≥6.5%. CIMT and QCS of the common carotid artery were assessed by the high-resolution B-mode ultrasound unit and an echo-tracking system integrated in an uhrasound unit respectively. Results (1) In categorical analyses, the mean level of CIMT was increased from 720 μm in subjects with a duration of diabetes mellitus ≤2 years to 770 μm in subjects with a duration of diabetes mellitus > 2 years and plasma HbAlc< 6.5% (P < 0.05), and to 800 μm in subjects with a duration of diabetes mellitus > 5 years and plasma HbAlc≥ 6.5 % (P < 0.01). The corresponding values for Q CS values were 4.5, 4.5 and 5.1 (P <0.05), respectively. (2) In multiple stepwise regression analyses, CIMT wassignificantly associated with the duration of diabetes mellitus, systolic blood pressure and serum concentration of total cholesterol, whereas QCS was significantly associated with age, duration of diabetes mellitus, smoking, HbAlc, systolic blood pressure (P < 0.05 or P < 0.01). Conclusion QCS is suitable in evaluating the impact of blood glucose on carotid artery, while CIMT is an appropriate tool for long-term follow-up of diabetic patients.

Objective To investigate the mutation screening of the GJB3,GJB2,mtDNA 1555 A>G and SLC26A4 gene in Hainan Pronive population with non-syndromic hearing impairment.Methods PCR were performed with one pair of primer in the coding sequence of GJB3,GJB2,mtDNA 1555 A>G and SLC26A4 gene.Bidirectional sequencing of PCR products was subsequently applied in 429 patients with hearing loss.Results 55 patients gene mutation of 429 patients were found. The point mutation in mtDNA was found in 5 patients (1.1 7%).1 5 5 5 A>G mutation of mtDNA was found in 4 patients. 1494 C>T mutation of mtDNA was found in one patients.GJB2 gene mutation was found in 25 patients (5.83%).235 del C mutation of GJB2 gene was found in 9 patients.235 del C/GJB2 299 del AT mutation was found in two patients.235 del C mutation was found in 10 patients.176 del 16 mutation was found in 4 patients.SLC26A4 mutation was found in 22 patients (5.13%).IVS7-2 A>G mutation of SLC26A4 was found in 4 patients.2168 A>G mutation of SLC26A4 was found in one patient.IVS7-2 A>G mutation was found in 12 patients.2168 A>G mutation was found in 5 patients.538 C>T mutation of GJB3 gene was found in 3 patient.IVS7-2A>G mutation and 2168 A>G muation of SLC26A4 gene was found in 4 of 22 EVA patients.Conclusion GJB2 gene and SLC26A4 gene have revealed responsible genes for Hainan deafness patients.

With the recognition of antineutrophil cytoplasmic antibodies (ANCA)-related vasculitis and widespread vaccination against viral hepatitis B, the prevalence of polyarteritis nodosa (PAN) varied considerably. In our study, patients diagnosed as polyarteritis nodasa (PAN)based on the 1990 American College of Rheumatology(ACR) criteria were reclassified using 2007 European Medicines Agency(EMA) algorithm modified by 2012 Chapel Hill Consensus Conference(CHCC) definitions, aiming to evaluate the new classification criteria for the diagnosis of PAN. A total of 113 PAN patients admitted to Peking Union Medical College Hospital from January 2002 to December 2018 were retrospectively analyzed, who were classified into three subtypes including 9 patients with cutaneous, 80 with classic and 24 Hepatitis B virus (HBV) associated PAN. All patients were reclassified according to 2007 EMA algorithm using CHCC 2012 definitions. As a result, 7 patients were diagnosed as microscopic polyangiitis(MPA) and 19 patients with unclassified vasculitis based on the new classification criteria. The diagnostic rate of PAN was gradually declined as the classification criteria of vasculitides was update. However, there are quite a few PAN patients in China, whom rheumatologists should pay attention to the early diagnosis and treatment.

Objective To explore the protective effects of a new military functional food NB-5 on psychological stress-induced oxidative stress .Methods Rat whiskers were completely removed to induce the oxidative stress , and the concen-trations of MDA and protein carbonyl in various organs were detected to study the damage to membrane lipid and protein . Rats were fed with NB-5 for 4 weeks, and the oxidative stress was induced by whisker cutting .Biochemical marks men-tioned above were detected to explore the protective effects of NB-5.Results and Conclusion Lipid and protein peroxida-tion occurred in the brain , heart, liver, spleen and kidney after whisker removal due to emotional stress , while the catalase ( CAT) activity decreased significantly in these organs except the spleen .In this experiment model , NB-5 showed a good free radical scavenging activity to reduce the lipid and protein peroxidation among whisker -cutting rats fed with NB-5 in ad-vance.So NB-5 can serve as a good food for soldiers in case of emergency incidents .

Objective:To standardize the naming of organ at risk (OAR) and target area during cervical cancer radiotherapy based on AAPM TG-263.Methods:After self-programming of Matlab software to implement the reading and resolution of radiotherapy structure files, the naming of each substructure was automatically output, recorded and restored. After naming all substructures, the structure names were classified by keywords. According to TG-263, a standard naming conversion table of OAR and target area was developed, and the classified structure names were standardized through procedures. Finally, the standardized named radiotherapy structure files were output and imported into the treatment planning system (TPS).Results:The radiation structure of 144 patients with cervical cancer was successfully transformed and displayed correctly in TPS. Before the transformation, the naming of OAR and target area lacked of uniform norms and standards, and the naming of the same structure significantly differed. After the transformation, 43 naming methods of OAR and 74 naming methods of the target area were unified into 20 and 8 naming methods, which were more convenient for staff understanding and communication.Conclusion:The standardization of cervical cancer radiotherapy structure naming can reduce the inconsistency of naming and provide reference for the standardized naming of pelvic tumors.

Objective To elucidate gene mutation and promoter methylation changes of p16 gene in pheochromocytomas (PHEO) and paragangliomas (PGL) and to assess its relation with tumor clinical characters. Methods A total of 34 tumors (20 PHEO, 14 PG L, 15 benign, 19 malignant) were collected.Direct sequencing of p16 gene after PCR was performed to analyze genetic alterations. Hypermethylation of p16 gene promoter CpG island was analyzed by methylation specific PCR(MSP). In addition, mRNA expression was detected by RT-PCR. Results Homozygous deletion and gene mutation were not observed in 34 PHEO and PGL. Aberrant methylation of p16 gene promoter CpG island was found in 35.3% (12 of 34 tumors, 3 PHEO, 9 PGL). The p16 promoter hypermethylation in PGL was significantly higher than PHEO (P=0. 005). The higher p16 promoter hypermethylation was associated with malignant behavior, tumor number, and younger age at presentation, but no statistical significance, due to the limited number of cases. The p16 mRNA expression in malignant cases was lower than in benign tumors(0.83±0.65 vs 1.12±0.81 ,P=0.278). Conclusion p16 gene homozygous deletion and mutation were not frequent in PHEO and PGL. The promoter hypermethylation is mainly attributed to inactivation of the p16 gene.

OBJECTIVEIn terms of the problems of now available dental machinable ceramics, a new type of calcium-mica glass-ceramic, PMC-I ceramic, was developed, and its machinability was compared with that of Vita MKII quantitatively. Moreover, the relationship between the strength and the machinability of PMC-I ceramic was studied.

METHODSSamples of PMC-I ceramic were divided into four groups according to their nucleation procedures. 600-seconds drilling tests were conducted with high-speed steel tools (Phi = 2.3 mm) to measure the drilling depths of Vita MKII ceramic and PMC-I ceramic, while constant drilling speed of 600 rpm and constant axial load of 39.2 N were used. And the 3-point bending strength of the four groups of PMC-I ceramic were recorded.

RESULTSDrilling depth of Vita MKII was 0.71 mm, while the depths of the four groups of PMC-I ceramic were 0.88 mm, 1.40 mm, 0.40 mm and 0.90 mm, respectively. Group B of PMC-I ceramic showed the largest depth of 1.40 mm and was statistically different from other groups and Vita MKII. And the strength of the four groups of PMC-I ceramic were 137.7, 210.2, 118.0 and 106.0 MPa, respectively.

CONCLUSIONThe machinability of the new developed dental machinable ceramic of PMC-I could meet the need of the clinic.