Objective To prepare in-house calibrator of apolipoprotein A1 and apolipoprotein B with the values are traceable to the WHO-International reference materials SP1-01 and SP3-07.Methods We participated in the Northwest lipid research laboratory (NWLRL) protocol for target value transfer of WHO-IFCC apo A1 and apoB reference materials, and evaluated of comparability of the measurement.The analyses were performed on a Olympus AU 400, apo A1 and apoB were determined by immunoturbidimetric endpoint method.Results Through 3-step study, the precision and accuracy of apo A1 and apoB determination of our company met the NWLRL criteria.Correlation coefficients between obtained and assigned values of NWLRL on 40 individual serum samples were 0.983 for apo A1, 0.987 for apoB, the average absolute bias were 2.7 % for apo A1, 3.0 % for apoB, respectively.Conclusion NWLRL issued the certificate to Zhongsheng Beikong Bio-Technology and Science Inc.for the in-house apo A1 and apoB calibrator values are traceable to the WHO- International reference materials.

Objective To construct human apolipoprotein O (apolipoprotein O, ApoO) expression vector and obtain recombinant fusion protein thioredoxin (Trx)-ApoO by pET prokaryotic expression system. Methods The ApoO gene fragment from the human liver cDNA library was amplified by PCR. The resulting product was cloned into pET-32a(+) vector and sequenced. The confirmed cDNA was cloned into plasmid E.coli DH10B and then transformed into E.coli BL 21 (DE3) where it was induced to express protein by isopropyl β-D-1-thiogalactopyranoside (IPTG).The fusion protein was purified by Ni-NTA resin. Results The ApoO gene was cloned by PCR and a 519 bp DNA fragment was shown on the agarose electrophoresis. The cloned gene was sequenced and demonstrated to have the same sequence as that of human ApoO gene in GenBank which justified a successful construction of recombinant plasmid. ApoO cDNA gene fragment was induced by IPTG, and a 34 kD recombinant fusion protein Trx-ApoO was tested on sodium dodecyl sulfate polyacrylamide (SDS-PAGE). Conclusion Human ApoO gene is successfully cloned and its recombinant fusion protein Trx-ApoO is expressed.

Objective To examine the application value of reticulocyte hemoglobin content (CHr)for diagnosing iron deficiency in premenopausal women.Methods The levels of CHr,hemoglobin (Hb), mean cellular volume(MCV),red cell distribution width (RDW) were measured on the ADVIA 120 (Bayer Diagnostics) automated hematology analyzer.Transferrin saturation (TS) and ferritin (SF) were measured on chemistry analyzer.Results CHr in iron deficiency without anemia group were significantly lower than that in the healthy control group (P＜0.01)and significantly higher than that in iron deficiency anemia group(P＜0.01).CHr in anemia of chronic disease group were significantly higher than that in iron deficiency anemia group(P＜0.01).Receiver operator characteristic curve (ROC) analysis in diagnosis of iron dificiency without anemia demonstrated that the area under the curve for CHr,SF,RDW,MCV,Hb were 0.872,0.798,0.721,0.713,0.677,respectively (P＜0.01).So CHr has a better overall sensitivity than SF,Hb,MCV and RDW in the diagnosis of iron deficiency without anemia.ROC also showed that the area under the curve for Hb,RDW,CHr,SF and MCV was 1.000,0.969,0.958,0.953 and 0.926,respectively (P＜0.01) in iron deficiency anemia.Conclusion CHr is the early and sensitive predictor of iron deficiency in premenopausal women,especially for the diagnosis of iron deficiency without anemia.

Objective To study the effect of interleukin 2(IL-2)of clinical dose range,on the proliferation of human peripheral blood T cells,with special emphasis on the number and functional phenotype of γδT cells.Methods Human peripheral blood mononuelear cells(PBMCs)were isolated by density gradient centrifugation and cultured for 2 weeks at different IL-2 concentrations.Ratio and phenotype of different T cell subpopulations before and after in vitro expansion were explored by immunofluorescence staining.Cell number was estimated by trypan blue staining and cell counting.Results Within the four functional phenotypes of Vδ1 as well as Vδ2 γδT cells,CD27~+cells(including CD27~+CD45RA~+and CD27~+CD45RA~-subsets)expressed lymphoid tissue homing receptor CCR7,whereas CD27~-cells(including CD27~-CD45RA~+and CD27~-CD45RA~-subsets)had the peripheral tissue homing potential.All the studied γδT functional subsets showed the expression of activity related receptors,and the ability of a rapid production of various amount of cytotoxicity related effectors following mitogen stimulation.Although IL-2 at high concentration suppressed the proliferation of CD4 T cells,it may promote the proliferation of γδT cells.The proliferated γδT cells were mainly CD27~-CD45RA~-effector cells.Conclusion IL-2 of the clinical dose range may promote proliferation of human peripheral blood γδT cells,which might have important biological significance in IL-2 based anti-tumor therapy.

Objective To study the prevalence of metabolic syndrome (MS) in patients with type 2 diabetes according to three different diagnostic criteria and the effects of accumulation of MS components on blood lipid profile and to explore possible causes of their increased morbidity of cardiovascular diseases. Methods Components of MS and blood lipid profile were observed cross-sectionally in 157 patients with type 2 diabetes and analyzed using recommended diagnostic criteria by the World Health Organization (WHO, 1999), the National Cholesterol Education Program-Adult Treatment PanelⅢ( NCEP-ATPⅢ, 2001) and the Chinese Diabetes Society under Chinese Medical Association (CDS, 2004). Results Prevalence of MS was 73. 9% , 49. 7% and 52. 9% , and that of dyslipidemia was 54. 8% , 61. 2% and 54. 8% , with the criteria by WHO, NCEP-ATPⅢand CDS, respectively. Dyslipidemia was included in the most common combinations of MS. Blood level of triglycerides increased significantly, that of high density lipoprotein cholesterol ( HDL-C) decreased significantly, and size of low density lipoprotein cholesterol (LDL-C) particles decreased with the increase of the number of MS components, with a more significant coefficient of correlation by the criteria of CDS than that by the other two criteria. Conclusions In patients with type 2 diabetes, prevalence of MS could be more than 50% , most of them complicated with dyslipidemia, one of the most common components of MS, including elevated triglycerides, decreased HDL-C and decreased size of LDL-C particles, which could easily cause cardiovascular disease. The criteria by CDS could reflect the change in blood lipid profile in Chinese population more objectively.

Objective Aortic dissection is accute in occurrence along with fast change , which is inclined to to induce psy-chotic symptoms and affect the lives of patients .The study was to investigate the effects of healthcare cooperative intervention on pa-tients with psychiatric symptoms secondary to aortic dissection . Methods We established medical cooperation organization structure to clarify the division of responsibility along with a quick assessment form , a process chart and a green channel for emergencey treat-ment.The above method was applied to intervene 32 patients with psychiatric symptoms secondary to aortic dissection and observe the diagnosis time , the time from diagnosis to surgery , the remission time of psychiatric symptoms , the incidence of adverse events and the length of stay. Results 32 patients were diagnosed within 30min followed by emergency surgery within 30min after diagnosis, which was 1-2 hours shorter than the previous diagnosis and surgery preparation time .5-21 d after surgery , psychiatric symptoms have been effectively controlled.Extubation, falling out of bed, self-injury or injury and other adverse events did not occur .No patient died, and the average length of stay was 13.5d, which was 1.75d shorter than the length of stay of previous similar patients . Conclusion The healthcare cooperation method can quickly identify patients with psychiatric symptoms secondary to aortic dissection , prevent adverse events and shorten the length of stay .

Objective:To investigate the Repetitive transcranial magnetic stimulation combined with biofeedback non motor symptoms on clinical effects and serum levels of IL-6,CRP and TNF-α of Parkinson's disease.Methods:84 cases of Parkinson's disease in our hospital were randomly divided into experimental group and control group,with 42 cases in each group.The control group were treated with EMG and EEG biofeedback,while the experimental group were treated with repetitive transcranial magnetic stimulation on the basis of the control group.Then the serum levels of C reactive protein (CRP) and interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α),the non motor symptoms questionnaire (NMSQuest),Parkinson's disease Sleep Scale (PDSS) and Hamilton Depression Scale (HAMD) score in the two groups were observed and compared before and after the treatment.Results:Compared with before treatment,the serum levels of CRP,IL-6 and TNF-α,the NMSQuest and HAMD in the two groups decreased after the treatment,while the PDSS score increased,and the differences were statistically significant (P＜0.05);Compared with the control group,the serum levels of CRP,IL-6 and TNF-α,NMSQuest and HAMD in the experimental group were lower,while the PDSS score was higher,and the differences were statistically significant (P＜0.05).Conclusion:Repetitive transcranial magnetic stimulation combined with biofeedback can reduce the serum levels of IL-6,CRP and TNF-α in patients with Parkinson's disease,which has better clinical effect on the non motor symptoms.

Objective To construct and analyze the model of the sleeve gastrectomy with modified jejunoileal bypass (SG/MJIB) on non-obese Goto - kakizaki (GK) rats. Methods GK rats were randomly divided into SG/MJIB, sham-SG/MJIB, pair-fed (PF) and controls group. Before and after surgery, the changes of weight, food intake, fasting glucose, oral glucose tolerance test (OGTT), insulin tolerance test (ITT), fasting plasma Insulin and the histological changes in islet were dynamically observed for 16 weeks. Results From the 4th week postoperative, the weight changes of SG/MJIB significantly decreased compared with sham-SG/MJIB and PF group (P＜0.01). Fasting glucose concentration of SG/MJIB animals was lower than sham-SG/MJIB, PF and controls group (P＜0.05). The OGTT of SG/MJIB rats was obviously improved compared with the sham-SG/MJIB, PF and controls group （P＜0.01). Two weeks after operation, glucose tolerance was better improved in SG/MJIB than preoperative, the area under the curve (AUC) of blood glucose concentration decreased by about 38.9% (P＜0.01). Postoperative insulin levels in SG/MJIB group were obviously lower than sham-SG/MJIB group throughout the experiment(P＜0.05). In the 16th week, the morph of pancreatic islet of SG/MJIB was obviously improved. In the SG/MJIB group, the number of positive β-cell and mature acinus was significantly increased, while sham surgery groups had no obvious changes as mentioned above. Conclusions SG/MJIB was directly linked to the reduction in glucose levels in GK rats, independently weight loss and caloric intake, and it can be served asa stable long-lasting hypoglycemic surgery model to research the mechanism for the treatment of type 2 diabetes.

OBJECTIVE: To establish a multiplex genotyping system of mtDNA SNP. METHODS: A multiplex analysis system of 16-plex mtDNA SNP loci was established with allele specific PCR and capillary electrophoresis genotyping technology. Fifty samples from unrelated Chinese Han individuals were typed with the multiplex system. The multiplex assay was validated by comparing with the direct sequencing method. RESULTS: The genotypes of all 50 samples were correctly determined by the multiplex system. The optimal genotypic graphs were obtained with an input DNA of 0.5-10 pg, and the typing results were completely consistent with those by direct sequencing method. CONCLUSION The established multiplex system by allele specific PCR has high sensitivity, operational simplicity and high accuracy. It provides an effective and high output method for mtDNA SNP typing.
Alleles ; DNA ; DNA, Mitochondrial ; Genotype ; Genotyping Techniques ; Humans ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVETo evaluate the cytotoxicity of gelatin/alginate hydrogel scaffolds prepared by 3D bioprinting in human dental pulp cells (HDPCs) and compare the cell adhesion and proliferation of the cells seeded in the biomaterial using two different methods.

METHODSHDPCs isolated by tissue block culture and enzyme digestion were cultured and passaged. Gelatin/alginate hydrogel scaffolds were printed using a bioplotter, and the cytotoxicity of the aqueous extracts of the scaffold material was tested in the third passage of HDPCs using cell counting kit-8. Scanning electron microscopy and trypan blue were used to assess the adhesion and proliferation of the cells seeded in the scaffold material at a low or high concentration.

RESULTSThe aqueous extract of the scaffolds at different concentrations showed no obvious cytotoxicity and promoted the proliferation of HDPCs. The scaffolds had a good biocompatibility and HDPCs seeded in the scaffold showed good cell growth. Cell seeding at a high concentration in the scaffold better promoted the adhesion of HDPCs and resulted in a greater cell number on the scaffold surface compared with low-concentration cell seeding after a 5-day culture (P<0.05).

CONCLUSIONGelatin