Objective:To evaluate the clinical performance of low-count platelet(PLT) samples for Mindray BC-6800 automatic hematology analyzer and compare the detection results between two sampling mode(automatic injection, manual injection).Methods: To evaluate the repeatability and contamination rate of low-count platelet samples depend on ICHC, CLSI and other standards; to choose 108 blood samples with thrombocytopenia and detect them in two kinds of detection mode (automatic and manual operation), and then evaluate the consistency of low-count PLT samples between the two modes.Results: The results of two detection modes for low-count platelet samples showed there were well repeatability between them. The CV% of those samples which PLT were upon 20×109/L can be assurance within 5% in two modes. The value and fluctuation of SD and range of those samples which PLT were under 20×109/L were smaller; the SD of two modes were 0.5 and 0.8, respectively; and the Range were 1 and 2, respectively. Both of the two modes were less than 1% in carryover rates, high PLT accounts has no influence on low accounts. The correlation coefficient was up to 0.9910 between the two modes by analyzing the 108 blood samples. The results demonstrate there were low deviation and well consistency between the two modes by analyzing medical decision level concentration point for them.Conclusion: Mindray BC-6800 has good clinical performance to detect low-count PLT samples and there are well consistency between automatic and manual operation. Clinical Laboratory chooses which suitable mode to detect samples may depend on self-situation.

Objective To create a MIN6 cell line overexpressing murine Reg3? cDNA and to investigate its cell viability character under low glucose concentrations.Methods The full-length murine Reg3? cDNA was inserted into the plasmid pcDNA3.1(-).Then MIN6 cells were transfected with the Reg3?-pcDNA3.1 and pcDNA3.1 vector alone to establish Reg3?-overexpression and empty vector MIN6 cell line.Reg3? protein in the low glucose concentration cell medium was detected by Western blot.Cell viability was evaluated by an MTT reduction conversion assay.Results Three Reg3?-overexpression MIN6 cells were confirmed by real-time PCR and Western blot.Reg3? expression was barely detectable in the cells transfected with the empty vector alone.In contrast,the levels of Reg3? mRNA and protein in three pcDNA-Reg3?-transfected clones were increased by an average 10-and 6-fold,respectively.Western blot also revealed Reg3? protein release into the culture medium.In MTT cell proliferation assay,compared to vector-transfection alone,three clones of Reg3?-overexpression MIN6 cells exhibited 2-fold increases in cell number over 7 days when cultured in the presence of 10 mL/L fetal bovine serum and 5.5 mmol/L glucose.Conclusion The Reg3?-overexpression MIN6 cells have been established.Reg3? protein was detectable in culture medium,supporting an endocrine action,and Reg3? overexpression promoted islet cell growth.

Objective To investigate the effect of saturated fatty acid palmitate acids(PA) action on viability,proliferation and apoptosis of murine insulinoma MIN6 cells,and the possible intracellular pathways activated by PA.Methods Cell viability was evaluated with MTT reduction conversion assay.The Annexin-Ⅴ/FITC cell death detection kit was used to monitor PA-induced apoptosis.(IAP) family(XIAP and cIAP-2),cyclin D1 and CDK4 were further detected using Western blot.Results ① PA of different concentration significantly inhibited the proliferation of MIN6 cells(P

Objective To explore the feasibility of using the quantitative reference extract of ginkgo leaf total lactones instead of single component reference for the quantitative assay of Ginkgo Folium.Methods HPLC-ELSD method was performed by using a Diamonsil C18 column (250 mm×4.6 mm,5 μm) with methanol-water as the mobile phase at the gradient elution mode.Flow rate was 1.0 mL·min-1.The parameters of ELSD detector were as follows,the drifit tube temperature was 105 ℃,and the flow rate of nitrogen(N2) was 3 L·min-1.Results The linear ranges of ginkgolide A,ginkgolide B,ginkgolide C,and bilobalide were 0.735-5.879 μg (r=0.999 6),0.404-6.060 μg (r=0.999 6),0.296-4.439 μg (r=0.999 6),and 1.001-6.006 μg (r=0.999 7),respectively.The recoveries and RSD of the four components were 95.6% (4.0%),97.3% (4.5%),99.3% (5.0%),and 100.4% (2.1%),respectively.Conclusion The quantitative reference extract of ginkgo leaf total lactones can be used as the substitute for the determination of terpene lactones.

A temperature-sensitive mutant strain was isolated after transposon mutagenesis with Tn5 and named MT54.PCR was carried out with primers designed according to the sequence of transposon,the PCR products showed that the MT54 carried transposon in the genome. The mutant grew well at 30℃in minimal medium(MM)containing p-nitrophenol(PNP)as sole carbon source,while it cannot grow at 37℃in the same medium,NO_2.detection results also proved that.Comparing the degradation rate of PNP and hydroquinone of MT54 and DLL-E4 at different temperature,it was speculated that the mutant site locate in the PNP degradation related genes.

Background Corneal collagen cross-linking (CXL) shows good clinical effects for keratoconus,and de-epithelized CXL appears to be benefit to the distribution and absorption of riboflavin in cornea stroma.However,de-epithelization of CXL will increase the infective risk and corneal healing time.It is very important to understand and control the affecting factors of corneal repair after de-epithelization of CXL.Objective This study was to evaluate the characteristics of corneal epithelial repair and analyze the relevant factors affecting corneal healing time after de-epithelized CXL.Methods A series-cases observational study was performed.De-epithelized CXL was performed on 77 eyes of 68 keratoconus patients in Shandong Eye Hospital from September 2013 to September 2015 under the approval of Ethic Committee of this hospital and informed consent of each patient.The age,corneal curvature,corneal thickness,breakup time of tear film (BUT),corneal front astigmatism (Astig) and epithelial healing time of the patients were recorded after surgery.The correlations between corneal epithelium healing time and above-mentioned factors were analyzed.Results De-epithelized CXL was smoothly finished in all the eyes.The corneal epithelium healing time was 2-12 days after surgery,with the average healing time 5 (4,6) days.The mean age,thickness at corneal thinnest point,minimal cornea curvature (Kf),maximal corneal curvature (Ks),corneal average curvature (Km) and Astig was 22.00 (18.00,25.00) years,436 (412,470) μm,47.40 (44.70,50.45) D,52.10 (49.00,54.55) D,50.00 (47.15,53.15) D and-3.30 (-5.45,1.70) D,respectively.Spearman rank correlation analysis showed significant negative correlations between corneal epithelium healing time and BUT or the thickness at corneal thinnest point (BUT:rs =-0.334,P =0.003;corneal thickness:rs =-0.417,P =0.000),and thesignificant positive correlations were found between corneal epithelium healing time and Km,Kf and Ks (Km:rs =0.449,P =0.000;Kf:rs =0.300,P =0.008;Ks:rs =0.432,P =0.000).There were no considerable correlarions between corneal epithelium healing time and age or Astig (age:rs =0.023,P =0.845;Astig:rs =-0.190,P =0.098).Multiple linear regression analysis were carried out to study the dependent variable and independent factors.Because of the multiple co-linearity between variables,this paper corrects the model by using ridge regression.There is significant negative correlation between BUT,corneal thickness and corneal healing time,respectively (both at P＜0.05),corneal curvature Km and Kf is positively correlated with corneal healing time (both at P ＜ 0.05).Conclusions The corneal thickness,Kf,Km,as well as BUT are influencing factors of epithelial healing after CXL.

?AIM: To evaluate the effects on recovering of corneal wound and postoperative discomfort of different methods for primary pterygium.?METHODS: Forty-seven cases ( 60 eyes ) of primary pterygium were excised under microscope with limbal epithelial transplantation, with sharp dissection ( 24 cases, 30 eyes, Group A) and blunt dissection (23 cases, 30 eyes, Group B).All cases were followed up for 1d to 1mo.?RESULTS: The recovering of corneal wound was better in Group B on 1st day and 3rd day after surgery.Pain, photophobia and tears, foreign body sensation were more serious in group A on 1st day after surgery with a statistically significant difference (P=0.005,0.015,0.012). Pain, photophobia and tears, foreign body sensation were more serious in Group A on 3rd day after surgery with a statistically significant difference ( P=0.019,0.018, 0.015).There was no statistically significant difference on 1wk and 1mo after surgery (P>0.05).? CONCLUSION: Compared with sharp dissection, primary pterygium excised with blunt dissection can significantly improve recovering of corneal wound and postoperative discomfort.

The application of structured education in patients with type 2 diabetes mellitus showed significant effect. This paper introduced relevant content of structured education, the application status quo of structured education in patients with type 2 diabetes mellitus at home and abroad, aiming to provide reference for exploring health education models of patients with diabetes mellitus and reduce patients' complications, thus improving their quality of life.

Objectives:To study the clinical significance of electron beam CT with three-dimensional reconstruction in evaluating acetabular fractures.　Methods:Twenty-six cases of acetabuar fractures were examined under X-ray and 3D reconstruction. Degrees of displacement were evaluated and classified for the selection of therapy.　Results:The 26 cases of acetabuar fractures were: 1 case of anterior column fracture,3 cases of posterior column fracture,11 cases of posterior wall fracture,4 cases of transverse fracture, and 7 cases of compound fracture. All the 26 cases were accurately evaluated by EBCT 3D image, according to the image, appropriate surgical treatment were applied to 15 cases, and 11 cases were treated nonoperatively.　Conclusions:EBCT 3D reconstruction could revealed the characteristics of acetabular fractures, which aided the evaluation and selection of therapy, increased the veracity of procedures and reduced the unnecessary operations.

AIM: To construct a lentiviral vector harboring RNAi sequence targeting human annexin A2 (ANX A2) and to investigate the functions of ANX A2 in antiphospholipid antibody (APL)-induced tissue factor (TF) expression in monocytes. METHODS: Four different short hairpin RNAs (shRNA) targeting ANX A2 gene were constructed and cloned into the pGCSIL-GFP vector. After identification with PCR and sequencing, the effective interference sequence was further determined by Western blotting analysis. The recombinant lentivirus LV-RNAi-ANX A2 harvested from 293T cells was transferred into THP-1 cells and the ANX A2 mRNA and protein expression on THP-1 cells were examined. The transferred-THP-1 cells were stimulated by APL/β_2GPI compound, and the TF mRNA and TF activity was assayed. RESULTS: The RNAi sequences targeting the human ANX A2 gene were successfully inserted into the lentiviral vector and the high-performance RNA interference sequence was sieved out. The recombinant lentivirus was harvested from 293T cells with a viral titer of 3×10~(12) TU/L. THP-1 cells infected with LV-RNAi-ANX A2 showed almost lockout of ANX A2 both at mRNA and protein levels. The TF expression was also significantly decreased in the transferred-THP-1 cells induced by APL/β_2GPI compound. CONCLUSION: The lentiviral vector constructed in the present study blocks the ANX A2 expression in THP-1 cells and further inhibits the TF expression induced by APL/β_2GPI compound, which indicates that ANX A2 do play an important role in APL-induced TF expression on monocytes.