Quality evaluation of cryoprecipitate coagulation factors from buffy coat-derived plasma at different time periods / 中国输血杂志

【Objective】 To investigate the quality of cryoprecipitates prepared from buffy coat-derived plasma of fresh whole blood at room temperature 20℃-24℃ isolated at different time periods, explore the optimal time for preparing cryoprecipitates, so as to improve the utilization rate of blood. 【Methods】 A total of 250 bags of whole blood collected by CPDA-1 and stored at 20℃-24℃ from October 2020 to December 2020 were randomly selected as the experimental group, and divided into groups A1 (0-8 h), A2 (8-10 h), A3 (10-12 h), A4 (12-14 h) and A5 (14-16 h) (with 50 bags in each group) according to the preparation time point. The upper-buff-coat plasma was separated and quickly frozen as the source for cryoprecipitates. Meanwhile, another 50 bags of fresh frozen plasma prepared within 0-16h after routine storage at 2℃-6℃ were randomly selected as the control group (group B), which was used as the raw plasma to make cryoprecipitate. Coagulation factor Ⅷ (Ⅷ factor) and fibrinogen (FIB) were detected, and the effect of different preparation time and different storage temperature on the content of factor Ⅷ and FIB and the pass rate were compared. 【Results】 In comparison to the control group, the Ⅷ factor content of groups A4 and A5 was significantly decreased, and the differences between groups A4, A5 and B were statistically significant (P0.05). The Factor Ⅷ content ≥60 IU/ bag prepared from buffy coat-derived plasma accounted for 96.4% (1.5 U) in the experimental group. 【Conclusion】 The buffy coat-derived plasma prepared within 12 h at 20℃-24℃ is suitable for preparing 2 U cryoprecipitate coagulation factor, while that prepared within 12-16 h is suitable for preparing 1.5 U cryoprecipitate coagulation factor.

Research and curriculum construction of International Classification of Diseases based on campus online open course / 中华医学教育探索杂志

Blended learning mode combines the advantages of traditional teaching and online learning and has become the important contents of teaching reform. This study systematically introduces the construction route and content of the blended learning course of International Classification of Diseases (ICD) based on campus online open course (COOC), including the front-end analysis in preparatory stage, curriculum design, teaching activities and teaching assessment, and preliminary shows the achievements of curriculum construction in order to improve teaching quality and training more excellent students.

Research progress on three-dimensional printed interbody fusion cage / 生物医学工程学杂志

Spinal fusion is a standard operation for treating moderate and severe intervertebral disc diseases. In recent years, the proportion of three-dimensional printing interbody fusion cage in spinal fusion surgery has gradually increased. In this paper, the research progress of molding technology and materials used in three-dimensional printing interbody fusion cage at present is summarized. Then, according to structure layout, three-dimensional printing interbody fusion cages are classified into five types: solid-porous-solid (SPS) type, solid-porous-frame (SPF) type, frame-porous-frame (FPF) type, whole porous cage (WPC) type and others. The optimization process of three-dimensional printing interbody fusion cage and the advantages and disadvantages of each type are analyzed and summarized in depth. The clinical application of various types of 3D printed interbody fusion cage was introduced and summarized later. Lastly, combined with the latest research progress and achievements, the future research direction of three-dimensional printing interbody fusion cage in molding technology, application materials and coating materials is prospected in order to provide some reference for scholars engaged in interbody fusion cage research and application.

Quality analysis of platelet concentrates prepared with optimized blood collection bags / 中国输血杂志

【Objective】 To evaluate the quality of platelet concentrates prepared by two different blood collection bags, so as to provide references for the development of high-quality platelet preparation. 【Methods】 Platelet concentrates were prepared using buffy coating from the whole blood collected by conventional and optimized single-use blood collection bags with leukoreduction filter, respectively. The volume of whole blood collected was 400 mL, and 60 bags in total. They were divided into group A (conventional collection bags, n=30), and the size of buffy coating pouch was 15 cm×12 cm; group B (optimized collection bags, n=30), and the size of buffy coating pouch was 11 cm×9 cm. 【Results】 There were significant differences between group A and group B in the amount of red blood cells contamination, platelet content, and platelet yielding rate (P<0.05), which were (2.62±0.57)×109/mL vs (1.37±0.35)×109/mL, (4.41±0.31)×1010/mL vs (6.21±0.63)×1010/mL, and (55.03±0.06)% vs (79.23±0.09)%, respectively. 【Conclusion】 The buffy coating pouch with the size of 11 cm×9 cm can produce better platelet concentrates, thus improves the safety and efficacy of clinical blood transfusion.

Analysis of risk factors of bile leakage in patients after laparoscopic common bile duct exploration and primary suture / 国际外科学杂志

Objective To explore the risk factors of bile leakage in patients with laparoscopic common bile duct exploration and primary suture for the purpose of providing clues for reducing occurrence of bile leakage.Methods The clinic data of 193 choledocholithiasis patients with laparoscopic common bile duct exploration and primary suture from October 2012 to March 2017 were retrospective analysed.All patients were divided into bile leakage group (23 patients) and non-bile leakage group (170 patients).Risk factors influencing the incidence of bile leakage were determined by analyzing 21 relevant factors with one-way analysis of variance and Logistic multivariate regression analysis.Count data and ordinal data was expressed as frequency or a percentage.Chi-square test was used to compare with groups of count data,rank-sum test was for comparison between groups of ordinal data,and Logistic regression was for multivariate analysis.Results Among all the patients,the incidence of bile leakage was 11.92％ (23/193).The results of univariate analysis revealed that cholangitis,jaundice,bile characteristics,muddy stone,number of stones,incarcerated stone,open and close peristalsis of duodenal papilla were correlated with bile leakage (x2/Z =2.537,2.122,81.834,50.709,13.242,26.958,90.207,P ＜0.05).The result of multivariate analysis revealed that bile characteristics,muddy stone,incarcerated stone,open and close peristalsis of duodenal papilla was correlated with bile leakage (Wals =14.002,8.899,6.577,5.582,P ＜0.05).Conclusion Bile characteristics,muddy stone,incarcerated stone,open and close peristalsis of duodenal papilla were main risk factors of bile leakage in patients with laparoscopic common bile duct exploration and primary suture.

Influence of transcriptional factor Sp1 on expression of bile acids transporters MRP3 and MRP4 / 局解手术学杂志

Objective To investigate the influence of transcriptional factor Sp1 on expression of bile acids transporters MRP3 and MRP4 so as to perfect the regulatory mechanism of MRP3 and MRP4 expression.Methods Transformed Sp1-overexpression and Sp1 siRNA plas-mids to HepG2 cell and obtained the stably cells line.Then the expression levels of bile acids transporters MRP3 and MRP4 were measured by RT-qPCR,and the change of protein levels were detected by Western blot.Results The stably cells line Sp1-OE-HepG2 and Sp1siRNA-HepG2 were successfully transformed.The mRNA expression and protein levels of MRP3 and MRP4 were significantly increased in Sp1-OE-HepG2 cells,among which the mRNA expression of MRP3 mRNA increased 2.8 times,the protein levels of MRP3 increased 3.0 times,and the mRNA expression and protein levels of MRP4 increased 3.2 times and 2.5 times respectively.Conversely,the mRNA expression and protein levels of MRP3 and MRP4 were decreased in Sp1 siRNA-HepG2 cells,among which the mRNA expression of MRP3 mRNA de-creased 52%,the mRNA expression of MRP4 mRNA decreased 58%,the protein levels of MRP3 decreased 57%,and the protein levels of MRP4 decreased 60%.Conclusion Transcriptional factor Sp1 could regulate the expression of bile acids transporters MRP3 and MRP4 in HepG2 cells.

Effect of glycyrrhizic acid on growth of Streptococcus mutans under acid environment in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the antibacterial activity of glycyrrhizic acid (GA) against Streptococcus mutans (S. mutans) under acidic environment in vitro.</p><p><b>METHODS</b>Working culture were prepared by inoculation of S. mutans into TPY broth followed by static incubation under anaerobic condition at 37 degrees C for 24 h. TPY broth was supplemented with three kinds density of GA (0.78, 1.57, 3.13 mg x mL(-1)), whose acidity was regulated to pH7.0, pH 5.5 and pH4.0. And the group of pH 7.0 was used as negative control. The growth of S. mutans was measured by A600 of bacteria suspension and counting colony forming unit (CFU). In addition, the survival rate of S. mutans was calculated.</p><p><b>RESULTS</b>In pH 5.5 groups, the survival rates of 0.78, 1.57 and 3.13 mg x mL(-1) GA groups were 60.96%, 60.27% and 45.58%, respectively, and in pH4.0 groups, the survival rates were 68.75%, 53.12% and 45.83%. In 0.78, 1.57 and 3.13 mg x mL(-1) GA groups, the survival rates of pH5.5 and pH4.0 were 52.25% and 39.05%, 74.39% and 43.11%, 86.38% and 55.30%, respectively.</p><p><b>CONCLUSION</b>GA could inhibit the growth of S. mutans under acidic environment, which the effect is improved as the acidity increased.</p>

Relationship between N-linked oligosaccharides in SEA module and proteolytic cleavage within C-terminal domain of rodent Muc3 / 第三军医大学学报

Objective The C-terminal domain of rodent Muc3 is proteolytically cleaved.This study is to explore the relationship between N-linked oligosaccharides in SEA module and the proteolytic cleavage within C-terminal domain of rodent Muc3.Methods Truncated rodent Muc3 C-terminal domains with complete SEA module(p20SEA) were produced by site-directed mutagenesis to insert a stop code in the required place.Proteins were detected by pulse/chase and immunoprecipitation method,or SDS/PAGE and Western blot.Inhibition of glycosylation of the expressed protein was performed by using tunicamycin.Results Muc3 C-terminal domain was posttranslationally cleaved to produce a V5-tagged 30 000 extracellular glycopeptide and a Myc-tagged 49 000 membrane-associated glycopeptide.Treatment with tunicamycin to transfected COS-1 cells led to the abundant production of 60 000 uncleaved and whole-length Muc3 C-terminal domain,the 30 000 N-terminal fragment shifted to 22 000 and 49 000 C-terminal fragment shifted to 41 000 after deglycosylation.The truncated Muc3 C-terminal domain containing complete SEA module but without the following residues led to production of 36 000 uncleaved and whole-length protein,and 30 000 cleaved product shifted to 22 000 after deglycosylation.Conclusion Proteolytic cleavage in both complete rodent C-terminal domain and complete SEA module without the following residues were partially inhibited by tunicamycin.

Influence of proteolytic cleavage blockage and N-linked oligosaccharide inhibition on the membrane targeting of rodent Muc3 C-terminal domain / 解放军医学杂志

Objective To explore the correlation between membrane targeting of rodent Muc3 C-terminal domain and proteolytic cleavage blockage within its SEA module and N-linked oligosaccharides inhibition.Methods COS-1 cells were transfected with three different expression vectors containing rodent Muc3 C-terminal domain,namely p20,p20t and p20s/a by lipofectAMINE reagent.Inhibition of N-glycosylation of the expressed protein was performed by using tunicamycin.The transfected COS-1 cells(fixed or unfixed) were detected by immunolocalization experiments(anti-V5 and anti-Myc antibody) for the protein expression.Results In fixed COS-1 cells,the expressed product of p20 transfectant detected using both anti-Myc and anti-V5 antibodies was found to localize in perinuclear position and on the plasma membrane.While in the unfixed cells,immunostaining was only confined on cell surface using anti-V5 antibody.The expressed product of p20t transfectant was detected by anti-V5 antibody to localize only in perinuclear region,as observed in a few fixed cells.The distribution of p20s/a fluorescence resembled that of p20 transfectant.Plasma membrane targeting of the non-glycosylated products due to tunicamycin treatment still occurred in transfected COS-1 cells and resembled the glycosylated products.Conclusions The blockage of proteolytic cleavage within C-terminal domain of rodent Muc3 and its inhibition of N-linked oligosaccharides in SEA module cannot affect its membrane targeting.The only apparent requirement for membrane targeting is the transmembrane and/or cytoplasmic tail segments which exist in the C-terminal domains of rMuc3.