Objective:To explore the application of online and offline mixed teaching reform in clinical laboratory hematology experimental teaching.Methods:The online and offline mixed teaching mode was used in the clinical laboratory hematology experimental teaching, and the online and offline mixed teaching was realized by establishing the basic teaching framework, reforming the teaching content, enriching the teaching resources, introducing the case teaching method, opening the form laboratory, and online examination mode.Results:The overall effect of teaching reform is evaluated through the form assessment, operation assessment, student interview and teacher interview.Conclusions:The online and offline mixed teaching reform changes the traditional teaching mode and conforms to the modern education and teaching mode. The experiment content is consistent with the teaching objectives. This mixed teaching reform could lighten teachers' teaching burden, promote teachers' teaching ability, and solve the shortage of clinical laboratory hematology experimental teaching resources. It is a new teaching mode to realize the transformation of teachers from traditional teaching to diversified teaching, and students from passive learning to active learning. So it is suitable for the application in clinical laboratory hematology experimental teaching.

目的:探讨开放数码互动形态实验室教学改革模式在临床血液学检验形态实验教学中应用的效果.方法:选取2016级与2015级医学检验技术本科学生作为研究对象.2016级学生形态教学采取开放数码互动形态实验室教学改革模式,2015级采取常规形态实验教学方法.期末通过形态考核和问卷调查两种方法评价效果.结果:2016级学生形态考核成绩高于2015级(P<0.0001).问卷调查结果显示,大部分学生认为开放数码互动形态实验室激发了学习兴趣和主动思考能动性,有利于提高细胞形态辨认能力和综合素质.结论:开放数码互动形态实验室可以提高学生识别骨髓细胞能力,教学改革效果明显.

目的 为探究铁摄取相关基因iucA在尿路致病性大肠埃希菌(UPEC)致病中的作用,利用UPEC模式菌株CFT073,通过 λRed同源重组方法构建iucA基因缺失株ΔiucA,同时构建回补株C-iucA.方法 测定A600绘制CFT073、ΔiucA和C-iucA菌株在LB液体培养基和无菌尿液中的增殖曲线.比较CFT073、ΔiucA和C-iucA菌株对人膀胱癌上皮细胞株5637体外黏附和侵袭能力.构建小鼠尿路感染模型,检测CFT073、ΔiucA和C-iucA菌株在膀胱的定植能力.结果 CFT073、ΔiucA和C-iucA菌株在LB液体培养基中增殖速率相似(P=0.153).ΔiucA在无菌尿液中的增殖速率低于CFT073(P=0.001),C-iucA的增殖速率较ΔiucA有所上升(P=0.005).ΔiucA对5637细胞的黏附和侵袭能力均低于CFT073(P=0.007、0.002).C-iucA的黏附和侵袭能力较ΔiucA有所上升(P=0.046、0.037).ΔiucA在小鼠膀胱的定植能力低于CFT073和C-iucA(P=0.002、0.017).结论 iucA基因可能通过促进UPEC增殖、黏附和侵袭以及在靶器官的定植能力在UPEC致病中发挥作用.

目的:了解肾移植术后肺部感染的发生情况、病原体分布以及相关危险因素.方法:对2017年4月至2019年4月在广州医科大学附属第二医院器官移植科接受肾移植手术共627例患者的临床资料进行回顾性分析.结果:随访期内,627例患者中有105例(16.75％)发生肺部感染,其中5例(4.76％)发生在移植后1个月内,61例(58.10％)发生在2～6个月内,39例(37.14％)发生在6个月以后.105例肺部感染患者病原体检测阳性者70例,样本类型以痰液和血液为主,占89.33％,咽拭子及胸水占10.67％.其中细菌感染54例次(55.10％)、真菌感染16例次(16.33％)、CMV病毒感染12例次(12.24％)、卡氏肺孢子菌感染1例次(1.02％)以及混合感染15例次(15.31％).铜绿假单胞菌、肺炎克雷伯菌、大肠埃希菌、嗜麦芽窄食单胞菌和金黄色葡萄球菌是位于前五位的病原体.单因素分析显示,患者有糖尿病史(P=0.044)、白细胞计数<4×109L-1(P=0.015)、出现肾功能延迟恢复(P=0.008)和发生急性排斥反应(P=0.037)是肾移植术后发生肺部感染的危险因素.经多因素分析发现,白细胞计数<4×109L-1(OR=2.499,95％CI1.321～4.726,P=0.005)、出现肾功能延迟恢复(OR=2.894,95％CI 1.128～7.429,P=0.027)和发生急性排斥反应(OR=2.233,95％CI 1.121～4.446,P=0.022)是肺部感染发生的独立危险因素.结论:术后2～6个月是肾移植术后肺部感染的高发时期.收集不同来源的样本和联合多种检测方法可以提高病原体的检出率.术后白细胞计数<4×109L-1、出现肾功能延迟恢复和急性排斥反应的患者是肾移植术后肺部感染的高危人群.

The flipped classroom teaching mode based on small private online course (SPOC) combines the advantages of online and offline learning, and is widely used in medical education. This research took the course of clinical hematology for undergraduates in four-year program of clinical laboratory technology from Guangzhou Medical University as an example to use a SPOC based flipped classroom teaching mode. The teaching effect was evaluated by examination test, questionnaire survey and interviews by teachers and students. The examination results showed that 15.4% (17/113) of the students gained excellent scores (≥90 points) and 92.9% (105/113) of the students passed the examination (≥60 points). There are 79.6% (90/113) gained experimental scores over 18 points (out of 20 points). Totally 72.1% (70/97) of the students believed that this teaching mode enhanced their capacity of self-regulated learning and 90.7%(88/97) of the students were satisfied with the case-oriented module with integrated teaching mode in the experimental course. In addition, 32 students (40 in total) and 10 teachers (12 in total) believed that the teaching effectiveness of flipped classroom teaching mode based on SPOC was better than that of the traditional learning. Eight teachers believed that this teaching mode was a great challenge to their arrangement and organization abilities in class. The flipped classroom teaching mode based on SPOC demonstrates a good effectiveness, which improves students' capacity of self-regulated learning and of resolving clinical problems. However, this teaching pathway needs strong capacity of teachers with both knowledge and skills to run in clinical training of undergraduates.

Objective:To compared the effects of sodium citrate anticoagulation, amikacin supplement, magnesium sulfate supplement and predilution method on the determination of platelets in EDTA-PTCP (Ethylene Diamine Tetraacetic Acid Dependent pseudobocytopenia).Methods:After confirming the patients with EDTA-PTCP, whole blood samples were collected simultaneously in EDTA-K2 and sodium citrate anticoagulated tubes. Amikacin (6.5 mg/mL) and magnesium sulfate (25 mg/mL) were added after aspiration of EDTA-K2 treated whole blood. Samples anticoagulated with EDTA-K2 and sodium citrate, supplemented with amikacin and magnesium sulfate were tested for their platelet counts at 0 h, 1 h, 2 h, 3 h and 4 h, respectively. Platelet aggregations were also confirmed by microscopic examination at 0 h, 1 h, 2 h, 3 h and 4 h, respectively. In addition, we collected the peripheral blood of patients with EDTA-PTCP for pre dilution mode testing and manual counting.Results:Thirty-three patients with EDTA-PTCP were confirmed in The Second Affiliated Hospital of Guangzhou Medical University from July 2019 to January 2020. Platelet aggregations disappeared in 3 of 20 sodium citrate treated samples. The platelet counts of sodium citrate group were higher than that in EDTA-K2 group (P＜0.05) but were lower than manual counting (P＜0.01). Platelet aggregations disappeared in 17 of 32 patients after adding amikacin into EDTA-K2 treated whole blood. There were statistical differences between amikacin supplement group and EDTA-K2 group (P＜0.01). There was statistical differences between amikacin supplement group and manual counting (P＜0.05). The platelet counts in magnesium sulfate supplement group did not show a significant increase within 4 hours. No significant differences were found between magnesium sulfate supplement group and EDTA-K2group (P＞0.05). The platelet counts of magnesium sulfate supplement group were much lower than manual counting (P＜0.01). The platelet counts in pre-dilution group showed a remarkably increase compared to EDTA-K2 group (P＜0.01). There were no significant differences in platelet counts between pre dilution and manual counting (P＞0.05).Conclusions:Sodium citrate anticoagulation and amikacin supplement were not always effective for correcting platelet counts in vitro. Amikacin supplement had better dissociative effects on platelet aggregations than sodium citrate anticoagulation. Magnesium sulfate supplement had a poor dissociation effect on correcting platelet counts. Pre-dilution was the most accurate method. When it is inconvenient to collect peripheral blood from the patient, amikacin supplement is recommended for venous blood.

目的 比较两种化学方法制备纳米金的增敏效果,找出相对合适的制备纳米金增敏金标免疫层析试验的方法.方法 以金标免疫层析试验检测血浆绒毛膜促性腺激素(HCG)为基础,用盐酸羟胺和抗坏血酸还原氯金酸制备纳米金颗粒,判断纳米金原位所形成的纳米颗粒颜色,检测其灵敏度.结果 盐酸羟胺还原法增敏血浆HCG金标免疫层析试验,相对增敏前检测灵敏度提高约16倍,且重复性较好.抗坏血酸还原法增敏血浆HCG金标免疫层析试验,相对增敏前检测灵敏度提高约4倍.结论 盐酸羟胺还原法制备纳米金对血浆HCG金标免疫层析试验增敏倍数较高,重复性较好,试验结果可靠,可推广使用.

目的 分析肾移植术后尿路感染(UTI)的发生率、病原体谱、多重耐药菌比例、相关危险因素及其对肾功能的影响.方法 回顾性分析2016年8月至2017年8月在广州医科大学附属第二医院器官移植科接受肾移植的308例患者的临床资料,对其在随访期内尿液培养、药敏试验、肾功能检查及其他临床资料进行总结分析.结果 在(504.84±99.78)d的回访期内,308例患者中有69例(22.40％)发生UTI 173例次,其中无症状菌尿90例次(52.02％),下尿路感染53例次(30.64％),上尿路感染30例次(17.34％).140例次(80.92％)UTI发生在术后6个月内.大肠埃希菌、肠球菌属及克雷伯菌属是位于前3位的病原体,其中产超广谱β-内酰胺酶(ESBLs)大肠埃希菌阳性率为55.07％,产ESBLs肺炎克雷伯菌阳性率为33.33％,未发现耐万古霉素肠球菌.女性、糖尿病、泌尿系统梗阻和双J管留置时间延长是UTI发生的独立危险因素.UTI组和非UTI组相比,肾功能并无明显变化.结论 肾移植术后UTI临床表现以无症状菌尿居多且具有较高的上尿路感染发生率,多重耐药菌阳性率的上升需要密切关注,女性、糖尿病、泌尿系统梗阻和长时间留置双J管的患者是移植术后UTI发生的高危人群.

骨髓细胞形态教学是临床血液学检验课程的重点和难点,教学中面临形态学的特殊性 、学时减少 、教学效率低 、共享性与互动性差等诸多问题.为适应移动终端设备在教育教学中的应用趋势,本研究小组将HTML5互动网页运用在骨髓细胞形态教学中,将抽象 、微观的骨髓细胞形态直观化与具体化的同时,实现了教学图像信息资料的共享性 、互动性 、再现性与重复性,真正满足骨髓细胞形态教学的各种需求,为骨髓细胞形态教学构建出了一种全新的 、多维动态的教学方式.

目的 对无症状菌尿(ASB)及症状性尿路感染(UTI)患者尿液中分离的大肠埃希菌进行系统分群、毒力基因及耐药性检测.方法 收集该院2014年5月至2016年3月从尿路感染患者尿液中分离的大肠埃希菌共145株,其中ASB组65株,UTI组80株.利用多重聚合酶链反应(PCR)对其进行系统分群及10种毒力基因表达检测;采用纸片扩散(K-B)法对其进行耐药性检测.结果 A SB组中,A、B1、B2、D群分别占10.77％ 、38.47％ 、43.07％ 、7.69％.U T I组中,A、B1、B2、D群分别占12.50％ 、10.00％ 、62.50％ 、15.00％,两组系统分群结果差异有统计学意义(P<0.05).ASB组黏附相关基因afa和sfa的表达率高于UTI组(P<0.05).UTI组细胞毒性相关基因细胞毒性坏死因子1(cnf1)和溶血素(hlyA),铁摄取相关基因fyuA和iutA的表达率高于ASB组(P<0.05).ASB组和UTI组对18种抗菌药物的耐药率及产超广谱β-内酰胺酶菌株阳性率差异无统计学意义(P>0.05).结论 ASB和UTI不同的系统分群特征,可能与其表现出不同的致病能力有关.ASB菌株毒力较UTI低,降低了其急性致病能力,但具有更高表达率的黏附相关基因,这可能有助于细菌实现持续定植.尿路感染大肠埃希菌的高耐药率必须引起临床重视.

OBJECTIVE:To express and purify polyphosphate kinase (PPK) from Proteus mirabilis and prepare the polyclonal antibody against PPK. METHODS:The antigenicity and hydrophobicity of PPK were analyzed using software. The N-terminal conservative sequence containing 309 amino acids was selected as the target peptide, and its corresponding gene sequence with modification based on prokaryotic cells-preferred codon was synthesized and inserted into plasmid pET28b(+). The constructed recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and induced with IPTG. The expressed fusion protein was purified using Ni-affinity chromatography. The purified protein was injected along with adjuvant in rabbits to prepare the polyclonal antibodies against PPK. RESULTS AND CONCLUSION:PPK fusion protein expressed by E. coli was purified successfully using Ni-affinity chromatography. ELISA result demonstrated that the harvested rabbit anti-sera against PPK had a high titer of 1:512 000, and Western blotting showed a good specificity of the antibody, which can be used further study of the role of PPK in the pathogenesis of Proteus mirabilis infection.

本文结合微信应用医学检验专业的教学实践，从理论教学、实验教学、PBL 教学、课后辅导几个方面分析了微信移动教学的可行性、优越性及目前教学实践中存在的问题，并提出了相应的解决方案，旨在利用微信平台为信息化教学开辟新的途径，并为医学院校教学改革提供实践参考。

目的 构建尿路致病性大肠埃希菌FYUA基因敲除株,比较野生株和敲除株之间生物学特性的改变.方法 利用λRed同源重组系统构建FYUA基因敲除株△FYUA;测定A600绘制CFT073和△FYUA在LB液体培养基和无菌尿液中的增殖曲线;平板菌落计数法比较CFT073和△FYUA菌落形成能力;利用96孔板结晶紫染色法比较CFT073和△FYUA生物膜形成能力.结果 成功构建CFT073 FYUA基因敲除株△FYUA;CFT073和△FYUA在LB液体培养基中具有相似的增殖曲线,△FYUA在无菌尿液中的增殖速率明显低于CFT073(P＜0.05);CFT073和△FYUA菌落形成能力无明显区别;△FYUA的生物膜形成能力低于CFT073(P＜0.05).结论 FYUA基因对于菌株在无菌尿液中的生长繁殖以及生物膜形成可能发挥重要作用.

随着素质教育的不断推进，单一的终结性考核方式已逐渐无法适应高等教育的改革。本文主要结合作者在临床检验基础课程教学过程中的经验和体会，对在教学中实施多元化考核体系的必要性及具体方式展开探讨。

Objective To explore the role of ppk1 gene in E.coli CFT073 strain during urinary tract infection (UTI). Methods C57BL/6 mouse models of acute UTI with the wild-type(CFT073) and ppk1 mutant (△pk1) infected, were used to compare the bacteria colonization and inflammation induction abilities of bladder tissues. Results In the mouse models, the △pk1 strain showed a significantly lower infection rate (73.3% vs 93.3%) and lower adhesion frequency of bladder (0.01%vs 0.5%) than those of the CFT073 strain. The expression of IL-6 and TNF-αwere reduced in the bladder of △pk1 infected group (P<0.05). Hematoxylin-Eosin tissue staining showed that the damage degree of bladders in △pk1 infected mice were less serious than the CFT073 infected mice. Conclusion ppk1 gene plays an important role in E.coli colonization to bladder and the inflammation induction ability.

Objective To explore the relationship between resistin rs1862513 with type 2 diabetes mellitus (T2DM ) in Guangdong area .Methods 182 cases of newly diagnosed T2DM in the Second Affiliated Hospital of Guangzhou Medical College from October 2010 to October 2011 were collected as the treatment group and contempo-raneous 191 subjects with healthy physical examination as the control group .The distribution frequency of rs1862513 locus C/G genotype and the logistic regression analysis results were compared .Results The genotype frequency of rs1862513 locus reached the genetic equilibrium with the group representative .Among them ,χ2 = 4 .645 ,P= 0 .098 , the difference of distribution frequency of rs1862513 locus C/G genotype had no statistical significance (P > 0 .05) ;the morbidity rate of GG homozygote was 1 .375 times of the mobidity rate of CC homozygote (OR = 1 .375 ,95% CI :0 .575 - 3 .285) .The difference of the risk degrees among genotypes had no statistical significance (P> 0 .05) .Conclu-sion rs1862513 genotype has no correlation with T 2DM in Guangdong area .

Objective:(1) To explore the difference of three kinds of adiponectin rs +16861194 (A / G ) genotype distribution in the type 2 diabetes patients and healthy people ; (2) To investigate the relationship of adiponectin rs+16861194 gene polymorphism with some biochemical indicators from T2DM and control.Methods:Selected 158 cases of type 2 diabetes patients as the experimental group, 132 cases of healthy people as the control group. The genotypes of adiponectin rs-16861194 in experimental group and the control group were detected; At the same time, fasting plasma glucose, triglyceride (TG), high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL) were measured seperately. Fasting insulin was measured by chemiluminescence and insulin resistance index was calculated.Results:①T2DM group and CON group adiponectin rs +16861194 genotype distribution were: AA (70.00%), AG (24.37%), G (5.63%) and AA (74.24%), AG (24.24%), GG (1.52%), the distribution showed no statistically significant difference (P> 0.05); ②Grouped by the genotype of adiponectin rs +16861194, the level of FBG, FIN, IRI, TG, CHOL, HDL-CH, LDL-CH showed no statistically significant difference (P> 0.05) between the genotypes in T2DM and CON group. ③ Compraring between T2DM group and CON group,.the level of FBG, FIN, IRI, TG, HDL in the AA type of T2DM group and CON group had significant difference (P<0.05), while the level of CHOL and LDL showed no statistically significant difference (P> 0.05) ; The level of FBG,IRI,TG,HDL in the AG+GG type of T2DM group and CON group had significant difference (P<0.05), while the level of FIN,CHOL,LOL showed no statistically significant difference (P> 0.05).Conclusions:(1) Three kinds of locus genotype distribution of rs +16861194 (A / G) had no significant difference in Type 2 diabetes patients and healthy people.(2) The genotype of adiponectin rs+16861194 had no effect on the level of FBG、FIN、IRI、TG、CHOL、HDL-CH、LDL-CH in type 2 diabetes patients and healthy people.

WPBL教学（网络式PBL教学）是将经典的PBL教学理念与网络技术有机结合而创造的一种新的教育模式。本文通过结合在医学检验专业中应用WPBL教学模式的教学实践，简述了WPBL教学实践的基本流程、WPBL教学模式较经典的PBL教学模式的优势以及WPBL教学过程中教师应该注意的几个问题。