在"新医科"背景下,云南地区医学发展迫切需要"五术"新型医学人才,医学细胞生物学作为医学专业基础核心课程,推进融入"五术"医学人才培养理念的"金课"建设势在必行.该文以"受体介导的胞吞"为例,以学生为中心进行融合整合式内容重构、课程思政、智慧教学、交互式模拟实验、项目式学习等创新性元素的混合式教学设计及实践.

课程思政就是要寓价值观引导于知识传授和能力培养之中,实现专业课与思政课同向同行,帮助学生塑造正确的世界观、人生观和价值观,这是人才培养的应有之义,更是必备内容.《医学细胞生物学》以细胞的生命现象和规律为研究对象,与人体各项生命活动密切相关,该课程是医学生专业基础必修课.该文以昆明医科大学为例,阐述《医学细胞生物学》课程思政建设路径探索,为医学类其他课程思政教育提供借鉴思路和经验.

目的 探究融入思政教育的速课混合式教学在全科医学专业《医学细胞生物学》课程中的实践和效果评价。方法 以昆明医科大 学2020级全科医学专业学生为研究对象，随机分为实验班和对照班。实验班级开展速课混合式教学，对照班级采用传统多媒体教学。通过考 试成绩分析和问卷调查，对实践效果进行评价。结果 成绩分析结果提示，该混合式教学显著提升了学生及时理解和掌握知识点的能力，同时 对于终结性考核也有一定帮助。学生主观评价也对该教学方式给予高度认可。 结论 本研究将思政教育融入专业课程，并采用混合式教学方 法，在全科医学实践中取得了很好的课程育人实效。

目的 探讨案例教学在医学遗传学课程教学的应用和效果.方法 选择昆明医科大学2018级儿科专业为实验班,2018级临床专业为对照班.在实验班实施案例教学,在对照班实施传统的课堂讲授式教学.通过考试成绩分析和问卷调查,对教学效果进行评价.结果 实验班的期末考试成绩明显高于对照班(P<0.0001).问卷调查结果显示多数学生对案例教学的效果满意;认为能帮助理解和灵活应用遗传病知识的占:93.04％;能提高解决问题能力的占:93.04％;能提高分析问题能力的占:95.65％;能提高学习能动性的占:94.78％.结论 案例教学适合医学遗传学课程,能提高教学质量.

为探讨PCNA基因在小鼠睾丸生后发育全过程中的表达规律,以16组出生后不同发育时期的昆明种正常小鼠睾丸组织为材料,在其石蜡组织切片上进行PCNA蛋白的免疫组化检测及PCNA基因的DNA-mRNA原位杂交检测.结果发现:PCNA蛋白仅在精原细胞及初级精母细胞中表达,其表达峰值出现在小鼠生后第11天;PCNA基因在初级精母细胞、次级精母细胞、圆形精子细胞以及少数精原细胞中均有表达,其中初级精母细胞中的表达信号相对强烈.上述结果表明,PCNA基因参与了小鼠睾丸发育过程中精原细胞及初级精母细胞的DNA复制过程,其mRNA表达和蛋白质表达存在着一定差异.

Objective To investigate the correlation of the polymorphisms of angiotensin Ⅱtype 1 receptor (AT1R)gene A1166C and angiotensinogen(AGT)gene M235T and blood biochemical indicators with essential hypertension(EH)in the male of Yi nationality in Yunnan province. Methods 92 hypertensive and 70 normotensive of Yi people were recruited in Yunnan province. Their basic information and blood biochemical indicators were obtained by questionnaire and standardized physical examination. The genes polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)technique. Results(1)For the male EH patients of Yi nationality,1166C and 235T allele frequencies were 0.038 and 0.772. No significant differences in allele frequency or genotype frequency were observed between the two groups(P > 0.05).(2)The levels of TC,LDL-C and TG in EH were significantly higher than those in controls(P < 0.05). Conclusion The AT1R A1166C and AGT M235T polymorphisms were not related to the pathogenesis of EH in the male of Yi people. However,the high levels of TC,TG and LDL-C might be the risk factors of EH.

Boysidia Ancey,1881 is a small group in Chondrininae and distributes in South and Southeast Asia. A total of 24 species were recorded in south Qinling Mountains,China. Among of these species,one was identified as a new record.Descriptions of this new species and other 8 species of Boysidia( Bensonenella) were given in the text. The type specimens of the new species were kept in Institute of Zoology,Chinese Academy of Sciences,and the Department of Cell Biology and Medical Genetics,Kunming Medical University.Key to subgenus of Boysidia( Boysidia),Boysidia( Bensonella),Boysidia( Paraboysidia) and Boysidia( Dasypupa)1. Columellar tooth with node-shaped on aperture deep,angular tooth and parietal tooth united into 2 crescent knifeshaped tooch ……………………………………………………………Boysidia( Dasypupa) Thompson and Dance,1983 Columellar tooth without node-shaped on aperture deep …………………………………………………………………22. Columellar tooth oblique ……………………………………………………………………………………………3 Columellar tooth not oblique,horizontal ………………………………………Boysidia( Paraboysidia) Pilsbry,1917 3. Angular tooth and parietal tooth united into 1 flake-shaped tooch,palatal tooth with hook-shaped………………………………………………………………………………………………………Boysidia( Bensonella) Pilsbry,1900 Angular tooth and parietal tooth independence,parallel ………………………………Boysidia( Boysidia) Pilsbry,1917Boysidia( Bensonella) qingliangfengensis sp. nov.( Fig. 1: A-E)Holotype: alt. 3. 40 mm,diam. 2. 10 mm; alt. of aperture 1. 15 mm,diam. of aperture 0. 75 mm. Collected from Longtangshan town,Linan city,Qingliangfeng Nature Reserve( 30°01u0027N,118°80u0027E),June 26,2013.Paratypes: 126 specimens. alt. 3. 30- 3. 45 mm,diam. 2. 00- 2. 20 mm; alt. of aperture 1. 10- 1. 20 mm,diam. of aperture 0. 700- 0. 815 mm. Collected from Longtangshan town,Linan city,Qingliangfeng Nature Reserve( 30°01u0027N,118°80u0027E),June 26,2013.Etymology: The name of the new species was based on Longtangshan town,Linan city,Qingliangfeng Nature Reserve from where the specimens were collected.Shell small,thin,luster,elongatedly conic. 5(1/2) whorls,moderately convex,spire long,the body whorl inflated,the bottom of body whorl not extended to aperture,and not form a bell-shaped. Apex blunt,embryo-whorls( 1-2(1/2)) increasedslowly,smooth,striae absent. Suture deep. Shell yellow brown or pale yellow in colour. Aperture elliptical,aperture margin thick,slightly expanded and not reflexed,with 16 rib-strias on the neck of aperture. With 14 teeth in the aperture: 1little nipple-formed infraparietal lamella,l large flake-formed parietal lamalla,1 big flake-formed angular lamella,1 small nipple-formed twin-lamella,2 small nipple-shaped parallel lamella on right aperture margin,2 small nipple-shaped suprapalaral plicae on the parallel lamella down,1 big nipple-shaped upper palatal plicae and 1 small nipple-formed interpalatal plicae,1 small nipple-shaped lower palatal plicae on aperture botton. 1 big nipple-formed basal plicae on the below part of columella margin nearly,1 small nipple-shaped columellar lamella on the columella margin above,1 small nipple-formed subcolumellar lamella on the columellar lamella down. Umbilicus big,with hole-formed,and the umbilicus was not covered by columella margin.Habitat: Boysidia( Bensonella) qingliangfengensis sp. nov.,as other Boysidia( Bensonella) species,usually lives on limestone areas of mountainous regions,in dark,moist habitats with bryophyte and lichen,in the cracks of the rocks or under the faller leaves.This new species is bigger than Boysidia( Bensonella) plicidens( Benson,1849),but differs obviously from the bigger size: the new species shell with whorls 5(1/2),alt. 3. 40 mm,diam. 2. 10 mm; alt. of aperture 1. 15 mm,diam. of aperture20. 75 mm. Aperture elliptical,aperture margin thick,slightly expanded and not reflexed,with 16 rib-strias on the neck of aperture,with 14 teeth on the aperture: 1 little nipple-formed infraparietal lamella,l large flake-formed parietal lamalla,1big flake-formed angular lamella,1 small nipple-formed twin-lamella,2 small nipple-shaped parallel lamella on right aperture margin,2 small nipple-shaped suprapalaral plicae on the parallel lamella down,1 big nipple-shaped upper palatal plicae and 1 small nipple-formed interpalatal plicae,1 small nipple-shaped lower palatal plicae on aperture botton. 1 big nipple-formed basal plicae on the below part of columella margin,1 small nipple-shaped columellar lamella on the columella margin above,1 small nipple-formed subcolumellar lamella on the columellar lamella down. The latter with 5 whorls( alt.2. 00 mm,diam. 1. 50) in small size,and the aperture irregular,subtriangular,having 9 folds teeth: 3 parietal lamellae,of which the upper two are long; one small dentiform columellar lamellae; five palatals,of which two subbasal plicae being minute. Basal margin has a callous outside. Umbilicus narrow.

详细记述了喇叭螺属Boysidia一新种,金平喇叭螺新种Boysidia(Bensonella)jinpingensis sp.nov..对新种的形态进行了详细描述,并与其近似种进行了对比和讨论.模式标本保存于昆明医科大学细胞生物学与医学遗传学系标本室.

目的 应用生物信息学分析软件预测结核分枝杆菌脂酰辅酶A合成酶(FadD3)氨基酸序列的结构与功能.方法 应用生物信息学在线工具NCBI (http://www.ncbi.nlm.nih.gov/)、Expasy(http://ca.expasy.org/)、SWISS-MODEL(http://swissmodel.expasy.org/)、客户端软件clustalx等进行多重序列比较、同源性分析、同源模建及蛋白功能活性位点分析;应用ProtParam软件(http://web.expasy.org/protparam)预测FadD3基因编码FadD3的蛋白理化性质;采用最小值进化法(the Minimum Evolution method)构建分子进化树. 结果 FadD3 (Mt-FACS)与其他物种的脂酰辅酶A合成酶(FACS)含相同的保守序列及催化活性位点,第174-185氨基酸残基是AMP结合部位,也是FadD3的保守区域;第422-497氨基酸残基是FACS与底物(脂酸)结合的部位,亦是结核分枝杆菌脂酰辅酶A合成酶的C末端.分子进化分析表明结核分枝杆菌脂酰辅酶A合成酶(Mt-FACS)与睾丸酮丛毛单胞菌脂酰辅酶A合成酶(Ct-FACS)的进化关系较近,与黑腹果蝇脂酰辅酶A合成酶(Dm-FACS)的进化关系较远. 结论 生物信息学预测提示FadD3是结核分枝杆菌脂类代谢及耐药性研究的潜在靶蛋白.

In order to investigate the relationship between Margarya melanioides distribution and sediment nutrient elements in the Dianchi Lake, 87 sampling points were selected where M.melanioides and sediment samples were collected in 2013. Diagram displays the sampling points where living M. melanioides were collected, and displays the location distribution and density of living M. melanioides. 8 sediment samples were selected and 16 nutrient elements (Organic matter, phosphorus, nitrogen, zinc, iron, manganese, magnesium, calcium, copper, sodium, po-tassium, arsenic, mercury, lead, cadmium, and chromium) were detected by chemical assay. Based on whether living M. melanioides was collected or not, 8 sediment samples were divided into two groups. 16 nutrient elements were analyzed using a completely random design for two samples t test between the two groups, and the results showed that only copper element showed significant difference between two groups (P<0.05). It is probably be-cause of that the growth of M. melanioides effectively absorbed the heavy metal elements, resulting in significant decrease in copper element content in lake sediment. The relationship between M. melanioides' growth and the nu-trient component in lake sediment was elucidated, which provides a scientific basis for lake pollution prevention and M. melanioides' population protection.

目的 建立田螺科已有种类的系统发育关系,探讨螺蛳(Margarya melanioides)的分类地位.方法 测定采自云南高原湖泊中的螺蛳、方形环棱螺和中华圆田螺的线粒体COI、16S rRNA基因序列,并结合GenBank中田螺科已有物种序列,计算螺蛳和其他主要类群的遗传距离,用最大简约法(MP)、最大似然法(ML)和贝叶斯法(BI)构建包含螺蛳的田螺科部分物种系统发育树.结果 COI基因第三位密码子转换已达到饱和.在COI基因片段数据集、COI第1,2位密码子数据子集、16S基因片段数据集中,螺蛳与圆田螺属的遗传距离最小.在MP法、ML法、BI法构建的系统发育树中,螺蛳和中华圆田螺聚为一支,我国的环棱螺属种类和非洲该属未聚在一起.结论 螺蛳与中华圆田螺的亲缘关系较近,线粒体COI联合16S基因片段可以弥补形态学在研究田螺科物种系统发育关系时的不足.

BACKGROUND:Anticancer drug and organic metal complexes wil form a new structure or a change in ion concentration, thus changing both the activity and toxicity to produce a synergistic effect. OBJECTIVE:To synthesize new high-efficient and low-toxic metal-fluorouracil complexes as anticancer drugs. METHODS:Copper, zinc and iron salts and fluorouracil were used to synthesize four copper, zinc and iron-fluorouracil complexes that were [Cu(5-Fu)2Cl2], [Cu(5-Fu)2(NO3)2], [Fe(5-Fu)3]SO4 and [Zn(5-Fu)2Cl2]. Preliminary chemical structures of the four complexes were confirmed by elemental analysis and mass spectrometry. Their inhibitory activity on human cancer cells, human leukemia cellline K562 and human colon cancer cellline HCT-116, was measured by MTT colorimetric assay. RESULTS AND CONCLUSION:[Cu(5-Fu)2Cl2], [Cu(5-Fu)2(NO3)2], [Zn(5-Fu)2Cl2] and [Fe(5-Fu)3SO4] were successful y synthesized. These four complexes at a mass concentration of 0.1-100 mg/L inhibited the proliferation of K562 and HCT-116 to different extents. The IC 50 values of these four complexes on K562 and HCT-116 cells were lower than those of fluorouracil, and their cytotoxicity was 1.5-7.8 times higher than that of fluorouracil. To conclude, copper/iron/zinc-fluorouracil complexes exhibit synergic inhibitory effects on cancer cellproliferation.

为探索FadD3在结核分枝杆菌胆固醇分解代谢中的作用,从结核分枝杆菌H37 Ra的全基因组中克隆了FadD3基因,并在大肠杆菌BL21中表达.根据NCBI公布的结核分枝杆菌全基因组序列设计一对引物,PCR扩增FadD3基因.PCR扩增产物与克隆载体pGEM3Zf(+)进行拼接,得到重组基因FadD3-pGEM3Zf(+),再转化到大肠杆菌DH5α得到克隆.PCR检测阳性克隆,再与表达载体pYUB28b拼接,得到重组质粒FadD3-pYUB28b.阳性重组质粒亚克隆到大肠杆菌BL21宿主菌进行自体诱导表达.经过表型筛选及鉴定分析,已成功构建了重组表达质粒FadD3-pYUB28b.SDS-PAGE和Western blotting证实,重组基因FadD3-pYUB28b在大肠杆菌BL21中有表达产物.实验结果表明,以pYUB28b为表达载体,FadD3重组基因在大肠杆菌表达体系于温度分别为18,28℃有包涵体形式的表达蛋白,在37℃无表达产物.

Objective To explore the role of student-centered teaching model in medical genetics teaching.Methods 205 students of Grade 2011 majored in clinical medicine in Kunming Medical University were divided into two groups.The teaching model selected by students was used in the experimental group to compare with the teaching model selected by teachers in the control group.A questionnaire survey and a theoretical course examination were used to assess the teaching effectiveness between the two groups.Results The test result indicated that the average score in the experimental group was significantly higher(P<0.05) than that in the control group and the questionnaire survey results showed that the majority of students were satisfied with the teaching model selected by students.Conclusion The student-centered teaching model can effectively improve the quality of medical genetics learning.

Objective To investigate the protein-protein interaction(PPI) between an important DNA base excision repair enzyme uracil-DNA glycosylase(StoUDG)and thioredoxin(StoTRX)in hyperthermophilic archaeon Sulfolobus tokodaii.Methods The bacterial two-hybrid reporter strain was co-transformed by pBT recombined with StoUDG gene and pTRG recombined with StoTRX gene.The strength of PPI was detected by observing the bacterial colony in selective plating medium.Results(1)The co-transformation of bacterial two-hybrid plasmids enabled reporter strain to resistant to chloramphenicol,tetracycline and kanamycin.(2)The reporter strain which contained pBT-TRX and pTRG-UDG plasmids grew slowly in selective plating medium without self-activation.Conclusion The results showed a weak interaction between StoUDG and StoTRX,which indicated that StoTRX played a role in UDG-dependent DNA base excision repair.

The AR42J-B13 rat pancreatic acinar cell line was used to identify pancreatic transcription factors and exogenous growth factors (GFs) that might facilitate the reprogramming of exocrine cells into islets. Adenoviruses were used to induce exogenous expression of the pancreatic transcription factors (TFs) Pdx1, MafA, Ngn3 and Pax4. Individually Pdx1, MafA and Pax4 had no effect on the expression of endocrine markers, whilst adeno-Ngn3 on its own increased the expression of Pax4, Ngn3 and NeuroD. In combination the four TFs had a significant effect on the expression of insulin 1 and 2 that was associated with a change in cell morphology from a rounded to a spindle-like shape. Amongst a range of growth factors, Betacellulin and Nicotinamide were shown to enhance the effects of the four TFs. The presence of adeno-Pax4 in the differentiation cocktail was important in limiting the expression of glucagon and in generating glucose sensitive insulin secretion. Further experiments asked whether the adenoviral TFs could be replaced by protein transduction domain (PTD)-containing TFs. The results showed that the PTD-TFs could mimic in part the effects of the adeno-TFs, but the resultant cells did not undergo the important morphological change associated with differentiation to endocrine lineages and levels of endogenous markers were very much lower. In summary, the results describe a cocktail of four TFs and two GFs that can be used to induce formation of glucose sensitive insulin secreting cells from ARJ42 cells, and demonstrate that it would be difficult to replace adenoviral transduction with PTD-TFS.

Although embryonic stem (ES) cell-derived hepatocytes have the capacity for liver engraftment and repopulation, their in vivo hepatic function has not been analyzed yet. We aimed to determine the metabolic function and therapeutic action of ES cell-derived hepatocytes after serial liver repopulations in fumaryl acetoacetate hydrolase knockout (Fah−/−) mice. Albumin expressing (Alb+) cells were obtained by hepatic differentiation of ES cells using two frequently reported methods. After transplantation, variable levels of liver repopulation were found in Fah−/− mice recipients. FAH expressing (FAH+) hepatocytes were found either as single cells or as nodules with multiple hepatocytes. After serial transplantation, the proportion of the liver that was repopulated by the re-transplanted FAH+ hepatocytes increased significantly. ES cell-derived FAH+ hepatocytes were found in homogenous nodules and corrected the liver metabolic disorder of Fah−/− recipients and rescued them from death. ES cell-derived hepatocytes had normal karyotype, hepatocytic morphology and metabolic function both in vitro and in vivo. In conclusion, ES cell-derived hepatocytes were capable of liver repopulation and correction of metabolic defects after serial transplantation. Our results are an important piece of evidence to support future clinical applications of ES cell-derived hepatocytes in treating liver diseases.

Protein transduction domains (PTDs), such as the HIV1-TAT peptide, have been previously used to promote the uptake of proteins into a range of cell types, including stem cells. Here we generated pancreatic transcription factors containing PTD sequences and administered these to endoderm enriched mouse embryonic stem (ES) cells under conditions that were designed to mimic the pattern of expression of these factors in the developing pancreas. The ES cells were first cultured as embryoid bodies and treated with Activin A and Bone morphogenetic protein 4 (BMP4) to promote formation of definitive endoderm. Cells were subsequently plated as a monolayer and treated with different combinations of the modified recombinant transcription factors Pdx1 and MafA. The results demonstrate that each transcription factor was efficiently taken up by the cells, where they were localized in the nuclei. RT-qPCR was used to measure the expression levels of pancreatic markers. After the addition of Pdx1 alone for a period of five days, followed by the combination of Pdx1 and TAT-MafA in a second phase, up-regulation of insulin 1, insulin 2, Pdx1, Glut2, Pax4 and Nkx6.1 was observed. As assessed by immunocytochemistry, double positive insulin and Pdx1 cells were detected in the differentiated cultures. Although the pattern of pancreatic markers expression in these cultures was comparable to that of a mouse transformed β-cell line (MIN-6) and human islets, the expression levels of insulin observed in the differentiated ES cell cultures were several orders of magnitude lower. This suggests that, although PTD-TFs may prove useful in studying the role of exogenous TFs in the differentiation of ES cells towards islets and other pancreatic lineages, the amount of insulin generated is well below that required for therapeutically useful cells.

Objective To observe the training effect of the family rehabilitation on patients with traumatic paraplegia.Methods In our hospital,from August 2005 to June 2011,43 patients with traumatic paraplegia were randomly divided into two groups:22 patients in the observation group and 21 patients in the control group.The family rehabilitation was applied intensively at the base of the regular nursing in the observation group,the rugular nursing was applied in the control group.Result The incidence of the complication in the observation group was 4.5%,it was significantly lower than that in the control group(38.1%)by the T-test(P0.05),there was a significance statistically.Conclusion The training effect of the family rehabilitation on the patients with traumatic paraplegia in the observation group is better than the control group,as it can reduce the incidence of complications and improve the activities of daily living.

The testic tissue samples of normal Kunming mice are taken at 17 different stages of postnatal development.Immunohistochemical technique is used on the paraffin tissue sections.The expression of Bax protein in mouse testis is examined throughout the postnatal development.The results show that Bax protein is expressed generally in the testis of mice from day 1 to day 57.Bax protein is localized in all mouse testicular cell types.These results suggest that Bax protein exists in spermatogenic cells during the postnatal development of mouse testis.However,Bax protein expresses in spermatogenic cells of the testis may not mean apoptosis to these cells.