体外培养的小鼠骨髓基质细胞对血细胞集落形成的支持

为探讨骨髓基质细胞在液体条件下支持造血的能力,对小鼠骨髓有核细胞体外培养,在液体培养基中形成基质细胞贴壁层,更换培养液,再种植骨髓有核细胞。结果在贴壁的基质细胞上形成了细胞集落。按细胞形态特征分为两类:一类为基质细胞集落,仅为偶见。另一类为造血细胞集落,数量较多,并且集落通过一定的结构与贴壁的基质细胞连接。结果提示,造血细胞集落的形成,需要有与骨髓基质细胞接触的微环境,并可对基质细胞的生长发育有一定的反作用。本文提出了造血细胞集落的液体培养法。     

快速培养纯化猪角朊细胞及复合羊膜体外培养实验研究

目的研究快速培养和纯化猪角朊细胞的方法及观察角朊细胞在脱细胞羊膜上的生长状况,为组织工程皮肤研究提供实验依据.方法采用加10%胎牛血清的人无血清角朊细胞培养基(defined keratinocyte-SFM,DKSFM)培养原代猪角朊细胞,分别用DKSFM(A组)、加5%胎牛血清的DKSFM(B组)、加10%胎牛血清的DKSFM(C组)培养传代猪角朊细胞,于接种后1、3、5和7 d观察猪角朊细胞的形态及生长曲线.利用猪角朊细胞与培养瓶黏附牢固的特点,用0.02%乙二胺四乙酸(ethylene diamine tetraacetic acid,EDTA)和0.05%胰蛋白酶分步消化,纯化细胞.将第2代猪角朊细胞接种在冻干脱细胞羊膜上,直接苏木素染色、石蜡切片、免疫组织化学染色等方法观察猪角朊细胞在脱细胞羊膜上的生长状况.结果采用加10%胎牛血清的DKSFM培养原代猪角朊细胞,细胞生长速度快,形态好,培养5 d铺满培养瓶底60%～70%.B组培养的传代猪角朊细胞生长速度较C组培养猪角朊细胞生长速度快.A组培养传代的角朊细胞,细胞生长缓慢.用0.02?TA和0.05%胰蛋白酶分步消化的方法纯化猪角朊细胞,可获得纯度为95%以上的猪角朊细胞.将第2代猪角朊细胞接种在脱细胞羊膜后12 d,可形成单层细胞,呈多角形、铺路石样排列;14 d和16 d细胞进一步密集,16 d细胞出现老化.结论 10%胎牛血清的DKSFM培养原代猪角朊细胞,5%胎牛血清的DKSFM培养传代猪角朊细胞,细胞生长速度快.用0.02?TA和0.05%胰蛋白酶分步消化的方法纯化猪角朊细胞,可以获得高纯度猪角朊细胞.脱细胞羊膜为猪角朊细胞提供良好的支架,接种培养后12 d,细胞形态最好.     

基于《辅行诀五脏用药法要》探讨乌梅丸论治情志病

乌梅丸载于《伤寒杂病论》厥阴篇,历代医家多认为其有温脏安蛔之功,临床应用范围较为局限,自清代医家柯琴提出其为厥阴病之主方后乌梅丸才被医家重视,其临床应用亦得以拓展。乌梅丸证之成因,实则归于厥阴风木气运失常,阴阳不相顺接,而非仅为蛔虫内扰所致。以《黄帝内经》“开阖枢”理论分析六经之功能,并结合《伤寒杂病论》原文可证“阴枢”实为厥阴,乌梅丸为厥阴病之主方,有顺接阴阳之功,故其可治疗厥阴枢机不利,寒热错杂之证。《辅行诀五脏用药法要》与《伤寒杂病论》二者同源,均参考了《汤液经法》所载内容,《辅行诀五脏用药法要》中补泻诸方及救逆方之组成有规律可寻,均依文中所载“汤液经法图”而成,与五行理论密切相关。该文通过探索“汤液经法图”组方规律及药物五行归属,以“汤液经法图”组方规律剖析乌梅丸,可证乌梅丸主要作用于肝、脾、心三脏,依黄帝内经五脏藏神理论,此三脏与情志调节密切相关,故以乌梅丸论治厥阴枢机不利、寒热错杂所致之情志病有据可循,文末亦列举概述近年乌梅丸治疗情志病的报道。该文为临床应用乌梅丸治疗情志病提供了思路及依据,扩展了其应用范围,古方亦能为今用。     

论医学生和谐就业伦理观的培养

结合目前医学生在就业过程中出现的不和谐伦理现象,分析了其主客观原因,提出了医学生和谐就业伦理观培养的现实途径.     

聚吡咯表面涂层对体外培养成骨细胞生长矿化能力的影响

目的：研究导电玻璃表面聚吡咯（Ppy）涂层对成骨细胞生长矿化能力的影响。方法：将成骨细胞接种于聚吡咯涂层材料表面，采用HE染色，Von kossa染色、SEM检测法观察成骨细胞在Ppy涂层表面的生长矿化能力。结果： 成骨细胞在ITO以及ITO表面聚吡咯涂层的基底上均能良好生长，并且可在聚吡咯涂层的表面形成矿化结节，经Von Kossa染色呈阳性反应。结论：聚吡咯涂层具有良好的生物相容性，适于作为骨内种植材料应用。     

体外扩增恒河猴自体内皮细胞人工血管移植研究（一）

目的:为恒河猴人工血管移植研究提供内皮化人工血管.方法;将恒河猴表浅静脉内皮细胞在体外培养13.89±1.36天;扩增培养的内皮细胞衬里于用纤维蛋白胶预衬的ePTEE人工血管,继续培养9天.结果:细胞数增加147.93±88.68倍.所培养的细胞为二倍体细胞,纯度99%.原代及传代细胞培养上清液中6-keto-PGFla和vWF含量无显著性差异.细胞种植后2小时和9天、人工血管腔面见一层均匀的基质,基质表面有一层连续的内皮细胞单层.内皮细胞紧密排列,呈梭状,形态饱满.细胞种植后9天,内皮细胞胞浆内微丝、微管数量明显增加,表明细胞骨骼成熟.结论:内皮化人工血管可用于置换恒河猴动脉.     

Prenatal diagnosis of Gaucher disease Assay of the ß-glucosidase activity in amniotic fluid cells cultivated in two laboratories with different cultivation conditions

Sixteen pregnancies at risk for Gaucher disease - six with the Norrbottnian form, one with a juvenile form with a similar clinical course to the patients from Norrbotten and nine with the infantile form - have been monitored by the assay of β-glucosidase activity in cultivated amniotic fluid cells with natural labelled glycosylceramide as substrate. Two methods of cultivation were compared in respect of their effect on the activity of lysosomal enzymes. No significant difference was found between the two marker enzymes, β-galactosidase and N -acetyl-β-glucosaminidase, but the β-glucosidase activity was significantly higher in the cells cultivated with one of the methods. In four of the pregnancies at risk, the β-glucosidase activity in the cultivated amniotic fluid cells was less than 5 % of that in the two control materials. These fetuses were regarded as affected with Gaucher disease and were aborted. Differentiation between controls and Gaucher heterozygotes was not possible in cultivated amniotic fluid cells. The diagnosis of Gaucher disease in the amniotic fluid cells was confirmed in three of the four cases by the assay of the β-glucosidase activity in the liver and brain of the aborted fetuses. The glucosylceramide content of the liver from two aborted fetuses was not augmented. The β-glucosidase activity was examined in seven placentas from pregnancies at risk for Gaucher disease and found to be in agreement with that in the cultivated amniotic fluid cells.     

Oriented migration and differentiation of neuroblasts in organ cultures of mouse embryonic spinal cord

During cultivation of transverse sections of the spinal cord of 12–14 day mouse embryos in Maximow's chambers differentiation of nerve and glial cells took place with the formation of the organotypical structure of the explant. In some cases migration of groups of bipolar neuroblasts was observed in the 3rd–4th week of cultivation outside the explant, along bundles of nerve fibers consisting of axons of mature nerve cells located in the central fragment of the cultivated tissue. The results show that nerve fibers growing out from the explant can guide migrating neuroblasts and can evidently induce their subsequent differentiation. The hypothesis is put forward that similar processes of migration of neuroblasts along tracts formed by axons can also take place in the developing brain and that they are followed by differentiation where these tracts terminate.Group for Culture of Nerve Tissue, Laboratory of Functional Synaptology, Institute of the Brain, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR B. N. Klosovskii.( Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 8, pp. 1002–1005, August, 1976.     

Comparison of Three Methods for Diagnosis of Cutaneous Leishmaniasis

Background

Leishmaniasis is one of the infectious parasitic diseases of highest incidence in the world. Cutaneous Leishmaniasis (CL) has long been reported in Shiraz, Southern Iran. There is a need to find a sensitive and specific method for treatment and control of the disease.

Methods

We have compared the sensitivity of the conventional methods microscopy and cultivation of lesion scrapes against PCR amplification of parasite kinetoplast DNA from these samples. The samples (n=219) were obtained from the patients clinically suspected of CL. The smears were stained with Giemsa for microscopy and cultured in Novy-Nicolle-McNeal (NNN) blood agar for promastigote growth. For PCR, the dry smears were scraped off the slides and DNA was extracted.

Results

The positive rates from 219 specimens were 76.71%, 50.68%, and 93.61% for microscopy, cultivation, and PCR, respectively. The highest correlation was found between PCR and microscopy method (P=0.014). In PCR assay, 95.61%, 3.9%, and 0.49% of the samples were identified as Leishmania major, L. tropica, and dermatropic L. infantum, respectively.

Conclusion

The PCR method appears to be the most sensitive for the diagnosis of CL and is valuable for identifying the other species of Leishmania with confusing dermatropic signs.     

来华医学学历留学生教育发展趋势初探

针对近年我国来华医学学历留学生教育持续发展现状，对周边国家医学教育需求、不同培养模式优劣以及培养质量标准化等进行了分析，探讨了来华医学学历留学生教育未来发展趋势。