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The Relationship between the Implantation of Ovarian Stimulated Embryos and the Expression of Leukemia Inhibitory Factor Protein in Mice Metrium

觀察受體鼠妊娠和胚胎著床情況,并檢測胚胎移植時小鼠子宮內膜中白血病抑制因子(Lif)表達水平,探討超排卵對小鼠胚胎著床潛能的影響。方法:建立超排周期胚胎和自然周期胚胎移植小鼠模型,比較妊娠率、胚胎著床率的差異及其與Lif蛋白的表達水平之間的關系。結果:超排卵周期受體組的妊娠率(20.00%)和胚胎著床率(8.33%)顯著低于自然周期組的妊娠率(55.00%)和胚胎著床率(35.00%)(P＜0.05)。自然周期胚胎和超排周期胚胎受體組內膜中Lif蛋白的表達水平相似(P＞0.05),妊娠受體組Lif蛋白的表達水平顯著高于未孕受體組(P＜0.05),但單胎妊娠和多胎妊娠受體組內膜中Lif蛋白的表達水平相似(P＞0.05)。結論:超排卵可能降低胚胎的著床潛能,Lif蛋白的表達水平與胚胎著床有關,但與著床胚胎的數目無比例關系。

CULTURE AND INDUCED MATURATION OF FETAL MOUSE LIVER CELLS ON POLY-L-LACTIC ACID SCAFFOLD

Objective To investigate the feasibility of fetal liver cells for liver tissue engineering, the supporting function of poly L lactic acid (PLLA) scaffold for fetal liver cells and the effects of oncostatin M (OSM), nicotinamide (NA) and dimethyl sulfoxide(DMSO) on growth and hepatic differentiation. Methods After three dimensional PLLA scaffolds having a porous structure were prepared by using NH 4HCO 3 particle, fetal liver cells obtained from E14.5 C57BL/6CrSlc murine embryos were inoculated in the scaffolds. Cells were cultured in Williams’E medium with or without OSM, NA and DMSO for 30 days. Changes in cell number, liver-specific function, and cellular morphology were observed. Results When compared with in monolayer culture, cell number and albumin secretion increased obviously in three-dimensional PLLA. Alburmin secretion increased slightly in OSM group of monolayer culture, but increased obviously in OSM groupo of PLLA culture and in OSM/NA/DMSO group of both monlayer and PLLA cultures. Conclusion The three-dimensional PLLA scaffold is a good supporting material for the cultivation of tetal liver cells. OSM, NA and DMSO remarkaly stimulated maturation of hepatic parenchymal cells in vitro in terms of morphology and liver-specific function.

IDENTIFICATION OF ECTOMESENCHYMAL STEM CELLS OF HUMAN FETAL FACIAL PROCESSES AND SPONTANEOUS DIFFERENTIATION TO SMOOTH MUSCLE CELLS

OBJECTIVE: To investigate the characteristic and phenotype of ectomesenchymal stem cells of human fetal facial processes and the procedure of spontaneous differentiation to smooth muscle cells. METHODS: The primary ectomesenchymal cells of E 50 human fetal facial processes were isolated by 2.5 g/L trypsin and cultured with DMEM/F 12 with 10(-6) U/L leukemia inhibitor factor(LIF). The morphology and growth rate were observed by inverted microscop. After being withdrawn LIF, the characteristic of cells were identified by immunohistochemistry and RT-PCR. Ultrastructure was observed by transmission electron microscope. RESULTS: The cultured cells displayed monolayer growth and were fibroblast-like with 2-4 processes. The cells were stainely positived for anti-human natural killer cell marker-1, Vimentin, S-100, neuron specific enolase, myoglobin and VIII factor, but negatively for glial fibrillary acidic protein, neural fiblament, alpha-SMA and cytokeratin in immunohistochemistry. Two days after being withdrawn the LIF, cells expressed alpha-SMA in protein and mRNA levels. The cells were rich in muscular filament-like structure and dense bodies under transmission electron microscope. CONCLUSION: Cultured cells are undifferentiated ectomesenchymal stem cells. The cells have the potential for differentiating spontaneously to smooth muscle cell.

兔胚胎全層視網膜移植至大鼠視網膜下的觀察

Effects of Short-term Fertilization versus Conventional Fertilization in Vitro Fertilization Cycles: A Meta-Analysis

ObjectiveTo systematically review the clinical effects of short-term and conventional fertilization for vitro fertilization-embryo transfer (IVF-ET). MethodsRandomized controlled trials (RCTs) about the clinical effects of short-term fertilization versus conventional fertilization for IVF-ET were searched in PubMed, The Cochrane Library (Issue 8, 2014), CBM, CNKI, WanFang Data and VIP from inception to August 2014. Two reviewers independently screened literature according to the inclusion and exclusion criteria, extracted data, and assessed methodological quality of included studies. Then meta-analysis was performed using RevMan 5.2 software. ResultsA total of six RCTs involving 1 373 patients were finally included. The results of meta-analysis indicated that:short-term fertilization was superior to conventional fertilization in increasing high quality embryo rates (OR=1.42, 95%CI 1.18 to 1.70, P=0.000 2) as well as clinical pregnancy rates (OR=1.67, 95%CI 1.33 to 2.09, P < 0.000 01). However, the two groups were alike in fertilization rates, polyspermy rates, and miscarriage rates. ConclusionCurrent evidence indicates that short-term fertilization is superior to conventional fertilization in increasing high quality embryo rates as well as clinical pregnancy rates. Due to limited quality and quantity of the included studies, the above conclusion should be verified by conducting more large-scale, high quality RCTs with long-term follow-up.

STUDY ON DIFFERENTIATION OF EMBRYONIC STEM CELLS INTO OSTEOBLAST IN VITRO INDUCING BY 1,25 (OH)2VD3

Objective To investigate the effect of 1,25(OH)2VD3 on differentiation of embryonic stem cells (ESCs) into osteoblasts. Methods Osteoblasts were isolated and cultured from calvarium of 2-day-old Kunming white mice, embryoid bodies (EBs) were prepared with modified zur Nieden method. EBs were divided into 4 groups according to different mediums: group A, as the control group, in which EBs medium contained no leukemia inhibitory factor; group B, in which EBs medium contained supplements of Vitamin C (VC, 50 μg/mL) and β-glycerophosphate (β-GP, 50 mmol/L); group C, inwhich EBs medium was the same as that of group B and 5 × 104 osteoblasts of 3rd passage were seeded into each well; group D, in which the medium contained supplements of VC (50 μg/mL), β-GP (50 mmol/L) and 1,25(OH)2VD(4 × 10-9 mol/L), and 5 × 104 osteoblasts of 3rd passage were seeded into each well. The ALP activity was determined by ALP reagent kit every 5 days. The RQ-PCR was performed to measure the mRNA expressions of osteocalcin (OCN). Al izarin red S staining was performed to count the bone nodules. Results The expression of ALP witnessed no obvious change in each group within 5 days after adherence of EBs, but increased gradually after 5 days. The expression of ALP in group D reached the peak at 20 days. Red nodules with clear outl ine and different sizes were evident by microscope. Al izarin red S staining testified the number of bone noudles in groups A, B, C and D was 20 ± 8, 18 ± 5, 31 ± 1 and 50 ± 1, respectively, indicating significant differences between groups C, D and groups A, B (P lt; 0.05), no significant difference between group A and group B (P gt; 0.05), and a significant difference between group C and group D (P lt; 0.05). The result of RQ-PCR showed that the mRNA expressions of OCN in groups A, B, C and D was 10.18 ± 1.17, 20.29 ± 1.03, 18.84 ± 4.07 and 32.15 ± 5.23, respectively, indicating significant differences between groups C, D and groups A, B (P lt; 0.05), no significant difference between group A and group B (P gt; 0.05), and a significant difference between group C and group D (P lt; 0.05). Conclusion The combined action of 1,25(OH)2VD(4 × 10-9 mol/L), VC, and β-GP can effectively promote the differentiation of the ESCs-derived osteoblasts.

利用畢博平臺進行組織學與胚胎學教學的探索和體會

摘要:在優質資源共享、促進信息化教學的時代背景下,許多高校開始建立基于Blackboard教學平臺的畢博網絡課程。結合組織胚胎學課程特點,我們就如何建立和在教學過程中如何充分利用該平臺進行《組織學與胚胎學》教學等問題進行了探索,為今后提高網絡教學質量、促進教學改革提供了經驗。

FETAL CADACVERIC MUTIPLE ORGANS PROCUREMENT FOR TRANSPLANTATION OF PANCREAS AND THYROID-PARATHUROID GLANDS(ONE CASE REPORT)

This is the first successful case expriences,a method of the procurement of the fetal cadavertic multiple argans for transplantation of the pancreas and thyroid-pararthyroid glands was produced. The liver,pancreas,duodenum,spleen,and both kidneys were harvested en bloc by a group of surgeons,and the right hem-ithyroid-parathyroid glands with pedicle of thd blood vessels wre removed by another group.The pancreas together with the spleen were transplanted to a patient having diabetes mellitus. The thyroid-parathyroid glands were given to another case with bypothyroidism and hypoparathyroidism.Both cases had good results.This method had dicreased the warm ischemia of the transplants,and could provide liver,pancreas,spleen,kidneys and thyroid-parathyroid glands to solve the problem of shortage of fetal organs.

INDUCED DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELL INTO ENDOTHELIALCELL IN VITRO

Objective To explore an optional condition to induce mouse embryonic stem cell(ESC) to differentiate into endothelial cells so as to provide seedcells for tissue engineered vascular. Methods The embryos from one pregnant 12.5days mouse was harvested to culture the mouse embryonic fibroblasts(MEF). The ESC was reanimated by common method, and used to cultured into embryoid body(EB) in vitro. The EB which was used to induce into endothelial cells was divided into two groups. The EB was cultured in the EB medium with 3ng/ml transforming growth factor β1, 50 ng/ml vascular endothelial cell growth factor and 1 μmol/L potent and selective inhibitor of activin receptorlike kinase receptors in experimental group. The EB was cultured in the EB medium in the control group. After 14 days, RTPCR and immunohistochemistry were used to detect vWF and CD34, to analyze the morphology and type of the differentiated cells fromESC. Results The primary MEF had a high proliferation activity. At the 3rdday, the fusion rate of MEF was about 90% with a fusiform shape. The cells was fusiform shape and arranged compactly with fullness of nucleus and 2-3 entoblasts. The 3rd5th generations EB was polygonal with fullness of cytoplasm and 3-4 entoblasts. ESC could maintain undifferentiated state, and the cells unit lookedlike bird nest with smooth margin; the cells was small at size and b refractivity with high rate of nuclein and rapid proliferation. At 3 days of dropculture, EB can seen grossly and at 3 days of suspension, large and transparent EBformed. EB was spread radiately with an intensive adhesion at the 2nd day. In experimental group, many round cells was differentiated around EB from the 4thday to the 7th day, and form tubular structures from the 10th day to the 14th day. The vWF and CD34 were expressed. In control group, EB could not form tubularstructures, and the vWF and CD34 were not expressed. Conclusion ESC can differentiate into endothelial cells under some conditions, and form vessellike structure under condition culture, which can provide sources of seed cells for tissue engineered vessel.

兒童頭頸部胚胎型橫紋肌肉瘤復發一例報道及文獻回顧

目的胚胎型橫紋肌肉瘤（ERMS）是一種幼兒、青少年較多見的的軟組織腫瘤。通過介紹典型病例及文獻復習，以總結ERMS的治療經驗。 方法報道1例腮腺咬肌區來源的頭頸部ERMS患兒，二次切除術后再次復發，腫瘤增長速度快，局部侵及面廣，累及區域淋巴結。復發后行3個周期TI（紫杉醇脂質體+異環磷酰胺）一線化學療法（化療）后給予適形調強放射治療（放療），觀察患兒治療后的療效。完善相關檢查，排除腫瘤遠處轉移后，行3個周期TI化療，化療結束2周后行適型調強放療，化療期間每周期及放療前后復查腫瘤原發灶CT，根據實體腫瘤RESEST version 1.1療效評價標準，測量腫瘤最大徑，比較其在治療前后及過程中的變化，進行療效評價。 結果患兒3個周期化療后療效評價部分緩解，放療結束后療效評價為完全緩解，治療后迄今無復發轉移征象。 結論ERMS復發后暫無標準治療方案，對于巨大頭頸部橫紋肌肉瘤，如果無法手術切除，可進行化療+放療綜合治療。新輔助化療采用TI方案局部腫瘤消退效果好。