体外分离培养人脐静脉内皮及内皮下间充质干细胞

目的:建立人脐静脉内皮及内皮下间充质干细胞体外培养和扩增的方法,探讨其生物学特性,建立间充质干细胞体外培养扩增体系。方法:实验于2006-04/2006-09在辽宁医学院解剖学实验室完成。解剖细胞培养室为无菌百级培养间。正常健康产妇顺产或剖宫产的新生儿脐带由辽宁医学院附属第一医院提供,产妇及其家属均知情同意,并经医院伦理委员会批准。实验方法:①体外分离和培养贴壁细胞:无菌条件下取正常健康产妇分娩或剖宫产脐带,将其用预热PBS充分洗涤去血渍后,从脐静脉一端插入留置针,用预热PBS冲净静脉腔血后,用止血钳夹闭另一端,注入经预热至37℃的Ⅰ型胶原酶,置于37℃水浴箱中消化,30min后放出胶原酶,并用PBS冲洗血管腔,收集消化液和冲洗液,400r/min离心10min,吸弃上清液,重悬于M199培养基(含体积分数为0.15的胎牛血清,2mmol/L谷氨z酰胺,2μg/L碱性成纤维细胞生长因子,100U/mL青霉素,100U/mL链霉素)。以5×108L-1密度接种于6孔培养板中,置于37℃、体积分数为0.05的CO2饱和湿度培养箱中培养,48h后全量换液,以后每3d全量换液。待细胞80%融合时,0.25%胰酶消化,按1×108L-1传代培养。②间充质干细胞生长曲线的测定:取传代培养细胞,按2×107L-1密度接种于24孔培养板内,每天取3孔,将细胞消化计数,连测8d,绘制间充质干细胞生长曲线。③间充质干细胞表面抗原检测:在24孔塑料培养板内放置无菌的盖玻片,每孔中种植108L-1第2代细胞悬液1mL。采用免疫细胞化学方法进行细胞表面抗原检测。结果:①间充质干细胞的形态学观察:接种的细胞48h后细胞完全贴壁生长,其镜下形态有呈椭圆形、多角形的内皮细胞以及呈梭形的成纤维样细胞,有的形成漩涡状生长的集落。②间充质干细胞生长曲线的分析:传代培养的潜伏期约为24～36h,细胞倍增时间约为30～36h,对数增殖期约为二三天,对数增殖期后第5天进入平台期。③间充质干细胞表面抗原特性:免疫细胞化学分析结果显示,间充质干细胞表面抗原cd166、cd44阳性,而vWF阴性,说明分离获得的细胞具有间充质干细胞的特点。结论:所建立的分离和培养方法可获取人脐静脉黏附细胞中一组独特的细胞群,具有间充质干细胞的生物学特性。     

The efficacy of subcutaneous recombinant human erythropoietin in the correction of phlebotomy-induced anemia in autologous blood donors

The efficacy of subcutaneous recombinant human erythropoietin (rhEPO) (500 U/kg; administered twice a week during the 3 weeks before surgery) in the recovery of preoperative hemoglobin concentrations within a 3- week period was studied in 40 patients, each of whom donated 2 units (900 mL) of blood for their own use before total hip replacement surgery. Twenty autologous blood donors received rhEPO (EPO group) and 20 were not treated (control group). The initial hemoglobin concentration (14.0 +/− 1.0 g/dL [140 +/− 10 g/L]) was completely recovered before surgery (14.0 +/− 1.6 g/dL [140 +/− 16 g/L]) in the EPO group, while a decrease from 13.8 +/− 1.1 to 12.2 +/− 1.3 g per dL (138 +/− 11 to 122 +/− 13 g/L) was observed in the control group. The preoperative reticulocyte count showed more than sixfold increase in the EPO group, whereas a twofold to threefold increase was found in the control group. Serum ferritin concentration fell to 42 +/− 29 micrograms per L in the EPO group and to 54 +/− 35 micrograms per L in the control group. The postoperative serum erythropoietin concentration in the EPO group was significantly lower than that in the control group, but it did not differ from the pretreatment value and was attended by a higher hemoglobin concentration after surgery. Only transient flu-like symptoms were mentioned by patients who were treated with rhEPO. Changes in blood pressure or platelet count or other adverse events were not observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Respiratory insufficiency in neuronopathic and neuropathic disorders

Twenty-nine patients with a neuronopathic or neuropathic disorder were referred for assessment of respiratory insufficiency between 1978 and 1994. Diagnoses included spinal muscular atrophy (6), chronic idiopathic demyelinating neuropathy (4), Vialetto-van Laere syndrome (3), hereditary motor and sensory neuropathy (3) and a miscellaneous group (5). We also describe seven patients with Guillain-Barre syndrome (GBS) who required long-term ventilatory support for over 6 months to 7 years after the initial illness. Respiratory insufficiency occurred as a consequence of respiratory muscle weakness, impaired bulbar function and restrictive lung defects. In some groups presentation was with progressive nocturnal hypoventilation culminating in acute respiratory failure. Five patients with GBS or chronic idiopathic demyelinating neuropathy were weaned from ventilatory support up to 18 months after the initial illness. The remaining 24 patients required continuous or nocturnal ventilatory support using intermittent positive-pressure ventilation (13), negative pressure ventilation (4), nasal-mask-delivered intermittent positive-pressure ventilation (4), nasal-mask-delivered continuous positive-pressure ventilation (3), mouthpiece-assisted ventilation by day (2) and rocking bed (1). None have been weaned from support after a period of ventilation ranging from one month to 10 years. Eight patients have subsequently died.      

Adaptive differentiation of murine lymphocytes. IV. (Responder × Nonresponder) F(1) T cells can be taught to preferentially help nonresponder,rather than responder, B cells

Responses to the synthetic terpolymer L-glutamic acid, L-lysine, L-tyrosine (GLT) in the mouse are controlled by H-2-1inked Ir-GLTgenes. (Responder × nonresponder) F(1) hybrid mice, themselves phenotypic responders, can be primed with GLT to develop specific helper cells capable of interacting with 2,4-dinitrophenyl hapten (DNP)-primed F(1) B cells in response to DNP-GLT. Unlike the indiscriminant ability of F(1) helper T cells for conventional antigens (i.e. not Ir gene-controlled), which can help B cells of either parental type (as well as F(1)) equally well, GLT-primed F(1) T cells can only provide help under normal circumstances for B lymphocytes of responder parent origin; they are unable to communicate effectively with nonresponder parental B cells (1, and the present studies). The present studies reveal, however, that the induction of a parental cell-induced allogeneic effect during priming of F(1) mice to GLT actually dictates the direction of cooperating preference that will be displayed by such F(1) helper cells for B cells of one parental type or the other. Thus, F(1) T cells, primed to GLT under the influence of an allogeneic effect induced by parental BALB/c cells, develop into effective helpers for nonresponder A/J B cells, but fail to develop effective helpers for responder BALB/c B cells, and vice-versa. In contrast, F(1) T cells, primed to GLT under the influence of an allogeneic effect induced by either parental type, display significantly enhanced levels of helper activity for B cells derived from F(1) donors. These results are interpreted to reflect the existence of two interdependent events provoked by the allogeneic effect: one event augments the differentiation of GLT-specific helper T cells belonging to the subset corresponding to the opposite parental type; this would explain the development of increased helper activity provided to partner B cells of opposite parental type (as well as of F(1) origin). The second event, we postulate, involves the production of responses against the receptors which normally self-recognize native cell interaction determinants; this form of anti-idiotype response is restricted against self- recognizing receptors of the same parental type used for induction of the allogeneic effect, hence explaining diminished helper activity of such F(1) cells for partner B lymphocytes of corresponding parental type.     

种植牙——人类的第三副牙齿

研究显示,若全口无牙,人的全身机能将下降40％～60％,佩戴假牙之后,全身机能也只能恢复到正常的81％-95％．种植牙的诞生为彻底解决缺牙带给患者咀嚼功能和语言发音障碍,影响面容美观和心理伤害等长期困扰其健康和生活质量问题提供了一个重要出路．伴随技术的成熟,种植牙也因此被誉为继乳牙、恒牙之后,人类的第三副牙齿．它是生命科学和材料与工程科学完美结合所诞生的产物,是近半个世纪以来口腔医学献给人类健康的一份厚礼．种植牙作为天然牙齿的替代体,人工牙根可以和牙床完全融合,     

ANDROGENETIC ALOPECIA IN MEN: THE SCALE OF THE PROBLEM AND PROSPECTS FOR TREATMENT

While the precise incidence of androgenetic alopecia is unknown, it is universally acknowledged to be the most common hair problem in men. Balding is generally associated with ageing; consequently, the desire to prolong a youthful appearance inevitably leads to demands for effective treatments. Further, changing attitudes in modern society have resulted in people becoming concerned about their appearance and less tolerant about conditions that might be alleviated by medical intervention. The importance of hair loss upon quality of life has been underestimated by the medical profession. Clinicians failing to accept hair loss as an important medical problem ignore the real distress suffered by a significant proportion of those affected. New options for treatment that selectively target the metabolic pathways involved in the balding process are showing promise. The first generation of such drugs, Propecia, is now available in some countries and other molecules are currently under development.     

Positron emission tomography detected thyroid incidentaloma

We describe the case of a 48-year-old woman who presented with a thyroid lesion incidentally detected on positron emission tomography/computed tomography for a suspicious lung lesion. Subsequent clinical examination and investigations revealed a 3cm nodule in the left lower pole of the thyroid. Fine needle aspiration was indeterminate for malignancy. A left hemithyroidectomy was performed and histology confirmed a benign thyroid adenoma with an incidental micropapillary carcinoma. The literature regarding the best management for thyroid incidentalomas remains uncertain and, as such, each patient must be managed on an individual basis.     

Intense pulsed light vs. pulsed‐dye laser in the treatment of facial acne: a randomized split‐face trial

Background Various laser and light therapy have been increasingly used for the treatment of acne vulgaris. Patients and methods Twenty patients with facial acne were treated using intense pulsed light (IPL) on one side of the face and pulsed dye laser (PDL) on the other to compare the efficacy and safety of IPL and PDL. Treatment was performed 4 times at 2‐week intervals. Treatment effectiveness was determined using lesion counts, acne severity, patient subjective self‐assessments of improvement, and histopathological examinations, which included immunohistochemical staining for transforming growth factor‐β (TGF‐β). Results Numbers of total acne lesions decreased following both treatments. For inflammatory lesions such as papules, pustules and nodules, IPL‐treated sides showed an earlier and more profound improvement than PDL‐treated sides. However, at 8 weeks after the 4th treatment, a rebound aggravation of acne was observed on IPL‐treated sides. On the contrary, PDL produced gradual improvements during the treatment sessions and these improvements lasted 8 weeks after the 4th treatment. Non‐inflammatory lesions as open and closed comedones also showed improvement following both treatments and PDL‐treated sides showed better improvement as the study proceeded. Histopathological examinations showed amelioration in inflammatory reactions and an increase in TGF‐β expression after both treatments, which were more prominent for PDL‐treated sides. Conclusion Both PDL and IPL were found to treat acne effectively, but PDL showed a more sustained effect. TGF‐β might play a key role in the resolution of inflammatory acne lesions.     

Non‐ablative 1550‐nm erbium‐glass and ablative 10 600‐nm carbon dioxide fractional lasers for acne scars: a randomized split‐face study with blinded response evaluation

Background Non‐ablative 1550‐nm erbium‐doped fractional photothermolysis systems (FPS) and 10 600‐nm carbon dioxide fractional laser systems (CO₂ FS) have been effectively used to treat scars. Objective We compared the efficacy and safety of single‐session treatments of FPS and CO₂ FS for acne scars through a randomized, split‐face, evaluator‐blinded study. Methods Eight patients with acne scars were enrolled in this study. Half of each subject’s face was treated with FPS and the other half was treated with CO₂ FS. We used a quartile grading scale for evaluations. Results At 3 months after the treatment, the mean grade of improvement based on clinical assessment was 2.0 ± 0.5 for FPS and 2.5 ± 0.8 for CO₂ FS. On each side treated by FPS and CO₂ FS, the mean duration of post‐therapy crusting and scaling was 2.3 and 7.4 days respectively and that of post‐therapy erythema was 7.5 and 11.5 days respectively. The mean VAS pain score was 3.9 ± 2.0 with the FPS and 7.0 ± 2.0 with the CO₂ FS. Conclusion We demonstrated the efficacy and safety of single‐session acne scar treatment using FPS and CO₂ FS in East Asian patients. We believe that our study could be used as an essential reference when choosing laser modalities for scar treatment.     

3，4－二羟基苯乙酮对老年大鼠血小板膜磷脂成分的影响

ＤＨＡＰ１００ｍｇ·ｋｇ－１·ｄ－１，ｐｏ，持续１０周，可使老龄大鼠血小板膜磷脂（ＰＬ）含量增加，膜磷脂与膜胆固醇比值（ＰＬ／Ｃ）相对增高；血小板磷脂成分——磷脂酰乙醇胺（ＰＥ）和磷脂酰胆碱（ＰＣ）含量也有明显增加（ｐ＜０．０１），但血小板膜总脂及膜胆固醇含量无明显变化（Ｐ＞０．０５）。本实验结果提示：ＤＨＡＰ对血小板功能的影响可能与其抑制血小板膜磷脂降解、改变血小板膜组分有关。