低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

转化生长因子β1短发夹RNA对白蛋白刺激人肾近曲小管上皮细胞细胞因子表达的影响

目的 研究pcDU6载体质粒介导的转化生长因子β1（TGF-β1）短发夹RNA（shRNA）对人血清白蛋白（HSA）致人肾近曲小管上皮细胞（HK2细胞）增殖，TGF-β1、结缔组织生长因子（CTGF）和纤连蛋白（FN）表达的影响，并探讨有关机制。方法 构建TGF-β1shRNA的pcDU6载体质粒，体外培养HK2细胞株。采用脂质体转染将表达TGF-β1shRNA的pcDU6质粒载体（pcDU6-A1-A2和peDU6-B1-B2）分别导人实验组细胞。用HSA（5g／L）刺激HK2细胞12h或24h。四甲基偶氮唑盐（MTF）比色法测定细胞增殖水平。RT-PCR半定量分析HK2细胞中TGF-β1、CTGF和FNmRNA的表达水平；双抗夹心酶联免疫吸附法检测HK2细胞培养液中TGF-β1及FN蛋白质水平。结果 HK2细胞在HSA刺激下，其TGF-β1、CTGF及FNmRNA的表达明显上调，培养液中TGF-β1和FN的蛋白质含量亦明显升高（P〈0．05）。与pcDU6空载体组比较，pcDU6载体质粒介导的TGF-β1shRNA干扰组TGF-β1、CTGF及FNmRNA的表达明显下调（P〈0．05）。TGF-β1shRNA转染HK2细胞后12h或24h，细胞培养液中TGF-β1和FN蛋白质含量明显下降，HK2细胞增殖被部分抑制（P〈0．05）。在细胞增殖、TGF-β1、CTGF及FN基因表达方面，TGF-β1shRNA干扰组组间比较，以及pcDU6空载体转染组与HSA刺激组比较，差异均无统计学意义。结论 peDU6载体质粒介导的TGF-β1shRNA能够明显抑制HSA刺激下HK2细胞增殖、TGF-β1、CTGF和FN基因的表达。HSA刺激HK2细胞增殖及CTGF和FN基因的过表达可能通过TGF-β1介导。     

高糖对大鼠系膜细胞肝细胞生长因子受体表达的影响及其机制研究

目的 观察高糖作用下大鼠系膜细胞(MsC)肝细胞生长因子（HGF）受体c-Met的表达，并探讨其机制和意义。 方法 用RT-PCR和Western 印迹方法检测高糖作用大鼠MsC的不同时间点（0、12、24、48、96 h）c-Met的表达。分别用光辉霉素A（mithramycin A）和SU11274抑制转录因子Sp1的DNA结合活性和阻断c-Met。用电泳迁移率改变实验(EMSA)观察Sp1与c-Met基因启动子的结合活性。以荧光探剂二氯双氢荧光素二乙酸酯(DCFH-DA)捕获细胞内活性氧。 结果 大鼠MsC的c-Met表达在高糖作用12、24和48 h都明显上升，96 h开始下降。光辉霉素A呈浓度依赖性抑制高糖作用下大鼠MsC的c-Met表达上调。大鼠MsC内Sp1与c-Met基因启动子的结合活性在高糖作用下明显增强。HGF及c-Met显著抑制高糖诱导的大鼠MsC内活性氧的增多。 结论 高糖作用下大鼠MsC的c-Met表达增强，其机制可能是通过Sp1介导。HGF-c-Met信号通路激活能抑制高糖所致大鼠MsC内的氧化应激反应。     

生长板软骨细胞促进骨髓基质干细胞血管内皮生长因子的表达

目的观察骨髓基质干细胞(BMSCs)在生长板软骨细胞旁分泌作用下血管内皮生长因子(VEGF)的表达规律及其与成骨分化的相关性。方法大鼠BMSCs与生长板软骨细胞进行间接共培养,培养终末期做细胞化学染色,定量测定碱性磷酸酶(ALP)活性,用RT-PCR方法半定量检测VEGFmRNA的表达。结果生长板软骨细胞持续高表达VEGF。BMSCs随共培养时间的延长,ALP活性升高,BMSCs的VEGF的表达也逐渐增强。培养液加入两种分泌型VEGF中和抗体后,VEGF表达趋势不变,ALP活性仍为升高趋势,也不影响培养终末期钙化结节的形成。培养终末期BMSCs的CD31和CD34均阴性。结论BMSCs成骨分化过程中VEGF的表达符合成骨细胞分化基因的表达规律,与成骨细胞特征性基因的表达趋势一致,体外条件共培养条件下,中和VEGF后并不能阻碍BMSCs的成骨分化。     

脊髓血管畸形血管内皮生长因子在脑脊液的表达

目的探讨脊髓血管畸形是否存在血管内皮生长因子(VEGF)表达,及其表达与临床特征之间是否存在相关性。方法在手术中留取56例脊髓血管畸形的脑脊液标本,其中动静脉畸形(SCAVM)23例,硬脊膜动静脉瘘(SDAVF)19例,海绵状血管瘤(CM)14例;以我院12例蛛网膜下腔阻滞病人脑脊液作为对照组,分别通过酶联免疫吸附方法(ELISA)测定脊髓血管畸形脑脊液中VEGF含量。结果SCAVM脑脊液中VEGF浓度为(277±66)pg/ml、SDAVF为(318±54)pg/ml、CM为(146±32)pg/ml均高于正常对照组(123±27)pg/ml。术前栓塞和出血均可导致脑脊液VEGF升高。结论VEGF与脊髓血管畸形的发生、发展存在一定关系,出血和栓塞均可影响VEGF的表达。     

Cigarette smoke toxicants alter growth and survival of cultured mammalian cells.

Our purpose was to determine the effects of six cigarette toxicants (pyridine, nicotine, 2-ethylpyridine, 3-ethylpyridine, p-cresol, and pyrazine) on three types of cultured mammalian cells (human umbilical vein endothelial cells [HUVECs], human microvascular endothelial cells [HMVECs], and NIH 3T3 cells) using a cell proliferation/survival assay. Synchronized cells were cultured in proliferation or survival medium containing various doses (10(-18)M-10(-2)M) of the tested chemicals. After 48 h, cells were counted using a hemacytometer. The no observable adverse effect level (NOAEL), lowest observable adverse effect level (LOAEL), and the efficacy were determined for each compound in the cell proliferation and survival assays. Pyridine and p-cresol did not show significant effects with any cell types, except at high doses. Derivitization of the pyridine ring altered its potency, especially when an ethyl group or nitrogen was added. In survival medium, nicotine stimulated proliferation of all three cell types at doses found in smoker's serum (10(-8)M-10(-7)M). For HUVEC and HMVEC, 2-ethylpyridine, 3-ethylpyridine, and pyrazine inhibited proliferation in proliferation medium and induced cell death in survival medium at attomolar and femtomolar doses. All chemicals, except pyridine and pyrazine, stimulated NIH 3T3 cell proliferation at low doses and induced cell death at high doses. LOAELs and efficacies revealed that endothelial cells from a developing organ (umbilical cord) were more sensitive to these chemicals than endothelial cells from an adult organ (lung). 3-Ethylpyridine and pyrazine, which induced cell death at low doses, are added to consumer products and should be subjected to further toxicological testing.     

Female patients complaining about hair loss: documentation of defective scalp hair dynamics with contrast-enhanced phototrichogram

BACKGROUND: The complaint of chronic hair loss frequently affects female subjects and there is little or no objective technology available in the general dermatology or even in the hair clinics to guide the observer in the management of the patient. The purpose of this report is to share the results of refined hair growth measurements that were collected in 92 female subjects complaining about hair loss. METHODS: Clinically they were classified as having a patterned hair loss according to Ludwig (L; n=50), diffuse hair loss (D; n=13) or no visible hair loss but complaining of hair shedding (N; n=29). Two scalp sites on the top of the head and one occipital site were investigated after clipping by close-up photography before and after a hair dye (contrast enhancement, CE). Forty-eight hours later a new photograph was taken after CE in view of phototrichogram analysis (CE-PTG). Finally a last hair clip was performed 30 days later and hair thickness and length determined for linear growth measurements (LHGR). RESULTS: Herein we confirm that the top of the head shows usually a higher hair density than occipital sites, a physiological observation that applies both to men and women. From the technological perspective, we also document that CE improves hair detection in all sites. Interestingly, in affected patients (L and D) the relative increase of hair counts after CE was much higher (range +22.4% to +28.3%) compared with apparently unaffected females (N; range +8.2% to +9.7%). This increase in hair counts was only due in part to the presence of less pigmented thinning hair (thickness less than 40 microm). Such thin hairs were found in statistically significantly higher proportions in younger patients with mildly severe (grade I) patterned alopecia (Ludwig: L). In other patients with hair loss and in more severe forms of patterned alopecia - especially in older patients - the thin hair is not detected in abnormal proportions. In all sites slower growth rates and decreased anagen percentages indicate a defective hair replacement programme distinguishing L patterns from diffuse hair loss and from apparently unaffected patients complaining of chronic hair loss. Globally, we also noted that increasing age is associated with significant regression of scalp hair (decreased hair counts, thinner hair and slower LHGR). CONCLUSION: On the basis of the present data together with female data from the literature and our own studies in male subjects, we suggest a three-step mechanism leading to hair loss 1.Shortening of growth phase the hair cycle with maintained thick hair, i.e. more frequent hair cycling that leads to more hair shedding. 2.Intermittent production of short thin hair, i.e. morphological evidence of miniaturisation. 3.Very occasional or almost no hair production, i.e. dormant follicles or irreversible follicular atrophy. Depending on the genetic background, hormonal microenvironment in the scalp and conditioning of individual hair follicle bio-responses, female and male patterned hair loss may end up into different phenotypes.     

Effect of Kang''''ai Injection(康艾注射液) on Serum Level of Soluble Interleukin-2 Receptor and Vascular Endothelial Growth Factor in Patients with Esophageal Carcinoma during Radiotherapy

Radiotherapy(RT)is an important ap-proach for treatment of esophageal carcinoma(EC),especially for patients who have missedthe chance of surgical section.The efficacy ofRTis not satisfactory for its1-year,3-year and5-year survival rates being merely50%,20%,and10%respectively(1).Researches in recent years showed thatbetter efficacy can be obtained by combiningRTwith Chinese herbal medicines for EC thatincapable to be resected.Kang ai Injection(康艾注射液,KAI)is a Chinese herbal prep-arati…     

G蛋白耦联受体激酶结合蛋白1通过调节细胞外调节激酶1/2的活性促进成骨细胞迁移

目的探索G蛋白耦联受体激酶结合蛋白1(GITI)在成骨细胞迁移中的作用,并分析其机理。方法通过Western blot方法检测GIT1蛋白在鼠的成骨细胞内的表达；用免疫荧光染色方法确定：在血小板衍生生长因子(PDGF)不刺激和刺激的条件下,GIT1和细胞外调节激酶1/2(ERK1/2)在成骨细胞内的位置；用共同免疫沉淀的方法测定GIT1和ERK1/2相互结合,并且用免疫荧光双染的方法确定这两种蛋白相互结合的位置；用包含GIT1-RNA发夹结构的腺病毒感染成骨细胞后,用免疫荧光染色方法确定磷酸化ERK1/2(pERK1/2)在成骨细胞内的位置,用划痕愈合法检测在PDGF刺激下的迁移能力。结果在成骨细胞内,PDGF刺激导致了GIT1和ERK1/2的相互结合,并且这种结合发生在成骨细胞的局部粘附内。包含GIT1-RNA发夹结构的腺病毒明显抑制了pERK1/2招募至成骨细胞局部粘附内以及PDGF所刺激的成骨细胞的迁移。结论在PDGF刺激下,GIT1招募pERK1/2至成骨细胞的局部粘附内,从而促进成骨细胞的迁移。     

血管内皮生长因子与妊娠高血压综合征发病的关系

目的　探讨血管内皮生长因子 (VEGF)在妊娠高血压综合征发病中的作用。方法　分别采用酶联免疫吸附试验检测 40例妊高征孕妇的血清VEGF水平 ,免疫组化检测胎盘及蜕膜组织VEGF及CD3 4表达情况 ,35例正常孕妇作对照。结果　①妊高征组孕妇的外周血VEGF水平及胎盘组织MVD明显低于正常妊娠组 (P <0 0 5 ) ;②两组胎盘绒毛滋养叶细胞和蜕膜组织中均有VEGF阳性表达 ,胎盘组织强阳性表达高于蜕膜。与对照组比较 ,其胎盘组织VEGF强阳性表达的轻度妊高征无显著性差异 ;而中度和重度妊高征与对照组相比 ,则明显降低 ,差异显著 (P <0 0 5 )。各组孕妇蜕膜组织中VEGF的表达强度和差异无显著性。③孕妇外周血VEGF水平与新生儿出生体重 (r =0 2 9,P <0 0 5 )和胎盘重量 (r =0 34,P <0 0 1)均存在直线正相关关系。结论　妊高征患者血清VEGF水平和胎盘组织MVD降低 ,胎盘组织VEGF表达明显下降 ,都可能在妊高征的发病中起一定的作用。