大鼠退变椎间盘中环氧化酶-2、缺氧诱导因子-1α的表达及其与血管生成的关系

目的探讨环氧化酶-2(COX-2)与缺氧诱导因子-1α(HIF-1α)在退变的大鼠椎间盘的表达相关性,以及它们与椎间盘内新生毛细血管生成的相关性。方法选取3个月龄的大白鼠20只,随机分为实验组与对照组。将实验组10只大鼠手术切除腰1-骶1棘突、关节突,棘上、棘间韧带,切断双侧竖棘肌,然后缝合切口。对照组不行手术。12周后,处死所有大鼠,立即取L4-5椎间盘。通过免疫组化方法检测两组实验动物椎间盘中COX-2、HIF-1α表达情况及微血管的密度(MVD)。并通过统计分析描述COX-2与HIF-1α之间的关系及它们分别与椎间盘微血管密度(MVD)之间的关系。结果实验组的髓核中HIF-1α、COX-2阳性细胞率明显增高,在对照组的髓核组织中极少出现阳性细胞。此外,实验组椎间盘中的微血管密度明显增加。用直线相关分析表明,实验组动物椎间盘髓核组织内HIF-1α、COX-2之间呈显著正相关关系(P<0.05,r=0.678)。经多元线性回归分析实验组椎间盘髓核组织中MVD值与HIF-1α的表达之间和MVD与COX-2的表达之间具有相关性(P均<0.05)。结论 1.大鼠椎间盘退变过程中HIF-1α、COX-2的表达有相互促进作用;2.退变椎间盘的MVD值与HIF-1α及COX-2的高表达具有相关关系。     

大鼠烧伤后库普弗细胞在促炎细胞因子产生中的作用

目的　观察大鼠严重烧伤后早期 ,库普弗细胞在肿瘤坏死因子α(TNFα)、白细胞介素(IL) 1β、IL 6产生中的作用。方法　观察 (1)烧伤血清对体外培养的大鼠库普弗细胞分泌TNFα、IL 1β、IL 6的刺激作用 ;(2 )烧伤后大鼠库普弗细胞的细胞因子mRNA表达变化 ;(3)应用库普弗细胞特异性抑制剂三氯化钆后 ,烧伤大鼠血浆内细胞因子含量变化。　结果　烧伤血清能刺激库普弗细胞释放TNFα、IL 1β、IL 6 ;大鼠烧伤后库普弗细胞TNFα、IL 1β、IL 6mRNA表达量显著升高 ;预先抑制库普弗细胞的活性 ,烧伤后血浆TNFα、IL 1β、IL 6水平均显著降低 ,分别为烧伤组的 34.71%、36 99%、33.70 %。结论　库普弗细胞是大鼠烧伤后血浆中TNFα、IL 1β、IL 6的主要来源     

Novel mutations in response to vitamin B6 in primary hyperoxaluria type 1 after only kidney transplantation: a case report

Recently, the mainstream curative treatment for primary hyperoxaluria type 1 (PH1) is combined liver and kidney transplantation, and only kidney transplantation is considered ineffective for most PH1 patients. Furthermore, vitamin B6 (B6) is the only permitted drug available for treatment. However, except for specific mutations such as G170R and F152I in gene AGXT, data of B6 effect on other mutations are lacking. Insufficient research has evaluated the efficacy of the combination of kidney transplantation and B6 treatment in the therapeutic strategy in PH1 patients. Here, we report a case of a 52-year-old male with frequent stone events and end-stage renal diseases (ESRD), and subsequently undergone kidney transplantation. Sudden rising of serum creatinine within two months after the transplantation. After gene sequencing, the mutations of A186V, R197Q, and I340M were presented in gene AGXT. Therefore, the patient was diagnosed with PH1. B6 administration was attempted during the period of waiting for liver transplantation. Four-week oral B6 therapy (50 mg tid) reduced the serum creatinine of the patient from 194 to 145 µmol/L, which revealed that the patient probably responded to B6 treatment. At the almost three-year follow-up, the patient’s serum creatinine remained reduced (130 µmol/L), without urinary oxalate excretion. In this case, we established a positive effect, even a beneficial result, of the use of B6 as a retrospective therapeutic choice in PH1 treatment after kidney transplantation.     

TNF-α McAb对急性失血性休克大鼠肝脏的作用

目的探讨肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)在急性失血性休克大鼠肝损伤中的作用及TNF-α单克隆抗体的保护作用。方法将24只雄性SD大鼠随机均分为三组:对照组(A组)、休克+乳酸林格氏液复苏组(B组)和休克+TNF-α单克隆抗体复苏组(C组)。B和C组大鼠通过股动脉放血,制作急性失血性休克动物模型;B组用乳酸林格氏液复苏,C组用含TNF-α单克隆抗体(3mg/kg)的乳酸林格氏液复苏,而A组在同等条件下不进行失血。分别检测各组大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、TNF-α水平和肝组织中MDA、SOD含量,并在光镜和电镜下观察各组大鼠肝组织的病理变化。结果 B、C组大鼠血清ALT、AST、TNF-α水平和肝组织中MDA含量较A组升高,SOD含量降低;C组ALT(343.63±35.61)U/L、AST(748.75±49.76)U/L、TNF-α(99.38±13.16)pg/mL、丙二醛(26.33±1.30)nmol/mgProt较B组ALT、AST、TNF-α、MDA含量降低,C组肝组织中SOD含量(510.14±47.44)U/mgProt较B组升高;在光镜、电镜下观察,C组肝组织损伤较B组减轻。结论 TNF-α可能是急性失血性休克肝损伤的重要因子之一,使用TNF-αMcAb复苏可以减轻失血性休克时大鼠肝损伤,具有一定的保护作用。     

Premature senescence and cellular phenotype transformation of mesangial cells induced by TGF-B1

Abstract

Background: Transforming growth factor-β1 (TGF-β1) is a polypeptide member of the transforming growth factor β superfamily of cytokines and performs many cellular functions. Its overexpression may lead to renal fibrosis. Aim: This study planed to investigate the effects of TGF-β1 on the cell cycle and phenotype of mesangial cells. Methods: Rat mesangial cells were cultured together with different concentrations (0, 1, 2, 5, and 10?ng/mL) of TGF-β1 for specified times from 0?min to 72?h. 0?ng/mL TGF-β1 and 0?min served as controls. Cell cycles were assessed by flow cytometry and α-smooth muscle actin expression (α-SMA) protein expression by western blot analysis. All data were presented as Mean?±?SD. Statistical analysis was performed by using one-way analysis of variance and correlation analysis. Results were considered significant at p?<?0.05. Results: After 15?min of co-culture with different concentrations of TGF-β1, the percentage of mesangial cells in G0/G1 phase was significantly elevated compared to the control (p?<?0.05). 12?h co-culture induced cell hyperplasia, 24?h co-culture obvious up-regulation of α-SMA (p?<?0.01) and one or two cells’ myofibroblast phenotype transition, and 36?h co-culture several cells’ phenotype transition. Correlation analysis prompted that the TGF-β1-induced premature aging was time-dependent (p?<?0.01). Conclusion: TGF-β1 may induce mesangial cells’ premature senescence and myofibroblast-like phenotype transformation time-dependently, which may contribute to the development of early stage of glomerulosclerosis.     

Beneficial Effect of Pentoxifylline on Cisplatin-Induced Acute Renal Failure in Rabbits

Pentoxifylline (PTX) has been reported to inhibit TNF-α production and prevent several types of acute renal failure. This study was undertaken to determine the effect of PTX on the cisplatin-induced acute renal failure in rabbits. Rabbits received a single injection of cisplatin (5 mg/kg, i.p.) with or without PTX pretreatment (30 mg/kg, i.v.). Alterations in renal function, apoptotic cell death, and TNF-α mRNA expression were measured at 24 or 48 h after cisplatin injection. Cisplatin caused an increase in BUN and serum creatinine levels, a reduction in GFR, and an increase in fractional Na⁺ excretion. Such changes were significantly attenuated by PTX pretreatment (30 mg/kg, i.p.) 30 min before and 24 h after cisplatin injection. Morphological evaluation showed that cisplatin injection induced diffuse proximal tubular necrosis and the effect was reduced by PTX pretreatment. Cisplatin induced apoptotic cell death in renal cortex and the effect was significantly prevented by PTX. Treatment of opossum kidney cells with cisplatin resulted in cell death, which was significantly prevented by PTX. The increase in lipid peroxidation and the decrease in renal blood flow induced by cisplatin were not affected by PTX. The expression of TNF-α mRNA was increased after cisplatin injection and the effect was inhibited by PTX pretreatment. These results suggest that cisplatin-induced acute renal failure in rabbits is associated with an induction of TNF-α-mediated apoptosis, and that PTX may exert a protective effect against cisplatin nephrotoxicity by inhibiting TNF-α production.     

地拉罗司对大鼠窄蒂缺氧皮瓣模型促进微血管生成的实验

目的 探索术前应用地拉罗司(deferasirox)对大鼠乒乓拍样狭长窄蒂缺氧皮瓣模型微血管生成的促进作用及与用药时间的相关性.方法 成年雄性SD大鼠40只,随机分为对照组(8只),实验组(32只,分为1、3、5、7d组,每组8只).实验组:地拉罗司100 mg/kg分别连续灌胃1、3、5、7d;对照组:生理盐水100 mg/kg连续灌胃7d,灌胃结束后,次日行乒乓拍样窄蒂任意型皮瓣成形术,均于术后第7天取材,应用免疫组织化学、Western印迹及实时荧光定量聚合酶链式反应(PCR)法检测CD34表达,应用实时荧光定量PCR检测低氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)的表达.结果 实验组较对照组皮瓣成活率明显提高,随着用药时间的增加,皮瓣微血管密度(MVD)、HIF-1α和VEGF均逐渐增高,7d达最高.结论 地拉罗司能诱导HIF-1α的分泌,促进VEGF产生,增加MVD表达,明显具有提高皮瓣对低氧的耐受,减少内皮细胞低氧后的坏死率,对缺血缺氧的窄蒂皮瓣具有较好的保护作用.     

Pulsed electromagnetic fields rapidly modulate intracellular signaling events in osteoblastic cells: comparison to parathyroid hormone and insulin.

Pulsed electromagnetic field (PEMF) devices are approved for the healing of bone nonunions, but there is a lack of understanding as to their mechanism of action at the cell and molecular level. Intermittent parathyroid hormone (PTH) therapy is currently utilized for treatment of osteoporosis, and is also being investigated for the purpose of augmenting fracture healing. Insulin and IGF-1 are also thought to play important anabolic roles in osteogenesis. In this report, signaling pathways activated by acute PTH or insulin treatments were compared to those activated by PEMF treatment in osteoblast-like cells. Some signaling molecules like the extracellular response kinases 1/2 (Erk1/2) and the cAMP response element binding protein (CREB) were activated by insulin and PTH, respectively, but not by PEMF treatment. Other signaling molecules like the insulin receptor substrate-1 (IRS-1), the S6 ribosomal subunit kinase, and the endothelial nitric oxide synthase (eNOS) were phosphorylated by PTH, insulin, and PEMF to the same relative extent and within the same time frame. IRS-1, eNOS, and S6 have been implicated in bone anabolism, and our results suggest that the anabolic effects of PEMF may be mediated, in part, through the activation of these proteins.     

去甲斑蝥素阻断αvβ6-ERK-ETS1信号通路抑制结肠癌细胞上皮-间质转化实验研究

目的:探讨去甲斑蝥素抑制结肠癌细胞上皮-间质转化的分子机制,为中药抗癌机制的研究奠定基础。方法:将HT-29结肠癌细胞经NCTD处理后,光镜下观察细胞形态学变化;流式细胞术检测处理前后细胞表面上皮表型及间质表型标志物的变化情况;Western Blotting分析常见调控细胞EMT过程信号传导通路中关键分子及其磷酸化状态的变化;转录因子活化技术观察核因子磷酸化状态的变化。结果:显微镜下观察发现,经NCTD处理后,结肠癌细胞EMT过程受到抑制;流式细胞术证实肿瘤细胞整合素αvβ6表达降低,同时上皮表型标志物表达升高,间质表型标志分子表达减少;Western Blotting分析常见调控细胞EMT过程信号传导通路中仅有ERK及p-ERK表达水平降低,其他关键蛋白无明显变化;转录因子活化技术发现利用si RNA技术干扰整合素αvβ6后,ERK下游的核转录因子中仅有ETS-1磷酸化水平改变,Western Blotting分析证实经NCTD处理后出现与干扰αvβ6表达造成的一致性改变。结论:去甲斑蝥素通过阻断αvβ6-ERK-ETS1信号通路抑制结肠癌细胞上皮-间质转化过程,从而发挥抗癌作用。     

辛伐他汀对大鼠肾缺血再灌注损伤中Wnt/JNK影响的研究

目的:探讨辛伐他汀(simvastatin SIM)对大鼠肾缺血再灌注损伤中Wnt/JNK的影响。方法:雄性wistar大鼠36只随机分为:假手术组、手术组、SIM干预组,每组12只,再将每组按取材时间不同分为6 h、48 h两个时相组。手术方法:假手术组开腹分离双侧肾动脉,但不夹闭,后关闭腹腔;手术组游离双侧肾动脉,动脉夹夹闭45 min后恢复灌注建立肾IRI模型;SIM干预组术前1周给大鼠灌胃SIM 20 mg.kg-1.d-1,其余处理同手术组。肾IRI后6 h、48 h,检测血清Scr值,肾组织行病理学观察及免疫组织化学法测定Wnt4、JNK1及其信号通路下游炎症因子TNFα的表达,并采用real time PCR法测定肾组织Wnt4 mRNA、JNK1 mRNA的表达。结果:肾组织病理学观察:假手术组肾组织形态结构基本正常;手术组肾小管上皮细胞有不同程度的肿胀、坏死和脱落、排列紊乱,管腔扩张,肾间质水肿、炎细胞浸润明显,以IRI后6 h为著;SIM干预组肾组织损伤程度较手术组明显减轻。Scr值测定:手术组Scr值较假手术组明显升高(P<0.01);SIM干预组Scr值较手术组有所下降(P<0.01);以IRI后6 h,Scr值较高(P<0.01)。免疫组化检测:假手术组肾小管上皮细胞Wnt4、JNK1、TNFα表达较弱;手术组IRI后肾小管上皮细胞胞浆内Wnt4、JNK1、TNFα表达均比假手术组增强(P<0.01),以6 h时为著(P<0.01)。SIM干预组Wnt4、JNK1、TNFα表达水平较手术组减弱(P<0.05)。real time PCR检测:假手术组肾小管上皮细胞Wnt4 mRNA、JNK1 mRNA表达较弱;手术组肾小管上皮细胞Wnt4 mRNA、JNK1 mRNA表达均较假手术组增强(P<0.01),以6 h为著(P<0.01);SIM干预组Wnt4 mRNA、JNK1 mRNA表达比手术组均有所下降(P<0.01)。相关性分析:JNK1与Wnt4、TNFα均成正相关(rs=0.94,rs=0.95);Wnt4 mRNA、JNK1 mRNA成正相关(rs=0.75);且均与Scr成正相关(rs分别为rs=0.93,rs=0.96,rs=0.94,rs=0.70,rs=0.35)。结论:辛伐他汀预处理可能通过抑制Wnt/JNK信号通路的表达,从而减轻肾IRI程度。     

结直肠癌患者手术前后血清中IL-6和NO水平的临床意义

目的探讨结直肠癌患者手术治疗前后血清中IL-6和NO水平变化及其临床意义。方法对40例结直肠癌患者分别采用酶联免疫分析法和化学比色法测定手术前后血清中的IL-6和NO水平,并与10名正常健康人作比较。结果结直肠癌患者手术前血清中IL-6水平高于正常人（P〈0.05）,NO水平明显低于正常人（P〈0.01）;经手术治疗2周后,患者血清IL-6水平较治疗前降低（P〈0.05）,而NO水平明显升高（P〈0.05）。结论结直肠癌患者血清中IL-6和NO水平的变化,对病情和预后判断具有重要的临床意义。     

A型肉毒毒素治疗挛缩性瘢痕

目的 探索A型肉毒毒素(botulinum toxin type A,BTXA)治疗挛缩性瘢痕的疗效.方法 选取26例挛缩性瘢痕患者,随机分为A型肉毒毒素组(BTXA组)和曲安奈德组(TAC组,对照组),注射药物治疗前测量各组患者瘢痕长轴长度,并于注射后再次测量其长度,1次/月,共6次,通过比较治疗前后差值评价药物疗效.切取各组瘢痕组织行免疫组织化学检测,观察α平滑肌肌动蛋白(α-SMA)及肌球蛋白-Ⅱ的表达情况.结果 药物作用1个月后,BTXA组较TAC组瘢痕挛缩程度明显减轻(P＜0.05),尤以6个月时差异最明显,BTXA组和TAC组瘢痕长轴长度差值分别为(1.23±0.42) cm和(0.56±0.33) cm.免疫组织化学结果显示,BTXA组瘢痕内α-SMA及肌球蛋白-Ⅱ表达较TAC组明显减少(P＜0.05).结论 A型肉毒毒素治疗挛缩性瘢痕操作简单、效果明显,值得推广应用.     

血清6-keto-PGE1α水平对监测盘源性腰痛的临床价值

目的对血清前列腺素E2（PGE2）代谢产物6-keto-PGE1α与退变椎间盘渗透性改变的相关性进行研究，探讨PGE2作为反映盘源性腰痛的特异性炎症指标的可能性。方法①收集20例盘源性腰痛分为治疗前和治疗后两组，分别测定两组血清标本前列腺素E2水平。②采用同期健康献向者10名，作为健康对照组。结果A组（盘源性腰痛患者治疗前）53．8470±13．763pg／ml，B组（盘源性腰痛患者治疗后）46．1025＋5．516pg／ml，A组明显高于B组及c组，差异具有显著性（P〈0．05）。对A、B两组资料进行配对t检验发现t=2．875，经双尾检验P〈0．05，盘源性腰痛患者治疗前后差异具有显著性意义。结论①前列腺素E2与盘源性腰痛炎症机制密切相关。②监测血清6-keto-PGE1α能很好的反映出椎间盘内的渗透性变化和盘内炎症水平。     

Glucose intolerance and hepatocellular carcinoma: recent findings for old diseases

In the last years, an increasing number of evidences on the influence of metabolic syndrome on the occurrence of hepatocellular carcinoma (HCC) have been developed. Type 2 mellitus diabetes (T2MD) has been found to increase the occurrence of primary liver tumors and to define a more aggressive carcinogenetic process. Furthermore, several preclinical and observational studies and a recent meta-analysis have shown that anti-diabetic drugs can modify the risk of HCC development in patients with T2DM. However, despite these evidences, underlying molecular mechanisms linking both pathological conditions have to be completely cleared yet. The study published by Gao et al. has found a possible molecular link between the two conditions, describing the predisposition to T2DM and HCC given by the haploinsufficiency of nuclear receptor coactivator 5 (NCOA5) in murine models. The authors have generated Ncoa5+/– (haploinsufficient) male mice and shown that 94% of male mutant mice developed HCC within 18 months of age, this in contrast with Ncoa5+/+ and Ncoa5+/– female mice. These results suggest that NCOA5 haploinsufficiency is linked to HCC development in male mice. Moreover, mutant male mice showed significantly elevated levels of fasting blood glucose and markedly decreased glucose tolerance and insulin sensitivity compared to Ncoa5+/+ littermates. This well-constructed work sheds light on the molecular link between T2DM and HCC and opens the way to further biological and clinical studies in the field of liver tumor prevention and treatment.     

Synergistic action of inflammation and lipid dysmetabolism on kidney damage in rats

In kidney disease, inflammation and lipid dysmetabolism are often associated together, however, the effect and mechanism of inflammatory mediators and lipid dysmetabolism on kidney damage is still unclear. In this study, Wistar rats were randomized into four groups: normal diet?+?saline (Group N), high-fat diet (HF)+?saline (Group HF), normal diet?+?adriamycin (Group ADR), HF?+?adriamycin (Group ADR?+?HF). After 10?weeks of feeding, rats in each group were randomly sacrificed. We found that the protein content of urine in ADR and ADR?+?HF groups were significantly higher than that of group N and HF while the serum levels of total protein and albumin in the ADR and ADR?+?HF groups decreased correspondingly. The serum levels of triglyceride, total cholesterol and low-density lipoprotein in the HF, ADR and ADR?+?HF groups increased. In the treatment groups, mesangial proliferation, matrix accumulation, tubular vacuolization, inflammatory cell infiltration and fat deposition were detected. These pathological changes were the most serious in the ADR?+?HF group. The expression of tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) were increased in each treatment group, especially in the ADR?+?HF group. Our results suggested that the inflammatory factors and abnormal lipid levels can activate the inflammatory response in kidney of the Wistar rats, and lead to a series of pathological changes in renal tissue, and inflammatory factors and lipid dysmetabolism can aggravate damage in the kidney.