ObjectiveTo evaluate the effect of leucocyte- and platelet-rich plasma (L-PRP) on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in treating avascular necrosis of the femoral head (ANFH) in rabbits. MethodsTwenty-four New Zealand white rabbits (4-6 months old, both genders, weighing 2.0-3.0 kg) were used for the establishment of bilateral ANFH models and divided into 4 groups (n=6). BMSCs were isolated from the bone marrow of iliac crest, cultured and identified. L-PRP was prepared by Landesberg method. Core decompression only (group A), core decompression and L-PRP implantation (group B), core decompression and BMSCs implantation (group C), and core decompression and implantation of BMSCs and L-PRP were performed in 4 groups. To evaluate bone formation and remodeling of the defects, X-ray photography was taken at 2, 4, and 8 weeks postoperatively. The modified Lane-Sandhu scoring system was used to evaluate the bone formation. Two rabbits were sacrificed at 2, 4, 8 weeks after operation to harvest the specimens for histological observation, new blood vessel count and new bone area ratio. ResultsThe observations of radiology and histology displayed different degrees of bone regeneration at bone defect sites in each group. At 2, 4, and 8 weeks postoperatively, the results of Lane-Sandhu X-ray photography scoring, new blood vessel count, and new bone area ratio showed that groups C and D were significantly better than groups A and B, group D was significantly better than group C. and group B was significantly better than group A (P<0.05). ConclusionThese findings demonstrate that L-PRP can promote osteogenic differentiation of BMSCs in treating ANFH in rabbits, and core decompression associated with BMSCs and L-PRP is an effective and feasible method to treat ANFH.
ObjectiveTo investigate the expressions of IL-10,tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in serum and lung tissue of COPD rats in order to elucidate the potential mechanism of airway inflammation. MethodsForty-five healthy adult male SD rats were randomly divided into a COPD model group (n=30) and a normal control group (n=15). The COPD rat model was established by intratracheal instillation of lipopolysaccharide (LPS) and exposure to cigarette smoke for 28 days. The concentrations of IL-10,TNF-α and IFN-γ in serum and lung tissue were measured by ELISA. ResultsTNF-α level of serum and lung tissue in the COPD model group increased significantly compared with the control group(P<0.05),while the levels of IFN-γ and IL-10 decreased significantly[serum:(44.68±8.67) ng/L vs. (75.96±10.59) ng/L;lung tissue:(64.55±9.03) ng/L vs. (94.06±8.71) ng/L,P<0.01]. The level of IL-10 in serum and lung tissue was negatively correlated with TNF-α (serum:r=-0.67,lung tissue:r=-0.80,P<0.01). The level of IL-10 in serum and lung tissue was positively correlated with IFN-γ (serum:r=0.64,lung tissue:r=0.72,P<0.01). The level of IL-10 in serum and lung tissue was negatively correlated with the percentage of neutrophils(serum:r=-0.70,lung tissue:r=-0.67,P<0.01). ConclusionIn COPD rats,down regulation of IL-10 plays an important role in regulation of airway inflammation.
目的 系统评价白细胞介素1β（IL-1β）与癫痫发作的相关性。 方法 2012年5月－2013年10月检索PubMed数据库（1985年1月－2013年7月）、Medline数据库（1985年1月－2013年7月）、中国知网（1998年1月－2013年7月）和万方数据库（1996年1月－2013年7月），并辅以谷歌学术搜索引擎进行手工检索。由两名研究者按照研究的纳入与排除标准进行病例对照研究的选择，对入选文献的质量参照“非随机对照临床试验质量评价标准与计分表”进行评价和计分，评估纳入研究的方法学质量并对有效的信息进行数据提取。然后采用RevMan 5.0统计软件进行Meta分析，以加权均数差（WMD）为效应指标。 结果 共纳入5个研究，包括82例癫痫发作的患者和471例正常对照。所纳入的研究未描述详细的抽样方法及其检测方法的特异性。Meta分析结果显示：癫痫发作72 h以内的患者与正常对照人群之间血浆IL-1β的浓度差异无统计学意义[WMD=0.04 pg/mL，95%CI（−0.07，0.16）pg/mL，P=0.46]。 结论 IL-1β可能没有直接参与癫痫的发作，而是通过与其他炎性因子之间的相互作用来实现。因此，需要进一步的研究来证明IL-1β在癫痫发作中的作用。
摘要:目的：探讨急性冠脉综合征患者血清IL-10/IL-6平及NFκB活性变化。方法：采用ELISA法检测45例急性冠脉综合征（ACS）患者,20例稳定性心绞痛（SAP）患者,20例非冠心病为对照者血清IL-10、IL-6水平；同时细胞免疫组化测定各组外周血单个核细胞NF-κB活性。结果: ACS组血清IL-6/IL-10 比值及NF-κB活性均高于SAP组及对照组(ACS: 1.69 ±0.53,0.32± 0.12;SAP: 1.06 ± 0.38,0.13 ±0.07;对照组: 0.92 ± 0.41,0.11±0.09, 均P＜0.05)。结论：炎症介质及抗炎症介质分泌失衡在急性冠脉综合征中发挥了重要作用。Abstract: Objective: To evaluate the clinical value of Theratio IL6/IL10Interleukin10(IL10)/ interleukin6(IL6) and Nuclear factorκappa B activation in acute coronary syndrome. Methods: Serum level of IL10, IL6 were measured for 45 cases patients of acute coronary syndrome (ACS), 20 cases patients of stable angina pectoris (SAP) and 20 cases patients without Coronary heart diseas（CHD）as control group by means of Enzyme linked immune absorption assay, while NFκB Activation measured by cell immunohistochemical method In Peripheral blood monouclear cell. Results： The ratio IL6/IL10 and Nuclear factorκappa B activation were significantly higher in patients with ACS (169±053,032±012) than in those of SAP (106±038,013±007) and Control (092±041,011±009)(Plt;005). Conclusion：there was Inflammatory imbalance between IL10 and IL6 in ACS, Inflammatory effects is important to devope to acute coronary syndrome
目的:比较镜检法和尿液分析仪检测白细胞的差异。 方法: 用两种方法测定900例尿液标本中的白细胞,并对其结果进行比较分析。结果:两种方法检测结果有明显差异(Plt;005),以镜检法为标准,尿液分析仪检测白细胞假阳性率为169%,假阴性率为158%。结论: 检测尿液白细胞时镜检法和尿液分析仪尽量结合应用,以减少假阴性率和假阳性率,保证检验结果的准确性。
目的:观察CCl4攻击小鼠致肝纤维化血清IL4、IL6、IL8的动态变化规律及其表达的影响。方法:将小鼠分为4组:雌、雄性小鼠对照组、雌、雄性小鼠体积分数为30%四氯化碳花生油组(皮下注射)。用ELISA及酶标仪方法检测小鼠血清IL4、IL6、IL8的含量及肝脏、脾脏的表达情况。结果: CCl4攻击可使小鼠血清IL6、IL8含量增加,IL4的表达进一步减少。结论:CCl4可能通过攻击小鼠IL4、IL6、IL8的表达参与抗炎反应过程,细胞因子在肝纤维化发病过程中起重要的作用。检测血清IL4、IL6、IL8浓度的变化可作为了解其与肝纤维化的关系。
Objective To construct recombinant adenovirus vector co-expressing human interleukin (hIL)-10 and green fluorescent protein (GFP) for study of the expression of genes of interest in vascular smooth muscle cells （VSMCs）. Methods hIL-10 cDNA was amplified from pUCm-T/hIL-10 cDNA using polymerase chain reaction (PCR), and cloned into shuttle plasmid pShuttle-IRES-hrGFP-1. Kanamycin resistance screeninged for recombinant plasmids, which were linealized with PmeⅠand transformed into BJ5183-AD-1 containing pAdEasy-1 by electroporation after determining the insert’s sequence correct by NotⅠ and XholⅠdigestion， sequencing and basic local alignment search tool (BLAST). Prepared recombinant adenovirus plasmids were transformed into XL10-Gold cells. Amplified plasmids were transfected to AD-293 cells for packaging after being linearized with PacⅠ. PCR was used to determine target gene; The titer of the recombinant adenovirus was measured. VSMCs were transfected by recombinant adenovirus and viewed under fluorescence microscope. hIL-10 concentration in transfected VSMCs supernant was measured by enzyme linked immune sorbent assay (ELISA). Results Recombinant shuttle plasmids contained interest gene. Recombinant adenovirus had 30 kb and 3 kb fragments after digestion with PacⅠ. PCR indicated that the recombinant adenovirus contained interest gene. The titer of recombinant adenovirus was 3×10１0 efu/ml. Transfected VSMCs had GFP expression and hIL-10 concentration in supernatant was 25 ng/106 cells. Conclusion The recombinant adenovirus co-expressing hIL-10 and GFP is successfully constructed and could effectively express in VSMCs, this lays the foundation for the gene therapy of vascular intimal hyperplasia.
Objective To construct a regulatable plasmid containing single chain fusion gene of murine interleukin-12 (mIL-12) which was regulated with mifepristone (RU486) and explore its expression in vitro. Methods The p40 and p35 subunit sequence of mIL-20 were respectively obtained from the plasmid GCp35Ep40PN by polymerase chain reaction (PCR) and they were cloned into pCA14 plasmid after introducing a linker by overlap PCR. The single chain mIL-12 gene was comfirmed by sequencing and subcloned into pRS-17 vector which contains RU486 regulator cassette. The positive clone named pRS-RUmIL-12 was identified by restriction endonuclease digestion and PCR. Lipofectamine 2000 was used to transfect the pRS-RUmIL-12 to HEK293 cells followed by manufacturer’s recommendations. The protein concentration of mIL-12 induced with RU486 in supernatant of the transfected HEK293 cells was measured by ELISA. Results The sequence of single chain mIL-12 what we obtained was the same as the expected result. The results of restriction endonuclease digestion and PCR showed that the RU486-inducible regulatory vector (pRS-RUmIL-12) was successfully constructed. No significant mIL-12 protein concentration in supernatant of HEK293 cells activation was measured without the inducer RU486, whereas higher concentration of the mIL-12 protein was observed in the presence of RU486. The relationship of concentration of the mIL-12 protein and RU486 was positive correlated under definite range. Conclusion A regulatable eukaryotic expression plasmid of mIL-12 single chain fusion gene was constructed, which could be used in the further research of gene regulation and gene therapy.
【Abstract】ObjectiveTo inquire into the regulative effect of interferon (IFN) on human leucocyte antigen-DR(HLADR) expression in human colonic cancer cells. MethodsThe expression of HLA-DR in 4 colonic cancer cell lines HCT-8，lovo，SW-480 and Ls-174-T was detected with immunohistochemical SP and ELISA method before and after being stimulated by different doses of α-IFN or γ-IFN. ResultsAll cell lines except lovo cell were HLA-DR positive before stimulation with α-IFN or γ-IFN. HLA-DR expression was enhanced on all of the 4 cell lines after stimulation, and it was correlated with γ-IFN and the dose of γ-IFN. The effect of γ-IFN was more obvious than that of α-IFN. ConclusionIFN can enhance HLA-DR expression in human colonci cancer cells and clinical therapy of IFN for colon cancer has a certain applying prospect.