云南拉祜族六个基因座的遗传多态性研究

目的：了解云南拉祜族６ 个短串联重复序列基因座遗传多态性分布。方法：应用复合扩增技术对ＣＳＦ１ＰＯ、ＴＰＯＸ、ＴＨ０１ 、Ｆ１３Ａ０１、ＦＥＳＦＰＳ和ｖＷＡ等６ 个ＳＴＲ基因座进行分析,采用高分辨率的聚丙烯酰胺凝胶电泳分离、银染法显影技术,对云南拉祜族上述６ 个ＳＴＲ基因座进行遗传多态性调查。结果：ＣＳＦ１ＰＯ基因座,观察到８ 个等位基因,２０ 个基因型;ＴＰＯＸ 基因座,观察到５ 个等位基因,１４ 个基因型;ＴＨ０１ 基因座,观察到６ 个等位基因,１７ 个基因型;Ｆ１３Ａ０１ 基因座,观察到６ 个等位基因,１３ 个基因型;ＦＥＳＦＰＳ基因座,观察到７ 个等位基因,１５ 个基因型;ｖＷＡ基因座,观察到７ 个等位基因,１７ 个基因型。结论：上述６ 个短串联重复序列基因座基因频率分布与Ｈａｒｄｙ Ｗｅｉｎｂｅｒｇ 平衡吻合良好。     

Peripheral blood CD45RO+T cells is a predictor of the effectiveness of neoadjuvant chemoradiotherapy in locally advanced rectal cancer

To investigate the relationship between the changes in circulating CD45RO⁺T lymphocyte subsets following neoadjuvant therapy for rectal cancer in patients with locally advanced rectal cancer.The clinicopathological data of 185 patients with rectal cancer who received neoadjuvant therapy in the General Surgery Department of Beijing Chaoyang Hospital affiliated to Capital Medical University from June 2015 to June 2017 were analyzed. Venous blood samples were collected 1 week before neoadjuvant therapy and 1 week before surgery, and the expression of CD45RO⁺T was detected by flow cytometry. The receiver operating characteristic curve analysis was used to determine the optimal cut-off point of CD45RO⁺ratio. Log-rank test and multivariate Cox regression were used to analyze the overall survival rate (OS) and disease-free survival rate (DFS) associated with CD45RO⁺ratio.Circulating CD45RO⁺ratio of 1.07 was determined as the optimal cut-off point and CD45RO⁺ratio-high was associated with lower tumor regression grade grading (P = .031), T stage (P = .001), and tumor node metastasis (TNM) stage (P = .012). The 3-year DFS and OS rate in the CD45RO⁺ratio-high group was significantly higher than that in the CD45RO⁺ratio-low group (89.2% vs 60.1%, P<.001; 94.4% vs 73.2%, P<.001). The multivariate Cox analysis revealed that elevated CD45RO⁺ratio was an independent factor for better DFS (OR, 0.339; 95% CI, 0.153–0.752; P = .008) and OS (OR, 0.244; 95% CI,0.082–0.726; P = .011).Circulating CD45RO⁺ratio could predict the tumor regression grade of neoadjuvant therapy for rectal cancer, as well as long-term prognosis. These findings could be used to stratify patients and develop alternative strategies for adjuvant therapy.     

依达拉奉治疗急性脑出血疗效观察及对血清细胞因子水平的影响

目的探讨依达拉奉注射液对急性脑出血患者神经功能及血液细胞因子IL-1、IL-8、TNF-α水平的影响。方法对126例首次发病且于起病24 h内入院的急性脑出血患者分为治疗组及对照组,对照组予常规治疗,治疗组在对照组治疗的基础上加用依达拉奉。2组均于入院时、入院第7天、第14天采外周血测IL-1、IL-8、TNF-α,同时神经功能评分,并进行对比分析。结果 2组患者血清细胞因子IL-1,IL-8,TNF-α浓度均随病程逐渐增高,但治疗组升高幅度低于对照组,于14 d时下降幅度明显大于对照组(P<0.05)。治疗后2组神经功能缺损评分均降低,与治疗前比较均有显著性差异(P均<0.05),且治疗组明显优于对照组(P<0.05)。结论依达拉奉可以改善患者神经功能,通过减轻细胞因子介导的炎症反应对脑组织实施保护作用。     

孟如治疗系统性红斑狼疮诊疗思路

系统性红斑狼疮（SLE）是一种自身免疫性慢性炎症崔结缔组织病,根据其临床表现,属中医痹病、阴阳毒、血证、水肿、丹疹、虚劳等症范畴。SLE西医治疗主要运用激素、免疫抑制剂、抗疟药、抗组织排异药物等。由于激素治疗的副作用,以及SLE难于治愈的情况,临床积极探讨中医药治疗方法。中医辨证分型主要有以下几型：热毒炽盛,阴虚内热,气阴两伤,风湿热痹,邪热伤肝,脾肾两虚。     

Adding Three Cycles of CAPOX after Neoadjuvant Chemoradiotherapy Increases the Rates of Complete Response for Locally Advanced Rectal Cancer

Background and Objectives: the total neoadjuvant chemoradiotherapy (TNT) includes different strategies, but the most appropriate model remains uncertain. The purpose of this retrospectively study was to evaluate the safety and pathological response in the consolidation chemotherapy model. Methods: patients with cT3/T4 or TxN + M0 rectal cancer that were receiving neoadjuvant chemoradiotherapy (CRT) (50 Gy with oral capecitabine)/TNT (CRT followed by three cycles of CAPOX) during September 2017 to September 2019 in our department were included. All of the patients were recommended to receive radical surgery. Results: a total of 197 patients were included. Eighty-one patients received CRT, while one hundred and sixteen patients received TNT. Nine patients did not undergo surgery because of the distant metastases (one patient (1.2%) in CRT group, two patients (1.7%) in TNT group) or a refusal of resection (two patients in CRT group, four patients in TNT group). The pathological complete response (pCR) rate was 32.7% in TNT compared with 12.8% in CRT (p = 0.002). There was no statistically significant difference in grade 3 acute toxicities of neoadjuvant treatment and surgical complications between the two groups. Conclusions: the consolidation chemotherapy model is safe for patients with locally advanced rectal cancer and it has a high pCR rate. The long-term follow-up is necessary to be evaluated in a future prospective, randomized trial.     

富血小板血浆联合骨髓间充质干细胞对兔股骨头坏死BMP-2/Smads通路的影响

目的研究富血小板血浆联合骨髓间充质干细胞对兔股骨头坏死BMP-2/Smads通路的影响。方法随机将40只新西兰兔随机分为模型组、富血小板血浆(3%)组、骨髓间充质干细胞(1×106/m L)组、富血小板血浆(3%)+骨髓间充质干细胞(1×106/m L)组。建立兔股骨头坏死模型,术后4w分别向各组股骨头内髓芯减压后注射富血小板血浆、骨髓间充质干细胞。另设空白组10只新西兰兔。空白组与模型组不注射任何物质作为对照。造模后第14周,取材。通过HE染色观察兔股骨头骨髓腔内的病理学及骨陷窝空缺率变化情况。酶联免疫(ELISA)法检测血清中血管内皮生长因子(VEGF)的含量。蛋白免疫印迹(Western blot)法检测股骨头中BMP-2、Smad1、Smad5、Runx2的蛋白水平的变化。结果与模型组相比,各治疗组骨髓腔内骨小梁变粗,脂肪细胞减少,骨细胞增多,骨陷窝空缺率降低(P0.05);各治疗组兔血清VEGF的含量增加,股骨头中BMP-2、Smad1、Smad5、Runx2的蛋白水平提高。结论富血小板血浆与骨髓间充质干细胞可调节BMP-2/Smads信号通路,促进股骨头骨细胞增殖,减少骨陷窝空缺率。     

运用CRISPR/Cas9技术构建PCSK9点突变家兔模型

目的：运用CRISPR/Cas9基因编辑技术构建前蛋白转化酶枯草溶菌素9（PCSK9）点突变家兔模型。方法：根据PubMed基因蛋白数据对人和兔的PCSK9蛋白功能区进行Blast分析,发现人PCSK9基因的386S（丝氨酸）氨基酸功能区与兔PCSK9基因的485S同源。根据家兔PCSK9基因的485S对应的碱基替换位置及序列分析结果设计3条单链向导RNA和1条单链寡核苷酸供体模板。将合成的单链向导RNA、Cas9 mRNA和单链寡核苷酸供体共同注射入家兔受精卵细胞质内并将胚胎移植入待孕母兔体内。对获得的F0代兔进行PCR、TA克隆、脱靶检测以鉴定PCSK9^S386A是否突变成功。利用获得的PCSK9^S386A基因点突变家兔进行繁殖,扩大群体。结果：共获得15只F0代兔,其中1只为PCSK9^S386A点突变纯合子,2只为PCSK9^S386A点突变杂合子,且该突变可以稳定遗传。结论：利用CRISPR/Cas9技术成功构建了PCSK9^S386A点突变家兔模型,为探究PCSK9功能减弱的分子机制,开发可靠、有效的诊断和治疗措施提供了良好的动物模型。     

Combined effects of TNF‐α, IL‐1β, and HIF‐1α on MMP‐2 production in ACL fibroblasts under mechanical stretch: An in vitro study

The dynamics between inflammatory factors, mechanical stress, and healing factors, in an intra‐articular joint, are very complex after injury. Injury to intra‐articular tissue [anterior cruciate ligament (ACL), synovium] results in hypoxia, accumulation of various pro‐inflammatory factors, cytokines, and metalloproteases. Although the presence of increased amounts of matrix‐metalloproteinases (MMP) in the joint fluid after knee injury is considered the key factor for ACL poor healing ability; however, the exact role of collective participants of the joint fluid on MMP‐2 activity and production has not been fully studied yet. To investigate the combined effects of mechanical injury, inflammation and hypoxia induced factor‐1α (HIF‐1α) on induction of MMP‐2; we mimicked the microenvironment of joint cavity after ACL injury. The results show that TNF‐α and IL‐1β elevate the activity of MMP‐2 in a dose‐ and time‐dependent manner. In addition, mechanical stretch further enhances the MMP‐2 protein levels with TNF‐α, IL‐1β, and their mixture. CoCl₂‐induced HIF‐1α (100 and 500 µM) also increases the levels and activity of MMP‐2. Mechanical stretch has a strong additional effect on MMP‐2 production with HIF‐1α. Our results conclude that mechanical injury, HIF‐1α and inflammatory factors collectively induce increased MMP‐2 production in ACL fibroblasts, which was inhibited by NF‐κB pathway inhibitor (Bay‐11‐7082). © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29: 1008–1014, 2011     

运用 CRISPR/Cas9技术构建 PCSK9 点突变家兔模型

目的：运用CRISPR/Cas9基因编辑技术构建前蛋白转化酶枯草溶菌素9（ PCSK9）点突变家兔模型。 方法：根据PubMed基因蛋白数据对人和兔的PCSK9蛋白功能区进行Blast分析，发现人 PCSK9基因的386S（丝氨酸）氨基酸功能区与兔 PCSK9基因的485S同源。根据家兔 PCSK9基因的485S对应的碱基替换位置及序列分析结果设计3条单链向导RNA和1条单链寡核苷酸供体模板。将合成的单链向导RNA、Cas9 mRNA和单链寡核苷酸供体共同注射入家兔受精卵细胞质内并将胚胎移植入待孕母兔体内。对获得的F0代兔进行PCR、TA克隆、脱靶检测以鉴定PCSK9 ^S386A是否突变成功。利用获得的PCSK9 ^S386A基因点突变家兔进行繁殖，扩大群体。 结果：共获得15只F0代兔，其中1只为PCSK9 ^S386A点突变纯合子，2只为PCSK9 ^S386A点突变杂合子，且该突变可以稳定遗传。 结论：利用CRISPR/Cas9技术成功构建了PCSK9 ^S386A点突变家兔模型，为探究PCSK9功能减弱的分子机制，开发可靠、有效的诊断和治疗措施提供了良好的动物模型。     

抑制核因子-κB对Toll样受体4在急性坏死性胰腺炎大鼠胰腺组织中表达的影响

目的：观察急性坏死性胰腺炎（ANP）大鼠胰腺组织中Toll样受体4（TLR4）的表达情况。并且应用核因子（NF）-κB活化抑制剂四氢化吡咯二硫代氨基甲酸酯（PDTC）进行干预，观察阻断NF-κB以后，TLR4在ANP大鼠胰腺组织中表达的变化，探讨ANP中TLR4与NF-κB的关系及NF-κB的活性对TLR4表达的影响。方法：72只SD大鼠随机分为假手术组、ANP组和PDTC预处理组。采用胆胰管内逆行注射5%牛磺胆酸钠溶液诱导大鼠ANP模型。PDTC预处理组在建立模型前1h按100mg&#183;kg^-1腹腔注射给药。分别于建模后3h、6h、12h将大鼠分批处死。观察胰腺病理，用免疫组化及WesternBlot检测胰腺组织TLR4与NF-κB表达变化，RT-PCR检测胰腺组织TNFα、IL-1β和IL-6mRNA表达。结果：同假手术组相比，ANP组胰腺组织TLR4、NF-κB表达及TNFα、IL-1β和IL-6mRNA表达明显升高（P〈0.05）；同ANP组相比，PDTC预处理组胰腺组织炎症明显减轻，胰腺组织TLR4、NF-κB表达及TNFα、IL-1β和IL-6mRNA表达明显降低（P〈0.05）。结论：TLR4和NF-κB在ANP大鼠胰腺组织中的表达水平明显增高，PDTC抑制ANP大鼠胰腺组织中NF-κB表达的同时，下调TLR4蛋白水平表达和TNFα、IL-1β和IL-6mRNA表达。TLR4和NF-κB是介导炎症反应的枢纽，因而它们在ANP发生发展中起重要作用。TLR4和NF-κB可能不是简单的上下游关系，其关系还需进一步研究。