Engraftable neural crest stem cells derived from cynomolgus monkey embryonic stem cells

Neural crest stem cells (NCSCs), a population of multipotent cells that migrate extensively and give rise to diverse derivatives, including peripheral and enteric neurons and glia, craniofacial cartilage and bone, melanocytes and smooth muscle, have great potential for regenerative medicine. Non-human primates provide optimal models for the development of stem cell therapies. Here, we describe the first derivation of NCSCs from cynomolgus monkey embryonic stem cells (CmESCs) at the neural rosette stage. CmESC-derived neurospheres replated on polyornithine/laminin-coated dishes migrated onto the substrate and showed characteristic expression of NCSC markers, including Sox10, AP2α, Slug, Nestin, p75, and HNK1. CmNCSCs were capable of propagating in an undifferentiated state in vitro as adherent or suspension cultures, and could be subsequently induced to differentiate towards peripheral nervous system lineages (peripheral sympathetic neurons, sensory neurons, and Schwann cells) and mesenchymal lineages (osteoblasts, adipocytes, chondrocytes, and smooth muscle cells). CmNCSCs transplanted into developing chick embryos or fetal brains of cynomolgus macaques survived, migrated, and differentiated into progeny consistent with a neural crest identity. Our studies demonstrate that CmNCSCs offer a new tool for investigating neural crest development and neural crest-associated human disease and suggest that this non-human primate model may facilitate tissue engineering and regenerative medicine efforts.     

我院药物代谢酶和药物作用靶点基因检测与精准药学服务实践与总结

目的回顾我院药物代谢酶和药物作用靶点相关基因检测与精准药学服务实践过程,总结经验,与同行分享。方法从准备工作、相关检测项目的确定,学术推广、项目优化和开展情况等方面详细阐述我院基因检测开展过程与精准药学服务情况。结果根据临床需求,我院已开展了以心脑血管药和抗精神病药为主的27种药物,26项检测,2018年全年位1587名患者提供个体化用药建议,受到临床医生和患者欢迎。结论基于代谢酶和药物作用靶点相关基因检测的个体化用药建议是临床药师参与精准治疗的重要途径,有助于医生和药师之间的沟通,有利于提高药学服务水平。     

Kinetics of phytohemaglutinin-induced IFN-γ and TNF-α expression in peripheral blood mononuclear cells from patients with chronic hepatitis B after liver transplantation

AIM: To study the association between host immunity and hepatitis B virus (HBV) recurrence after liver transplantation. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from 40 patients with hepatitis B and underwent orthotopic liver transplantation (OLT) before and 2, 4, 8 wk after surgery. After being cultured in vitro for 72 h, the levels of INF-gamma and TNF-alpha in culture supernatants were detected with ELISA. At the same time, the quantities of HBV DNA in serum and PBMCs were measured by real time PCR. RESULTS: The levels of INF-gamma and TNF-alpha in PBMC culture supernatants decreased before and 2, 4 wk after surgery in turns (INF-gamma 155.52+/-72.32 ng/L vs 14.76+/-9.88 ng/L vs 13.22+/-10.35 ng/L, F = 6.946, P = 0.027 < 0.05; TNF-alpha 80.839+/-46.75 ng/L vs 18.59+/-17.29 ng/L vs 9.758+/-7.96 ng/L, F = 22.61, P = 0.0001 < 0.05). The levels of INF-gamma and TNF-alpha were higher in groups with phytohemagglutinin (PHA) than in those without PHA before surgery. However, the difference disappeared following OLT. Furthermore, INF-gamma and TNF-alpha could not be detected in most patients at wk 4 and none at wk 8 after OLT. The HBV detection rate and virus load in PBMC before and 2, 4 wk after surgery were fluctuated (HBV detected rate: 51.4%, 13.3%, 50% respectively; HBV DNA: 3.55+/-0.674 log10 copies/mL vs 3.00+/-0.329 log10 copies/mL vs 4.608+/-1.344 log10 copies/mL, F = 7.582, P = 0.002 < 0.05). HBV DNA in serum was 4.48+/-1.463 log10 copies/mL before surgery and <10(3) copies/mL after OLT except for one with 5.72 x 10(6) copies/mL 4 wk after OLT who was diagnosed as HBV recurrence. The levels of INF-gamma and TNF-alpha were lower in patients with a high HBV load than in those with a low HBV load (HBV DNA detected/undetected in PBMCs: IFN-gamma 138.08+/-72.44 ng/L vs 164.24+/-72.07 ng/L, t = 1.065, P = 0.297 > 0.05, TNF-alpha 80.75+/-47.30 ng/L vs 74.10+/-49.70 ng/L, t = 0.407, P = 0.686 > 0.05; HBV DNA positive/negative: IFN-gamma 136.77+/-70.04 ng/L vs 175.27+/-71.50 ng/L, t = 1.702, P = 0.097 > 0.05; TNF-alpha 75.37+/-43.02 ng/L vs 81.53+/-52.46 ng/L, t = 0.402, P = 0.690 > 0.05). CONCLUSION: The yielding of INF-gamma and TNF-alpha from PBMCs is inhibited significantly by immunosuppressive agents following OLT with HBV load increased, indicating that the impaired immunity of host is associated with HBV recurrence after OLT.     

Updated Options for Liver-Limited Metastatic Colorectal Cancer

Liver metastases from colorectal cancer (CRC) are common in patients presenting with an initial diagnosis of metastatic disease or at the time of recurrence. Without treatment, patients with metastatic disease have a poor prognosis. Surgical resection of the metastases might provide long-term benefit.; however, the size, number, or location of the metastases can limit the ability to perform a resection. The use of chemotherapy, both systemic and via hepatic artery infusion, in patients undergoing surgery for liver metastases from CRC has augmented the long-term survival benefits and even the cure obtained in some patients with surgery. Chemotherapy might also convert a portion of patients with initially unresectable liver metastases to resectable. A growing body of literature is helping to define the role of chemotherapy for potentially resectable liver metastases and for initially unresectable liver metastases. The introduction of newer agents such as oxaliplatin and irinotecan, and targeted agents such as cetuximab and bevacizumab, has led to meaningful improvements in response rates and survival over those previously achieved with 5-fluorouracil. Further trials are needed to refine the use of chemotherapy and targeted agents in the management of patients with liver metastases.     

偏瘫患者平衡功能的定量评定:BPM和BBS的对照研究

目的:分析脑卒中、脑外伤患者(病例组)和健康人(对照组)两组受试对象在平衡功能方面的差异,为临床康复提供依据。方法:病例组和对照组各为21例。对每一位受试者先给予一次平衡测试仪(balanceperformancemonitor,BPM)测试,BPM测试分别在睁眼及闭眼状态下各测试两次,取其均值分析。BPM测试后稍微休息再进行Berg平衡量表(Bergbalancescale,BBS)评定。结果:病例组BPM的重心分布、摆动轨迹长和摆动面积三个指标较对照组明显增大;BBS测试总分、静态总分和动态总分的得分与对照组比较均明显降低,且动态平衡功能的改变较静态平衡功能的改变更明显。两组之间BPM和BBS的各个测试指标均有显著性差异(P<0.01)。结论:①BPM和BBS是评定平衡功能的理想工具,BBS较BPM的应用更为方便。②与同年龄组的健康人相比,脑卒中、脑外伤患者的平衡控制能力降低,尤以动态平衡功能减弱明显     

Association of Tumor Necrosis Factor-α-863C/A Gene Polymorphism with Chronic Obstructive Pulmonary Disease

The aim of this study was to investigate genetic effects on the pathogenesis of chronic obstructive pulmonary disease (COPD). The study was conducted as a prospective case–control study in a medical center in southern Taiwan. The patient group consisted of 145 male patients with smoking-related COPD and a control group of 139 resistant smokers from July 2004 to September 2009. We compared allele and genotype frequencies of three tag single nucleotide polymorphisms (SNP) of the TNF-α gene promoter region at −308, −863, and −1031 in all subjects. We also analyzed the influence of each genetic variant on pulmonary function parameters, body mass index (BMI), serum TNF-α levels, and outcomes among heavy smokers with or without COPD. COPD patients had a significantly lower A allele frequency (9.7 vs. 15.1%, OR = 0.6, p = 0.048, false discovery rate q = 0.144) and a significantly lower A carrier genotype frequency (19.3 vs. 30.2%, OR = 0.52, p = 0.042, q = 0.135) than resistant smokers. The −863 CA genotype was associated with a better FEV₁/FVC ratio (79 vs. 71.5%, p = 0.034), and higher BMI (24.9 vs. 23.6 kg/m², p = 0.048). In addition, COPD patients with the −1031 C carrier genotype had higher serum TNF-α levels (20.9 vs. 16.2 pg/ml, p = 0.01). BMI (hazard ratio = 0.84, 95% CI = 0.74–0.96, p = 0.008) was the only independent predictor for mortality. The TNF-α −863 A allele may confer a degree of resistance to the susceptibility to and muscle wasting of COPD among heavy smokers.     

利培酮治疗帕金森病合并精神障碍的临床研究

目的:探讨利培酮治疗帕金森病(PD)合并精神障碍的疗效及不良反应。方法:收集PD合并精神障碍患者268例,其中阳性症状为主者93例,阴性症状为主者175例,皆符合CCMD-3和ICD-10的诊断标准,两者进行对照,观察共12周。采用阴性症状量表(SANS)和阳性症状量表(SAPS)评价患者的精神病性症状的疗效。采用副反应量表(TESS)评定不良反应。结果:利培酮对幻觉等阳性症状改善明显,对意志缺乏、兴趣缺乏等症状改善不明显,部分患者运动障碍较前加重。结论:利培酮对治疗PD幻觉等阳性精神症状的患者疗效肯定,并且是安全的。     

经骨折椎单节段复位固定治疗创伤性胸腰椎骨折的临床疗效

目的评价经骨折椎单节段椎弓根螺钉复位固定治疗创伤性胸腰椎骨折的临床疗效。方法采用单节段椎弓根螺钉固定治疗创伤性胸腰椎骨折132例。按AO分型,A型117例,B型8例,C型7例。通过测量骨折椎楔变指数（wI）、骨折节段矢状指数（SI）评价影像学效果,并通过Oswestry功能障碍指数评价临床疗效。结果所有患者手术顺利,手术用时平均87min,术中出血量平均136ml。随访128例,平均随访时间30．4个月（11～54个月）,全部骨性融合,除1例出现螺钉松动外,无内固定失败。术前骨折椎WI和S1分别为（42．96±11．51）％和13．06°±4．13°,术后WI和S1分别为（21．78±7．66）％和4．47°±2．28°,与术前比较差异具有统计学意义（P〈0．01）。除2例外,最后一次随访时未见明显矫正丢失（P〈0．05）。Oswestry功能障碍指数较术前获得明显改善（P〈0．05）。结论经骨折椎单节段椎弓根螺钉复位固定融合术具有创伤小、用时短、出血少、脊椎运动功能单位丢失减少等优点,可用于治疗部分创伤性胸腰椎骨折。     

17β-雌二醇对新生牛血清诱导的血管平滑肌细胞增殖反应的影响及其机制

目的：探讨雌甾-1,3,5（10）-三烯-3,17β-二醇（17β-雌二醇）对新生牛血清诱导血管平滑肌细胞（VSMCs）增殖反应的影响及其机制。方法：用新生牛血清刺激培养的VSMCs增殖,MTT法检测17β-雌二醇对新生牛血清诱导的VSMCs增殖的影响,并用Western blot检测caveolin-1蛋白,诱导型一氧化氮合酶（iNOS）及NO在此过程中的变化。结果：与对照组比,新生牛血清可促进VSMCs增殖（P〈0.05）,17β-雌二醇作用24 h后能明显抑制上述作用（P〈0.05）;17β-雌二醇预处理可抑制新生牛血清诱导的VSMCs中caveolin-1,iNOS蛋白表达下降及NO释放（P〈0.05）;17β-雌二醇受体阻断剂1-{4-[2-（n,n-二甲氨基）乙氧基]苯}-1,2-二苯-1-丁烯（他莫昔芬）预处理可部分逆转17β-雌二醇的抑制作用（P〈0.05）。结论：17β-雌二醇通过其受体可抑制新生牛血清诱导的VSMCs增殖,此过程与上调caveolin-1蛋白表达,激活iNOS,增加NO释放有关。