胚胎干细胞来源造血细胞的细胞表型和功能分析

【目的】 分析经卵黄囊(YS)、胎肝(FL)和骨髓来源(BM)的基质细胞条件培养液(SCCM)诱导产生的胚胎干细胞(ES)来源造血细胞的细胞表型与功能差异。【方法】 制备YS-SCCM、 FL-SCCM及BM-SCCM,将3种基质细胞条件培养液分别加入ESE 14.1细胞分化培养体系培养7 d,通过对分化ESE14.1细胞造血发育表面标志FLK-1、Integrin-α4(整联蛋白-α4、Sca-1(干细胞抗原-1)、CD34的检测、体外高增殖潜能集落形成细胞(HPP-CFC)分析及体内脾集落形成单位(CFU-S)检测,评价3种基质细胞条件培养液对ESE14.1细胞体外造血发育的调控作用。【结果】 经FL-SCCM诱导的EB细胞Flk-1、Integrinα4+ 和Sca-1+ 细胞均高于YS-SCCM和BMSC-CM诱导组,分别为3.03%、2.9%和13.74%;经BMSC-CM诱导产生的CD34+ 细胞比例最高,为1.07% 。经FLSC-CM或BMSC-CM诱导产生的造血细胞其HPP-CFC产率明显高于对照组,分别为7.4个/105细胞(P < 0.01)和5.8个/105细胞(P < 0.05);经FLSC-CM或BMSC-CM诱导产生的造血细胞其CFU-S产率亦明显高于对照组,分别为8.5个/5 × 105细胞和6.75个/5 × 105细胞(P < 0.001)。【结论】 YS-SCCM、 FL-SCCM及BM-SCCM均可诱导ESE14.1向造血细胞分化,FL-SCCM和BM-SCCM造血定向诱导效率较高,所产生的细胞具备造血细胞的正常功能,FL-SCCM诱导产生的造血细胞原始程度高于BM-SCCM诱导产生的造血细胞。     

蒙药材胡德日奈都日吉德的质量标准初步研究

目的:建立蒙药胡德日奈都日吉德的质量标准。方法:参照《中华人民共和国药典》(四部)2015年版相关方法,对胡德日奈都日吉德杂质、水分、灰分、浸出物进行检测;采用硅胶G薄层板,以大戟二烯醇为对照,以石油醚(60~90℃)-乙酸乙酯(10∶1)为展开剂,用10%硫酸乙醇溶液显色,对胡德日奈都日吉德进行鉴别;采用紫外分光光度法,以大戟二烯醇为对照,在545 nm波长下对胡德日奈都日吉德的活性成分萜类化合物进行定量分析。结果:胡德日奈都日吉德三批样品杂质在0.69%~0.75%之间,平均值为0.72%,RSD为4.26%;水分含量在7.68%~7.74%之间,平均值为7.72%,RSD为0.03%;总灰分0.20%~0.22%之间,平均值为0.21%,RSD为2.86%;酸不溶性灰分在0.21%~0.23%之间,平均值为0.22%,RSD为4.82%;浸出物1.24~1.35之间,平均值为1.31%,RSD为4.64%。在供试品的色谱中,与对照品色谱相应的位置上,显示相同颜色的斑点。总萜在0.6720~5.3790 mg/L呈良好的线性关系,该方法平均回收率为97.48%,RSD为0.478%,3批样品总萜含量在25.67~35.94 mg/g之间,均含量为31.03 mg/g。结论:根据实验结果,制定蒙药胡德日奈都日吉德药材(按干燥品计算)中杂质、水分、总灰分、酸不溶性灰分分别不得过1%,10.0%,0.4%、0.4%;水溶性浸出物不得少于1.0%;总萜含量不得少于20 mg/g。本办法可用于蒙药胡德日奈都日吉德药材的质量控制。     

自拟加味二仙汤联合克罗米芬治疗肾阳虚型无排卵性不孕症

我们自2008年1月～2009年1月采用中西医结合治疗肾阳虚型无排卵性不孕症30例,并与单用西药治疗的30例作对照,疗效较好,现报道如下.     

Neonatal side effects of maternal labetalol treatment in severe preeclampsia

Objective

Labetalol is often used in severe preeclampsia (PE). Hypotension, bradycardia and hypoglycemia are feared neonatal side effects, but may also occur in (preterm) infants regardless of labetalol exposure. We analyzed the possible association between intrauterine labetalol exposure and such side effects.

Study design

From 1 January 2003 through 31 March 2008, all infants from mothers suffering severe PE admitted to one tertiary care center were included. Severe PE was defined according to the International Society for the Study of Hypertension in Pregnancy (ISSHP) criteria. Infants exposed to labetalol in utero (labetalol infants) were compared with infants, who were not exposed to labetalol (controls). Neonatal records were reviewed for hypotension (RR < mean gestational age in weeks), bradycardia (heartrate < 100/min) and hypoglycaemia (glucose < 2.7 mmol/L) in the first 48 postnatal hours.

Results

Of 109 infants, 55 had been exposed to labetalol, whereas 54 were not (controls). Gestational age at delivery and birthweight were similar in both groups (31.8 vs. 32.8 weeks (p = 0.06) and 1510 vs. 1639 grams (p = 0.25), respectively for the labetalol vs. control group). Hypotension occurred significantly more in conjunction with labetalol exposure (16, (29.1%) vs. 4 (7.4%); p = 0.003), irrespective of the route of administration. Patent ductus arteriosus (PDA) was present in 9 (56%) of hypotensive labetalol infants compared to 1 (24%) infant in the hypotensive control group (NS). In a multivariate regression model, labetalol exposure, the need for intubation and PDA appeared independently associated with hypotension (P < 0.001). Hypoglycemia occurred in 26 (47.3%) of labetalol infants and in 23 (42.6%) of control infants (p = 0.62). Bradycardia occurred in 4 (7.3%) of labetalol infants and in 1 (1.9%) of control infants (p = 0.18). Hypoglycemia was more common in premature infants (n = 45 (48,9%) vs. n = 4 (23.5%), p = 0.05) in both labetalol and control infants.

Conclusion

Hypotension is more common after maternal labetalol exposure, regardless of the dosage and route of administration. The need for intubation and the presence of a PDA also play a role. Hypoglycemia is a very common finding in this population and is merely related to prematurity and independent of labetalol exposure as was the incidental occurrence of bradycardia. These findings on the neonatal side effects of maternal labetalol treatment in preeclampsia underline the importance of frequent blood glucose and blood pressure measurements in the first days of life, especially in intubated preterm infants with a PDA.     

Lack of positive allosteric modulation of mutated α1S267I glycine receptors by cannabinoids

Loss of inhibitory synaptic transmission within the dorsal horn of the spinal cord plays a key role in the development of chronic pain following inflammation or nerve injury. Inhibitory postsynaptic transmission in the adult spinal cord involves mainly glycine. Ajulemic acid and HU210 are non-psychotropic, synthetic cannabinoids. Cannabidiol is a non-psychotropic plant constituent of cannabis sativa. There are hints that non-cannabinoid receptor mechanisms of these cannabinoids might be mediated via glycine receptors. In this study, we investigated the impact of the amino acid residue serine at position 267 on the glycine-modulatory effects of ajulemic acid, cannabidiol and HU210. Mutated α₁S267I glycine receptors transiently expressed in HEK293 cells were studied by utilising the whole-cell clamp technique. The mutation of the α₁ subunit TM2 serine residue to isoleucine abolished the co-activation and the direct activation of the glycine receptor by the investigated cannabinoids. The nature of the TM2 (267) residue of the glycine α₁ subunit is crucial for the glycine-modulatory effect of ajulemic acid, cannabidiol and HU210. An investigation of the impact of such mutations on the in vivo interaction of cannabinoids with glycine receptors should permit a better understanding of the molecular determinants of action of cannabinoids.     

Decision-making,knowledge, and psychosocial outcomes in pediatric siblings identified to donate hematopoietic stem cells

Purpose: To (a) describe the decision-making experience and psychosocial outcome of sibling hematopoietic stem cell (HSC) donors, and (b) to determine the feasibility of completing a prospective and longitudinal assessment of HSC sibling donors at a single institution.

Design: A mixed-methods approach was utilized.

Sample and methods: 12 potential siblings HSC donors aged 10–21?years completed various psychological measures and participated in semi-structured interviews at three time points in the donation experience: pre-donation, within 1?week after the harvest procedure, and six months post-donation. Caregivers also completed parent-proxy measures.

Findings: Qualitative analysis indicated donors want to make their own decision about donation but may not be given the option or may feel that there is no choice given their limited awareness of alternative options. Donors felt well prepared for the donation procedure but demonstrated a poor understanding of possible recipient outcomes. A minority of donors endorsed emotional distress prior to and after donation; however, this was not linked to recipient health. Forty percent of donors felt that they had inadequate support following their donation. Small sample size restricted quantitative data analysis.

Conclusions and implications: Utilizing a donor advocate offers opportunity to work with donors to encourage decision-making tied to ideals rather obligation, increase education about possible recipient outcomes, and offer support at key times, such as when a recipient dies. Future research should include prospective multi-site studies.     

西施泰与吡柔比星联合灌注减少膀胱灌注化疗所致膀胱并发症的多中心临床研究

目的 验证西施泰膀胱灌注减少非肌层浸润性膀胱癌TURBt术后灌注化疗并发症的疗效和安全性.方法 对120例符合入选/排除标准患者进行多中心、随机、空白对照的疗效和安全性临床研究.患者均先行TURBt,之后观察组联合灌注吡柔比星和西施泰,对照组仅灌注吡柔比星.以膀胱疼痛直观模拟分级(VAS)评分为主要疗效指标,以血尿、膀胱刺激症状为次要疗效指标,严密观察不良事件、实验室检查、治疗前后生命体征的变化,评价其疗效和安全性. 结果 2组患者人口学特征与基线资料相似,具有可比性.观察组治疗前后VAS评分差值和改善率分别为2.24±1.70与(92.92±14.76)%,对照组分别为0.70±1.82与(20.59±87.34)%,2组比较差异均有统计学意义(P＜0.01);2组VAS总分变化值差异也有统计学意义,并且2组VAS评分灌注前后改善率的比较从访视2开始差异就有统计学意义.观察组尿频次数从治疗前的(9.06±4.09)次减少至(6.69±2.89)次,对照组从(8.85±3.32)次增加至(10.15±4.40)次,组间比较差异有统计学意义(P＜0.01).观察组夜尿次数从治疗前的(2.88±1.74)次减少至(1.47±1.62)次,对照组从(3.22±2.30)次减少至(2.91±1.73)次,差异亦有统计学意义(P＜0.01).2组治疗前后尿急、排尿困难及血尿方面的改善不明显,未发现与西施泰有关的不良事件. 结论西施泰与化疗药物联合灌注能明显改善膀胱灌注化疗患者VAS评分状况,迅速、持续缓解患者的膀胱疼痛,并改善患者尿频与夜尿症状,提高患者生活质量.VAS评分高的患者改善效果更明显.西施泰与化疗药联合灌注安全性与临床耐受性良好.

Abstract：

Objective To verify the efficacy and safety of intravesical instillation of Cystistat in reducing complications caused by intravesical chemotherapy after TUR-BT in non-muscle invasive bladder cancer patients. Methods One hundred and twenty patients who met the inclusion/exclusion criteria were enrolled into this multi-centered, randomized and blank controlled clinical study. Selected patients were randomized into the observation group and control group. TUR-BT was carried out in both groups followed by pirarubicin (THP) and Cystistat intravesical instillation in the observation group, and THP intravesical instillation alone in control group. Visual analog scale (VAS) was used as the primary efficacy variable. The secondary efficacy variables were assessments of hematuria and bladder irritation symptoms. Adverse events, laboratory tests and changes of vital signs before and after treatment were strictly observed during observation to evaluate the efficacy and safety of Cystistat.Results Demographics and baseline characteristics were comparable in both groups. The differences and the improvement rate of VAS score in the 2 groups were significant, both P＜0.01. The changes of VAS score and the improvement rate before and after treatment were (2. 24±1.70) and (92. 92±14.76) % in observation group and (0. 70±1.82) and (20. 59±87.34)% in control group respectively. According to the covariance analysis, there were significant differences in changes of VAS score between the observation group and the control group. Also, the improvement rate of VAS score was significant from visit 2. The urine frequency decreased from 9.06±4.09 to 6. 69±2.89 in observation group and increased from 8. 85±3. 32 to 10. 15±4.40 in control group, P＜0.01. There were also significant differences in changes of nocturia before and after treatment between these two groups (P＜0.01), the nocturia decreased from 2. 88±1.74 to 1. 47±1.62 in observation group and 3. 22±2.30 to 2.91±1.73 in control group, respectively. The changes of WHO assessment for hematuria,urgency and dysuria were not significantly different between the 2 groups. No Cystistat related adverse event was observed. Conclusions Cystistat combined instillation can significantly improve the VAS score of patients with chemotherapeutic agent instillation. Relief of bladder pain, frequency and nocturia are more rapidly and more durable in Cystistat combined instillation group. The improvement is more effective in patients with a high VAS score. Cystistat instillation with chemotherapeutics agents is both well tolerated and safe.     

同种异基因造血干细胞移植急性移植物抗宿主病小鼠模型的制作

目的 制作同种异基因造血干细胞移植急性移植物抗宿主病(GVHD)小鼠模型.方法 以C57BL/6( H-2b)小鼠为供者,Balb/c( H-2d)小鼠为受者,进行同种异基因骨髓移植.设立全身照射(TBI)对照组(4只)、GVHD组(10只)、单纯骨髓移植组(10只)及正常对照组(4只).TBI对照组仅进行致死性TBI,TBI后不进行骨髓移植;GVHD组于TBI前5d开始饮用含320 mg/L庆大霉素和250 mg/L红霉素的饮用水,移植当天以60Co γ射线行一次性TBI,总剂量8.0Gy,TBI后5h内每只小鼠经尾静脉输注C57BL/6小鼠骨髓细胞2×106个+脾细胞1×107个;单纯骨髓移植组预处理与GVHD组相同,每只小鼠经尾静脉输注C57BL/6小鼠骨髓细胞2×106个.移植后观察小鼠的精神状态、活动能力、体位改变、皮毛、体重和大便等,记录每只小鼠的存活时间,计算存活率,并绘制生存曲线.濒死小鼠的皮肤、肝脏、小肠和骨髓行病理检查.结果 TBI对照组小鼠的存活时间为(9.0±0.7)d,GVHD组为(32.0±3.2)d,单纯骨髓移植组为(17.5±1.6)d,3组间两两比较,存活时间的差异均有统计学意义(P＜0.01).TBI对照组病理检查显示造血功能衰竭.GVHD组于移植后第10～13天出现急性GVHD表现,其皮肤、肝脏和小肠组织的病理表现均符合Ⅰ～Ⅱ度急性GVHD改变,单纯骨髓移植组也于移植后第10～13天出现GVHD表现,但其GVHD表现和组织学改变明显轻于GVHD组,仅为0～Ⅰ度GVHD.结论 Balb/c小鼠经致死性TBI后移植同种异基因小鼠骨髓细胞+脾细胞可成功制作稳定的急性GVHD模型.     

海洋微生物实验室培养

目前,超过95%的海洋微生物尚不能被培养,这造成了海洋微生物活性物质资源获取上的瓶颈问题.本文通过分析传统微生物培养面临的难题,在微生物实验室培养新技术的基础上,借鉴陆栖微生物工业化发酵生产的经验,进一步探讨如何开发海洋微生物,为微生物工业化生产新药开辟新道路提供参考.     

1101株病原菌耐药性分析

目的了解本医院2009年1月～2010年6月临床主要病原菌分布以及对常用抗菌药物的耐药情况。方法采用纸片扩散法进行抗菌药物敏感性实验,WHONET5.3软件分析耐药性监测数据。结果 1 101株临床主要分离病原菌中革兰阳性菌282株（25.6%）,革兰阴性菌819株（74.4%）,肠杆菌科中大肠埃希氏菌和肺炎克雷伯菌其中产ESBLS菌株的检出率分别为37.9%和27.3%,对碳青烯酶类抗菌药物高度敏感,耐甲氧西林金黄色葡萄球菌和耐甲氧西林凝固酶阴性葡萄球菌的检出率分别为27.7%和9.2%。结论开展连续的耐药性监测,可为指导临床合理使用抗菌药物控制感染提供有力依据。