微滴式数字PCR和实时荧光定量PCR对新型冠状病毒肺炎患者不同时间血便尿中核酸检测结果初步探讨

目的 分析微滴式数字PCR(droplet digital PCR, ddPCR)和实时荧光定量PCR(quantitative real-time PCR,qPCR)的核酸检测结果,比较两种方法检测各类样本的差异性,为改进新型冠状病毒核酸检测方案提供数据支持。 方法 利用ddPCR和qPCR技术对已经确诊的3例新型冠状病毒肺炎患者发病不同时间的全血、尿液、粪便共22份标本进行新型冠状病毒核酸检测。 结果 两种方法对人保守区域基因扩增结果一致:全血标本信号最强,尿液次之,粪便最少;ddPCR在1份全血,1份尿液,5份粪便中检出ORF-1ab和N基因的阳性微滴,qPCR仅在3份粪便中检出上述基因,漏检的3个标本基因拷贝数平均浓度为128 copies/ml;ddPCR在发病<5、5~15、>15 d的各类标本中都有检出,qPCR检出以中晚期为主;重症病例用ddPCR均可测到阳性微滴,qPCR检测的各类标本均为阴性;轻症病例的各类标本中qPCR只有粪便核酸检测阳性,ddPCR检出率高于qPCR。 结论 ddPCR可以有效克服qPCR 灵敏度不足的难题,是对qPCR 的有益补充,尤其是针对病毒载量比较低的血液、尿液和可疑的粪便或肛拭子标本,适用于早期感染的判断及患者治愈后出院诊断。     

Intrathecal catheterisation after observed accidental dural puncture in labouring women: update of a meta-analysis and a trial-sequential analysis

BackgroundOur meta-analysis from 2013 showed that inserting a catheter intrathecally after an observed accidental dural puncture can reduce the need for epidural blood patch in labouring women requesting epidural analgesia. We updated our conventional meta-analysis and added a trial-sequential analysis (TSA).MethodsA systematic literature search was conducted to identify studies that compared inserting the catheter intrathecally with an epidural catheter re-site or with no intervention. The extracted data were pooled and the risk ratio (RR) and 95% confidence interval (95%CI) for the incidence of post-dural puncture headache (PDPH) was calculated, using the random effects model. A contour-enhanced funnel plot was constructed. A TSA was performed and the cumulative Z score, monitoring and futility boundaries were constructed.ResultsOur search identified 13 studies, reporting on 1653 patients, with a low risk of bias. The RR for the incidence of PDPH was 0.82 (95%CI 0.71 to 0.95) and the RR for the need for epidural blood patch was 0.62 (95%CI 0.49 to 0.79); heterogeneity of both analyses was high. The TSA showed that the monitoring or futility boundaries were not crossed, indicating insufficient data to exclude a type I error of statistical analysis. Contour-enhanced funnel plots were symmetric, suggesting no publication bias.ConclusionsConventional meta-analyses showed for the first time that intrathecal catheterisation can reduce the incidence of PDPH. However, TSA did not corroborate this finding. Despite increasing use in clinical practice there is no firm evidence on which to base a definite conclusion.     

早期十全大补汤联合肠内营养乳剂治疗胃癌术后(气血两虚证)喂养不耐受临床评价

目的探讨早期使用十全大补汤联合肠内营养乳剂(TP)治疗胃癌术后(气血两虚证)发生喂养不耐受(FI)的影响因素及对营养指标、中医证候积分的影响。方法回顾性分析术后早期行十全大补汤联合TP治疗的80例胃癌术后(气血两虚)患者的病历资料,根据是否出现FI分为耐受组(34例)和不耐受组(46例)。FI的相关影响因素进行单因素及多因素分析,并观察FI对患者营养指标、中医证候积分的影响。结果单因素分析显示,患者术后第1天下床活动时间、开始肠内营养(EN)的时间、使用营养泵、早期灌肠与FI的发生密切相关(P <0. 05);多因素Logistic回归分析显示,患者第1天下床活动时间≥2 h[OR=0. 022,P=0. 001,95%CI(0. 002,0. 223)]、使用营养泵[OR=0. 021,P=0. 000,95%CI(0. 003,0. 162)]是FI发生的独立危险因素;术后10 d,耐受组患者白蛋白(ALB)、血红蛋白(Hb)升高水平优于不耐受组(P <0. 05),中医证候积分显著低于不耐受组(P <0. 05)。结论胃癌术后(气血两虚证)患者早期给予十全大补汤联合TP治疗开始后,患者第1天下床活动时间不短于2 h、使用营养泵能有效减少FI的发生,并改善了患者的营养状态,减轻了中医临床症状。     

TLR9在不同胎龄新生儿脐血中的表达变化

目的 通过观察早产儿不同胎龄Toll样受体9（TLR9）的表达，探讨早产儿免疫功能低下的机制。方法 采集2010年7月至2014年6月在上海市嘉定区妇幼保健院产科出生的活产新生儿的脐血229份，按胎龄分为4组，28~31周组，31~34周组，34~37周组，≥37周组，采用流式细胞术和实时荧光定量PCR方法，分别检测其TLR9的蛋白和mRNA表达情况，了解其与胎龄之间的关系，并分析mRNA和蛋白表达间的相关性。结果 TLR9阳性细胞率在28~31周组，31~34周组，34~37周组，≥37周组分别为（15.93±6.23）%，（11.63±6.70）%，（13.66±6.88）%，（20.51±12.06）%；其在胎龄28~31周较高，至31~34周逐渐下降至最低，两组差异有统计学意义（P<0.05）；34~37周后TLR9阳性细胞率表达逐渐升高，至≥37周达最高，两胎龄组比较，差异具有统计学意义（P<0.05）。31~37周间新生儿脐血TLR9阳性细胞率与胎龄呈正相关（r=0.273，P=0.006）。TLR9 mRNA表达在28~31周组，31~34周组，34~37周组，≥37周组分别为（4.95±3.44）%，（8.89±8.49）%，（13.91±10.92）%，（7.19±7.11）%；其在28~36周逐渐升高，与胎龄呈正相关（r=0.355，P< 0.001）。≥37周TLR9 mRNA表达量下降，该值虽高于28~31周，但差异无统计学意义（P>0.05）。相关性分析表明，同胎龄时期同样本新生儿的TLR9 mRNA和TLR9阳性细胞率之间存在负相关（r=-0.227，P=0.011）。结论 TLR9阳性细胞率和TLR9 mRNA表达在不同胎龄组新生儿间有差异，TLR9阳性细胞率表达在31~37周间随着胎龄的增加而增加，TLR9 mRNA在28~36周间随着胎龄的增加而增加。     

Efficacy of a weight loss program prior to robot assisted radical prostatectomy in overweight and obese men with prostate cancer

BackgroundObesity in prostate cancer patients is associated with poor prostate-cancer specific outcomes. Exercise and nutrition can reduce fat mass; however, few studies have explored this as a combined pre-surgical intervention in clinical practice.PurposeThis study examined the efficacy of a weight loss program for altering body composition in prostate cancer patients prior to robot assisted radical prostatectomy (RARP).MethodsA retrospective analysis of 43 overweight and obese prostate cancer patients, aged 47–80 years, who completed a very low-calorie diet (~3000–4000 kJ) combined with moderate-intensity exercise (90 min/day) prior to RARP. Whole body and regional fat mass (FM) and lean mass (LM) were assessed by dual-energy x-ray absorptiometry pre- and post-program. Body weight, waist circumference, and blood pressure were assessed weekly, with surgery-related adverse effects recorded at time of surgery and follow-up appointments.ResultsWith a median of 29 days (IQR: 24–35days) on the program, patients significantly (p < 0.001) reduced weight (−7.3 ± 2.9 kg), FM (−5.0 ± 2.6 kg), percent body fat (−3.1 ± 2.5%), trunk FM (−3.4 ± 1.8 kg), LM (−2.4 ± 1.8 kg), and appendicular LM (−1.2 ± 1.0 kg). Lower weight, FM, percent FM, trunk FM, and visceral FM were associated with less surgery-related adverse effects (r_s = 0.335 to 0.468, p < 0.010). Systolic and diastolic blood pressure were reduced (p < 0.001) by 15 ± 22 and 8 ± 10 mmHg, respectively over the weight loss intervention.ConclusionUndertaking a combined low-calorie diet and exercise program for weight loss in preparation for RARP resulted in substantial reductions in FM, with improvements in blood pressure, that may benefit surgical outcomes.     

Experimental determination of the recombination correction factor kS for SNC 125c,SNC 350p and SNC 600c ionization chambers in pulsed photon beams

Accurate ionization chamber measurements of the absorbed dose to water require the correction of incomplete collection of charges created within the chamber volume. According to current dosimetry protocols such as the TRS-398 or the DIN 6800-2, incomplete charge collection is accounted for by the correction factor k_s, which can be determined numerically or experimentally. The method proposed by Burns & McEwen (Phys. Med. Biol., 1998) was used in this study to determine the coefficients γ and δ used for the calculation of the correction factor k_s of three ionization chambers, the SNC 125c, the SNC 600c and the SNC 350p (all Sun Nuclear Corp., Melbourne, Florida) for an absorbed dose to water range of 0.2 mGy to 1.6 mGy per pulse in pulsed photon beams. The shift of the effective point of measurement from the reference point Δz and the correction factor k_r were determined for the SNC 350p according to the draft DIN 6800-2:2019-07.