腔镜下Soave根治术与开腹改良Soave术治疗长段型先天性巨结肠患儿的疗效比较及对应激反应和控便功能的影响

摘要 目的：比较腔镜下Soave根治术与开腹改良Soave术治疗长段型先天性巨结肠（HD）患儿的疗效,观察两种术式对应激反应和控便功能的影响。方法：选取我院2017年4月～2020年9月期间收治的长段型HD患儿88例,根据手术方式的不同分为开腹组和微创组,例数分别为43例和45例。对比两组围术期指标、应激反应指标、控便功能和并发症发生情况。结果：微创组的术中失血量少于开腹组,手术时间、胃肠功能恢复时间、禁食时间、住院时间短于开腹组（P<0.05）,两组肠管切除长度组间对比无统计学差异（P>0.05）。两组患儿术后1 d心率（HR）、平均动脉压（MAP）较术前升高,血氧饱和度（SpO₂）较术前下降,但微创组HR、MAP低于开腹组,SpO₂高于开腹组（P<0.05）。两组患儿术后1年大便性状、排便次数、污粪、需要治疗（灌肠、药物、尿布）评分及Heikkinen总分均较术前升高,且微创组高于开腹组（P<0.05）。微创组的近期并发症总发生率和远期并发症总发生率均低于开腹组（P<0.05）。结论：与开腹改良Soave术相比,采用腔镜下Soave根治术治疗长段型HD患儿可缩短手术时间、禁食时间、住院时间、胃肠功能恢复时间,减少手术创伤,减轻机体应激反应,改善患儿控便功能,同时还可降低并发症发生率,效果较好。     

Spatio-temporal changes in germination and radical elongation of barley seeds tracked by proteome analysis of dissected embryo, aleurone layer, and endosperm tissues

Germination of barley is accompanied by changes in water-soluble seed proteins. 2-DE was used to describe spatio-temporal proteome differences in dissected seed tissues associated with germination and the subsequent radicle elongation. Protein identification by MS enabled assignment of proteins and functions to the seed embryo, aleurone, and endosperm. Abundance in 2-DE patterns was monitored for 48 different proteins appearing in 79 gel spots at 8 time-points up to 72 h post imbibition (PI). In embryo, a beta-type proteasome subunit and a heat shock protein 70 fragment were among the earliest proteins to appear (at 4 h PI). Other early changes were observed that affected spots containing desiccation stress-associated late embryogenesis abundant and abscisic acid (ABA)-induced proteins. From 12 h PI proteins characteristic for desiccation stress disappeared rapidly, as did a putative embryonic protein and an ABA-induced protein, suggesting that these proteins are also involved in desiccation stress. Several redox-related proteins differed in spatio-temporal patterns at the end of germination and onset of radicle elongation. Notably, ascorbate peroxidase that was observed only in the embryo, increased in abundance at 36 h PI. The surprisingly early changes seen in the protein profiles already 4 h after imbibition indicate that germination is programmed during seed maturation.     

Eye lens exposure to medical staff during endoscopic retrograde cholangiopancreatography

The paper presents a study of the radiation doses to eye lens of medical staff during endoscopic retrograde cholangiopancreatography (ERCP) procedures performed in a busy gastroenterology department. For each procedure the dose equivalent to the eye, exposure time, dose rate, Kerma Area Product and fluoroscopy time were recorded. Measurements were performed for a period of two months in four main positions of the operating staff, and then extrapolated to estimate annual doses. The fluoroscopy time per ERCP procedure varied between 1.0 min and 28.8 min, with a mean value of 4.6 min. The calculated mean eye dose per procedure varied between 34.9 μSv and 93.3 μSv. The results demonstrated that if eye protection is not used, annual doses to the eye lens of the gastroenterologist performing the procedure and the anesthesiologist can exceed the dose limit of 20 mSv per year.     

全胸腔镜肺叶切除联合淋巴结清扫术治疗老年非小细胞肺癌患者的临床研究

目的:了解全胸腔镜肺叶切除联合淋巴结清扫术治疗老年非小细胞肺癌病人的临床效果。方法:选取我院2009年2月到2011年2月接受全胸腔镜肺叶切除联合淋巴结清扫术治疗的老年非小细胞肺癌患者共102例,其中高龄组(≧70岁)24例;非高龄组(70岁)78例。观察两组手术时间、手术中的出血量、术后的住院时间、术后的输血量、胸液总量、胸管放置时间、术后并发症、止痛药使用的次数等情况,并对两组患者进行随访了解其早期生存情况。结果:两组手术时间、术中出血量、术后输血量、术后住院时间、胸液的总量、胸管放置的时间、止痛药的使用次数、术后的并发症、平均的淋巴结数和淋巴结转移情况比较均无统计学差异(P0.05)。非高龄组1年生存率为98.1%,2年生存率为80.3%,高龄组1年生存率为95.3%,2年生存率为73.1%,非高龄组与高龄组早期生存状况比较无统计学意义差异(P0.05)。结论:采用全胸腔镜肺叶切除联合淋巴结清扫术治疗非小细胞肺癌病人,与非高龄病人进行比较,高龄病人能获得同样的治疗疗效和早期预后,所以,可在临床上采纳。     

Random dissection to select for protein split sites and its application in protein fragment complementation

To identify protein split sites quickly, a selection procedure by using chloramphenicol acetyl transferase (CAT) as reporter was introduced to search for folded protein fragments from libraries generated by random digestion and reassembly of the target gene, which yielded an abundant amount of DNA fragments with controllable lengths. Experimental results of tryptophan synthase alpha subunit (TSα) and TEM‐1 β‐lactamase agreed well with what the literature has reported. The solubility of these fragments correlated roughly with the minimum inhibitory concentrations of the CAT fusions. The application of this dissection protocol to protein fragment complementation assay (PCA) was evaluated using aminoglycoside‐3′‐phosphotransferase I (APH(3′)‐I) as a model protein. Three nearly bisectional sites and a number of possible split points were identified, and guided by this result, four novel pairs of fragments were tested for complementation. Three out of four pairs partially restored the APH activity with the help of leucine zippers, and a truncated but active APH(3′)‐I (Δ1–25) was also found. Finally, the weakly active APH(3′)‐I‐(1–253)NZ/CZ (254–271) containing a short 18 residue tag was further improved by error‐prone PCR, and a best mutant was obtained showing a fourfold improvement after just one round of evolution. These results demonstrate that protein random dissection based on the CAT selection can provide an efficient search for protein breakage points and guide the design of fragments for protein complementation assay. Furthermore, more active fragment pairs can be achieved with the classical directed evolution approach.     

Compartmental fasciotomy and isolating a muscle from neighboring muscles interfere with myofascial force transmission within the rat anterior crural compartment

Muscles within the anterior crural compartment (extensor digitorum longus, EDL; tibialis anterior, TA; and extensor hallucis longus, EHL) and within the peroneal compartment were excited simultaneously and maximally. All muscles were kept at constant length with the exception of EDL, for which muscle length was changed by moving its proximal tendon. Active and passive force was measured at proximal as well as distal EDL tendons and at the combined distal tendons of TA and EHL (TA+EHL). In the initial experimental condition, a difference (F(proximal) > F(distal)) in EDL force, amounting to 0-14% of proximal force, was confirmed for most EDL lengths. This is interpreted as a clear proof of extramuscular myofascial force transmission, as no significant EDL length effects could be shown on TA+EHL force. Repeated measurements were confirmed to cause marked changes of both proximal and distal length-force characteristics, such as a shift of the whole ascending limb of the active curve, including optimum length, to higher lengths without decreasing optimum force, and decreasing active force at low lengths (by approximately 57%). Repeated measurements also lowered proximal and distal EDL passive force (by up to 35%). The proximo-distal difference in passive as well as active EDL force was decreased, but persisted. At most lengths, this difference for active force amounted to a constant fraction (14%) of proximal force. TA+EHL force was not affected significantly. Subsequently, acute effects of experimental surgical alterations were studied: The first manipulation was full lateral fasciotomy of the anterior crural compartment that caused a further decrease in active force at the proximal EDL but not at the distal EDL tendon. Passive forces showed no further significant changes. The proximo-distal EDL active force difference decreased to 0-5% of proximal force. After fasciotomy, TA+EHL force increased by 30%. This was interpreted as evidence of increased intramuscular and decreased extramuscular myofascial force transmission. The second manipulation was full isolation of EDL from TA+EHL, but not from extramuscular connective tissues, which caused a further decrease of the EDL proximo-distal force differences, indicating a stiffening effect of the presence of TA+EHL on the extramuscular matrix. For EDL active force the difference was no longer significantly different from zero. In contrast, for EDL passive force the proximo-distal force difference persisted. It is concluded that extramuscular myofascial force transmission is an important feature of the anterior crural compartment. The magnitude of this force transmission requires that it be considered in analysis of muscular function.     

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging

The Drosophila brain and visual system are widely utilized model systems to study neuronal development, function and degeneration. Here we show three preparations of the brain and visual system that cover the range from the developing eye disc-brain complex in the developing pupae to individual eye and brain dissection from adult flies. All protocols are optimized for the live culture of the preparations. However, we also present the conditions for fixed tissue immunohistochemistry where applicable. Finally, we show live imaging conditions for these preparations using conventional and resonant 4D confocal live imaging in a perfusion chamber. Together, these protocols provide a basis for live imaging on different time scales ranging from functional intracellular assays on the scale of minutes to developmental or degenerative processes on the scale of many hours.     

Endoscopic ultrasound-guided fine needle aspiration (EUS-FNA): experience of an academic centre in the USA

S. Zhang, D. V. S. DeFrias, R. Alasadi and R. Nayar
Endoscopic ultrasound‐guided fine needle aspiration (EUS‐FNA): experience of an academic centre in the USA Objectives: Endoscopic ultrasound‐guided fine needle aspiration (EUS‐FNA) has become widely accepted as an effective modality for obtaining tissue for primary diagnosis and staging. We have been using EUS‐FNA since July 2001 and herein we summarize our experience over a 5‐year period. Methods: A computer‐based search for in‐house EUS‐FNA was performed in the pathology database from July 2001 to October 2006. To calculate the sensitivity, specificity and accuracy of EUS‐FNA, the cytology diagnosis was compared with the surgical follow‐up. Results: A total of 951 EUS‐FNAs were performed during the study period and included 279 pancreatic solid lesions, 186 pancreatic cyst lesions, 249 lymph node aspirations, 111 gastrointestinal (GI) tract submucosal lesions, and 126 miscellaneous lesions. EUS‐FNA had a very high sensitivity and accuracy for solid pancreatic lesions (94.7 and 97.7%, respectively), low sensitivity and accuracy but high specificity (47, 64.8 and 95%, respectively) for cystic lesions. Cyst fluid carcinoembryonic (CEA) levels were significantly higher in mucinous neoplasms than non‐neoplastic cysts. EUS‐FNA also had very high sensitivity and specificity for detecting metastatic carcinoma in lymph nodes (95 and 100%, respectively). GI submucosal spindle cell tumours were further classified with immunohistochemical stains performed either on a cell block or a core biopsy obtained via EUS guidance. Conclusions: EUS‐FNA has a very high sensitivity and accuracy for pancreatic solid lesions, but the sensitivity for cystic lesions is generally low. Cyst fluid chemical analysis for CEA is helpful, but the overlap between mucinous neoplasm and non‐neoplastic cysts is significant. Recognizing GI contamination is important and immunohistochemical stains are useful for GI submucosal spindle cell lesions.