实时影像融合的超声虚拟导航技术联合射频消融术治疗原发性肝癌合并门静脉癌栓患者的疗效及对血清Bax、Cyfra21-1的影响

摘要 目的：探讨实时影像融合的超声虚拟导航技术联合射频消融术治疗原发性肝癌合并门静脉癌栓患者的疗效及对血清BCL-2同源的水溶性相关蛋白（Bax）、细胞角蛋白19片段（Cyfra21-1）的影响。方法：选择本院2017年1月到2021年4月收治的原发性肝癌合并门静脉癌栓患者82例作为研究对象,根据1：1随机数字表法将患者分为虚拟导航组与对照组各41例,虚拟导航组给予实时影像融合的超声虚拟导航技术联合射频消融术治疗,对照组给予单纯超声引导联合射频消融术治疗。结果：虚拟导航组的进针次数、融合时间、布针时间少于对照组（P<0.05）;虚拟导航组治疗后3个月的胆汁瘤、肝脓肿、膈肌损伤、肺部感染等并发症发生率为4.9 %,低于对照组的29.3 %（P<0.05）。虚拟导航组治疗后3个月的总有效率为82.9 %,高于对照组的51.2 %（P<0.05）。两组治疗后3个月的血清谷丙转氨酶（ALT）、谷草转氨酶（AST）水平低于治疗前,虚拟导航组低于对照组（P<0.05）。两组治疗后的血清Bax、Cyfra21-1含量低于治疗前,虚拟导航组低于对照组（P<0.05）。结论：实时影像融合的超声虚拟导航技术联合射频消融术治疗原发性肝癌合并门静脉癌栓能降低血清Bax、Cyfra21-1含量,改善患者的肝功能,提高消融效率,还可减少并发症的发生,最终提高患者的总体治疗效果。     

术前TFF1、HSP70、ASPH与原发性肝癌患者手术切除术后早期复发的关系研究

摘要 目的：研究术前三叶因子1（TFF1）、热休克蛋白70（HSP70）、天冬氨酸-天冬酰胺β羟化酶（ASPH）与原发性肝癌（PHC）患者手术切除术后早期复发的关系。方法：选取2018年1月～2019年12月我院收治的83例PHC患者,均行手术切除治疗,术后均随访2年,根据是否复发分为复发组49例以及未复发组34例。比较两组术前TFF1、HSP70、ASPH水平差异,收集患者基线资料,采用多因素Logistic回归分析PHC患者手术切除术后早期复发的危险因素,采用受试者工作特征（ROC）曲线分析术前TFF1、HSP70、ASPH水平预测PHC患者手术切除术后早期复发的效能。结果：复发组术前TFF1、HSP70、ASPH水平均高于未复发组（P＜0.05）。复发组肿瘤直径≥5 cm、肿瘤数目为多发、有血管侵犯的患者比例高于未复发组（P＜0.05）。多因素Logistic回归分析结果显示：肿瘤直径≥5 cm、多发肿瘤、血管侵犯及术前TFF1、HSP70、ASPH高水平是PHC患者手术切除术后早期复发的危险因素（均OR＞1,P＜0.05）。ROC曲线分析结果显示：术前TFF1、HSP70、ASPH联合检测预测PHC患者手术切除术后早期复发的曲线下面积为0.815,明显高于上述三项指标单独检测的0.704、0.713、0.707。结论：术前TFF1、HSP70、ASPH与PHC患者手术切除术后早期复发密切相关,联合检测三项指标水平可能有助于预测PHC患者手术切除术后早期复发。     

Mutational analysis of CYP1B1 (rs56010818) variant in primary open angle glaucoma (POAG) affected patients of Pakistan

BackgroundPrimary open angle glaucoma (POAG) occurs due to the discrepancies in the angle of anterior chamber characterized by the alterations in intraocular pressure, optic nerves head changes and central loss of visual field. In molecular research, CYP1B1 mutations modulates an integral role in association with glaucoma. Current study was undertaken to reveal the homozygous and heterozygous patterns of CYP1B1 c.1169 G > A variant (rs56010818) in POAG patients of Pakistan.MethodsAfter consent, total n = 88 POAG patients undergone through standard ophthalmological investigations before their recruitment in this study. The blood samples were utilized for DNA isolation. The genotyping of CYP1B1 c.1169 G > A variant was carried out by Sanger sequencing. The mutational patterns and its association with clinical variables were demonstrated by statistical and bioinformatic tools.ResultsIt was evident that the frequencies of heterozygous G/A and homozygous mutants A/A genotypes were higher in males (36.5%, 7.7%) than females (30.6%, 2.8%) of POAG population. Furthermore, the juvenile patients exhibit high manifestation of carrier genotype (66.6%) in comparison to adult patients (31.7%). The results also indicated the significant relationship of intraocular pressure with homozygous mutant A/A genotype of CYP1B1 variant in POAG patients (p < 0.05).ConclusionsOur study provided the mutational data of CYP1B1 R390H variant and the patterns of homozygosity and heterozygosity along with clinical associations. Overall, this study revealed the genetic predisposition of CYP1B1 c.1169 G > A variant in the patients of POAG in Pakistan. The findings could be helpful for genetic screening and in-depth understanding of underlying causes in the pathogenesis of POAG.     

树鼩原代皮肤成纤维细胞的分离培养技术

建立稳定的树鼩(Tupaiabelangeri)皮肤成纤维细胞的体外培养体系,可为有关此类细胞的实验和疾病树鼩细胞模型提供技术支持。取树鼩大腿内侧皮肤用组织块贴壁法和胶原酶Ⅰ消化法分离皮肤细胞,胰蛋白酶差别消化法纯化细胞;用MEM(10%FBS)完全培养基和含低血清生长添加物(LSGS)的培养基培养细胞;免疫荧光和蛋白印迹法鉴定细胞,并测定细胞的生长、冻存和复苏特性。经树鼩皮肤细胞分离效果比较,胶原酶消化法比组织块贴壁法更适合用于树鼩原代皮肤细胞分离;对分离及冻存复苏后细胞生长状况观察比较发现,添加了LSGS的MEM培养基更利于细胞存活、生长;细胞形态观察、免疫荧光和蛋白印迹检测鉴定所分离的细胞为树鼩皮肤成纤维细胞。成功建立了树鼩原代皮肤细胞的分离、纯化方法,并优化了该细胞的培养条件。     

树鼩原代小胶质细胞的分离培养、纯化与鉴定

本文旨在建立树鼩(Tupaia belangeri)小胶质细胞原代培养及纯化的方法,为利用新型实验动物树鼩进行相关研究工作提供实验材料。将新生树鼩大脑皮质机械分离,皮质组织块用胰蛋白酶消化后制成细胞悬液;培养9～10 d后,分别采用直立手拍法、温和胰酶消化法以及恒温振荡法分离纯化树鼩小胶质细胞,通过差速贴壁进一步纯化。荧光显微镜下,利用小胶质细胞的特异性标记物CD11b抗体进行鉴定。结果显示,小胶质细胞分离培养第3天时呈静息状态,表现为梭形、杆状、分支状等不规则形态。细胞免疫荧光CD11b呈阳性。不同纯化方法细胞免疫荧光并计数显示,直立手拍法所获得的细胞产量明显高于恒温振荡法(P 0.05),细胞阳性率( 96%)明显高于温和胰酶消化法( 90%,P 0.05)。直立手拍法可获得产量及纯度高的树鼩原代小胶质细胞。     

Progressive retinal ganglion cell loss in primary open-angle glaucoma is associated with temperature circadian rhythm phase delay and compromised sleep

Advanced primary open-angle glaucoma (POAG) is characterized by progressive retinal ganglion cell complex (RGCC) damage that may cause subsequent disruption of the circadian rhythms. Therefore, we evaluated circadian body temperature (BT) rhythm and sleep characteristics of 115 individuals (38 men and 77 women) diagnosed with POAG. GLV (global loss volume; %), a measure of RGCC damage, was estimated by high-definition optical coherence tomography, and RGC functional ability was assessed by pattern electroretinogram amplitude (PERGA). Depending on dynamics of POAG progression criteria, two groups were formed that were distinctively different in GLV: Stable POAG group (S-POAG; GLV = 5.95 ± 1.84, n = 65) and Progressive POAG group (P-POAG; GLV = 24.27 ± 5.09, n = 50). S-POAG and P-POAG groups were not different in mean age (67.61 ± 7.56 versus 69.98 ± 8.15) or body mass index (24.66 ± 3.03 versus 24.77 ± 2.90). All subjects performed 21 around-the-clock BT self-measurements during a 72-h period and kept activity/sleep diaries. Results showed pronounced disruption of circadian physiology in POAG and its progression with increasing severity of the disease. The daily mean of BT was unusually low, compared to age-matched controls. Moreover, our results revealed distinctive features of BT circadian rhythm alterations in POAG development and POAG progression. S-POAG is associated with lowered BT circadian rhythm robustness and inter-daily phase stability compared to controls. In the P-POAG group, the mean phase of the circadian BT rhythm was delayed by about 5 h and phases were highly scattered among individual patients, which led to reduced group mean amplitude. Circadian amplitudes of individuals were not different between the groups. Altogether, these results suggest that the body clock still works in POAG patients, but its entrainment to the 24-h environment is compromised. Probably because of the internal desynchronization, bedtime is delayed, and sleep duration is accordingly shortened by about 55 min in P-POAG compared to S-POAG patients. In the entire POAG cohort (both groups), later sleep phase and shorter mean sleep duration correlate with the delayed BT phase (r = 0.215; p = 0.021 and r = 0.322; p = 0.0004, respectively). An RGCC GLV of 15% apparently constitutes a threshold above which a delay of the circadian BT rhythm and a shortening of sleep duration occur.     

Nitric oxide plays a central role in determining lateral root development in tomato

Nitric oxide (NO) is a bioactive molecule that functions in numerous physiological processes in plants, most of them involving cross-talk with traditional phytohormones. Auxin is the main hormone that regulates root system architecture. In this communication we report that NO promotes lateral root (LR) development, an auxin-dependent process. Application of the NO donor sodium nitroprusside (SNP) to tomato (Lycopersicon esculentum Mill.) seedlings induced LR emergence and elongation in a dose-dependent manner, while primary root (PR) growth was diminished. The effect is specific for NO since the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (CPTIO) blocked the action of SNP. Depletion of endogenous NO with CPTIO resulted in the complete abolition of LR emergence and a 40% increase in PR length, confirming a physiological role for NO in the regulation of root system growth and development. Detection of endogenous NO by the specific probe 4,5-diaminofluorescein diacetate (DAF-2 DA) revealed that the NO signal was specifically located in LR primordia during all stages of their development. In another set of experiments, SNP was able to promote LR development in auxin-depleted seedlings treated with the auxin transport inhibitor N-1-naphthylphthalamic acid (NPA). Moreover, it was found that LR formation induced by the synthetic auxin 1-naphthylacetic acid (NAA) was prevented by CPTIO in a dose-dependent manner. All together, these results suggest a novel role for NO in the regulation of LR development, probably operating in the auxin signaling transduction pathway.Abbreviations CPTIO 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide - DAF-2 DA 4,5-Diaminofluorescein diacetate - LR Lateral root - NAA 1-Naphthylacetic acid - NO Nitric oxide - NPA N-1-Naphthylphthalamic acid - PR Primary root - SNP Sodium nitroprusside     

Regulation of primary spinal neuron lineages after deletion of a major progenitor

Vertebrate embryos are able to reconstitute the body plan when early blastomeres are deleted, but it is not known whether this is accomplished by cells local to the lesion or by a readjustment of the entire pattern of the embryo. We distinguished between these two possibilities by studying which embryonic cells change primary spinal neuronal fates after deletion of a major spinal cord progenitor. After ablation of the V1.2 blastomere of the 16-cell Xenopus embryo, the spinal cord contained normal numbers of Rohon-Beard neurons and primary motoneurons, indicating that the remaining blastomeres numerically reconstituted these populations. Using lineage-tracing techniques we revealed a global response: 10 out of the 15 remaining blastomeres significantly changed the number of one or both neuronal types they produced. This widespread response indicates that position in the early embryo plays an important role in regulating the production of primary spinal neurons. However, not all cells are influenced solely by position; a vegetal cell transplanted into the position of the deleted V1.2 did not take on the neuronal fate of its new position. Thus, restitution of pattern relies on a combination of positional cues and intrinsic fate restrictions.     

Mass spectrometric strategy for primary structure determination of N-terminally blocked peptides

The mass spectrometric strategy including three steps is presented for primary structure determination of the N-terminally blocked peptides. First, the C-terminal sequencing is performed by using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry coupled with carboxypeptidase Y digestion. Then, the peptide is cleaved according to the obtained C-terminal sequence information and the resulting peptides are identified by mass spectrometry and Edman degradation after fractionation by reverse-phase chromatography. Finally, the N-terminal fragment is sequenced by tandem mass spectrometry. The strategy was successfully applied to the sequence determination of two novel N-terminally blocked peptides named EAFP1 and EAFP2.     

Mitochondria and autoimmunity in primary biliary cirrhosis

Primary biliary cirrhosis is an enigmatic autoimmune liver disease that predominantly affects women and is characterized by antimitochondrial antibodies and specific destruction of small bile ducts. Interestingly, patients with this disease not only have high titer antibodies to mitochondria, but also highly directed, liver-specific CD4 and CD8 cells directed at the same mitochondrial autoantigens. These mitochondrial autoantigens are all members of the 2-oxo dehydrogenase complex family and include the E2 component of pyruvate dehydrogenase as the major autoantigen. Moreover, the epitopes recognized by CD4, CD8 T cells and autoantibody, are all directed within the same region, namely the lipoyl domain of pyruvate dehydrogenase complex-E2. All cells in the body have mitochondria but there appear to be specific destruction of biliary cells. We believe that this specific destruction is secondary to a highly directed mucosal response that focuses on biliary cells because of the involvement of a polymeric immunoglobulin receptor, the presence of immunoglobulin A in mucosal secretions, and the unique apoptotic properties of biliary epithelium.