腹腔镜肝癌切除术联合脾脏切除对肝癌合并脾功能亢进的早期效果分析

目的探讨腹腔镜肝癌切除术联合脾脏切除对肝癌合并脾功能亢进的早期疗效。 方法回顾性分析2016年1月至2019年5月接受腹腔镜手术治疗的54例肝癌合并脾功能亢进患者临床资料。将肝癌切除同时行脾脏切除者纳入切脾组(25例),仅行肝癌切除术保留脾脏者纳入保脾组(29例)。临床数据使用统计学软件SPSS 24.0分析,术中术后各项指标、肝功能及免疫指标采用( ±s)表示,独立样本t检验;术后并发症等组间比较采用χ²检验。P<0.05为检验标准。 结果两组患者均成功完成手术,无中转开腹。切脾组手术时间明显较保脾组更长(P＝0.003),术中出血量及术后住院时间差异无统计学意义(P>0.05)。术后1周,两组患者外周血WBC、PLT均较术前明显升高,且切脾组显著高于保脾组(P<0.05);两组患者肝功能指标ALT、AST、Tbil均较术前明显升高,保脾组较切脾组更高(P<0.05);术后免疫功能指标切脾组CD4、CD4/CD8显著升高,而CD8显著降低,且明显优于保脾组(P<0.05)。切脾组及保脾组患者早期并发症发生率分别为16.0%及17.2%,两组间差异无统计学意义(P>0.05)。 结论腹腔镜肝癌切除联合脾脏切除治疗肝癌合并脾功能亢进安全可行,降低了手术创伤,早期疗效满意。     

A photo-switchable and thermal-enhanced fluorescent hydrogel prepared from N-isopropylacrylamide with water-soluble spiropyran derivative

Herein, a photo-switchable and thermal-enhanced fluorescent hydrogel has been fabricated from N-isopropylacrylamide (NIPAAm) with a mixture of water-soluble acryloyl-α-cyclodextrin/acryloyl-α-cyclodextrin-spiropyran (acryloyl-α-CD/ acryloyl-α-CD-SP) as cross-linkers. The physical properties, photochromic properties, and fluorescent behavior of the hydrogel were characterized. The fluorescence emission of the hydrogel can be reversibly switched ‘on/off’ by UV/visible light irradiation, and meanwhile the fluorescence intensity can be enhanced by increasing the temperature above the volume phase transition temperature (VPTT) of the hydrogel. The hydrogel also shows spatiotemporal fluorescent behavior, excellent cytocompatibility, and fatigue resistance in photochromic and photo-switchable fluorescent behaviors.     

Prevalence,genotypes, and risk factors of <Emphasis Type="Italic">Enterocytozoon bieneusi</Emphasis> in Asiatic black bear (<Emphasis Type="Italic">Ursus thibetanus</Emphasis>) in Yunnan Province,Southwestern China

Microsporidiosis is an important zoonotic disease, even leading to severe diarrhea. However, no information about prevalence and genotypes of Enterocytozoon bieneusi infection in Asiatic black bears in southwestern China is available. The objectives of the present study were to investigate the prevalence of E. bieneusi and to characterize their genotypes using the nested PCR amplification of the internal transcribed spacer region of the ribosomal RNA gene cluster and multilocus sequence typing (MLST). The overall prevalence of E. bieneusi was 19.75% (80/405) and the rate of E. bieneusi in Xishuangbanna (33.33%) was significantly higher than that in any other regions (Honghe, 17.65%; Dehong, 13.04%; Kunming, 0; P?=?0.01). Sequence analysis revealed that 4 known genotypes (D, n?=?2; SC02, n?=?10; SC01, n?=?5; and CHB1, n?=?4) and 13 novel genotypes (designed MJ1–MJ13) were identified. When 17, 5, 14, and 34 sequences at loci MS1, MS3, MS4, and MS7 via MLST analyses, representing 4, 4, 5, and 10 genotypes, respectively, were completed, one multilocus genotype (MLG novel-ABB1) was identified. This is the first report of E. bieneusi in Asiatic black bear in Yunnan province, Southwestern China. The results indicated the potential zoonotic risk of this parasite through the Asiatic black bear in this region and provided foundation data for preventing and controlling E. bieneusi infection of many other animals and humans in these regions.     

Cbl Ubiquitin Ligases Control B Cell Exit from the Germinal-Center Reaction

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A novel approach for rapid high-throughput selection of recombinant functional rat monoclonal antibodies

Background

Most monoclonal antibodies against mouse antigens have been derived from rat spleen-mouse myeloma fusions, which are valuable tools for purposes ranging from general laboratory reagents to therapeutic drugs, and yet selecting and expressing them remains a time-consuming and inefficient process. Here, we report a novel approach for the rapid high-throughput selection and expression of recombinant functional rat monoclonal antibodies with different isotypes.

Results

We have developed a robust system for generating rat monoclonal antibodies through several processes involving simultaneously immunizing rats with three different antigens expressing in a mixed cell pools, preparing hybridoma cell pools, in vitro screening and subsequent cloning of the rearranged light and heavy chains into a single expression plasmid using a highly efficient assembly method, which can decrease the time and effort required by multiple immunizations and fusions, traditional clonal selection and expression methods. Using this system, we successfully selected several rat monoclonal antibodies with different IgG isotypes specifically targeting the mouse PD-1, LAG-3 or AFP protein from a single fusion. We applied these recombinant anti-PD-1 monoclonal antibodies (32D6) in immunotherapy for therapeutic purposes that produced the expected results.

Conclusions

This method can be used to facilitate an increased throughput of the entire process from multiplex immunization to acquisition of functional rat monoclonal antibodies and facilitate their expression and feasibility using a single plasmid.

     

Persistent reduction in sialylation of cerebral glycoproteins following postnatal inflammatory exposure

Background

The extension of sepsis encompassing the preterm newborn’s brain is often overlooked due to technical challenges in this highly vulnerable population, yet it leads to substantial long-term neurodevelopmental disabilities. In this study, we demonstrate how neonatal neuroinflammation following postnatal E. coli lipopolysaccharide (LPS) exposure in rat pups results in persistent reduction in sialylation of cerebral glycoproteins.

Methods

Male Sprague-Dawley rat pups at postnatal day 3 (P3) were injected in the corpus callosum with saline or LPS. Twenty-four hours (P4) or 21?days (P24) following injection, brains were extracted and analyzed for neuraminidase activity and expression as well as for sialylation of cerebral glycoproteins and glycolipids.

Results

At both P4 and P24, we detected a significant increase of the acidic neuraminidase activity in LPS-exposed rats. It correlated with significantly increased neuraminidase 1 (Neu1) mRNA in LPS-treated brains at P4 and with neuraminidases 1 and 4 at P24 suggesting that these enzymes were responsible for the rise of neuraminidase activity. At both P4 and P24, sialylation of N-glycans on brain glycoproteins decreased according to both mass-spectrometry analysis and lectin blotting, but the ganglioside composition remained intact. Finally, at P24, analysis of brain tissues by immunohistochemistry showed that neurons in the upper layers (II–III) of somatosensory cortex had a reduced surface content of polysialic acid.

Conclusions

Together, our data demonstrate that neonatal LPS exposure results in specific and sustained induction of Neu1 and Neu4, causing long-lasting negative changes in sialylation of glycoproteins on brain cells. Considering the important roles played by sialoglycoproteins in CNS function, we speculate that observed re-programming of the brain sialome constitutes an important part of pathophysiological consequences in perinatal infectious exposure.

     

补骨脂素对骨质疏松小鼠骨折修复的影响

目的观察补骨脂素对骨质疏松小鼠骨折修复的影响。方法 20只雌性C57BL/6小鼠在建立骨质疏松模型后,随机分为2组:模型组和补骨脂素组,每组10只。所有小鼠在右侧股骨中段建立横形骨折,骨折术后第1天,补骨脂素组按20 mg/kg/d体重量灌胃,每天一次,连续21天,模型组以等剂量生理盐水灌胃。术后21天处死,骨折标本行钼靶X线、组织学和生物力学等检测方法。结果骨折术后第21天,补骨脂素组较模型组骨痂密度变高、骨折线模糊,两组骨痂生成面积比较无明显差异。补骨脂素组软骨细胞数量较模型组明显减少,成骨细胞增生较活跃,软骨骨化形成编织骨。补骨脂素组破骨细胞密度更稀疏,2组中破骨细胞形态和细胞核数量见明显差异。补骨脂素组最大载荷比模型组明显升高。结论补骨脂素作为一种植物雌激素能有效改善骨质疏松小鼠骨痂愈合质量和生物力学性能。     

Autophagy protects C. elegans against necrosis during Pseudomonas aeruginosa infection

Autophagy, a conserved pathway that delivers intracellular materials into lysosomes for degradation, is involved in development, aging, and a variety of diseases. Accumulating evidence demonstrates that autophagy plays a protective role against infectious diseases by diminishing intracellular pathogens, including bacteria, viruses, and parasites. However, the mechanism by which autophagy regulates innate immunity remains largely unknown. Here, we show that autophagy is involved in host defense against a pathogenic bacterium Pseudomonas aeruginosa in the metazoan Caenorhabditis elegans. P. aeruginosa infection induces autophagy via a conserved extracellular signal-regulated kinase (ERK). Intriguingly, impairment of autophagy does not influence the intestinal accumulation of P. aeruginosa, but instead induces intestinal necrosis. Inhibition of necrosis results in the survival of autophagy-deficient worms after P. aeruginosa infection. These findings reveal a previously unidentified role for autophagy in protection against necrosis triggered by pathogenic bacteria in C. elegans and implicate that such a function of autophagy may be conserved through the inflammatory response in diverse organisms.Autophagy, a well-conserved lysosomal pathway that involves the degradation of cytoplasmic components, plays important roles in a broad diversity of the biological processes, ranging from development, senescence, and lifespan extension, to cancer (1, 2). In addition, autophagy has a prominent role in resistance to bacterial, viral, and protozoan infection in metazoan organisms (3–6). Autophagy is unique in its capacity to sequester invading bacteria, and target these pathogens for lysosomal degradation, thus providing a mechanism for the elimination of intracellular microorganisms. For example, after the pathogenic bacterium Streptococcus pyogenes (group A Streptococcus) enters human epithelial cells, the bacterium in the cytoplasm is sequestered in autophagosome-like compartments and degraded upon fusion with lysosomes. In contrast, Streptococcus exits freely into the cytoplasm of autophagy-deficient Atg5^−/− cells that lack autophagic ability (3). In phagocytic cells, such as macrophages, Mycobacterium tuberculosis resides intracellularly in the phagosome and blocks phagolysosome biogenesis (4). Induction of autophagy by physiological or pharmacological factors promotes mycobacterial colocalization with the autophagosomes and results in a decreased viability of intracellular mycobacteria (4, 7). In addition to pathogen clearance, accumulating evidence suggests that autophagy is associated with other aspects of immunity and inflammation (8–11). For instance, lack of autophagy in macrophages results in the accumulation of dysfunctional mitochondria, which, in turn, promotes secretion of proinflammatory factors IL-1β and IL-18 (8, 10), suggesting that autophagy regulates inflammation responses by suppressing the secretion of immune mediators.The genetically tractable model host Caenorhabditis elegans provides a useful tool to study the innate immune system and the mechanism of host–pathogen interactions (12). In C. elegans, the autophagy machinery serves as an essential component of the host defense by degrading intracellular pathogens. Inhibition of autophagy in C. elegans leads to a significant increase in accumulation of the microsporidian Nematocida parisii, an intracellular pathogen of worms, whereas activation of autophagy by knockdown of let-363, the worm ortholog of TOR, causes a dramatic decrease in pathogen load (13). When the pathogenic bacterium Salmonella typhimurium invades the intestinal epithelial cells of worms, the bacterium is targeted to lysosomes for degradation in wild-type (WT), but not in autophagy-deficient animals (5). Besides S. typhimurium, several clinically relevant bacteria, such as Pseudomonas aeruginosa (14) and Staphylococcus aureus (15), also induce autophagy in C. elegans. Although inhibition of autophagy by knockdown of vps-34, a gene crucial for early steps of autophagy, reduces the survival of worms after S. aureus infection, vps-34 RNAi does not affect intestinal pathogen load. Thus, the role for autophagy in defense against S. aureus is probably not due to clearance of the pathogen, implicating that other mechanisms of host defense might exist.So far, the following major signaling cascades have been identified as involved in C. elegans defense against pathogenic bacteria: the p38 mitogen-activated protein kinase (MAPK) PMK-1 (16), DAF-16 (17), the extracellular signal-regulated kinase (ERK) MAPK MPK-1 (14, 18), the protein kinase D DKF-2 (19), the G protein-coupled receptor FSHR-1 (20), and the G protein Gqα EGL-30 (21). Here, we investigated the function of autophagy in innate immune responses to P. aeruginosa PA14 in C. elegans. Furthermore, we investigated the role of the signaling pathways related to innate immunity in the activation of autophagy and discovered that the ERK signaling was involved in autophagy during P. aeruginosa PA14 infection. Finally, our results show that autophagy protects worms against necrosis-induced organismal death triggered by P. aeruginosa.     

Female breast cancer statistics of 2010 in China: estimates based on data from 145 population-based cancer registries

Background

The aim of the study is to provide incidence and mortality data of female breast cancer at national level of China in 2010.

Methods

A total of 145 population-based cancer registries submitted qualified cancer incidence and mortality data to National Cancer Registration Center of China. Based on the qualified cancer registries’ data, we estimated the overall breast cancer incidence and mortality data of China in 2010 and reported breast cancer statistics by age and geographical area.

Results

The estimated number of female breast cancer cases was about 208 thousand. The crude incidence rate, age-standardized rate by China and World population were 32.43 per 100,000, 25.89 per 100,000 and 24.20 per 100,000, respectively. The incidence rates were higher in urban area than in rural area. And the incidence rates in Eastern area and Middle area were similar and higher that those in Western areas. The estimated number of female breast cancer death in 2010 of China was about 55.5 thousand. The crude, age-standardized mortalities by China population and World population were 8.65 per 100,000, 6.56 per 100,000 and 6.36 per 100,000, respectively. The mortality rates by geographical area had similar pattern to the incidence rates.

Conclusions

Breast cancer is still a major health burden for Chinese women especially in urban area. Prevention strategies such as weight control, high-quality screening and diagnosis may help control the disease.