血管周细胞在表达PDGF-B的实体瘤中诱发造血

背景与目的肿瘤组织持续生长需要红细胞充分供氧。在各类病理生理情况下,成年哺乳动物可在主要造血器官骨髓之外发生髓外造血。然而,肿瘤组织能否造血是完全未知的。大量证据表明,在肿瘤微环境中,血管周细胞保留了祖细胞特性,可分化成其他细胞类型。在此,我们探究了肿瘤组织内周细胞是否能调节造血及其作用机制。方法使用鼠源周细胞进行全基因组表达谱分析以测试周细胞是否可向红细胞分化。使用NG2-CreERT2:R26R-tdTomato小鼠对血管周细胞进行在体遗传示踪。使用流式细胞术、单细胞测序和集落形成实验对分选细胞进行鉴定。使用定量聚合酶链反应(quantitative polymerase chain reactionq,PCR)、酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)、细胞磁珠分选和免疫染色检查红细胞生成素(erythropoietin,EPO)的表达。构建骨髓移植小鼠模型以研究骨髓对肿瘤红细胞生成的影响。结果表达谱显示,在PDGF-B处理后,NG2+周细胞表现出造血干祖样特征,并向红系谱系分化。同时,PDGF-B可靶向癌症相关成纤维细胞以产生促红细胞生成的关键激素EPO。通过遗传示踪及流式分析,我们定义了NG2+细胞衍生的造血亚群。进一步的单细胞测序和集落形成实验结果显示,在PDGF-B刺激下,肿瘤中的NG2+细胞形成了与典型的骨髓造血干细胞不同的红系祖细胞。结论本研究提出了肿瘤组织内造血的新概念,并阐明肿瘤周细胞分化为红细胞的分子机制。靶向肿瘤造血作为一种新的治疗理念,可能对肿瘤治疗产生深远影响。

FAT10 level in human gastric cancer and its relation with mutant p53 level,lymph node metastasis and TNM staging

AIM:To investigate the role of FAT10 and mutant p53 in the pathogenesis,severity and prognosis of gastric cancer.METHODS:FAT10,mutant p53 mRNA and protein levels were measured by reverse transcription(RT)-PCR and immunohistochemistry in gastric cancer tissue(n = 62),tumoradjacent tissue(n = 62) and normal gastric tissue(n = 62).Relation of FAT10 and mutant p53 expression with clinicopathological features and clinical outcomes of gastric cancer patients were analyzed.RESULTS:The FAT10,mutant p53 mRNA and protein levels were signif icantly higher in gastric cancer than in its adjacent and normal tissue.The FAT10 and mutant p53 levels in gastric cancer tissue were significantly correlated with lymph node metastasis and tumor,nodes,metastasis(TNM) staging.Moreover,the high FAT10 level was associated with the overall survival rate of patients.Multivariate Cox-proportional hazards model analysis showed that mRNA and protein levels of FAT10 and mutant p53,lymph node metastasis,distant metastasis and TNM stage were the independent prognostic factors for gastric cancer.CONCLUSION:FAT10 may be involved in gastric carcinogenesis,and is a potential marker for the prognosis of gastric cancer patients.FAT10 and mutant p53 may play a common role in the carcinogenesis of gastric cancer.

Hepatitis E virus chimeric DNA vaccine elicits immunologic response in mice

AIM: To construct the plasmid pcHEV23 containing fragments of HEV ORF2 and ORF3 chimeric gene and to assess its ability to elicit specific immunologic response in mice.METHODS: The gene encoding the structural protein of HEV ORF2 fragment and full-length ORF3 was amplified by PCR. The PCR products were cloned into an eucaryotic expression plasmid pcDNA3. The resulting plasmid pcHEV23 was used as a DNA vaccine to inoculate BALB/c mice intramuscularly thrice at a dose of 100 or 200 μg.Mice injected with empty pcDNA3 DNA or saline served as control and then specific immune responses in the mice were detected.RESULTS: After 2-3 times of inoculation, all mice injected with pcHEV23 had anti-HEV IgG seroconversion and specific T lymphocyte proliferation. The lymphocyte stimulation index in the group immunized with pcHEV23(3.1±0.49) was higher than that in the control group (0.787±0.12, P＜0.01). None in the control group had a detectable level of anti-HEV IgG.CONCLUSION: DNA vaccine containing HEV ORF2 and ORF3 chimeric gene can successfully induce specific humoral and cellular immune response in mice.

Effects of propofol on ammonium chloride-exposed astrocyte morphology and aquaporin-4 expression

Ammonia induces astrocyte swelling, which is strongly associated with overexpression of aquaporin-4. However, the mechanisms by which ammonia induces astrocyte swelling, and subsequently upregulating aquaporin-4 expression, remain unknown. In the present study, astrocytes were cultured in vitro and exposed to ammonium chloride （NH4CI）, followed by propofol protein kinase C agonist, or antagonist, respectively. Astrocyte morphology was observed by light microscopy, and aquaporin-4 expression was detected by western blot analysis. Results showed that propofol or protein kinase C agonist significantly attenuated the degree of NH4CI-induced astrocyte swelling and inhibited increased aquaporin-4 expression. Propofol treatment inhibited aquaporin-4 overexpression in cultured astrocyte induced by NH4CI; protein kinase C pathway activation is potentially involved.

Preoperative intestinal stent decompression with primary laparoscopic surgery to treat left-sided colorectal cancer with obstruction: a report of 21 cases

Objective: This work aimed to study the safety and efficacy of preoperative intestinal stent decompression combined with laparoscopic surgery to treat left-sided colorectal cancer with obstruction (LCCO). Methods: Retrospective analysis was conducted on data obtained from 21 LCCO patients admitted to The First Affiliated Hospital of Zhejiang Chinese Medicine University during March 2008 and December 2011. To remove the intestinal obstruction, preoperative intestinal stent placement under colonoscopic guidance was performed. Approximately 7 to 10 days after the operation, laparoscopic radical surgery of colorectal cancer was conducted. Results: Among the 21 cases studied, laparoscopic surgery was successful in 20 patients. Emergent laparotomy was conducted in one patient because of tumor invasion in the ureter. The duration of the operation ranged from 180 to 320 min, and the average time was 220 min. The recovery time for bowel function ranged from 2 to 5 days with an average time of 3 days. Postoperative infection of the incision occurred in one case. No anastomotic leakage was observed in any of the cases. Conclusion: Preoperative intestinal stent decompression, combined with primary stage laparoscopic surgery, is a safe and effective method for the treatment of LCCO.

Effect of Sterilization Methods on Cell Binding Activity of Surface-immobilized Fibronectin

The effect of sterilization methods on biological activity of fibronectin on the surface of biomaterials was elaborated in the present study. Sterile protein- modified biomaterials were fabricated by microfilter filtration and UV irradiation, respectively. UV irradiation altered the conformation of surface- adsorbed fibronectin and further affected the attachment, morphology and biological function of endothelial cells. However, microfUter filtration did not to change the normal conformation of fibronectin, or the proliferation and biological function of endothelial cells, indicating that microfilter filtration sterilization is the most suitable method for protein-substrate.

Aneurysm Clipping and Outcome for Hunt &Hess Grade 4, 5 Subarachnoid Hemorrhage—A Literature Review

“Subarachnoid Hemorrhage is non-traumatic nasty bleeding into the subarachnoid area, the territory between the arachnoid and the piamater of the central nervous system showing prompt developing signs of neurological sequelae”. It is one among the neurological emergencies which is a very distressing cerebrovascular disease with complicated mechanisms that risks brain perfusion and its function, having higher morbidity and mortality rates. Its mortality rate is still ranged between 8.3% and 66.7%, with noticeable regional variations, beside recent advances in treatment approaches. The incidence of SAH among the population of 2 - 22.5/100,000 was reported with a minimum of 60% of aneurysm ruptures occurring amid ages of 40 and 60 years with 3:2 male:female ratio. The rupture risks for unruptured aneurysms are increased by the issues like present smokers, larger size of aneurysm, and amid young population. The surgical treatment decision should be contemplated upon factors such as aneurysm’s size, aneurysm’s location, patient’s illness history, and surgeon’s operative experiences. Latest technical progresses in imaging techniques, increased consideration of illness history, more awareness of incidences of aneurysms and use of micro neurosurgery, have raised the chance for detection of subarachnoid hemorrhage (SAH) and possible better outcomes with surgical management. Factors that may affect outcome include age, size and site of aneurysm, interval between ictus and surgery, CT Fisher Grade & Hunt and Hess Grading earlier to surgery, & Glasgow Coma Scale at the while of discharge. The studies here support the wide spread concept that surgical clipping of SAH for Hunt and Hess Grade 4, 5 SAH, which is also considered as poor Grade SAH stipulates an effective treatment if done earlier provides better outcome.

Perivascular localized cells commit erythropoiesis in PDGF-B-expressing solid tumors

Background:Tumors possess incessant growth features,and expansion of their masses demands sufficient oxygen supply by red blood cells(RBCs).In adult mammals,the bone marrow(BM)is the main organ regulating hematopoiesis with dedicated manners.Other than BM,extramedullary hematopoiesis is discovered in various pathophysiological settings.However,whether tumors can contribute to hematopoiesis is completely unknown.Accumulating evidence shows that,in the tumor microenvironment(TME),perivascular localized cells retain progenitor cell properties and can differentiate into other cells.Here,we sought to better understand whether and how perivascular localized pericytes in tumors manipulate hematopoiesis.Methods:To test if vascular cells can differentiate into RBCs,genome-wide expression profiling was performed using mouse-derived pericytes.Genetic tracing of perivascular localized cells employing NG2-CreERT2:R26R-tdTomato mouse strain was used to validate the findings in vivo.Fluorescence-activated cell sorting(FACS),single-cell sequencing,and colony formation assays were applied for biological studies.The production of erythroid differentiationspecific cytokine,erythropoietin(EPO),in TME was checked using quantitative polymerase chain reaction(qPCR),enzyme-linked immunosorbent assay(ELISA,magnetic-activated cell sorting and immunohistochemistry.To investigate BM function in tumor erythropoiesis,BM transplantation mouse models were employed.Results:Genome-wide expression profiling showed that in response to plateletderived growth factor subunit B(PDGF-B),neural/glial antigen 2(NG2)+perivascular localized cells exhibited hematopoietic stem and progenitor-like features and underwent differentiation towards the erythroid lineage.PDGF-B simultaneously targeted cancer-associated fibroblasts to produce high levels of EPO,a crucial hormone that necessitates erythropoiesis.FACS analysis using genetic tracing of NG2+cells in tumors defined the perivascular localized cell-derived subpopulation of hematopoietic cells.Single-cell sequencing and colony formation assays validated the fact that,upon PDGF-B stimulation,NG2+cells isolated from tumors acted as erythroblast progenitor cells,which were distinctive from the canonical BM hematopoietic stem cells.Conclusions:Our data provide a new concept of hematopoiesis within tumor tissues and novel mechanistic insights into perivascular localized cell-derived erythroid cells within TME.Targeting tumor hematopoiesis is a novel therapeutic concept for treating various cancers that may have profound impacts on cancer therapy.

Characteristics of Agraphia in Chinese Patients with Alzheimer＇s Disease and Amnestic Mild Cognitive Impairment

Background： Patients with Alzheimer＇s disease （AD） manifest progressive decline in writing abilities. Most studies on agraphia in AD have been performed in the alphabetic system, such as English. However, these findings may not be applicable to other written language systems. The unique features of the Chinese written script could affect the patterns of agraphia in Chinese AD patients. The aim of this study was to explore the features of writing errors in Chinese patients with AD and amnestie mild cognitive impairment （a-MCI）, as well as to study the relationship between their writing errors and neuropsychological functions. Methods： In this study, we performed an observational study in a group of subjects including 17 AD patients, 14 patients with a-MCI, and 16 elderly healthy controls. We analyzed the writing errors in these subjects and also studied the relationship between their writing errors and neuropsychological functions. Results： Our study showed that in patients whose mother tongue is Chinese, writing ability was comparatively well preserved in the MCI phase but significantly impaired when the disease progressed to the stage of AD. The writing errors showed corresponding increase with the severity of cognition decline, both in the types of errors and rate of occurrence. Analysis of the writing errors showed that word substitution and unintelligible words were the most frequent error types that occurred in all the three study groups. The occurrence rate of unintelligible words was significantly higher in the AD group compared with the a-MCI group （P = 0.024） and control group （P = 0.018）. In addition, the occurrence rates of word substitution were also significantly higher in AD （P = 0.013） and a-MCI groups （P = 0.037） than that of control group. However, errors such as totally no response, visuospatial impairment, paragraph agraphia, ideograph, and perseverative writing errors were only seen in AD group. Besides, we also found a high occurrence rate of visuoconstructional errors （13.3%） in our AD group. Conclusions： Our study confirmed that agraphia is an important feature in patients with AD. The writing error profile in patients whose native language is Chinese was unique compared to patients using the alphabetic language system.