miRNA-7 inhibits proliferation of human nasopharyngeal 5-8F carcinoma cells via EGFR/PI3K/Akt pathway

AIM:To investigate the relationship of microRNA-7 (miRNA-7) over-expression and epidermal growth factor receptor (EGFR)/phosphatidylinositol kinase-3 (PI3K)/protein kinase B (PKB, also called Akt) pathway in human nasopharyngeal carcinoma 5-8F cells. METHODS:The 5-8F cells were transfected with miRNA-7 mimics (carrying by Lipofectamine 2000). The expression of miRNA-7 was detected by real-time PCR. The cell proliferation was analyzed by CCK-8 assay. The cell colony-forming capability was determined by cell colony formation test. The expression of EGFR/PI3K/Akt at mRNA and protein levels was examined by real-time PCR and Western blotting. RESULTS:The expression level of miRNA-7 was significantly increased in 5-8F cells compared with negative control (NC) group and control group (P<0.01). The proliferation of NPC 5-8F cells was decreased extremely after tansfected with the miRNA-7 mimics (P<0.01), so did the result of the cell colony-formation test. The expression of EGFR/PI3K/Akt at mRNA and protein levels was significantly down-regulated compared with NC group and control group (P<0.01). CONCLUSION: Over-expression of miRNA-7 significantly inhibits the proliferation and colony-forming ability of nasopharyngeal carcinoma 5-8F cells by down-regulation of EGFR/PI3K/Akt pathway.     

Resveratrol inhibits chondrosarcoma via mitochondrial and PI3K/Akt signaling pathways

AIM：To investigate the inhibitory effects of resveratrol on chondrosarcoma and the relation with mitochondrial and PI3K/Akt pathways. METHODS：Chondrosarcoma SW1353 cells were treated with resveratrol at concentrations of 25, 50 and 100 μmol/L for the time intervals of 24 h, 48 h and 72 h. The viability and apoptosis of the SW1353 cells in the presence or absence of resveratrol were analyzed by CCK8 assay and Hoechst 33258 staining, respectively. The protein levels of Bcl-2, Bax, activated caspase-3, Akt and p-Akt were detected by Western blotting. The cell migration ability was determined by wound scratch assay. RESULTS：Exposure of the cells to resveratrol resulted in a decrease in the cell viability in a dose- and time-dependent manner (P<0.05). visible nuclei with apoptotic characteristics in resveratrol group were observed. The protein levels of activated caspase-3 and Bax were increased, and Bcl-2 and p-Akt were decreased compared with control group. The total Akt were not significantly changed. Resveratrol also significantly reduced the migration of tumor cells. CONCLUSION：Resveratrol induces apoptosis of chondrosarcoma, which plays a role of part through mitochondrial and PI3K/Akt signaling pathways.     

Influence of high-mobility group box 1 on proliferation of neural stem cell in pre-infarction cortex of focal cerebral ischemia/reperfusion model rats

AIM：To investigate the influence of high-mobility group box 1 (HMGB1) on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia/reperfusion model rats. METHODS：Male SD rats (n=48) were randomly divided into sham group, ischemia/reperfusion (I/R) group, RNA interference group and negative interference group. The rat middle cerebral artery was blocked to establish focal cerebral I/R model (ischemia for 1 h and reperfusion for 7 d). Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain. The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB1/GFAP and Western blotting. The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labeling of BrdU/nestin. RESULTS：The protein expression of HMGB1 in I/R group was much higher than those in sham group (P<0.05). RNA interference effectively inhibited the HMGB1 expression (P<0.05). Double labeled BrdU/nestin positive cells in I/R group were more than that in sham group (P<0.05). The double labeled BrdU/nestin positive cells were significantly decreased in RNA interference group (P<0.05). CONCLUSION：Focal cerebral ischemia/reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB1 protein level.     

人工催熟日本鳗鲡精子的显微和超微结构

对经人工催产获得的日本鳗鲡（Anguilla japonica）精子的显微和超微结构进行了观察。光镜下观察，日本鳗鲡精子有两种形态结构，一种精子的细胞核为圆形或近 似圆形，这精子较小，细胞核长径为(2.57±0.62) μm，短径为(2.11±0.59) μm，鞭毛长度为(38.35±7.71) μm；另外一种精子的细胞核为“眉形”或“新月形”，精子较大，细胞核长径为(7.66±1.09) μm，短径为(2.54±0.46) μm，鞭毛长度为(38.35±9.02) μm。透射电镜观察结果显示：圆形精子头部的顶端无顶体，植入窝位于细胞核底端的中间，由细胞核向内凹陷而成，呈一沟状，其走向与精子的长轴平行。中心粒复合体位于植入窝内。细胞核的下端有2～3个线粒体。基体的头端呈筒状，由电子致密物构成。基体的尾端分裂成9束。尾部从袖套腔中伸出且细长，轴丝始端与基体的尾端相连，微管多呈电子致密状，轴丝为典型的“9+2”结构。圆形精子为日本鳗鲡正常成熟的精子。“眉形”或“新月形”精子头部细胞核弯曲，在弯曲处有一圆形的球状物，球状物内含有线粒体和中心粒复合体，轴丝为“9+0”结构。这种精子可能存在发育缺陷或没有达到完全正常成熟。     

Enhancing mucosal immunity in mice by recombinant adenovirus expressing major epitopes of porcine circovirus-2 capsid protein delivered with cytosine-phosphate-guanosine oligodeoxynucleotides

A recombinant replication-defective adenovirus expressing the major epitopes of porcine circovirus-2 (PCV-2) capsid protein (rAd/Cap/518) was previously constructed and shown to induce mucosal immunity in mice following intranasal delivery. In the present study, immune responses induced by intranasal immunization with a combination of rAd/Cap/518 and cytosine-phosphate-guanosine oligodeoxynucleotides (CpG ODN) were evaluated in mice. The levels of PCV-2-specific IgG in serum and IgA in saliva, lung, and intestinal fluids were significantly higher in the group immunized with rAd/Cap/518 and CpG ODN than animals immunized with rAd/Cap/518 alone. The frequencies of IL-2-secreting CD4⁺ T cells and IFN-γ-producing CD8⁺ T cells were significantly higher in the combined immunization group than mice immunized with rAd/Cap/518 alone. The frequencies of CD3⁺, CD3⁺CD4⁺CD8^-, and CD3⁺CD4^-CD8⁺ T cells in the combined immunization group were similar to that treated with CpG ODN alone, but significantly higher than mice that did not receive CpG ODN. PCV-2 load after challenge in the combined immunization group was significantly lower than that in the phosphate-buffered saline placebo group and approximately 7-fold lower in the group treated with CpG ODN alone. These results indicate that rAd/Cap/518 combined with CpG ODN can enhance systemic and local mucosal immunity in mice, and represent a promising synergetic mucosal vaccine against PCV-2.     

Tissue-specific expression of the NOD-like receptor protein 3 in BALB/c mice

Activation of the innate immune system requires recognition of pathogen-associated molecular patterns, such as NOD-like receptors. The NOD-like receptor protein 3 (NLRP3) inflammasome is involved in induction of the pro-inflammatory cytokine, IL-1β, and subsequent inflammatory responses. NLRP3 inflammasome plays important roles in the inflammatory and innate immune responses associated with autoimmune/inflammatory syndrome. However, analysis of the tissue distribution and expression profiles in BALB/c mice is still incomplete. In this study, we investigated the tissue distribution and expression pattern of NLRP3 in BALB/c mice to further elucidate its function in innate immunity in this commonly used laboratory animal model. NLRP3 mRNA expression levels and tissue distribution of the protein were investigated by real-time quantitative PCR and immunohistochemical analyses, respectively. NLRP3 mRNA expression was higher in the kidney and inguinal lymph nodes than in other tissues. Cytoplasmic expression of NLRP3 was detected in the epithelial reticular cells of the spleen and thymus, lymphocytes in the inguinal lymph nodes, cardiac muscle cells, cerebral cortex neurons, alveolar macrophages, renal tubule cells and liver sinusoidal endothelial cells. The results of this study will assist investigators in interpreting site-specific functions and roles of NLRP3 in inflammatory responses.     

H7N9亚型禽流感对辽宁省畜牧业的影响及其对策

禽流感已成为国际上广泛关注的公共卫生事件,因此,加强对流感状况的认识和了解将有助于人们有效地防控流感。本文对禽流感病毒的流行现状及对辽宁省畜牧业的影响进行综述,旨在为有效防制禽流感提供理论依据。     

Helminth parasites of freshwater fishes from Cuatro Ciénegas,Coahuila, in the Chihuahuan Desert of Mexico: inventory and biogeographical implications

As part of an ongoing inventory of the helminth parasites of freshwater fishes in Mexico, 570 individual fish were collected between Apr 2008 and Oct 2011 in 26 localities along the Cuatro Ciénegas region in Coahuila State, northern Mexico. Seventeen species of hosts, mostly corresponding to Nearctic freshwater elements, were studied. A total of 8324 individual worms were collected during this survey, representing 25 species of helminths, of which 9 were digeneans, 3 monogeneans, 3 acanthocephalans, 9 nematodes and 1 cestode. Most of the records in this checklist represent new host or locality records. The information provided in this checklist may be helpful for our understanding of the biodiversity and historical biogeography of this host–parasite system, because in the Cuatro Ciénegas region occur a Nearctic freshwater fish fauna, along with Neotropical and endemic elements, and from a biogeographical point of view, this may represent a transitional area.     

Diagnostic imaging and pacemaker implantation in a domestic goat with persistent left cranial vena cava

Difficulty was encountered with the insertion of a right atrial pacing lead via the left jugular vein during lead and pacemaker implantation in a clinically normal goat as part of an ongoing rapid atrial pacing – induced atrial fibrillation research project. Fluoroscopic visualization of an abnormal lead advancement path prompted angiographic assessment which revealed a persistent left cranial vena cava (PLCVC) and prominent coronary sinus communicating with the right atrium. Angiography facilitated successful advancement and securing of the pacing lead into the right side of the interatrial septum. Cardiac magnetic resonance imaging/magnetic resonance angiography (MRI/MRA) allowed further characterization of this rare venous anomaly. Even though PLCVC has been reported once in a goat, to the authors' knowledge this is the first report to include MRI/MRA characterization of PLCVC and prominent coronary sinus with successful cardiac pacemaker implantation using the PLCVC.     

An intranuclear bacilliform virus associated with near extirpation of Austropotamobius pallipes Lereboullet from the Nant watershed in Ardéche, France

White-clawed crayfish, Austropotamobius pallipes, were endemic to the Nant watershed, Ardéche, France, until they were extirpated by epizootic mortality at the beginning of the twentieth century. A. pallipes were successfully reintroduced to the Nant watershed in the middle of the twentieth century. However, epizootic mortality was observed in the Nant watershed in the summer of 2000 during which time A. pallipes was extirpated from downstream regions. Dead and moribund crayfish were again detected in several episodes in summer 2001 and by October the range of A. pallipes was reduced to the headwaters of just one of the three streams in the watershed. Water quality for the watershed in summer 2001 was appropriate for crayfish habitation. Bacteriology and mycology on A. pallipes collected during several of the mortality episodes in 2001 failed to reveal a cause. However, histopathology revealed a high occurrence of intranuclear eosinophilic inclusions in hepatopancreatocytes of A. pallipes . The nuclei were hypertrophic and contained bacilliform virions consisting of a cylindrical nucleocapsid surrounded by a trilaminar envelope. Virions in section were approximately 63 × 258 nm and nucleocapsids were approximately 52 × 225 nm. It is unclear whether the intranuclear bacilliform virus was the cause of the mortality episodes or was a contributor to a disease complex involving one or several other undetected pathogens.