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Colorectal cancer (CRC) is one of the most common malignancies and most of the patients diagnosed with advanced CRC have unsatisfactory treatment effect and poor prognosis. The purpose of this study was to investigate the effect of CCNI2 on the development of CRC. In this sutdy, immunohistochemical staining was used to detect CCNI2 expression levels in clinical samples, meanwhile, the Kaplan‐Meier survival analysis was conducted. Celigo cell counting assay was used for screening shCCNI2s. QPCR and WB were performed to verify knockdown efficiency of CCNI2. Cell proliferation, colony formation, cell cycle, apoptosis, and mechanism investigation of CCNI2 knockdown were investigated by MTT assay, colony formation assay, fluorescence‐activated cell sorting, and human apoptosis antibody array, respectively. Otherwise, the mouse model of CCNI2 knockdown was also constructed. The results of immunohistochemical staining and qPCR indicated that CCNI2 had a high expression level in the CRC tissues and cell lines. Kaplan‐Meier survival analysis manifested that the high expression of CCNI2 suggested poor prognosis. The expression of CCNI2 was significantly reduced by CCNI2‐siRNAs, and the downregulated expression level of CCNI2 inhibited CRC cell proliferation and colony formation, arrested cell cycle in G2 phase, as well as promoted cell apoptosis. The various indexes of solid tumor in mice models indicated that CCNI2 knockdown could suppress the growth of CRC tumor. Based on the comprehensive analysis of the above results, CCNI2 was contributed to the progression of CRC and could serve as a prognostic marker for CRC., The influence of CCNI2 knockdown on CRC in vivo. Fluorescence images showed the growth of tumors in mice models

Long noncoding RNA (lncRNA) regulates the proliferation and migration of human retinal endothelial cells, as well as retinal neovascularization in diabetic retinopathy. Based on similarities between the pathogenesis of retinopathy of prematurity (ROP) and diabetic retinopathy, lncRNA may also play a role in ROP. Seven-day-old mice were administered 75 ± 2% oxygen for 5 days and normoxic air for another 5 days to establish a ROP model. Expression of lncRNA and mRNA in the retinal tissue of mice was detected by high-throughput sequencing technology, and biological functions of the resulted differentially expressed RNAs were evaluated by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses. The results showed that compared with the control group, 57 lncRNAs were differentially expressed, including 43 upregulated and 14 downregulated, in the retinal tissue of ROP mice. Compared with control mice, 42 mRNAs were differentially expressed in the retinal tissue of ROP mice, including 24 upregulated and 18 downregulated mRNAs. Differentially expressed genes were involved in ocular development and related metabolic pathways. The differentially expressed lncRNAs may regulate ROP in mice via microRNAs and multiple signaling pathways. Our results revealed that these differentially expressed lncRNAs may be therapeutic targets for ROP treatment. This study was approved by the Medical Ethics Committee of Shengjing Hospital of China Medical University on February 25, 2016 (approval No. 2016PS074K).

The mouse model of oxygen induced retinopathy is suitable for the study of various retinal neovascularization diseases, including retinopathy of prematurity. The maternally expressed gene 3 (MEG3) has been demonstrated to have an inhibitory effect on diabetic retinopathy. In this study, we investigated the role of MEG3 overexpression in oxygen-induced retinopathy in mice. The results showed that MEG3 overexpression effectively inhibited the production of retinal neovascularization in oxygen-induced retinopathy mice. It acts by down-regulating the expression of phosphoinositide 3-kinase, serine/threonine kinase, and vascular endothelial growth factor and pro-inflammatory factors. MEG3 overexpression lentivirus has a future as a new method for the clinical treatment of retinopathy of prematurity. The animal experiments were approved by the Animal Ethics Committee of Shengjing Hospital of China Medical University, China (approval No. 2016PS074K) on February 25, 2016.

Intravitreal injections of anti-vascular endothelial growth factor (VEGF) have become increasingly popular in the treatment of ocular diseases. However, few studies have determined the efficiency of unilateral intravitreal anti-VEGF injection in the fellow eye. Herein, we performed a study to investigate the drug concentration in fellow eyes and venous serum after unilateral intravitreal injection of conbercept into rabbit eyes. This is an experimental animal study. Thirty male New Zealand rabbits (60 eyes) were used. One eye of each rabbit was intravitreally injected with 0.5 mg of conbercept. Both eyes from six rabbits were enucleated on days 1, 3, 7, 14, and 30. Conbercept concentrations were measured in the serum, aqueous humor, and vitreous humor. We found conbercept was detected in the fellow eyes and serum of rabbits. Conbercept concentrations in the vitreous humor of the fellow eyes increased from 74.11 ng/ml on day 1 to 246.69 ng/ml on day 3 and then declined to 69.11 ng/ml after 30 days. The concentration in the aqueous humor peaked on day 1 with a concentration of 244.82 ng/ml and declined to 40.13 ng/ml after 30 days. The maximum conbercept concentrations in the aqueous humor and vitreous humor of fellow eyes were similar, which were 0.2 and 1.3% of those of the injected eye, respectively. A peak concentration of 102.49 ng/ml was achieved in the venous serum 1 day after intravitreal injection of conbercept, which was 0.08 and 0.5% of those of the maximum conbercept concentrations in the vitreous humor and aqueous humor of the injected eye, respectively, and 41.5 and 41.8% of the maximum conbercept concentrations in the vitreous humor and aqueous humor of the non-injected eye, respectively. In conclusion, after intravitreal injection of 0.5 mg of conbercept into rabbit eyes, very small amounts of conbercept were detected in the fellow non-injected eyes and venous serum.

Having a limited number of VH segments, cattle rely on uniquely long DH gene segments to generate CDRH3 length variation (3 ~ 70 aa) far greater than that in humans or mice. Bovine antibodies with ultralong CDRH3s (> 50 aa) possess unusual structures and abilities to bind to special antigens. In this study, we replaced most murine endogenous DH segments with bovine DH genes, generating a mouse line termed B-DH. The use of bovine DH genes significantly increased the length variation of CDRH3 and consequently the Ig heavy chain repertoire in B-DH mice. However, no ultralong CDRH3 was observed in B-DH mice, suggesting that other factors, in addition to long DH genes, are also involved in the formation of ultralong CDRH3. The B-DH mice mounted a normal humoral immune response to various antigens, although the B cell developmental paradigm was obviously altered compared with wild type mice. Additionally, B-DH mice are not predisposed to the generation of autoantibodies despite the interspecies DH gene replacement. The B-DH mice reported in this study provide a unique model to answer basic questions regarding the synergistic evolution of DH and VH genes, VDJ recombination, and BCR selection in B cell development.

Object: Understanding hub genes associated with gastric adenocarcinoma (GC) development could lead to effective advances to diagnose and treat the diseases. In order to discover possible signal pathways and hub genes for the disease, we utilized the bioinformatics tools to analyze its mechanism.Method: The gene chip of GSE7997was download from the GEO Datasets. Expression data of gastric cancer and its adjacent normal tissues were compared and the DEGs were acquired. The clusterProfiler and KEGG.db R packages were used for the analysis of its gene ontology process and KEGG pathways. What’s more, the PPI network was constructed by the STRING website. The hub genes were acquired by the plugin of the Cytohubba in Cytoscape. Finally, these genes were examined by the TCGA datasets and potential drugs were explored by Connectivity map.Results: The up regulated DEGs was mainly associated with the process of an extracellular matrix organization, an extracellular matrix organization, Collagen catabolic process, and multicellular organismal catabolic process. The down regulated DEGs have mainly associated with the process of digestion， cellular response to zinc ion， digestive system process, and organic hydroxy compound metabolic process. The up regulated DEGs was mainly located on PI3K-AKTsignal pathways, human papillomavirus infection，Cytokine-cytokine receptor interaction, and focal adhesion process. The down regulated genes were mainly associated with protein digestion and absorption, mineral absorption, and the pancreatic secretion. Cytohubba had found hub genes of COL1A2, COL5A1, COL4A1, COL5A2, COL6A3, COL11A1, SERPINH1, FN1, and down-regulated COL2A1.These genes were associated with the process of transforming growth factor, extracellular matrix, cell adhesion, wound healing and so on. As examined by the TCGA datasets, these altered genes were associated with overall survivaland no disease progress time (PConclusion: As mentioned above, we got 9 hub genes and tested by the TCGA datasets of gastric adenocarcinoma. They were associated with overall survival and disease free progressive time in gastric cancer patients. The bioinformatical analysis of the disease may enhance the understanding of the mechanism of disorders.

Background: Endophthalmitis caused by Mycobacterium fortuitum as a complication of cataract surgery is rare in China and globally. This study aimed to report the clinical features, surgical treatment strategies, antibiotic susceptibilities and treatment outcomes in patients with endophthalmitis caused by Mycobacterium fortuitum. Patients and methods: A case series of 15 patients (15 eyes) with post-cataract endophthalmitis caused by Mycobacterium fortuitum treated at the Ophthalmology Clinic of Shengjing Hospital between June 2008 and June 2017 was reviewed retrospectively. Information about Mycobacterium fortuitum culture results, clinical features, surgical procedures, and final visual acuity were obtained. Results: The vitreous samples were all positive. These eyes underwent total capsulectomy and intraocular lens explantation and vitrectomy combined with intravitreal and intravenous injections of antibiotics. Finally, 14 eyes had controlled inflammation and retained eyeballs. One eye had panophthalmitis and enucleation of the eyeball. Presenting visual acuity ranged from counting fingers to no light perception. Final visual acuity was better than 20/400 in 4 of 15 eyes (26.67%), 20/400 to hand motions in 7 of 15 eyes (46.67%), and light perception to no light perception in 4 of 15 eyes (26.67%) at 6-month follow-up visit. Conclusion: Mycobacterium fortuitum might be an etiologic agent in postoperative endophthalmitis. The Mycobacterium fortuitum endophthalmitis might be treated with early vitrectomy and targeted antibiotic therapy approach.

Objective To explore the genetic causes of Herlyn-Werner-Wunderlich syndrome (HWWS) using whole-exome sequencing. Design Retrospective genetic study. Setting Academic medical center. Patient(s) Twelve patients with HWWS. Intervention(s) Whole-exome sequencing was performed for each patient. Sanger sequencing was used to confirm the potential causative genetic variants. In silico analysis and American College of Medical Genetics and Genomics guidelines were used to classify the pathogenicity of each variant. Main Outcome Measure(s) Rare sequence variants associated with mullerian duct development and renal agenesis were identified and included in subsequent analyses. Result(s) A total of 11 variants were identified in 10 of 12 patients (83.3%) and were considered to constitute a molecular genetic diagnosis of HWWS. These 11 variants were related to 9 genes: CHD1L, TRIM32, TGFBR3, WNT4, RET, FRAS1, FAT1, FOXF1, and PCSK5. All variants were heterozygous and confirmed by Sanger sequencing. The changes included one frameshift variant, one splice-site variant, and eight missense variants. All of the identified variants were absent or rare in Genome Aggregation Database East Asian populations. One of the 11 variants (9.1%) was classified as a pathogenic variant according to the American College of Medical Genetics and Genomics guidelines, and 8 of the 11 variants (72.7%) were classified as variants of uncertain significance. Conclusion(s) To our knowledge, this is the first report of the genetic causes of HWWS. Renal agenesis-related genes, such as CHD1L, TRIM32, RET, and WNT4, may be associated with HWWS. Identification of these variants can not only help us understand the etiology of HWWS and the relationship between reproductive tract development and urinary system development, but additionally improve the level of genetic counseling for HWWS.

The aryl hydrocarbon receptor (AhR) is a ligand-dependent-activated transcriptional factor that regulates the metabolism of xenobiotic and endogenous compounds. Recent studies have shown that AhR is a novel master regulator of the mucosal immune system, including lungs and intestine. To elucidate the role of AhR in chronic severe asthma, AhR wild-type and knockout mice (AhR-/- ) were sensitized and challenged with ovalbumin for 4 weeks. To uncover the underlying mechanisms, inflammatory cells profile and cytokines production were analyzed in bronchial lavage fluid (BALF) and lung tissue. Compared to wild-type mice, AhR-/- mice had exacerbated asthma symptoms, including airway inflammation, mucus production, airway hyperresponsiveness, and airway remodeling. BALF monocytes, neutrophils, eosinophils, and lymphocytes were all enhanced in OVA-immunized AhR-/- mice. In OVA-immunized AhR-/- mice, T helper (Th) 17 cell-specific cytokine IL-17A, as well as airway remodeling factors, including epithelial-mesenchymal transition (EMT) markers and vascular endothelial growth factor (VEGF), were all enhanced in lung tissue. Moreover, human cohort studies showed that AhR gene expression in bronchial epithelial cells decreases in severe asthma patients. Loss of AhR leads to worsening of allergic asthma symptoms, indicating its importance in maintaining normal lung function and mediating disease severity.

During past few decades, studies have demonstrated that particulate matter (PM) is the most serious environmental pollutant in industrial countries which mainly contributes for increasing prevalence of chronic respiratory inflammatory diseases. Healthy food supplements to prevent the inflammatory diseases are common and show notable effects. The effects of the extract of Pyrus nivalis, a common fruit and herbal medicine in Taiwan, on PM-induced airway inflammation in mice were investigated by feeding the extract orally for 21 days. Results obtained from lung histology and bronchial alveolar lavage fluid (BALF) cell profile showed that oral feeding of P. nivalis extract did not affect the airway inflammation. However, it increased the phagocytic activity in BALF cells and induced M1-dominant macrophage differentiation in lungs. Our study showed that extract of P. nivalis might present the beneficial and therapeutic potential for clearance of PM and pathogens in airway. PRACTICAL APPLICATIONS: Pyrus nivalis is a common fruit and also used extensively in Chinese herbology. The pharmacological effects of P. nivalis have been reported in ancient Chinese Medical literature and known to possess anti-asthma and anti-cough properties as well as to enhance function of lungs. In this study, we found that orally feeding the extract of P. nivalis did not induce airway inflammation and affect particulate matter (PM)-induced inflammatory cells infiltration in lungs but increased phagocytosis with or without PM treatment which might indicate its therapeutic potential for clearance of PM and pathogens in airway.

Background Patients positive for anti-glutamic acid decarboxylase 65 (GAD65) antibodies have attracted increasing attention. Their clinical manifestations are highly heterogeneous and can be comorbid with tumors. Currently, there is no consensus on the therapeutic regimen for anti-GAD65-associated neurological diseases due to the clinical complexity, rarity and sporadic distribution. We reported six anti-GAD65 autoimmune encephalitis (AE) patients who received intravenous methylprednisolone (IVMP) or immunoglobulin (IVIG) or both. Then, we evaluated the therapeutic effect of both by summarizing results in previous anti-GAD65 AE patients from 70 published references. Results Our six patients all achieved clinical improvements in the short term. Unfortunately, there was no significant difference between IVMP and IVIG in terms of therapeutic response according to the previous references, and the effectiveness of IVMP and IVIG was 45.56% and 36.71%, respectively. We further divided the patients into different subgroups according to their prominent clinical manifestations. The response rates of IVMP and IVIG were 42.65% and 32.69%, respectively, in epilepsy patients; 60.00% and 77.78%, respectively, in patients with stiff-person syndrome; and 28.57% and 55.56%, respectively, in cerebellar ataxia patients. Among 29 anti-GAD65 AE patients with tumors, the response rates of IVMP and IVIG were 29.41% and 42.11%, respectively. There was no significant difference in effectiveness between the two regimens among the different subgroups. Conclusion Except for stiff-person syndrome, we found that this kind of AE generally has a poor response to IVMP or IVIG. Larger prospective studies enrolling large numbers of patients are required to identify the optimal therapeutic strategy in the future.

Plant vacuolar H+-transporting inorganic pyrophosphatase (V-PPase; EC 3.6.1.1) is a crucial enzyme that exists on the tonoplast to maintain pH homeostasis across the vacuolar membrane. This enzyme generates proton gradient between cytosol and vacuolar lumen by hydrolysis of a metabolic byproduct, pyrophosphate (PP i ). The regulation of V-PPase at protein level has drawn attentions of many workers for decades, but its mechanism is still unclear. In this work, we show that AVP1, the V-PPase from Arabidopsis thaliana, is a target protein for regulatory 14-3-3 proteins at the vacuolar membrane, and all twelve 14-3-3 isoforms were analyzed for their association with AVP1. In the presence of 14-3-3ν, -µ, -ο, and -ι, both enzymatic activities and its associated proton pumping of AVP1 were increased. Among these 14-3-3 proteins, 14-3-3 µ shows the highest stimulation on coupling efficiency. Furthermore, 14-3-3ν, -µ, -ο, and -ι exerted protection of AVP1 against the inhibition of suicidal substrate PP i at high concentration. Moreover, the thermal profile revealed the presence of 14-3-3ο improves the structural stability of AVP1 against high temperature deterioration. Additionally, the 14-3-3 proteins mitigate the inhibition of Na+ to AVP1. Besides, the binding sites/motifs of AVP1 were identified for each 14-3-3 protein. Taken together, a working model was proposed to elucidate the association of 14-3-3 proteins with AVP1 for stimulation of its enzymatic activity.

Mating disruption of Ostrinia furnacalis (Guenée) (Lepidoptera: Crambidae) with its sex pheromone has not been commonly used in NE China due to a lack of information about optimal sex pheromone dosages and the density of release points required in the field. During 2014-2016, first, the two active pheromone ingredients were evaluated in the laboratory alone at ca. 2.5-5.0 mg, or in combination at 0.2-6.0 mg, to disrupt male O. furnacalis mating behaviors. Then, mating disruption areas, with radii of 8.0 m, were determined with those same dosages in corn, an orchard, and soybean fields by comparing male captures in sentinel traps in the control plots with those in corresponding disruption treatments. Finally, 6.0 (F30) and 0.2 mg (Fs) dosages were used in fields at 20-640 and 200-6,400 release points/ha. We found that ≧6.0 mg of the binary pheromone mixture, or ca. 5.0 mg of either of the two single components, completely disrupted mating behaviors, and F30 of the binary mixture provided a 200-m2 disruption area, with at least 50% capture reductions. At a density of 60-640 and 600-6,400 points/ha in a corn field, F30 and Fs dosages provided 90% mating disruption, leaf protection, and ear protection. The dispenser densities and inverse male catches in traps tended to follow a noncompetitive mechanism of mating disruption. Since 85% disruption of mating with 200-400 0.02 mg release points/ha was obtained, that level is recommended as the choice in future NE China O. furnacalis IPM programs.

Background Bladder cancer was a malignant disease in patients, our research aimed at discovering the possible biomarkers for the diseases. Results The gene chip GSE31684, including 93samples, was downloaded from the GEO datasets and co-expression network was constructed by the data. Molecular complex detection(MCODE) was used to identify hub genes. The most significant cluster including 16 genes: CDH11, COL3A1, COL6A3, COL5A1, AEBP1, COL1A2, NTM, COL11A1, THBS2, COL8A1, COL1A1, BGN, MMP2, PXDN, THY1, and TGFB1I1 was identified. After annotated by BiNGO, they were suggested associated with collagen fibril organization and blood vessel development. In addition, the Kaplan Meier curves were obtained by UALCAN. The high expression of THY1, AEBP1, CDH11, COL1A1, COL1A2, COL11A1, MMP2, PXDN, BGN, COL5A1, COL8A1, and TGFB1I1 indicated poor prognosis of the patients(P

Catalyzed by a Cinchona-based thiourea, the [3+2] cycloadditions of barbiturate-based olefins with 3-isothiocyanatooxindoles proceeded readily, and furnished dispirobarbiturates in excellent chemical yields with excellent diastereo- and enantioselectivities. The absolute configuration of dispirobarbiturates was firmly confirmed by an X-ray single crystal structure analysis. A reaction mechanism was proposed to account for the enantioselective formation of dispirobarbiturates.

In this study, the dissipation and enantioselective degradation of trichlorfon (TF) enantiomers in mariculture pond water was investigated under various conditions using a chiral HPLC method. The results showed that the TF enantiomers dissipated following first-order kinetics in the mariculture pond water under natural, dark, sterile, and dark and sterile conditions. S-(+)-TF degraded more rapidly than R-(−)-TF under natural and dark conditions, whereas the dissipation of TF enantiomers in mariculture pond water was non-enantioselective under sterile, and dark and sterile conditions. Light and microbial activity played important roles in the dissipation of the TF enantiomers in mariculture pond water, and the TF enantiomers degraded faster at higher temperatures. Microbial activity was found to be responsible for the enantioselectivity under these conditions. The results of the study can be used to evaluate the environmental risk of TF at the enantiomer level.

The white-backed planthopper, Sogatella furcifera (Hemiptera, Delphacidae), is one of the most devastating rice pests. For a better control strategy, various genetic studies have been conducted using reverse-transcription quantitative real-time polymerase chain reaction (qRT-PCR). The appropriate application of qRT-PCR requires reliable endogenous controls; however, studies on this aspect of the white-backed planthopper are lacking. In the present study, nine commonly used reference genes, elongation factor 1-α (EF1-α), polyubiquitin (UB), ribosomal protein S18 (RPS18), actin 1 (ACT), α-1 tubulin (TUB), glyceraldehyde-3-phosphate (GAPDH), ribosomal protein L9 (RPL9), ribosomal protein L10 (RPL10), and 18S ribosomal RNA (18S), were evaluated by qRT-PCR for their expression stability under four different experimental conditions (different developmental stages, acquisition of Southern rice black-streaked dwarf virus (SRBSDV), different tissues, and different temperature stress). These results were analyzed using four software programs (geNorm, NormFinder, BestKeeper, and the delta Ct method) and a Web-based comprehensive tool RefFinder to compare and rank candidate reference genes. According to the results of RefFinder analysis, which integrates the abovementioned four software programs, TUB was ranked as the most suitable reference gene at different developmental stages and under different temperature stress, and GAPDH and RPL9 showed the highest stability for acquisition of SRBSDV and different tissues, respectively. These results will provide a solid foundation for future gene expression study on the white-backed planthopper, and also will give aids in establishing a standardized qRT-PCR procedure for other related insects.

Despite advances in management, bladder cancer remains a principal cause of cancer‑associated complications. Tumor necrosis factor receptor superfamily member 14 (TNFRSF14) is dysregulated in certain types of cancer; however, limited data are available on the expression and function of TNFRSF14 in bladder cancer. In the present study, the aim was to evaluate the expression and biological functions of TNFRSF14 in bladder cancer. Firstly, the expression levels of TNFRSF14 in bladder cancer tissue were examined using The Cancer Genome Atlas (TCGA) database. Secondly, reverse transcription‑quantitative polymerase chain reaction was utilized to investigate the expression levels of TNFRSF14 in the T24, SW780 and EJ‑M3 bladder cancer cell lines. Transfection and Cell Counting kit‑8 (CCK‑8) assay was used to evaluate whether TNFRSF14 overexpression or silencing would have an effect on cell proliferation of T24 and EJ‑M3 cells. In addition, TNFRSF14‑induced apoptotic cells were identified using Annexin V‑fluorescein isothiocyanate and propidium iodide staining. Western blot analysis was used to detect proteins associated with the phosphatidylinositol 3‑kinase pathway. According to the TCGA dataset, the expression levels TNFRSF14 were decreased in bladder cancer tissue compared with in normal control samples. Patients with bladder cancer exhibiting low expression levels of TNFRSF14 had a worse prognosis compared to those with high expression levels of TNFRSF14. Overexpression of TNFRSF14 in T24 cells led to increased apoptosis and inhibited cell proliferation in vitro. Western blotting demonstrated that TNFRSF14 overexpression increased the expression levels of caspase3‑p17 in T24 cells, but significantly decreased the expression levels of phosphorylated (p)‑protein kinase B (AKT) and P70 S6 kinase (P70). TNFRSF14 silencing in EJ‑M3 cells enhanced cell growth, inhibited cell apoptosis, increased the expression levels of p‑AKT and P70, and decreased the expression levels of caspase3‑p17. In conclusion, TNFRSF14 may serve a tumor suppressive role in bladder cancer by inducing apoptosis and suppressing proliferation, and act as a novel prognostic biomarker for bladder cancer.

Fibroblasts are the major effector cells of skin wound healing. Adipose‑derived stem cells can differentiate into fibroblasts under certain conditions. In the present study, it was hypothesized that adipose‑derived stem cells (ADSCs) could be induced by the adipose extracellular matrix (ECM) to differentiate into fibroblasts in order to promote skin wound healing. First, flow cytometry was used to detect the ratio of fibroblasts and relative expression of the fibroblast markers cytokeratin 19 (CK19) and vimentin in ADSCs. Then, the effect of the adipose ECM during the differentiation of ADSCs into fibroblasts was investigated by detecting the total amount of collagen fibers and degree of fibrosis, and the proliferation and cell cycle of differentiated fibroblasts, using the MTT assay and flow cytometry analysis respectively. Finally, a mouse skin wound model was established and treated with PBS, ADSC suspension or ECM + ADSCs to compare wound healing rate and expression of collagen I and collagen III by immunohistochemistry. Following induction of ADSCs with the adipose ECM, more fibroblasts were found, expression of CK19 and vimentin increased, and a greater degree of fibrosis occurred, which revealed the positive effect of the adipose ECM on the differentiation of ADSCs into fibroblasts. In addition, the induced fibroblasts had enhanced proliferation activity, with more cells in the S phase and fewer in the G2/M phase. The in vivo experiment indicated that the ECM produced by the ADSCs had a faster wound healing rate and increased expression of collagen I and collagen III compared with mice injected with PBS or ADSCs alone, which verified that ADSCs induced by the adipose ECM had a positive effect on skin wound healing. The present study demonstrated that the adipose ECM in combination with ADSCs may be a novel therapeutic target for the repair of skin injury, due to the ability of the adipose ECM to induce the differentiation of ADSCs into fibroblasts and to facilitate the wound healing process.

The white-backed planthopper, Sogatella furcifera, is a phloem sap feeder that secretes watery and gelling saliva during feeding. In this study, we identified the major proteins in watery saliva of S. furcifera by shotgun LC-MS/MS analysis combined with transcriptomic analysis. A total of 161 proteins were identified, which were divided into 8 function categories, including enzymes, transporter, calcium ion binding protein, salivary sheath protein, cytoskeleton protein, DNA-, RNA-, and protein-binding or regulating proteins, other non-enzyme proteins and unknown proteins. Gene expression pattern of 11 secretory proteins were analyzed by real time quantitative-PCR. We detected the mucin-like protein, which had a unique expression level in salivary gland, most likely to be a candidate effector involved in regulation of plant defense. This study identified the watery saliva component of S. furcifera and it provided a list of proteins which may play a role in interaction between S. furcifera and rice.

Retinal neovascularization (RNV) is a characteristic pathological finding of retinopathy of prematurity (ROP). Cysteine-rich 61 [Cyr61, also known as CCN family member 1 (CCN1)] has been reported to mediate angiogenesis. The aim of the present study was to investigate the mechanisms of CCN1/Cyr61-phosphoinositide 3-kinase (PI3K)/AKT signaling in ROP. The contribution of CCN1 to human umbilical vein endothelial cell (HUVEC) proliferation and apoptosis under hypoxic conditions was determined using a cell counting kit‑8 (CCK-8) and Annexin V/propidium iodide (PI) staining, respectively, as well as using siRNA targeting CCN1 (CCN1 siRNA). The cells exposed to hypoxia were also treated with the PI3K/AKT inhibitor, LY294002. In addition, mouse pups with oxygen-induced retinopathy (OIR) were administered an intravitreal injection of CCN1 siRNA. RNV was assessed by magnesium-activated adenosine diphosphate-ase (ADPase) staining. RT-qPCR, western blot analysis, immunofluorescence staining and immunohistochemistry were used to detect the distribution and expression of CCN1, PI3K and AKT. Exposure to hypoxia increased the neovascularization clock hour scores (from 1.23±0.49 to 5.60±0.73, P

This study investigated the effects of planting ages on microbial biomass and community diversity in kiwifruit orchard soils through cultured-based microorganism, microbial biomass and Biologanalyses. Our results showed that: (1) Counting of soil cultivable microorganism showed that the amountof bacteria increased firstly and then decreased with the increasing of planting age. The amount of fungi increased after planting kiwifruit and made the soil fungous. However, actinomyces did not change obviously. (2) Microbial biomasses C and N both increased at first, and then decreased obviously with planting ages. (3) Biolog analyses showed that there were significant differences in carbon utilization and soil microbial community functional diversities among kiwifruit orchard and wasteland soils. AWCD, Shannon index, Simpson index and McIntosh index were highest in five aged and lowest in eleven aged kiwifruit orchard soils. Relative utilization value of carbon sources showed that soil microbial communities tended more conducive to using carbohydrate and amino acids and less conducive to using polyphenols and polyamines with planting ages. The principal component analyses showed that there were significant differences in carbon utilizations. Significantly differences in PC1 were found among different aged kiwifruit orchard soils and the differences were mostly caused by carbohydrate, amino acids and polymers, which affecting soil microbial community under kiwifruit planting mostly. Nevertheless, uncultivated and cultivated kiwifruit soils were separated in PC2. We concluded that soil microbial biomass reduced and microbial community diversities decreased under long-term kiwifruit planting, and this might be the main reason that kiwifruit yield decrease and quality decline.

Treatment of micro-polluted source water is receiving increasing attention because of environmental awareness on a global level. We isolated and identified aerobic denitrifying bacteria Zoogloea sp. N299, Acinetobacter sp. G107, and Acinetobacter sp. 81Y and used these to remediate samples of their native source water. We first domesticated the isolated strains in the source water, and the 48-h nitrate removal rates of strains N299, G107, and 81Y reached 33.69%, 28.28%, and 22.86%, respectively, with no nitrite accumulation. We then conducted a source-water remediation experiment and cultured the domesticated strains (each at a dry cell weight concentration of 0.4 ppm) together in a sample of source water at 20-26 °C and a dissolved oxygen concentration of 3-7 mg/L for 60 days. The nitrate concentration of the system decreased from 1.57 ± 0.02 to 0.42 ± 0.01 mg/L and that of a control system decreased from 1.63 ± 0.02 to 1.30 ± 0.01 mg/L, each with no nitrite accumulation. Total nitrogen of the bacterial system changed from 2.31 ± 0.12 to 1.09 ± 0.01 mg/L, while that of the control system changed from 2.51 ± 0.13 to 1.72 ± 0.06 mg/L. The densities of aerobic denitrification bacteria in the experimental and control systems ranged from 2.8 × 10(4) to 2 × 10(7) cfu/mL and from 7.75 × 10(3) to 5.5 × 10(5) cfu/mL, respectively. The permanganate index in the experimental and control systems decreased from 5.94 ± 0.12 to 3.10 ± 0.08 mg/L and from 6.02 ± 0.13 to 3.61 ± 0.11 mg/L, respectively, over the course of the experiment. Next, we supplemented samples of the experimental and control systems with additional bacteria or additional source water and cultivated the systems for another 35 days. The additional bacteria did little to improve the water quality. The additional source water provided supplemental carbon and brought the nitrate removal rate in the experimental system to 16.97%, while that in the control system reached only 3.01%, with no nitrite accumulation in either system. Our results show that aerobic denitrifying bacteria remain highly active after domestication and demonstrate the applicability of such organisms in the bioremediation of oligotrophic ecosystems.

The white-backed planthopper, Sogatella furcifera (Horvath), is currently the only confirmed vector of Southern rice black-streaked dwarf virus (SRBSDV), which causes severe rice production losses in China. In this study, an absolute quantification qPCR method was used to detect viral gene mRNA expression levels at different developmental stages of white-backed planthoppers fed SRBSDV-infected rice plants. A comparison of viral copy numbers of the SRBSDV S10 gene at the same developmental stage indicated that the white-backed planthopper had higher viral copy numbers when the virus was acquired at the earlier developmental stages. The adult-stage white-backed planthoppers that had acquired the virus at the first-second nymphal stage displayed significantly higher viral titers than white-backed planthoppers that acquired the virus at the third-fourth nymphal stage, at the fifth nymphal stage, and at the adult stage. The fifth nymphal stage white-backed planthoppers that acquired the virus at the first-second nymphal stage displayed higher viral copy numbers than fifth nymphal stage white-backed planthoppers that acquired the virus at the third-fourth nymphal stage and at the fifth nymphal stage. The highest viral load value appeared in the middle adult stage. The annual immigration characteristics of white-backed planthoppers would be beneficial for the dispersal of SRBSDV because this virus could be transmitted far away following the migration of vigorous planthoppers. Therefore, investigating the change in the viral load at different life stages of SRBSDV-positive individuals is required to develop more effective control of the spread of SRBSDV in the field.

The quality of soil is affected by its microbiological composition, which may change over time. This study investigated differences in the soil quality during the maturation of apple orchards. Soil nutrients, soil enzymes, community-level physiological profiles (CLPP), and terminal restriction fragment length polymorphism (T-RFLP) profiles were determined in soil samples from four apple orchards (aged 3, 7, 15, and 23 years) and a neighboring wasteland. The orchard soil organic matter, total nitrogen, invertase, and urease levels increased initially as the apple orchards matured, before decreasing, while the soil total phosphorus accumulated. The total potassium levels were significantly lower in the orchards than the wasteland (P

Ethanol-soluble proteins (EP) were obtained from spotless smoothhound (Mustelus griseus) muscle, and the EP antioxidant hydrolysates (E-AMH) were prepared using papain. Five antioxidant peptides were isolated from the E-AMH using ultrafiltration and consecutive chromatographic methods including ion-exchange chromatography, gel filtration chromatography, and RP-HPLC, and their sequences were identified as Gly-Ala-Ala (217.3 Da), Gly-Phe-Val-Gly (378.5 Da), Gly-Ile-Ile-Ser-His-Arg (682.0 Da), Glu-Leu-Leu-Ile (486.7 Da), and Lys-Phe-Pro-Glu (519.6 Da). All of these purified peptides exhibited high scavenging activity on hydroxyl radical (IC50 1.6337 mg/ml, 0.8944 mg/ml, 0.0769 mg/ml, 0.1173 mg/ml, and 0.1510 mg/ml, respectively), ABTS radical (IC50 1.7541 mg/ml, 1.3055 mg/ml, 0.3493 mg/ml, 0.3274 mg/ml and 0.4645 mg/ml, respectively), and superoxide radical (IC50 0.6714 mg/ml, 0.3082 mg/ml, 0.1508 mg/ml, 0.1851 mg/ml, and 0.2053 mg/ml, respectively). These five purified peptides also effectively inhibited the autooxidation in linoleic acid model system. The antioxidant activities of these purified peptides were attributed to their amino acid compositions, sequences, and hydrophobicities. These results suggested that these five peptides isolated from the E-AMH of spotless smoothhound muscle were potent antioxidants and might be effectively used as food additives and pharmaceutical agents.

Homodimeric proton-translocating pyrophosphatase (H+-PPase; EC 3.6.1.1) maintains the cytoplasmic pH homeostasis of many bacteria and higher plants by coupling pyrophosphate (PPi) hydrolysis and proton translocation. H+-PPase accommodates several essential motifs involved in the catalytic mechanism, including the PPi binding motif and Acidic I and II motifs. In this study, 3 intrinsic tryptophan residues, Trp-75, Trp-365, and Trp-602, in H+-PPase from Clostridium tetani were used as internal probes to monitor the local conformational state of the periplasm domain, transmembrane region, and cytoplasmic domain, respectively. Upon binding of the substrate analog Mg-imidodiphosphate (Mg-IDP), local structural changes prevented the modification of tryptophan residues by N-bromosuccinimide (NBS), especially at Trp-602. Following Mg-Pi binding, Trp-75 and Trp-365, but not Trp-602, were slightly protected from structural modifications by NBS. These results reveal the conformation of H+-PPase is distinct in the presence of different ligands. Moreover, analyses of the Stern-Volmer relationship and steady-state fluorescence anisotropy also indicate that the local structure around Trp-602 is more exposed to solvent and varied under different environments. In addition, Trp-602 was identified to be a crucial residue in the H+-PPase that may potentially be involved in stabilizing the structure of the catalytic region by site-directed mutagenesis analysis.

Genetic variation and patterns of genetic differentiation of the rice stem borer, Chilo suppressalis (Lepidoptera: Pyralidae), from the South China were analyzed using 6 microsatellite markers and two partial mtDNA ( cox1 and cox2 ) regions. All of the 6 microsatellite loci were polymorphic in the studied seven populations. The allelic richness per population ranged between 5.67 and 14.00, and average H E and H O values were 0.6246-0.8329 and 0.2634-0.6061, respectively. As the mitochondrial genome is a single genetic locus, we only present results for the concatenated data set ( cox1 plus cox2 gene sequences, 513 bp). The concatenated data showed high level of genetic diversity and there are 23 variable polymorphic sites among the 513 sites in concatenated data. Nearly all of (20 of 21) pairwise F ST comparisons among populations showed genetic differentiation with moderate to high pairwise F ST values based on microsatellite markers. However, for the mtDNA data, most of the seven populations did not show significant differentiation with other populations. The differences of population differentiation obtained with the two different genetic markers could be mainly attributed to the different mutation rates of microsatellite and mtDNA. There was not genetic structure existed in these studied populations based on microsatellite loci and mtDNA data. The analysis based on network, mismatch distribution, Tajima's D and F S indicated that the studied populations were from the recent same ancestor or the same refuge and followed by a sudden demographic expansion condition.

The white-backed planthopper (WBPH), Sogatella furcifera (Horvath), is one of the serious rice pests because of its destructive feeding. The salivary glands of the WBPH play an important role in the feeding behaviour. Currently, however, very little is known about the salivary glands at the molecular level. We sequenced the salivary gland transcriptome (sialotranscripome) of adult WBPHs using the Illumina sequencing. A total of 65,595 transcripts and 51,842 unigenes were obtained from salivary glands. According to annotations against the Nr database, many of the unigenes identified were associated with the most studied enzymes in hemipteran saliva. In the present study, we identified 32 salivary protein genes from the WBPH sialotranscripome, which were categorized as those involved in sugar metabolism, detoxification, suppression of plant defense responses, immunity-related responses, general digestion, and other phytophagy processes. Tissue expression profiles analysis revealed that four of 32 salivary protein genes (multicopper oxidase 4, multicopper oxidase 6, carboxylesterase and uridine phosphorylase 1 isform X2) were primarily expressed in the salivary gland, suggesting that they played putative role in insect-rice interactions. 13 of 32 salivary protein genes were primarily expressed in gut, which might play putative role in digestive and detoxify mechanism. Development expression profiles analysis revealed that the expression level of 26 of 32 salivary protein genes had no significant difference, suggesting that they may play roles in every developmental stages of salivary gland of WBPH. The other six genes have a high expression level in the salivary gland of adult. 31 of 32 genes (except putative acetylcholinesterase 1) have no significant difference in male and female adult, suggesting that their expression level have no difference between sexes. This report analysis of the sialotranscripome for the WBPH, and the transcriptome provides a foundational list of the genes involved in feeding. Our data will be useful to investigate the mechanisms of interaction between the WBPH and the host plant.

Toll-like receptors (TLRs) are involved in the progression of ischemic brain injury and hence vascular dementia; however, the underlying mechanisms are largely unknown. Here, we have investigated the interrelationship between stress-responsive heme oxygenase (HO)-1 isoenzyme and TLR4 during chronic brain hypoperfusion. The right unilateral common carotid artery occlusion was performed by ligation of the right common carotid artery in C57BL/6J mice. The brain cortex or hippocampus was removed for western blotting and confocal immunofluorescence analysis. The link between HO-1 and TLR4 was further examined by silencing TLR4 and pharmacological inhibition of HO-1 in primary cultured cortical neurons. Cognitive dysfunction and decrease in cerebral blood flow in mice were observed 4 weeks after the occlusion. Our data further show that common carotid artery occlusion induced an increase in TLR4 and HO-1 protein levels. Although the administration of CoPP (10 mg/kg), HO-1 agonist, improved the cognitive dysfunction in a mice model of occlusion, western blot analysis in primary cultured cortical neurons showed that HO-1 was upregulated after lipopolysaccharide treatment; this was partially abolished by the TLR4 siRNA interference. The flow cytometry analysis showed that pharmacological inhibition of HO-1 by ZnPP (100 μM) further exaggerated lipopolysaccharide-induced neuronal cell death. Hence, stress-responsive HO-1 isoenzyme participates in TLR4-induced inflammation during chronic brain ischemia. The pharmacological manipulation of TLR4 or the HO-1 antioxidant defense pathway may represent a novel treatment strategy for neuronal protection in vascular dementia.

Sepia ink oligopeptide (SIO) is a tripeptide extracted from Sepia ink. To test the hypothesis that SIO inhibits prostate cancer by inducing apoptosis, the effects of SIO on the proliferation of three human prostate cancer cell lines were examined using a CCK-8 assay. SIO significantly inhibited the proliferation of DU-145, PC-3 and LNCaP cells in a time- and dose-dependent manner. Flow cytometry studies showed that exposing DU-145, PC-3 and LNCaP cells to 5, 10, or 15 mg/mL SIO for 24 h increased the percentage of the early-stage apoptotic cells from 11.84% to 38.26% (DU-145), 22.76% to 39.96% (PC-3) and 5.05% to 16.11% (LNCaP), respectively. In addition, typical morphologic changes were observed in the cells with acridine orange/ethidium bromide staining. SIO treatment induced strong S and G2/M phase cell cycle arrest in a dose-dependent manner in DU-145 and LNCaP. In contrast, SIO treatment induced strong Sub G1 and G0/G1 phase cell cycle arrest in a dose-dependent manner in PC-3. SIO exposure for 24 h decreased the expression of the anti-apoptotic protein Bcl-2 and increased the expression of the apoptogenic protein Bax. Moreover, the Bax/Bcl-2expression ratio was increased. Concurrently, the expression of caspase-3 was upregulated. These data support our hypothesis that SIO has anticarcinogenic properties.

Liver failure is associated with dyshomeostasis of efflux transporters at the blood-brain barrier (BBB), which contributes to hepatic encephalopathy. In this study we examined whether breast cancer resistance protein (BCRP), a major efflux transporter at the BBB, was altered during liver failure in rats. Rats underwent bile duct ligation (BDL) surgery, and then were sacrificed after intravenous injection of prazosin on d3, d7 and d14. The brains and blood samples were collected. BCRP function at the BBB was assessed by the brain-to-plasma prazosin concentration ratio; Evans Blue extravasation in the brain tissues was used as an indicator of BBB integrity. The protein levels of BCRP in the brain tissues were detected. Human cerebral microvessel endothelial cells (HCMEC/D3) and Madin-Darby canine kidney cells expressing human BCRP (MDCK-BCRP) were tested in vitro. In addition, hyperbilirubinemia (HB) was induced in rats by intravenous injection of unconjugated bilirubin (UCB). BDL rats exhibited progressive decline of liver function and HB from d3 to d14. In the brain tissues of BDL rats, both the function and protein levels of BCRP were progressively decreased, whereas the BBB integrity was intact. Furthermore, BDL rat serum significantly decreased BCRP function and protein levels in HCMEC/D3 cells. Among the abnormally altered components in BDL rat serum tested, UCB (10, 25 μmol/L) dose-dependently inhibit BCRP function and protein levels in HCMEC/D3 cells, whereas 3 bile acids (CDCA, UDCA and DCA) had no effect. Similar results were obtained in MDCK-BCRP cells and in the brains of HB rats. Correlation analysis revealed that UCB levels were negatively correlated with BCRP expression in the brain tissues of BDL rats and HB rats as well as in two types of cells tested in vitro. UCB elevation in BDL rats impairs the function and expression of BCRP at the BBB, thus contributing to hepatic encephalopathy.

The pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), a cotton pest probably native to Indo-Pakistan, invaded China at the beginning of the 20th century. Chinese P. gossypiella have been assumed to be the result of indiscriminate introductions from Pakistan and America by transport of cotton seed. We tested this long-held hypothesis and genotyped a total of 527 individuals from 14 sites at 13 microsatellite loci. We analyzed these data with traditional statistics as well as with Bayesian methods. The loci were, for the most part, highly polymorphic. The allelic richness of Chinese populations at six loci was greater than those of the Pakistani and American populations. Significant deficits of heterozygotes were recorded for all 14 populations, and null alleles were the most probable factor contributing to these deficits. Pairwise FST estimates showed that there was significant differentiation among the pooled Chinese, Pakistani, and American populations, and there was structure within most of the Chinese populations. The Bayesian analysis revealed that the combined Chinese, American, and Pakistani populations formed separate clusters, and the nine Chinese populations were divided into two clusters. Allelic frequency distributions showed that private and shared alleles within Chinese P. gossypiella were derived only partly from the Pakistani and American populations. The microsatellite-based genetic analyses suggested that the Chinese P. gossypiella populations originated from multiple sources.

To determine the role of endoplasmic reticulum (ER) stress and autophagy in apoptosis induced by bortezomib in human cervical cancer-derived HeLa cells and CaSki cells. Bortezomib treatment activated apoptosis, evidenced by increased expression of cleaved caspase-3 and cleaved PARP in both HeLa cells and CaSki cells. Bortezomib also induced the loss of the mitochondrial membrane potential, increased the level of ER stress-associated proteins GRP78, ATF4, and CCAAT-enhancer-binding protein homologous protein, and affected the expression of autophagy-related proteins; increasing the levels of LC3-II and ATG5–ATG12 and decreasing the level of p62. When we combined bortezomib with the ER stress activator tunicamycin, or autophagy inhibitors 3-methyladenine or chloroquine, cell growth inhibition and apoptosis were markedly enhanced. Bortezomib activates apoptosis signaling, and activation of ER stress and inhibition of autophagy enhances the cytotoxicity of bortezomib, suggesting that these combination treatments may be potential chemotherapy strategies for treating cervical cancer.

Hypoxia is a key factor in the pathogenesis of angiogenesis, and cysteine‑rich 61 (CCN1), an angiogenic factor, is involved in the development of pathological angiogenesis. The aim of the present study was to investigate the mechanism of CCN1 RNA interference (RNAi)‑induced inhibition of hypoxia‑induced pathological angiogenesis in endothelial cells. Human umbilical vein endothelial cells (HUVECs) were cultured under hypoxic conditions in vitro. The effects of inhibiting phosphoinositide 3‑kinase (PI3K)/Akt signaling using LY294002 were investigated in hypoxic HUVECs. The proliferation and apoptosis of HUVECs under hypoxia were assessed using CCN1 RNAi. The CCN1‑PI3K/Akt‑vascular endothelial growth factor (VEGF) pathway was analyzed under hypoxic conditions using reverse transcription‑quantitative polymerase chain reaction and western blotting. CCN1 RNAi inhibited the proliferation and induced the apoptosis of the HUVECs under hypoxia, with hypoxia significantly increasing the mRNA and protein expression levels of CCN1, Akt and VEGF. By contrast, CCN1 RNAi reduced the mRNA and protein expression levels of CCN1, Akt and VEGF in the HUVECs (P

MicroRNAs (miRNAs) are small (18 ~ 25 nt) noncoding single-stranded RNA molecules that act as negative regulators of gene expression and modulating the stability and/or the translational efficiency of target messenger RNAs. Studies have shown that miRNAs control diverse aspects of brain disease. Recently, several studies have suggested that miRNAs alter the response to ischemia reperfusion injury and regulate the expression of various key elements in cell survival and apoptosis. This review article gives a brief overview of some miRNAs (miR-15a/b, miR-21, miR-29b/c, miR-124, miR-145, miR-181, miR-200 family, miR-338, miR-422a, miR-497, and miR let 7 family) in cerebral ischemia reperfusion injury. Although miRNAs could be potential therapeutic targets for the treatment of ischemia reperfusion injury, their safety and other limitations need further confirmation.

Ruditapes philippinarum is a member of the Veneridae family of marine bivalve molluscs. RPOI‑1 (Ruditapes philippinarum oligopeptide) is a tetrapeptide that can be extracted from Ruditapes philippinarum by means of enzymolysis. This study showed that RPOI‑1 strongly inhibits proliferation and induces apoptosis in DU‑145 human prostate cancer cells. When cells were treated with varying concentrations of RPOI‑1, significant inhibition of proliferation was detected by an MTT assay, and sub‑G1 and G2/M phase cell cycle arrest was observed using flow cytometric (FCM) analysis. Furthermore, morphological changes characteristic of apoptosis and an increase in the proportion of apoptotic cells were observed using double sequential acridine orange/ethidium bromide staining, FCM analysis and transmission election microscopy. FCM studies showed that exposing DU‑145 cells to 10, 20 and 30 mg/ml RPOI‑1 for 24 h increased the percentage of cells in the early‑stages of apoptotis in a dose‑dependent manner, with the numbers rising from 3.01% in the control group to 13.40% in the group treated with the highest dose.

Osteoporosis is a bone disease causing impaired bone strength. It is characterized by increased osteoclast formation or enhanced bone resorption, leading to an increased risk of fragility fractures. Its prevalence increases with age. The advent of an aging population suggests that progressively more individuals will develop this disease in the aging population. A number of drugs for the prevention and treatment of osteoporosis act by inhibiting bone resorption. However, the effectiveness of osteoporosis treatment in clinical practice is limited. Since the osteoclast is the only cell in the body that is capable of resorbing bone, understanding its biology will be necessary for developing a new therapeutic approach for osteoporosis. Recently, it was discovered that the receptor activator of nuclear factor kappaB (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system is an important signal transduction pathway that regulates osteoclast formation. The binding of OPG to RANKL inhibits the binding between RANKL and RANK; this, in turn, prevents osteoclast precursors from differentiating and fusing to form mature osteoclasts. Therefore, the inhibition of the RANK/RANKL pathway inhibits osteoclast formation, differentiation, activation, and bone resorption. A potential clinical antiresorptive therapy can be developed by using an anti-RANKL monoclonal antibody, such as denosumab, that binds to RANKL with high affinity and specificity and blocks RANKL-RANK interactions.

We developed eight polymorphic microsatellite markers for the green leaf bug, Lygus lucorum Meyer-Dur. The number of observed alleles per locus in 48 individuals ranged from three to 13, whereas the observed and expected heterozygosities ranged from 0.133 to 0.810 and from 0.236 to 0.886, respectively. Cross-species amplifications were conducted both within and between generas, and the amplification results indicated that these primers enriched for L. lucorum Meyer-Dur are species-specific.

Thirteen polymorphic microsatellite markers were developed from an enriched partial genomic library for the pink bollworm, Pectinophora gossypiella (Saunders), one of the most important cotton pests in the world. The total number of alleles ranged from two to 12 for a sample of 50 individuals and the expected heterozygosity at these loci ranged from 0.042 to 0.830. Although the presence of null alleles in some loci is suspected, these polymorphic markers are likely to provide useful tools for the population genetic studies of this species.

A set of 10 polymorphic di- and trinucleotide microsatellite loci were developed for the forestry pest insect, masson pine moth, Dendrolimus punctatus Walker . The expected heterozygosity at these loci ranges from 0.285 to 0.859, and the observed allele numbers from five to 19. Cross species amplification of these loci in four other congeneric pine moth species indicates variable levels of loci conservation and thus cross-applicability. Therefore, the microsatellite loci reported here should be useful for population genetic and other related studies in the masson pine moth and other closely related species.

Triple-negative breast cancer (TNBC) is a particular type of breast cancer which is characterized by its biological aggressiveness, worse prognosis, and lack of prognostic markers or therapeutic targets in contrast with hormonal receptor-positive and human epidermal growth factor receptor 2-positive (HER2+) breast cancers. We aimed to evaluate survivin and epidermal growth factor receptor (EGFR) expression and their prognostic value and determine their relationships with the clinicopathological parameters of TNBC. A total of 136 patients who had undergone a resection of primary TNBC were enrolled at the Third Affiliated Hospital of Harbin Medical University from March 2003 to September 2005. Expression of ER, PR, HER2, EGFR, and survivin was assessed by immunohistochemistry. The association of TNBC and other clinicopathological variables and the prognostic value of survivin and EGFR expression were evaluated. Survivin was expressed in 62 (45.6 %) cases and EGFR was expressed in 82 (60.3 %) cases. Survivin expression was associated with menopausal status (P = 0.011), tumor size (P = 0.037), and lymph node status (P = 0.001). EGFR expression was associated with menopausal status (P = 0.029), lymph node status (P = 0.004), P53 expression (P = 0.001), Ki-67 expression (P = 0.028), and lymphatic vascular invasion (P = 0.037). A multivariate analysis demonstrated that tumor size (hazard ratio (HR) 1.587, 95 % confidence interval (CI) 1.081–2.330, P = 0.018 for disease-free survival (DFS); HR 1.606, 95%CI 1.096–2.354, P = 0.015 for overall survival (OS)), lymph node status (HR 2.873, 95%CI 1.544–5.344, P = 0.001 for DFS; HR 2.915, 95%CI 1.553–5.471, P = 0.001 for OS), tumor grade (HR 1.914, 95%CI 1.218–3.007, P = 0.005 for DFS; HR 1.983, 95%CI 1.228–3.203, P = 0.005 for OS), EGFR (HR 3.008, 95%CI 1.331–6.792, P = 0.008 for DFS; HR 3.151, 95%CI 1.374–7.226, P = 0.007 for OS), and survivin (HR 1.573, 95%CI 1.087–2.277, P = 0.016 for DFS; HR 1.607, 95%CI 1.088–2.374, P = 0.017 for OS) were of prognostic significance for disease-free and overall survival. We draw a conclusion from the present study that survivin and EGFR expression are useful prognostic markers of TNBC and might be useful for molecular targeting therapy of TNBC treatment.

The Chinese crested ibis, Nipponia nippon, is a critically endangered bird. The current population of this species has developed from four wild individuals rediscovered in 1981. Given its conservation status, there is considerable interest in assessing the genetic diversity and individual relatedness in this species. For this purpose, a set of eight polymorphic di- or trinucleotide microsatellite loci was developed for the crested ibis. The expected heterozygosity at these loci ranges from 0.01 to 0.50, with less than four alleles being observed at individual loci, a reflection of the serious population bottleneck experienced by this species.

We developed 12 polymorphic microsatellite markers for the rice stem borer Chilo suppressalis (Walker). These loci were screened for 96 individuals from eight populations across China. The total number of alleles ranged from 3 to 33 and the expected heterozygosity at these loci ranged from 0.131 to 0.671. These molecular markers will be useful for fully investigating the population structure and host preference of C. suppressalis at fine spatial scales.

In order to find out the infectious rate of hepatitis C virus (HCV) among chronic liver diseases, we investigated antibodies against hepatitis C virus and HCVRNA by method of ELISA, RIBA and RT-PCR in 410 patients with chronic liver disease. The prevalence of HCV infection was found to be 4%. Whereas the positive rate of HBsAg and HBV-DNA of these cases was 69% and 58%, respectively. There is no statistical significance between HCV infectious rate of patients with positive and negative HBsAg. The relative low infectious rate of HCV infection among chronic hepatic diseases indicates that HBV infection plays a more important role in causing chronic hepatitis than that of HCV. Thus, special emphasis should be paid to the preventive and therapeutic measures against hepatitis B in China.

ASC (Antidepressant-Sensitive Catalepsy) mice, bred for a high predisposition to catalepsy, are characterized by depression-like behavior and decreased immune responses. Chronic administration of fluoxetine, which is a selective serotonin reuptake inhibitor antidepressant widely used in clinical practice, to mice of this strain weakened catalepsy and normalized the number of rosette-forming cells in the spleen. In mice of the parental cataleptic strain CBA/Lac, fluoxetine had no effect on the level of catalepsy or the immune response. Analysis of the effects of fluoxetine on the functional activity of 5-HT(1A) and 5-HT(2A) receptors, and the expression of 5-HT(1A) receptor genes in the frontal cortex and midbrain and 5-HT(2A) receptors in the frontal cortex, as well as the tryptophan hydroxylase-2 and the serotonin transporter genes in the midbrain showed that the antidepressant had no effect on these parameters in ASC mice, but decreased the functional activity of 5-HT(2A) receptors in CBA/Lac mice. The possibility that the actions of fluoxetine on catalepsy and the immune response in mice with depression-like states are mediated via other serotoninergic mechanisms is discussed.

Yu Di,1 Yiou Zhang,2 Hongwei Yang,1 Aiyuan Wang,1 Xiaolong Chen11Department of Ophthalmology, Shengjing Affiliated Hospital of China Medical University, Shenyang, People’s Republic of China; 2Graduate School, China Medical University, Shenyang, People’s Republic of ChinaBackground: CCN1 (also called Cyr 61) is an extracellular matrix signaling molecule that has been implicated in neovascularization through its interactions with several endothelial integrin receptors. The roles of vascular endothelial growth factor (VEGF) in angiogenesis are well described. The aim of this study was to investigate the signal transduction mechanism of CCN1–PI3K/Akt–VEGF in retinopathy of prematurity (ROP), and the effects of CCN1 knockdown on ROP.Methods: The oxygen-induced retinopathy (OIR) model was established in C57BL/6J mice exposed to a high concentration of oxygen. Retinas were obtained from the normoxia, OIR, OIR control (treated with scramble siRNA) and OIR treated (with CCN1 siRNA) groups. Retinal neovascularization (RNV) was qualitatively analyzed with ADPase staining and quantitatively analyzed by counting neovascular endothelial cell nuclei at postnatal day 17 when RNV reached a peak. mRNA level and protein expression of CCN1, p-Akt, and VEGF were measured by real-time PCR and Western blotting, and located with immunohistochemistry.Results: CCN1 depletion resulted in less neovascularization clock hour scores in the number of preretinal neovascular cells compared with the OIR treated group (1.28±0.83 versus 4.80±0.82; and 7.12±2.50 versus 23.25±2.35, respectively, both P

Background Phosphatase of regenerating liver (PRL) family is classified as class IVa of protein tyrosine phosphatase (PTP4A) that removes phosphate groups from phosphorylated tyrosine residues on proteins. PRL phosphatases have been implicated in a number of tumorigenesis and metastasis processes and are highly conserved. However, the understanding of PRL expression profiles during embryonic development is very limited. Results In this study, we demonstrated and characterized the comprehensive expression pattern of Drosophila PRL, amphioxus PRL, and zebrafish PRLs during embryonic development by either whole mount immunostaining or in situ hybridization. Our results indicate that Drosophila PRL is mainly enriched in developing mid-guts and central nervous system (CNS) in embryogenesis. In amphioxus, initially PRL gene is expressed ubiquitously during early embryogenesis, but its expression become restricted to the anterior neural tube in the cerebral vesicle. In zebrafish, PRL-1 and PRL-2 share similar expression patterns, most of which are neuronal lineages. In contrast, the expression of zebrafish PRL-3 is more specific and preferential in muscle. Conclusions This study, for the first time, elucidated the embryonic expression pattern of Drosophila, amphioxus, and zebrafish PRL genes. The shared PRL expression pattern in the developing CNS among diverse animals suggests that PRL may play conserved roles in these animals for CNS development.