[Cohort study of efficacy and safety of polatuzumab vedotin combined with immunochemotherapy in patients with relapse/refractory diffuse large B cell lymphoma].

Objective: To investigate the efficacy and tolerability of Polatuzumab vedotin+rituximab±bendamustine (Pola-(B)R) in relapse/refractory diffuse large B cell lymphoma (R/R DLBCL) patients. Methods: The clinical data of 21 patients enrolled in Chinese Pola compassionate use program (CUP) in 4 centers from November 2019 to August 2020 were collected. There were 15 males and 6 females, and the median age was 56 years (ranged 25-76 years). Of the patients, 10 cases received Pola-BR regimen and the other 11 received Pola-R. Their clinical features, regimens, efficacy, and adverse events (AEs) were retrospectively analyzed. Results: Twenty-one patients with at least one efficacy evaluation were included. At data analysis cut-off point (12 Aug. 2020), the best overall response (BOR) rate was 81.0% (17/21) and the complete response (CR) rate was 19.0% (4/21). Kaplan-Meier survival estimation was performed, at a median follow-up of 54 days, three patients (14.3%) had disease progressed, and 18 patients (85.7%) were censored; the median progression-free survival (mPFS) was estimated to be 148 days. The incidence of adverse effects (AEs) of any grade was higher in Pola-BR group than Pola-R group (80.0% vs 63.6%). However, the incidence of grade 3-4 AEs were close in the two groups (30.0% vs 29.3%). The most common hematological toxicities were thrombocytopenia (28.6%, 6/21), neutropenia (28.6%, 6/21) and anemia (14.3%, 3/21), respectively. One patient with pneumonia and 1 patient with hemophagocytic syndrome recovered after symptomatic treatment. No peripheral neuropathy of grade≥2 was observed. Conclusions: The preliminary data suggested that, for heavily treated Chinese R/R DLBCL, the Pola-(B)R regimen still achieves promising efficacy and tolerable safety.

[Value of PET/MRI in the diagnosis of malignant pleural effusion in comparison with PET/CT].

Objective: To investigate the diagnostic value of PET/MRI for malignant pleural effusion (MPE), and compare its diagnostic difference with PET/CT. Methods: The data of 57 patients with suspected MPE admitted into Union Hospital of Tongji Medical College of Huazhong University of Science and Technology from October 2017 to January 2020 was analyzed. A total of 53 patients were included in the prospective study, and the whole body PET/CT and thoracic PET/MRI were performed on them respectively. Two physicians used a blind method to evaluate the morphological features of PET/CT and PET/MRI images, delineate the region of interest (ROI), obtain the maximum standard uptake value (SUVmax) of the ROI in the PET/CT and PET/MRI images. The target-to-background ratio (TBR) of the lesion was calculated. The diffusion-weighted imaging (DWI) characteristics of the pleura in PET/MRI images were analyzed. Taking pathological diagnosis as the gold standard, the diagnostic effect of PET/CT and PET/MRI on MPE were evaluated. Results: The 53 patients who were finally included were (62.8±1.7) years old, consisting of 31 males. Pathological results showed that 41 cases were MPE and 12 cases were benign pleural effusion (BPE). There were no statistical differences in age, gender and smoking history between the two groups (P>0.05). Bland-Altman analysis showed that the SUVmax of pleural lesions by PET/MRI was higher than that by PET/CT (6.4±0.6 vs 5.3±0.5, P<0.001). The TBR of PET/MRI was higher than that of PET/CT (2.2±0.2 vs 1.8±0.2, P<0.001). The sensitivity, specificity, and accuracy of PET/MRI in the diagnosis of MPE by combining imaging features such as SUVmax and DWI of pleural lesions were 75.6%, 100%, and 81.1%, respectively. The sensitivity, specificity, and accuracy of PET/CT combined with SUVmax and imaging features of pleural lesions in the diagnosis of MPE were 85.4%, 83.3%, and 77.4%, respectively. There was no statistically significant difference between PET/MRI and PET/CT in the area under the curve (AUC) for diagnosing MPE (0.934 vs 0.873, P>0.05). Conclusions: PET/MRI and PET/CT have the equivalent diagnostic efficiency for MPE. However, PET/MRI shows higher SUVmax and TBR for pleural lesions, and has specific pleural DWI imaging characteristics, which is worthy of further clinical research.

[Promoting effect of cyclin D1 overexpression on proliferation and epithelial mesenchymal transition of cervical squamous cell carcinoma SiHa cells].

Objective: To study effects of cyclin D1 overexpression on the proliferation and differentiation of cervical squamous cell carcinoma SiHa cells and to investigate related signaling molecules. Methods: Primers were designed to amplify the full length of cyclin D1 gene and cyclin D1 gene was amplified by PCR for constructing pcDNA3.1 plasmid vector. The construct was then transfected into SiHa cells, and the cells with stable overexpression of cyclin D1 were established, cyclin D1 gene and protein expression were detected by RT-PCR and Western blot, respectively. Cell growth curve was documented by MTT assay. CK7, E-cadherin, vimentin, Snail gene and protein expression in transfected cells were detected by RT-PCR and Western blot. RT-PCR was used to detect the mRNA expression of proliferation and differentiation-related genes like CDK4, CDK2, p21, p27, cyclin E, Rb, E2F, E6/E7 and Ki-67. After synchronization of cells, RT-PCR was used to detect of cyclin D1 and p21 mRNA expression at different time points of the cell cycle. Results: The G-3 cells with cyclin D1 overexpression were successfully established. The growth curve and Ki-67 mRNA expression accelerated in G-3 cells.Vimentin and Snail expression significantly increased at both gene and protein levels, while E-cadherin, CK7 gene and protein expression significantly decreased, indicating epithelial mesenchymal transitionoccurred in G-3 cells.Meanwhile, mRNA expression of cyclin D1, CDK4, CDK2, p21, p27, cyclin E, E2F and Rb increased, while E6/E7 and p16 showed no significant change. The expression trends of p21 and cyclin D1 were almost identical with fluctuation at different time points in the cell cycle. Conclusions: Overexpression of cyclin D1 induced by gene transfection promotes proliferation and epithelial mesenchymal transition in SiHa cells.The process is accompanied by up-regulation of CDK4, CDK2, p21, p27 and cyclin E genes.p21 expression increases synchronously with cyclin D1, suggesting a regulatory role in epithelial mesenchymal transition by affecting expression of vimentin in G-3 cells.

[Molecular mechanisms of resistance to phosphatidyl inositol 3-kinase inhibitors in triple-negative breast cancer cells].

OBJECTIVE: To explore the molecular mechanisms of resistance to phosphatidyl inositol 3-kinase (PI3K) inhibitors in triple-negative breast cancer (TNBC) cells. METHODS: HCC70 cells (TNBC) were transfected with siFZD7, siWANT5B or siGSK3 using lipofectamine 2000 transfection reagent. The expression levels of key proteins of WNT/ß-catenin and PI3K/AKT/mTOR pathways were determined by Western blot analysis. After HCC70, MCF-7 (ER-positive) and SK-BR3 (HER2-positive) cells were treated with PI3K/AKT/mTOR inhibitors, the inhibition rates of cell proliferation were measured by MTT assay, and half maximal inhibitory concentrations (IC50) were calculated. The altered activities of WNT/ß-catenin and PI3K/AKT/mTOR proteins were detected by Western blot and luciferase report gene assay, respectively. The nuclear translocation of ß-catenin protein was examined by immunofluorescence assay. Xenograft nude mouse model was used to evaluate the tumorigenicity of breast cancer cells treated with BKM120 in vivo. The expression levels of p-LRP6, p-4EBP1 and ß-catenin proteins in the tumor tissues were determined by immunohistochemical staining. RESULTS: The expression levels of FZD7, WANT5B and GSK3 proteins were significantly reduced in the HCC70 cells transfected with the target siRNAs. Meanwhile, the activity of WNT/ß-catenin was enhanced and PI3K/AKT/mTOR pathway was inhibited. PI3K/AKT/mTOR inhibitors suppressed MCF-7 and SK-BR3 cell proliferation. The IC50 of GDC-094, BKM120, XL147, perifosine, everolimus, and BEZ235 in MCF-7 cells were 0.46 mmol/L, 1.44 mmol/L, 4.34 mmol/L, 11.35 µmol/L, 53.71 µmol/L and 12.87 µmol/L respectively, and 0.63 mmol/L, 0.58 mmol/L, 3.74 mmol/L, 13.22 µmol/L, 60.00 µmol/L and 11.38 µmol/L in the SK-BR3 cells, respectively. The results of luciferase report gene assay showed that the luciferase activities in HCC70, MCF-7 and SK-BR3 cells treated with BKM120 were 1.75±0.05, 1.13±0.02 and 0.43±0.01, respectively. The luciferase activities in HCC70 and SK-BR3 cells were significantly different from that of the control cells (1.00±0.02, P<0.05). The immunohistochemical analysis showed that BKM120 inhibited mTOR activity, and the enhanced WNT/ß-catenin activity reversed the phenotype of inhibitory mTOR induced by BKM120. BKM120 suppressed the tumorigenic ability of MCF-7 and SK-BR3 cells in vivo, but had no effect on cultured HCC70 cells. The immunohistochemical analysis showed nuclear translocation of ß-catenin protein and increased expression level of p-LRP-6 protein in transplanted tumor tissues from HCC70 cells treated with BKM120, increased the level of p-LRP-6 protein, and no changes of p-4EBP1 protein expression. However, no nuclear translocation of ß-catenin protein and no decrease of p-LRP6 and p-4EBP1 protein levels in the transplanted tumor tissue of MCF-7 cells after treatment with BKM120. CONCLUSIONS: The triple-negative breast cancer HCC70 cells have drugs-resistance to PI3K inhibitors. The WNT/ß-catenin signaling pathway may regulate the PI3K/AKT/mTOR pathway, therefore, inducing the drug-resistance of TNBC cells to PI3K inhibitors.

[The risk of incident gastric cancer for populations with different precancerous gastric lesions: a prospective follow-up study].

Objective: To provide evidence for optimizing the screening strategy for gastric cancer (GC), we evaluated the risk of incident GC for individuals with different precancerous gastric lesions in a prospective cohort study. Methods: Based on the National Upper Gastrointestinal Cancer Early Detection Program launched in Linqu, Shandong, a high-risk area of gastric cancer in China, we included a total of 14 087 subjects diagnosed with different gastric lesions stages by endoscopic screening from 2012 to 2018. Study subjects were prospectively followed up until December 31, 2019. The incidence of GC during the follow-up was ascertained by repeated endoscopic examinations, cancer, death registry reports, and active follow-up of study subjects and was confirmed by reviewing medical records extracted from the hospital information management system. The Poisson regression model was applied to calculate the relative risk (RR) and 95%CI for GC occurrence among subjects with different gastric lesions. Results: Among 14 087 subjects with different gastric lesions as determined by their first endoscopic examination in 2012-2018, 7 608 (54.00%) had a global diagnosis of superficial gastritis (SG), 2 848 (20.22%) had chronic atrophic gastritis (CAG), 3 103 (22.03%) had intestinal metaplasia (IM), and 520 (3.69%) had low-grade intestinal neoplasia (LGIN). During the follow-up, 109 subjects were diagnosed with GC, including 63 with high-grade intestinal neoplasia (HGIN) and 46 with invasive GC. Compared to subjects having normal gastric mucosa or SG, those with CAG (RR=3.85, 95%CI: 2.04-7.28), IM (RR=5.18, 95%CI: 2.79-9.60), and LGIN (RR=19.08, 95%CI: 9.97-36.53) had significantly increased risk of progression to GC. Individuals with these gastric lesions had an elevated risk of developing HGIN and invasive GC. For subjects with LGIN, the RR was 22.96 (95%CI: 9.71-54.27) for developing HGIN and 14.64 (95%CI: 5.37-39.93) for developing invasive GC. Subgroup analyses found that all age group subjects with LGIN diagnosed during the initial endoscopic examination had a significantly increased risk of developing the GC. Conclusions: Our large-scale prospective study on a high-risk area of GC showed that most residents aged 40-69 years had gastric lesions of different stages. Subjects with more advanced gastric lesions had a significantly increased risk of progression to GC.

Synergistic mechanisms of retigeric acid B and azoles against Candida albicans.

AIMS: To clarify the underlying synergistic antifungal mechanisms of retigeric acid B (RAB) in combination with azoles against Candida albicans. METHODS AND RESULTS: Increased accumulation of rhodamine 123 in C. albicans was measured by both spectrophotometric method and flow cytometry. The inhibitory properties to the drug efflux of C. albicans were determined spectrophotometrically. The decreased cellular ergosterol synthesis was measured using its unique spectrophotometric absorbance profile, and the downregulation expression levels of CDR1 and ERG11 were detected by real-time reverse transcription polymerase chain reaction. Transmission electron microscopy investigation found the wrinkled cell membrane and the impaired cell wall. CONCLUSIONS: RAB synergizes the antifungal effect of azoles against C. albicans by inhibiting efflux pump activity, targeting the ergosterol biosynthesis pathway and increasing the fluidity for the resulted ergosterol depletion. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigating the mechanism of the synergy between RAB and azoles against C. albicans will help us to uncover the antifungal roles of this lichen-derived triterpene acid and find its possible clinical applications in overcoming fungal resistance.

[Imported falciparum malaria in Suzhou City: a report of two cases in 2019].

The demographic features, epidemiology, diagnosis and treatment of two cases with falciparum malaria imported into Suzhou City in 2019 were reported. These findings indicate a risk of imported malaria in visitors besides high prevalence in migrant labors, and much attention should be paid to children that are at a high risk of infections.

Surface tension measurements of surface freezing in liquid normal alkanes.

Surface tension measurements reveal surface freezing in liquid n-alkanes. A solid monolayer of molecules is found to exist up to 30 degrees C above the bulk freezing point. This surface phase exists only for carbon numbers 14 n

Effects of dietary inclusion of Lactobacillus and inulin on growth performance, gut microbiota, nutrient utilization, and immune parameters in broilers.

The effects of dietary Lactobacillus (BCRC 16092) and inulin on growth performance, intestinal microflora, mineral utilization, and tissue mineral contents were evaluated in broilers. The experiment was conducted using 1,152 one-day-old broilers randomly distributed to 9 treatments in a factorial arrangement (3 × 3) using 3 levels of inulin (0, 1, and 2%) and 3 levels of Lactobacillus addition (108, 109, and 1010 CFU/kg). Broilers (1 D of age; 8 replicates per treatments and 16 broilers per replicate) with an initial body weight of 48.36 ± 0.21g were evaluated for 42 D. A 4-D mineral digestibility trial was conducted during the final week of the experiment. The results showed that Lactobacillus supplementation can increase average daily gain and nutrient digestibility and improve feed/gain in broilers (P < 0.05). Moreover, Lactobacillus and inulin supplementation increased the numbers of Lactobacillus and Bifidobacteria, increased serum concentration of IgG and IgA, and decreased the numbers of Escherichia coli and pH in ileum and cecum. The present study demonstrated Lactobacillus and inulin fed to broilers has a positive effect on gut microbiota, growth and nutrient utilization, immune system, and mineral metabolism.

Effect of LGR4 on synovial cells and inflammatory factors in rats with traumatic osteoarthritis.

OBJECTIVE: Traumatic arthritis is one of the most common diseases in orthopedics. LGR4 is involved in bone formation and bone development. However, the role of LGR4 in synovial cells of rats with traumatic osteoarthritis has not been reported. MATERIALS AND METHODS: Sprague Dawley (SD) rats were randomly divided into the control group and model group. The Real Time-Polymerase Chain Reaction (RT-PCR), Western blot, and Enzyme-Linked Immunosorbent Assay (ELISA) were used to analyze the expression of LGR4 in synovial tissue and synovial fluid. Synovial cells were isolated and cultured, followed by transfection of LGR4-pcDNA3.1 plasmid into cells. Cell proliferation was analyzed by MTT and EdU assay, and the Caspase-3 activity was assessed using the Caspase-3 activity kit. The secretion of the inflammatory factors interleukin-1 (IL-1), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) was detected by ELISA. NF-κB signaling pathway changes were evaluated by the Western blot. RESULTS: In the model group, LGR4 mRNA expression in synovial tissue was significantly decreased, and the secretion of LGR4 in the synovial fluid was significantly decreased compared with the control group (p<0.05). LGR4 protein expression in the synovial membrane in the model group tissue was reduced. The transfection of LGR4-pcDNA3.1 plasmid into synovial cells promoted the LGR4 expression, inhibited the proliferation of synoviocytes, increased the Caspase-3 activity, the secretion of IL-1, TNF-α, and IL-6, as well as the decreased expression of NF-κB with a statistical significance, compared with the control group (p<0.05). CONCLUSIONS: LGR4 expression is reduced in the rat model of traumatic osteoarthritis. The upregulation of LGR4 expression can inhibit the secretion of the inflammatory factors and inhibit the proliferation of the synovial cells by regulating NF-κB signaling pathway, which may alleviate the development of the joint inflammation.

Sulfated polysaccharides and immune response: promoter or inhibitor?

Sulfated polysaccharides, which frequently connect to core protein, are expressed not only on cell surface but also throughout the extracellular matrix. Besides providing structural integrity of cells, sulfated polysaccharides interact with a variety of sulfated polysaccharides-binding proteins, such as growth factors, cytokines, chemokines and proteases. Sulfated polysaccharides play two-edged roles, inhibitor and promoter, in immune response. Some sulfated polysaccharides act as the immunosuppressor by blocking inflammatory signal transduction induced by proinflammatory cytokines, suppressing the activation of complement and inhibiting the process that leukocytes adhere to and pass through endothelium. On the contrary, the interaction between immune cells and sulfated polysaccharides produced by bacteria, endothelial cells and immune cells initiate the occurrence of immune response. It promotes the processes of recognizing and arresting antigen, migrating transendothelium, moving into and out of immune organ and enhancing the proliferation of lymphocyte. The structure of sulfated polysaccharides, such as molecular weight and sulfated sites heterogeneity, especially the degree of disaccharide sulfation, position of the sulfate moiety and organization of sulfated domains, may play critical role in their controversial effects. As a consequence, the interaction between sulfated polysaccharides and sulfated polysaccharide-binding proteins may be changed by modifying the structure of sulfated polysaccharides chains. The administration of drug targeting sulfated polysaccharide-protein interaction may be useful in treating inflammatory related diseases.

New strategy of antiangiogenic therapy for hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a hypervascular tumor, and tumor progression and prognosis is associated with angiogenesis. Extracellular matrix remodeling and inflammation play important roles in hepatocarcinogenesis. Some ingredients of extracellular matrix such as endostatin and sulfated polysaccharide, some immunomodulatory agents and cox-2 inhibitor suppress the angiogenesis of HCC. Because vasculogenic mimicry is associated with high tumor grade, some differentiation agents are used to inhibit antiagiogenesis. Besides suppressing the proliferation directly, somatostatin inhibits angiogenesis to suppress growth indirectly. Copper chelator prevents copper from functioning as a cofactor in angiogenesis. The renin-angiotensin system is frequently activated in patients with chronic liver diseases. Perindopril, an angiotensin converting enzyme inhibitor, inhibits angiogenesis by reducing vascular endothelial growth factor (VEGF) production. Kinase inhibitors of VEGF and epidermal growth factor receptors are expected to be of benefit for some patients. Following transarterial embolisation and/or resection, antiangiogenic therapy could prevent the recurring and metastasis. Hypoxia enhances the proliferation, suppresses the differentiation and apoptosis, and induces multidrug resistance of HCC. Because antiangiogenic therapies induce hypoxia, it should be borne in mind the side affects of antiangiogenic therapy. Because long-acting antiangiogenent are needed to control cancer, it needs more clinical studies to confirm the drug resistance of antiangiogenetic therapy.

An FGFR inhibitor converts the tumor promoting effect of TGF-ß by the induction of fibroblast-associated genes of hepatoma cells.

Tumors consistently mimic wound-generating chronic inflammation; however, why they do not heal like wounds with fibrotic scars remains unknown. The components of the tumor microenvironment, such as transforming growth factor ß (TGF-ß) and fibroblast growth factors (FGFs), may account for this phenomenon. Tumor formation involves continuous activation of the FGF pathway, whereas the repair of tissue injury is a self-limiting process accompanied with controlled activation of the FGF pathway. In the tumor microenvironment TGF-ß increases the secretion of FGFs, further promoting the malignant biological properties of tumors. However, during wound healing, sufficient TGF-ß together with moderate FGFs lead to matrix deposition and the formation of fibrotic scars. In the present study, TGF-ß1 combined with AZD4547, an FGF receptor (FGFR) inhibitor, transformed hepatoma cells into less malignant fibroblast-like cells with respect to morphology, physiological properties, and gene expression profiles. In vivo experiments showed that TGF-ß1 combined with AZD4547 not only inhibited tumor growth but also promoted tumor parenchyma fibrosis. Our results indicate that FGFR inhibitor treatment converts the effect of TGF-ß on the hepatocellular carcinoma cells from tumor promotion into tumor inhibition by enhancing the induction effect of TGF-ß on some fibroblast-associated genes. Converting human liver cancer cells into less malignant fibroblast-like cells and inducing tumor parenchyma cell fibrosis provides an alternative strategy for limiting tumor progression.

Sulfated oligosaccharides and tumor: promoter or inhibitor?

The topic about the role of sulfated oligosaccharides in carcinogenesis and progression of tumor remains controversial. The present review aims to evaluate the role of sulfated oligosaccharides in carcinogenesis and progression of tumor. The modification of sulfated oligosaccharides, especially chondroitin sulfate and heparan sulfate, is an important event in carcinogenesis and is correlated with the degree of differentiation. Enhance of chondroitin sulphate may promote carcinogenesis, while enhance of heparin sulphate may promote metastasis. Resistance of antiproliferation activity of sulfated oligosaccharides may contribute to the aberrant behavior of the cancer cell. Some researches supported that sulfated proteoglycan on the cell surface may enhance metastasis; while some soluble sulfated oligosaccharides could suppress metastasis. Thus, sulfated oligosaccharides play double roles, promoter or inhibitor, in carcinogenesis and tumor progression. Four topics about the correlation between sulfated oligosaccharides and carcinogensis and progression are very interesting and must be identified: whether the modified sulfated oligosaccharides have a different effect from the unmodified sulfated oligosaccharides; whether different sulfate oligosaccharides have the different action; whether the function of sulfated proteoglycan on the cell surface is different from that of soluble sulfated oligosaccharides; whether the function of sulfated oligosaccharides in primary tumor is different from that in metastasis tumor. Further data on the long-term safety of sulfate oligosaccharides for cancer patients are therefore required to allow overall risk-benefit assessments.

Cell-matrix talks: effects on differentiation of hepatocellular carcinoma.

Different event is a process that is dependent on stimulation of extracellular signals, signal transduction and gene express. Malignant transformation of hepatocytes may occur in cirrhosis, which is the result of extracellular matrix (ECM) remodeling. ECM could affect and maintain the differentiated phenotype of hepatocytes by regulating liver transcription factors. Moreover, ECM remodeling is correlated with dedifferentiation of hepatocellular carcinoma (HCC). Integrin-matrix adhesion system and E-cadherin/catenin adhesion complex mediate the cell-matrix interaction through focal adhesion kinase, extracellular-signal-regulated kinases and beta catenin/Wnt pathway. The different event of HCC compared with the reversion of abnormal cell-matrix interaction. New drugs that are power for regulating cell-matrix interaction may represent a novel therapeutic strategy for HCC.

Effects of sulfated polysaccharides on tumour biology.

Sulfated polysaccharides can act not only as anticoagulants but also as tumour inhibitors. Recent studies suggest that sulfated polysaccharides could affect tumour cells directly. Sulfated polysaccharides could inhibit the metastasis and proliferation of tumour cells by binding to growth factors and cell adhesion molecules. Moreover, sulfated polysaccharides could inhibit heparanase, which cleaves heparan sulfate chains of heparan sulfate proteoglycans and cause release of growth factors sequestered by heparan sulfate chains. Some sulfated polysaccharides can induce apoptosis and differentiation of tumour cells, but the mechanism is uncertain. In addition, sulfated polysaccharides can enhance the innate and adaptive immune response for tumour cells. Thus, the anti-tumour mechanism of sulfated polysaccharides can be explained, at least partly, through the effects on tumour biology directly.

Orientation-dependent nanostructure arrays based on anisotropic silicon wet-etching for repeatable surface-enhanced Raman scattering.

Repeatable fabrication of sensitive plasmonic substrates through a simple procedure has become a major challenge for SERS-based sensing and imaging. Herein, a new class of high-performance SERS substrates, including pyramid, ridged-hexagon, and quasi-triangle nanostructures, is successfully fabricated based on the nanosphere lithography technique and anisotropic wet etching. Using the wafer-scale Cr-hole array as the etching mask, cavity-templates of various configurations are fabricated by the orientation-dependent wet etching technique, from where the nanostructure arrays are finally peeled-off. The anisotropic wet etching on (100), (110), and (111) silicon wafers has been systematically studied at the nanoscale revealing the formation mechanism of these cavity-templates. The peeled-off nanostructure arrays provide high-density tips and/or gaps (about 2.5 × 10(7) mm(-2)) and thus facilitate the generation of "hot spots". The distribution of the electromagnetic field is visualized by the finite difference time domain calculation. And the calculation results are validated by SERS characterization. The SERS enhancement factors of these substrates are in the order of 10(6)-10(7), with the maximum enhancement factor of 1.32 × 10(7) yielded by the ridged-hexagon arrays. The proposed nanostructure arrays present excellent homogeneity and reproducibility (with the largest relative standard deviation of 16.43%) for the reason that the SERS-active substrates are peeled-off from an identical template. The cost-effective fabrication, high sensitivity, good homogeneity and well-performed reproducibility demonstrate that these orientation-dependent NSs are good candidates for SERS-based in vitro and in situ detection and biosensing.

Haematopoiesis in molluscs: A review of haemocyte development and function in gastropods, cephalopods and bivalves.

Haematopoiesis is a process that is responsible for generating sufficient numbers of blood cells in the circulation and in tissues. It is central to maintenance of homeostasis within an animal, and is critical for defense against infection. While haematopoiesis is common to all animals possessing a circulatory system, the specific mechanisms and ultimate products of haematopoietic events vary greatly. Our understanding of this process in non-vertebrate organisms is primarily derived from those species that serve as developmental and immunological models, with sparse investigations having been carried out in other organisms spanning the metazoa. As research into the regulation of immune and blood cell development advances, we have begun to gain insight into haematopoietic events in a wider array of animals, including the molluscs. What began in the early 1900's as observational studies on the morphological characteristics of circulating immune cells has now advanced to mechanistic investigations of the cytokines, growth factors, receptors, signalling pathways, and patterns of gene expression that regulate molluscan haemocyte development. Emerging is a picture of an incredible diversity of developmental processes and outcomes that parallels the biological diversity observed within the different classes of the phylum Mollusca. However, our understanding of haematopoiesis in molluscs stems primarily from the three most-studied classes, the Gastropoda, Cephalopoda and Bivalvia. While these represent perhaps the molluscs of greatest economic and medical importance, the fact that our information is limited to only 3 of the 9 extant classes in the phylum highlights the need for further investigation in this area. In this review, we summarize the existing literature that defines haematopoiesis and its products in gastropods, cephalopods and bivalves.

Copper bioavailability, blood parameters, and nutrient balance in mink.

A 3 × 3 + 1 factorial experiment was conducted based on a completely randomized design to evaluate the effects of different sources of copper on plasma metabolites, nutrient digestibility, relative copper bioavailability, and retention of some minerals in male mink. Animals in the control group were fed a basal diet, which mainly consisted of corn, fish meal, meat and bone meal, and soybean oil, with no copper supplementation. Mink in the other 9 treatments were fed the basal diet supplemented with Cu from reagent-grade copper sulfate (CuSO4), tribasic copper chloride (TBCC), or copper methionine (CuMet). Copper concentrations of the experimental diets were 50, 100, and 150 mg Cu/kg DM. Blood samples were collected via the toe clip at the end of study (d 42) to determine blood hematology and blood metabolites. A metabolism trial of 4 d was conducted during the last week of experimental feeding. There was a linear (P < 0.01) effect of dose of Cu on plasma Cu concentrations, ceruloplasmin concentration, and Cu-Zn superoxide dismutase activity. A linear response to Cu dose was noted for fat (P < 0.05) digestibility. Supplemental dose of Cu linearly increased (P < 0.05) liver Cu and decreased (P < 0.05) liver Zn level but did not alter liver Fe. The concentration of liver Cu of the mink fed with TBCC and CuMet diets was greater (P < 0.05) than that fed CuSO4. Compared with CuSO4 (100%), relative bioavailability values of TBCC were 104 and 104%, based on serum ceruloplasmin and liver copper, respectively, and relative bioavailability values of CuMet were 130 and 111%. CuMet and TBCC are more bioavailable than CuSO4. In conclusion, the relative bioavailability of CuMet obtained in this study was greater than that of CuSO4 and TBCC. Dose of Cu had an important effect on the regulating ceruloplasmin concentration, Cu-Zn superoxide dismutase activity, and the digestion of dietary fat in mink.

Role of nitric oxide in vascular hyper-responsiveness to norepinephrine in hypertensive Dahl rats.

OBJECTIVE: To determine whether the abnormal vascular responses observed in salt-sensitive hypertension are caused by an impairment in vascular nitric oxide function. DESIGN: Isometric tension was measured in aortic rings isolated from Dahl salt-sensitive and salt-resistant rats fed a regular-salt (0.4% NaCl) or a high-salt (8% NaCl) diet, with and without inhibition of endogenous nitric oxide synthesis. METHODS AND RESULTS: Systolic arterial pressure, measured weekly by the tail-cuff method, increased markedly in DS rats with a high-salt diet but did not increase in the other groups. In aortic rings, norepinephrine evoked dose-dependent contractions which were significantly increased in rings from DS rats with a high-salt diet Pretreatment with Nomega-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, increased the norepinephrine-induced contraction in all groups and abolished differences in contractile responses between high-salt DS rats and the other groups. Acetylcholine induced endothelium-dependent relaxation, which was significantly depressed in high-salt DS rats. L-NAME attenuated the acetylcholine-induced relaxation in all groups and abolished the difference in relaxation response between high-salt DS rats and the other groups. Sodium nitroprusside-induced relaxation was significantly depressed in high-salt DS rats. CONCLUSIONS: Vascular hypercontractile responses to norepinephrine in DS hypertensive rats can, in part, be explained by an impairment in endothelial nitric oxide production.