Optimization of medium composition for the production of exopolysaccharides from Phellinus baumii Pilát in submerged culture and the immuno-stimulating activity of exopolysaccharides

Optimization of medium composition for the production of exopolysaccharides (EPS) from Phellinus baumii Pilát in submerged culture and the immuno-stimulating activity of EPS were carried out. Firstly, the medium components having significant effect on EPS production were screened out to be glucose, yeast extract and diammonium oxalate monohydrate by using a 2⁽⁷⁻³⁾ fractional factorial design. Secondly, the concentrations of the three factors were optimized using central composite design in response surface methodology. As results, a quadratic model was found to fit for EPS production, and the optimal medium composition was determined as following (g/l): 34.12 glucose, 4 peptone, 5.01 yeast extract, 0.88 diammonium oxalate monohydrate, 0.75 MgSO₄ and 1 KH₂PO₄ and 0.0075 thiamine (VB₁). A yield of 2.363 ± 0.04 g/l for EPS was observed in verification experiment. Finally, EPS from P. baumii Pilát was found to have direct immuno-stimulating activity in vitro on splenocyte proliferative response and acid phosphatase activity in peritoneal macrophages in a dose-dependent manner.     

Effect of meal size on excess post-exercise oxygen consumption in fishes with different locomotive and digestive performance

Effects of feeding on pre-exercise VO₂ and excess post-exercise oxygen consumption (EPOC) after exhaustive exercise were investigated in sedentary southern catfish, active herbivorous grass carp, omnivorous crucian carp, and sluggish omnivorous darkbarbel catfish to test whether feeding had different effects on EPOC and to compare EPOC in fishes with different ecological habits. For fasting fish, the pre-exercise and peak post-exercise VO₂ were higher and recovery rates were faster in crucian carp and grass carp compared to those of darkbarbel catfish and southern catfish. EPOC magnitudes of grass carp and southern catfish were significantly larger than those of crucian carp and darkbarbel catfish. Feeding had no significant effect on peak post-exercise VO₂, recovery rate, and EPOC magnitude in grass carp. Both the pre-exercise and peak post-exercise VO₂ increased with meal size, while the EPOC magnitude and duration decreased significantly in the larger meal size groups of crucian carp and southern catfish. In darkbarbel catfish, both the pre-exercise and peak post-exercise VO₂ increased with meal size, but the VO₂ increment elicited by exercise was larger in feeding groups compared with the fasting group. These results suggest that (1) the characteristics of the post-exercise VO₂ profile, such as peak post-exercise VO₂ and recovery rate, were closely related to the activity of fishes, whereas the EPOC magnitude was not and (2) the effects of feeding on EPOC were more closely related to the postprandial increase in VO₂.     

Ca2+和K+对拟南芥幼苗镉毒害的缓解作用

该文探讨了外源钙(Ca)或钾(K)处理对不同程度的镉(Cd)胁迫(0–80 μmol·L^–1)下拟南芥(Arabidopsis thaliana)幼苗的生长和生理特性的影响。综合Ca和K对不同浓度Cd胁迫下拟南芥幼苗生长、根长以及生物量的影响情况, 表明各浓度Cd胁迫下外源Ca²⁺的最适缓解浓度均为10 mmol·L^–1; 而K⁺的最适缓解浓度在低浓度(20和40 μmol·L^–1)和高浓度(60和80 μmol·L^–1)Cd胁迫下分别为10 mmol·L^–1和20 mmol·L^–1。在低浓度Cd胁迫下, 添加适宜浓度的Ca²⁺或K⁺后幼苗可溶性蛋白和丙二醛(MDA)含量以及超氧化物歧化酶(SOD)活性相比未添加Ca和K的对照组无显著变化, 而过氧化物酶(POD)活性和总酚、类黄酮、花色素苷, 酸溶性硫醇化合物、谷胱甘肽(GSH)、植物螯合肽(PCs)的含量均下降; 高浓度Cd处理下, 添加适宜浓度的Ca²⁺或K⁺后幼苗的SOD活性升高, POD活性降低, 可溶性蛋白、MDA、总酚、类黄酮、花色素苷、酸溶性硫醇化合物、GSH以及PCs的含量也均低于对照组。在各浓度Cd胁迫下, 添加外源Ca或K均使拟南芥幼苗根部细胞DNA损伤减弱, 表现为TT嘧啶二聚体的累积量显著减少(P<0.05)。以上结果表明, 在Cd胁迫(尤其是高浓度Cd胁迫)下, 外源Ca或K通过调节酚类、金属螯合物质的代谢水平以及提高拟南芥的抗氧化能力来缓解Cd对拟南芥幼苗的毒害效应, 缓解细胞DNA损伤。该研究结果不仅能够为深入探讨Ca和K对缓解重金属毒害的分子机理提供实验依据, 而且为Ca和K应用于重金属污染的防治提供参考。     

Challenge in pathologic diagnosis of Alport syndrome: evidence from correction of previous misdiagnosis

Background

Pathologic studies play an important role in evaluating patients with Alport syndrome besides genotyping. Difficulties still exist in diagnosing Alport syndrome (AS), and misdiagnosis is a not-so-rare event, even in adult patient evaluated with renal biopsy.

Methods

We used nested case–control study to investigate 52 patients previously misdiagnosed and 52 patients initially diagnosed in the China Alport Syndrome Treatments and Outcomes Registry e-system.

Results

We found mesangial proliferative glomerulonephritis (MsPGN, 26.9%) and focal and segmental glomerulosclerosis (FSGS, 19.2%) were the most common misdiagnosis. FSGS was the most frequent misdiagnosis in female X-linked AS (fXLAS) patients (34.8%), and MsPGN in male X-linked AS (mXLAS) patients (41.2%). Previous misdiagnosed mXLAS patients (13/17, 76.5%) and autosomal recessive AS (ARAS) patients (8/12, 66.7%) were corrected after a second renal biopsy. While misdiagnosed fXLAS patients (18/23, 78.3%) were corrected after a family member diagnosed (34.8%) or after rechecking electronic microscopy and/or collagen-IV alpha-chains immunofluresence study (COL-IF) (43.5%) during follow-up. With COL-IF as an additional criterion for AS diagnosis, we found that patients with less than 3 criteria reached have increased risk of misdiagnosis (3.29-fold for all misdiagnosed AS patients and 3.90-fold for fXLAS patients).

Conclusion

We emphasize timely and careful study of electronic microscopy and COL-IF in pathologic evaluation of AS patients. With renal and/or skin COL-IF as additional criterion, 3 diagnosis criteria reached are the cutoff for diagnosing AS pathologically.

     

Transcriptome differences in the hypopharyngeal gland between Western Honeybees (Apis mellifera) and Eastern Honeybees (Apis cerana)

Background

Apis mellifera and Apis cerana are two sibling species of Apidae. Apis cerana is adept at collecting sporadic nectar in mountain and forest region and exhibits stiffer hardiness and acarid resistance as a result of natural selection, whereas Apis mellifera has the advantage of producing royal jelly. To identify differentially expressed genes (DEGs) that affect the development of hypopharyngeal gland (HG) and/or the secretion of royal jelly between these two honeybee species, we performed a digital gene expression (DGE) analysis of the HGs of these two species at three developmental stages (newly emerged worker, nurse and forager).

Results

Twelve DGE-tag libraries were constructed and sequenced using the total RNA extracted from the HGs of newly emerged workers, nurses, and foragers of Apis mellifera and Apis cerana. Finally, a total of 1482 genes in Apis mellifera and 1313 in Apis cerana were found to exhibit an expression difference among the three developmental stages. A total of 1417 DEGs were identified between these two species. Of these, 623, 1072, and 462 genes showed an expression difference at the newly emerged worker, nurse, and forager stages, respectively. The nurse stage exhibited the highest number of DEGs between these two species and most of these were found to be up-regulated in Apis mellifera. These results suggest that the higher yield of royal jelly in Apis mellifera may be due to the higher expression level of these DEGs.

Conclusions

In this study, we investigated the DEGs between the HGs of two sibling honeybee species (Apis mellifera and Apis cerana). Our results indicated that the gene expression difference was associated with the difference in the royal jelly yield between these two species. These results provide an important clue for clarifying the mechanisms underlying hypopharyngeal gland development and the production of royal jelly.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-744) contains supplementary material, which is available to authorized users.     

Overexpression of EMMPRIN is associated with lymph node metastasis and advanced stage of non-small cell lung cancer: a retrospective study

Background

Previous studies show that overexpression of EMMPRIN involved in the malignant biological behavior of tumors. This investigation was to disclose the expression status of EMMPRIN in non-small cell lung cancer (NSCLC) and its clinical value for the diagnosis of NSCLC.

Methods

The expression of EMMPRIN was examined using immunohistochemistry and enzyme-linked immunosorbent assay. The clinical value of EMMPRIN was evaluated by drawing a receiver operating characteristic (ROC) curve.

Results

NSCLC tissues and serum exhibited higher expression levels of EMMPRIN than the normal control (p?<?0.05), and the expression of the EMMPRIN was significantly associated with lymphatic invasion and advanced stage of NSCLC (p?<?0.05). ROC curve suggested that the threshold level of serum EMMPRIN for distinguishing NSCLC from control group was 80.3 pg/mL, and displayed a sensitivity of 97.22% and a specificity of 95%. And higher EMMPRIN expression in serum and tissues appeared to be risk factors for NSCLC development (risk ratio =1.56 and 1.1).

Conclusion

Overexpression of EMMPRIN was associated with lymphatic metastasis and advanced stage of NSCLC and test of serum EMMPRIN contributes to the NSCLC diagnosis.

     

Influence of Al3+ on the titer of spiramycin and effective components in fermentor

Spiramycin is a multicomponent antibiotic, and different components have different antibacterial activities. In Streptomyces spiramyceticus 16-10-2, spiramycin II and spiramycin III (SPMII and SPMIII) are the main components, while spiramycin I (SPMI) needs to be controlled below 12%. Based on this, the influences of Al³⁺ on total spiramycin titer and components were investigated in this work. Those experiments were mainly performed in 15?L fermentor and Al³⁺ made a great improvement in spiramycin titer. The optimal adding concentration and adding time of Al³⁺ were 0.32?g/L at 12?hr. Under this condition, spiramycin titer was increased by 19.51% compared with the control. Moreover, the percentage of SPMII and SPMIII was increased by 7.14%. At the same time, the time of mycelia autolysis was lengthened. In addition, the specific activities of acetyl-CoA synthetase, acetate kinase, acetylphosphotransferase, and acylating enzyme were much higher than those of control. The content of acetic acid and succinic acid was beyond 3 and 4.5 times than that of control, respectively.     

The Effects of Co-transplantation of Olfactory Ensheathing Cells and Schwann Cells on Local Inflammation Environment in the Contused Spinal Cord of Rats

Inflammatory response following spinal cord injury (SCI) is important in regulation of the repair process. Olfactory ensheathing cells (OECs) and Schwann cells (SCs) are important donor cells for repairing SCI in different animal models. However, synergistic or complementary effects of co-transplantation of both cells for this purpose have not been extensively investigated. In the present study, we investigated the effects of co-transplantation of OECs and SCs on expression of pro- or anti-inflammatory factor and polarization of macrophages in the injured spinal cord of rats. Mixed cell suspensions containing OECs and SCs were transplanted into the injured site at 7 days after contusion at the vertebral T10 level. Compared with the DMEM, SC, or OEC group, the co-transplantation group had a more extensive distribution of the grafted cells and significantly reduced number of astrocytes, microglia/macrophage infiltration, and expression of chemokines (CCL2 and CCL3) at the injured site. The co-transplantation group also significantly increased arginase⁺/CD206⁺ macrophages (IL-4) and decreased iNOS⁺/CD16/32⁺ macrophages (IFN-γ), which was followed by higher IL-10 and IL-13 and lower IL-6 and TNF-α in their expression levels, a smaller cystic cavity area, and improved motor functions. These results indicate that OEC and SC co-transplantation could promote the shift of the macrophage phenotype from M(IFN-γ) to M(IL-4), reduce inflammatory cell infiltration in the injured site, and regulate inflammatory factors and chemokine expression, which provide a better immune environment for SCI repair.