The Art of Nanoparticle Design: Unconventional Morphologies for Advancing Luminescent Technologies.

The advanced design of rare-earth-doped (RE-doped) fluoride nanoparticles has expanded their applications ranging from anticounterfeiting luminescence and contactless temperature measurement to photodynamic therapy. Several recent studies have focused on developing rare morphologies of RE-doped nanoparticles. Distinct physical morphologies of RE-doped fluoride materials set them apart from contemporary nanoparticles. Every unusual structure holds the potential to dramatically improve the physical performance of nanoparticles, resulting in a remarkable revolution and a wide range of applications. This comprehensive review serves as a guide offering insights into various uniquely structured nanoparticles, including hollow, dumbbell-shaped, and peasecod-like forms. It aims to cater to both novices and experts interested in exploring the morphological transformations of nanoparticles. Discovering new energy transfer pathways and enhancing the optical application performance have been long-term challenges for which new solutions can be found in old papers. In the future, nanoparticle morphology design is expected to involve more refined microphysical methods and chemically-induced syntheses. Targeted modification of nanoparticle morphology and the aggregation of nanoparticles of various shapes can provide the advantages of different structures and enhance the universality of nanoparticles.

Engineering Visible to Near-Infrared Luminescence through a Selective Doping Strategy for High-Performance Temperature Sensing.

Luminescence nanothermometers have garnered considerable attention due to their noncontact measurement, high spatial resolution, and rapid response. However, many nanothermometers employing single-mode measurement encounter challenges regarding their relative sensitivity. Herein, a unique class of tunable upconversion (UC) and downshifting (DS) luminescence covering the visible to near-infrared range (400-1700 nm) is reported, characterized by the superior Tm3+, Ho3+, and Er3+ emissions induced by efficient energy transfer. The outstanding negative thermal expansion characteristic of ScF3 nanocrystals has been found to guide excitation energy toward the relevant emitting states in the Yb3+-Ho3+-Tm3+-codoped system, consequently resulting in remarkable near-infrared III (NIR-III) luminescence at ∼1625 nm (Tm3+:3F4 → 3H6 transition), which in turn presents numerous opportunities for designing multimode ratiometric luminescence thermometry. Furthermore, by facilitating phonon-assisted energy transfer in Er3+-Ho3+-codoped systems, the luminescence intensity ratio (LIR) of 4I13/2 of Er3+ and 5I6 of Ho3+ in ScF3:Yb3+/Ho3+/Er3+ exhibits a strong temperature dependence, enabling NIR-II/III luminescence thermometry with superior thermal sensitivity and resolution (Sr = 0.78% K-1, δT = 0.64 K). These findings not only underscore the distinctive and ubiquitous attributes of lanthanide ion-doped nanomaterials but also hold significant implications for crafting luminescence thermometers with unparalleled sensitivity.

Toward Ultra-High Sensitivity Optical Thermometers and Bright Yellow LEDs Based on Phonon-Assisted Energy Transfer in Rare Earth-Doped La2ZnTiO6 Double Perovskite.

Double perovskites, a class of ceramic oxides with unique crystal structures and diverse physical properties, show promise for various technological applications including solar cells, photodetectors, and light-emitting diodes (LEDs). Despite limited research on rare earth-doped double perovskites, leveraging their ultrahigh luminous efficiency to achieve bright yellow LED emission and addressing energy transfer challenges between Yb3+ and Nd3+ ions in double perovskite La2ZnTiO6 with moderate phonon energy are explored in this work. Through phonon-assisted energy transfer, an ultrasensitive optical thermometer covering a wide temperature range is developed by utilizing the different temperature responses of Er3+ emission in the visible light region and Nd3+ emission in the near-infrared region based on the luminescence intensity ratio (LIR). All the results demonstrate that the rare earth (Yb-Er, Yb-Nd, and Yb-Nd-Er)-doped La2ZnTiO6 phosphors can be effectively utilized for ultrabright LED illumination and ultrahigh sensitivity self-calibrated temperature sensing. This research underscores the significance of phonon-assisted energy transfer in improving material properties and provides valuable insights for the advancement of multifunctional materials.

Simulation of the pollution abatement behavior of regional metal-related enterprises based on the interactive perspective of industrial agglomerations and emission reduction effects.

A machine learning method was used to process a multiagent information database to study the spatial distribution characteristics of agglomerations of metal-related enterprises and to analyze the spatial and temporal differentiation characteristics of pollution reduction in metal-related enterprises. Based on the spatial distribution of enterprises and a simulation of their pollution reduction behaviors, the layout of 380 enterprises sample is optimized, and the direction of industrial transfer is planned to give full play to the pollution reduction effect of enterprise agglomeration. The results showed that (1) the metal-related enterprises in the Chang-Zhu-Tan urban agglomeration have obvious spatial heterogeneity and are mainly distributed in the district of Changsha, the Qingshuitang Industrial Zone, Liling city and the Qibaoshan Industrial Zone of Liuyang city, while the metal-related enterprises in Shaoshan city, Zhuzhou County and Liling city are scattered. (2) The pollution emission behaviors of enterprises differ in time and space, and the pollution concentrations are highest in industrial parks such as Qingshuitang and Zhubu Port. (3) There is an interactive relationship between the degree of enterprise agglomeration and the pollution reduction effect. The spatial positive coupling degree between the concentration of metal-related enterprises and the degree of metal-related pollution is significant, accounting for 94.96% of the study area. Low pollution-high agglomeration areas, high pollution-low agglomeration areas, high pollution-high agglomeration areas, and low pollution-low agglomeration area account for 1.01%, 4.03%, 2.87%, and 92.09% of the study area, respectively. Finally, based on the new development concept of dual circulation and the theory of a two-oriented society in the new era, the paper puts forward suggestions and policies for the sustainable development of industrial transfer.

Solubility and Bioavailability Enhancement of Oridonin: A Review.

Oridonin (ORI), an ent-kaurene tetracyclic diterpenoid compound, is isolated from Chinese herb Rabdosia rubescens with various biological and pharmacological activities including anti-tumor, anti-microbial and anti-inflammatory effects. However, the clinical application of ORI is limited due to its low solubility and poor bioavailability. In order to overcome these shortcomings, many strategies have been explored such as structural modification, new dosage form, etc. This review provides a detailed discussion on the research progress to increase the solubility and bioavailability of ORI.

Preparation of oridonin nanocrystals and study of their endocytosis and transcytosis behaviours on MDCK polarized epithelial cells.

Context: Oridonin (ORI) has obvious anticancer effects, but its solubility is poor. Nanocrystal (NC) is a novel nano-drug delivery system for increasing bioavailability for ORI. However, the endocytosis and transcytosis behaviours of oridonin nanocrystals (ORI-NCs) through epithelial membrane are still unclear.Objectives: ORI-NCs were prepared and characterized. The in vitro cytotoxicity and endocytosis and transcytosis process on Madin-Darby canine kidney (MDCK) monolayer were investigated.Materials and methods: Anti-solvent precipitation method was adopted in preparation of ORI-NCs. Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) were adopted to explore crystallography of ORI-NCs. Sulforhodamine B (SRB) method was used to test the inhibition effect on proliferation of MDCK cells. Quantitative analysis by HPLC was performed to study the endocytosis and transcytosis of ORI-NCs and ORI bulk drug, and the process was observed by confocal laser spectrum microscopy (CLSM) and flow cytometry.Results: The particle size of ORI-NCs was about 274 nm. The crystallography form of ORI was not changed after prepared into NCs. The dissolution rate of ORI-NCs was higher than pure ORI in 120 min. At higher concentrations (34, 84 and 135 µg/mL), ORI-NCs significantly reduced the cell viability compared with free ORI (p < 0.05, p < 0.01). ORI-NCs demonstrated higher endocytosis in MDCK cells than free ORI (p < 0.01). In the transport process, ORI-NC was taken up into cells in an intact form, and excreted out from basolateral membrane of polarized epithelial cells in an intact form. The internalization and transmembrane amount increased as a function of time.Conclusions: ORI-NCs transported through the MDCK monolayers in an intact form.

Towards core-shell engineering for efficient luminescence and temperature sensing.

Research on the core-shell design of rare earth-doped nanoparticles has recently gained significant attention, particularly in exploring the synergistic effects of combining active and inert shell layers. In this study, we successfully synthesized 8 types of spherical core-shell Na-based nanoparticles to enhance the efficiency of core-shell design in upconversion luminescence and temperature sensing through the strategic arrangement of inert and active layers. The most effective upconversion luminescence was observed under 980 nm and 808 nm laser excitation using NaYF4 inert shell NaYF4:Yb3+, Er3+@ NaYF4 and NaYF4@ NaYF4:Yb3+, Nd3+ core-shell nanostructures. Moreover, the incorporation of the NaYbF4 active shell structure led to a significant increase in relative sensitivity in ratio luminescence thermometry. Notably, the NaYF4:Yb3+, Nd3+, Er3+@ NaYbF4 core-shell structure demonstrated the highest relative sensitivity of 1.12 %K-1. This research underscores the crucial role of inert shell layers in enhancing upconversion luminescence in core-shell structure design, while active layers play a key role in achieving high-sensitivity temperature detection capabilities.

MicroRNA-455-3p accelerate malignant progression of tumor by targeting H2AFZ in colorectal cancer.

Colorectal cancer (CRC) becomes the second leading cause of cancer-related deaths in 2020. Emerging studies have indicated that microRNAs (miRNAs) play a key role in tumorigenesis and progression. The dysfunctions of miR-455-3p are observed in many cancers. However, its biological function in CRC remains to be confirmed. By sequencing serum sample, miR-455-3p was found to be up-regulated in CRC patients. RT-qPCR demonstrated that the miR-455-3p expression was both higher in the serum and tumor tissues of CRC patients. Furthermore, it indicated that miR-455-3p had the ability in promoting cell proliferation, suppressing cell apoptosis, and stimulating cell migration. In vivo experiments also showed that miR-455-3p promoted tumor growth. Additionally, H2AFZ was proved as the direct gene target of miR-455-3p by dual-luciferase assay. Taken together, miR-455-3p functioned as a tumor promoter in CRC development by regulating H2AFZ directly. Thus, it has enormous potential as a biomarker in the diagnosis of CRC.

Clinical characteristics and prognosis in systemic lupus erythematosus-associated pulmonary arterial hypertension based on consensus clustering and risk prediction model.

BACKGROUND: Pulmonary arterial hypertension (PAH) is a severe complication of systemic lupus erythematosus (SLE). This study aims to explore the clinical characteristics and prognosis in SLE-PAH based on consensus clustering and risk prediction model. METHODS: A total of 205 PAH (including 163 SLE-PAH and 42 idiopathic PAH) patients were enrolled retrospectively based on medical records at the First Affiliated Hospital of Zhengzhou University from July 2014 to June 2021. Unsupervised consensus clustering was used to identify SLE-PAH subtypes that best represent the data pattern. The Kaplan-Meier survival was analyzed in different subtypes. Besides, the least absolute shrinkage and selection operator combined with Cox proportional hazards regression model were performed to construct the SLE-PAH risk prediction model. RESULTS: Clustering analysis defined two subtypes, cluster 1 (n = 134) and cluster 2 (n = 29). Compared with cluster 1, SLE-PAH patients in cluster 2 had less favorable levels of poor cardiac, kidney, and coagulation function markers, with higher SLE disease activity, less frequency of PAH medications, and lower survival rate within 2 years (86.2% vs. 92.8%) (P < 0.05). The risk prediction model was also constructed, including older age at diagnosis (≥ 38 years), anti-dsDNA antibody, neuropsychiatric lupus, and platelet distribution width (PDW). CONCLUSIONS: Consensus clustering identified two distinct SLE-PAH subtypes which were associated with survival outcomes. Four prognostic factors for death were discovered to construct the SLE-PAH risk prediction model.

Treatment of Ulcerative Colitis by Cationic Liposome Delivered NLRP3 siRNA.

Purpose: The abnormal activation of NLRP3 inflammasome is related to the occurrence and development of ulcerative colitis (UC). However, the ideal drug and delivery system remain important factors limiting the targeting of NLRP3 inflammasome in UC therapy. Gene therapy by delivering siRNA is effective in treating various diseases. Therefore, delivering siNLRP3 using an ideal vector for UC treatment is necessary. Materials and Methods: Nanoparticles delivering siNLRP3 were developed based on cationic liposome (CLP/siNLRP3). Their ability to inhibit NLRP3 inflammasome activation was monitored using Western blot (WB) and Enzyme-linked Immunosorbent Assay (ELISA). The ASC oligomerization in LPS-primed peritoneal macrophages (PMs) was detected by WB and immunofluorescence. Moreover, we assessed the role of CLP/siNLRP3 on dextran sodium sulfate (DSS)-induced UC by examining NLRP3 levels, pro-inflammatory cytokines expression, and disease-associated index (DAI). Flow cytometry (FCM) was used to detect the contents of macrophages and T cells. Finally, we assessed the safety of CLP/siNLRP3. Results: The prepared CLP was spherical, with a small particle size (94 nm) and low permeability. The CLP could efficiently protect siNLRP3 from degradation and then deliver siNLRP3 into PMs, inhibiting NLRP3 inflammasome activation. Also, the CLP/siNLRP3 could inhibit the secretion of mature IL-1ß and IL-18 from PMs, thereby achieving a favorable anti-inflammation effect. In vivo, CLP/siNLRP3 could effectively alleviate intestinal injury in UC mice, which was attributed to down-regulating levels of IL-1ß and IL-18, inhibiting infiltration of macrophages and other immune cells, and the polarization of M1 macrophages. Finally, pathological testing of tissue sections and blood biochemical tests showed no significant toxic effects of CLP/siNLRP3. Conclusion: We introduced a prospective approach for the efficient delivery of siRNA in vitro and in vivo with high safety and stability, which was found to have great potential in treating NLRP3-driven diseases in an RNA-silencing manner.

An update on the role of tumor necrosis factor alpha stimulating gene-6 in inflammatory diseases.

At present, the occurrence and development of inflammatory diseases are closely related to the abnormal changes of the content and function of many cytokines. At the same time, targeting related cytokines to prevent and treat diseases has also achieved good results. Therefore, it is very important to explore the role of various cytokines in inflammatory diseases. As an inflammation related protein, Tumor necrosis factor alpha stimulating gene-6 (TSG-6) has attracted more and more attention. In the process of disease, it's like a double-edged sword, showing different responses. It is constitutively expressed in some tissues with high metabolic activity and barrier protection. The diversity of its functions depends on the binding of TSG-6 with a variety of ligands, including matrix molecules, autoimmune regulatory factors and growth factors, participating in extracellular matrix remodeling and regulating protease network. This paper reviews the structure, biological function and research progress of TSG-6 in inflammatory diseases, in order to provide reference for drug development in the future.

TRIP13, identified as a hub gene of tumor progression, is the target of microRNA-4693-5p and a potential therapeutic target for colorectal cancer.

Colorectal cancer (CRC) is one of the digestive tract malignancies whose early symptoms are not obvious. This study aimed to identify novel targets for CRC therapy, especially early-stage CRC, by reanalyzing the publicly available GEO and TCGA databases. Thyroid hormone receptor interactor 13 (TRIP13) correlated with tumor progression and prognosis of patients after several rounds of analysis, including weighted gene correlation network analysis (WGCNA), and further chosen for experimental validation in cancer cell lines and patient samples. We identified that mRNA and protein levels of TRIP13 increased in CRC cells and tumor tissues with tumor progression. miR-4693-5p was significantly downregulated in CRC tumor tissues and bound to the 3' untranslated region (3'UTR) of TRIP13, downregulating TRIP13 expression. DCZ0415, a small molecule inhibitor targeting TRIP13, induced anti-tumor activity in vitro and in vivo. DCZ0415 markedly suppressed CRC cell proliferation, migration, and tumor growth, promoted cell apoptosis, and resulted in the arrest of the cell cycle. Our research suggests that TRIP13 might play a crucial role in CRC progression and could be a potential target for CRC therapy.

Exploration of the Shared Gene Signatures and Molecular Mechanisms Between Systemic Lupus Erythematosus and Pulmonary Arterial Hypertension: Evidence From Transcriptome Data.

Background: Systemic lupus erythematosus (SLE) is an autoimmune disease that can affect multiple systems. Pulmonary arterial hypertension (PAH) has a close linkage with SLE. However, the inter-relational mechanisms between them are still unclear. This article aimed to explore the shared gene signatures and potential molecular mechanisms in SLE and PAH. Methods: The microarray data of SLE and PAH in the Gene Expression Omnibus (GEO) database were downloaded. The Weighted Gene Co-Expression Network Analysis (WGCNA) was used to identify the co-expression modules related to SLE and PAH. The shared genes existing in the SLE and PAH were performed an enrichment analysis by ClueGO software, and their unique genes were also performed with biological processes analyses using the DAVID website. The results were validated in another cohort by differential gene analysis. Moreover, the common microRNAs (miRNAs) in SLE and PAH were obtained from the Human microRNA Disease Database (HMDD) and the target genes of whom were predicted through the miRTarbase. Finally, we constructed the common miRNAs-mRNAs network with the overlapped genes in target and shared genes. Results: Using WGCNA, four modules and one module were identified as the significant modules with SLE and PAH, respectively. A ClueGO enrichment analysis of shared genes reported that highly activated type I IFN response was a common feature in the pathophysiology of SLE and PAH. The results of differential analysis in another cohort were extremely similar to them. We also proposed a disease road model for the possible mechanism of PAH secondary to SLE according to the shared and unique gene signatures in SLE and PAH. The miRNA-mRNA network showed that hsa-miR-146a might regulate the shared IFN-induced genes, which might play an important role in PAH secondary to SLE. Conclusion: Our work firstly revealed the high IFN response in SLE patients might be a crucial susceptible factor for PAH and identified novel gene candidates that could be used as biomarkers or potential therapeutic targets.

IBSP, a potential recurrence biomarker, promotes the progression of colorectal cancer via Fyn/ß-catenin signaling pathway.

Colorectal cancer (CRC) is a frequently occurring digestive system cancer and postoperative tumor metastasis and recurrence are the main reasons for the failure of CRC treatment. The aim of this study was to identifying and validating key genes associated with metastatic recurrence of CRC. RNA expression of three datasets (GSE17538, GSE32323, and GSE29623) was used for biomarker discovery. We identified integrin-binding sialoprotein (IBSP) as a candidate biomarker which was validated in three clinical cohorts (GSE41258, GSE21510, and GSE39582) and our clinical specimens. The results suggested that IBSP expression significantly increased at mRNA and protein levels among CRC cases, which was associated with metastatic recurrence, metastasis, high risk of recurrence, and poor survival in CRC. Consistent results were obtained in CRC cells. The relative level of serum IBSP evidently increased among CRC patients relative to normal controls, and downregulated after operation. As suggested by gene set enrichment analysis (GSEA), the IBSP level was associated with cell-matrix adhesion in CRC. Functional experiments in vitro showed that IBSP promoted the growth and aggressiveness of CRC, and the potential mechanism by which IBSP promoted carcinogenesis of CRC was the abnormal activation of Fyn/ß-catenin signaling pathway. To sum up, findings in the present work indicate that IBSP can serve as the candidate biomarker for the diagnosis, treatment, and prognosis of CRC.

Extraction, structure, pharmacological activities and drug carrier applications of Angelica sinensis polysaccharide.

Angelica sinensis polysaccharide (ASP) is one of the main active components of Angelica sinensis (AS) that is widely used in traditional Chinese medicine. ASP is water-soluble polysaccharides, and it is mainly composed of glucose (Glc), galactose (Gal), arabinose (Ara), rhamnose (Rha), fucose (Fuc), xylose (Xyl) and galacturonic acid (GalUA). The extraction methods of ASP include hot water extraction and ultrasonic wave extraction, and different extraction methods can affect the yield of ASP. ASP has a variety of pharmacological activities, including hematopoietic activity, promoting immunity, antitumor, anti-inflammatory, antioxidant, anti-aging, anti-virus, liver protection, and so on. As a kind of natural polysaccharide, ASP has potential application as drug carriers. This review provides a comprehensive summary of the latest extraction and purification methods of ASP, the strategies used for monosaccharide compositional analysis plus polysaccharide structural characterization, pharmacological activities and drug carrier applications, and it can provide a basis for further study on ASP.

DNA barcoding and phylogenetic analysis of bagrid catfish in China based on mitochondrial COI gene.

The family Bagridae is a collection of species widely distributed in Africa and Asia with a high diversity of morphology. The species identification and phylogenetic relationship in this family have been confused and controversial. In order to explore the effectiveness of DNA barcoding in species identification of Bagridae, sequences of mitochondrial cytochrome c oxidase I (COI) gene of 20 species in four genera of Bagridae were used to analyse barcoding gap and to reconstruct phylogenetic relationship. Both the barcoding gap and the phylogenetic tree analysis showed that the COI gene-based DNA barcoding is an effective molecular technique for most species recognition of Chinese Bagridae. However, the rapid speciation and incomplete lineage sorting may affect the accuracy of DNA barcoding in species identification in certain species, and adding additional genes, such as nuclear gene, may help to achieve accurate identification of these species. The phylogenetic tree showed that the monophyly of genera Pelteobagrus, Leiocassis and Pseudobagrus did not exist, which supports that the species of genera Pelteobagrus, Pseudobagrus and Leiocassis distributed in China should be revised into one genus.

BGN and COL11A1 Regulatory Network Analysis in Colorectal Cancer (CRC) Reveals That BGN Influences CRC Cell Biological Functions and Interacts with miR-6828-5p.

PURPOSE: We explored specific expression profiles of BGN and COL11A1 genes and studied their biological functions in CRC using bioinformatics tools. PATIENTS AND METHODS: A total of 68 pairs of cancer and non-cancerous tissues from CRC patients were enrolled in this study. Methods we used in this articles including: qRT-PCR, Western blot analysis, ELISA, GO and KEGG regulatory network analysis, tumor infiltration, luciferase reporter-based protein and etc. RESULTS: According to The Cancer Genome Atlas (TCGA) data, BGN and COL11A1 expression levels were significantly higher in CRC patient samples than in samples from healthy controls. Moreover, levels were much higher in late-stage CRC than in early-stage disease, warranting evaluation of these genes as CRC prognostic biomarkers. Subsequently, qRT-PCR, Western blot analysis, and ELISA results obtained from analyses of CRC cells, tissues, and patient sera aligned with TCGA results. GO and KEGG regulatory network analysis revealed BGN- and COL11A1-associated genes that were functionally related to extracellular matrix (ECM) receptor pathway activation, with transcription factor genes RELA and NFKB1 positively associated with BGN expression and CEBPZ and SIRT1 with COL11A1 expression. Meanwhile, BGN and COL11A1 expression were separately and significantly correlated to tumor infiltration by six immune cell types. Additionally, kinase genes PLK1 and LYN appeared to be downstream targets of differentially expressed BGN and COL11A1, respectively. In addition, the expression of PLK1 mRNA was down-regulated while BGN was down-regulated. Finally, BGN effects on CRC cell proliferation, cycle, apoptosis, invasion, and migration were studied using molecular biological methods, including luciferase reporter-based protein analysis, qRT-PCR, and Western blot results, which revealed that miR-6828-5p may regulate BGN expression. CONCLUSION: We speculate that the use of BGN and COL11A1 as CRC biomarkers would improve CRC staging, while also providing several novel targets for use in the development of more effective CRC treatments.

Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris.

Yeast Pichia pastoris is a widely used system for heterologous protein expression. However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. PDGF-BB secreted by P. pastoris is very heterogeneous and contains multiple isoforms. We demonstrated that PDGF-BB was O-glycosylated during the secretion process and detected putative O-glycosylation sites using glycosylation staining and immunoblotting. By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. In addition, the unglycosylated PDGF-BBΔGly mutant exhibited the immunogenicity comparable to that of the wild-type form. Furthermore, antiserum against PDGF-BBΔGly also recognized glycosylated PDGF-BB, indicating that protein immunogenicity was unaltered by glycosylation. These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications.