Endoplasmic reticulum stress-related genes as prognostic and immunogenic biomarkers in prostate cancer.

BACKGROUND: The metastasis and aggressive nature of prostate cancer (PCa) has become a major malignancy related threat that concerns men's health. The efficacy of immune monotherapy against PCa is questionable due to its lymphocyte-suppressive nature. METHOD: Endoplasmic reticulum stress- (ERS-) and PCa-prognosis-related genes were obtained from the Molecular Signatures Database and the Cancer Genome Atlas database. The expression, prognosis and immune infiltration values of key genes were explored by "survival R package", "rms", "xCELL algorithm", and univariate-multivariate Cox and LASSO regression analyses. The "consensus cluster plus R package" was used for cluster analysis. RESULT: As ERS-related genes, ERLIN2 and CDK5RAP3 showed significant expressional, prognostic and clinic-pathologic values. They were defined as the key genes significantly correlated with immune infiltration and response. The nomogram was constructed with T-stage and primary treatment outcome, and the risk-prognostic model was constructed in the following way: Riskscore = (- 0.1918) * ERLIN2 + (0.5254) * CDK5RAP3. Subsequently, prognostic subgroups based on key genes classified the high-risk group as a pro-cancer subgroup that had lower mutation rates of critical genes (SPOP and MUC16), multiple low-expression immune-relevant molecules, and differences in macrophages (M1 and M2) expressions. Finally, ERLIN2 as an anti-oncogene and CDK5RAP3 as a pro-oncogene were further confirmed by cell phenotype assays and immunohistochemistry. CONCLUSION: We identified ERLIN2 and CDK5RAP3 as ERS-related genes with important prognostic and immunologic values, and classified patients between high- and low-risk subgroups, which provided new prognostic markers, immunotherapeutic targets, and basis for prognostic assessments.

Case Report: Detection of Treponema phagedenis in cerebrospinal fluid of a neurosyphilis patient by metagenomic next-generation sequencing.

Treponema phagedenis, a human commensal spirochete, has been reported world-wide as a key factor in the pathogenesis of bovine digital dermatitis. Here we report a case of T. phagedenis sequence detection in the cerebrospinal fluid (CSF) of a patient. The patient was diagnosed with neurosyphilis, and T. phagedenis was detected as the only microorganism in his CSF by metagenomic sequencing. The patient went through a round of penicillin therapy previously (2.4 million units of Benzathine Penicillin intramuscularly once a week for three weeks) that did not resolve the symptoms; after the diagnosis of neurosyphilis he was treated with Penicillin G Sodium 4.0 million units q4h intravenous for 14 days then his symptoms resolved. To the best of our knowledge, T. phagedenis has never been reported to be detected in a human's CSF before. This was also the first time it was detected by metagenomic next-generation sequencing. We propose that more etiological tests should be performed including culture and sequencing for more patients with syphilis, which will contribute to a deeper understanding of the pathogenicity of the spirochete.

Immune-pyroptosis-related genes predict the prognosis of kidney renal clear cell carcinoma.

BACKGROUND: Kidney renal clear cell carcinoma (KIRC) is a common cancer of the adult urological system. Recent developments in tumor immunology and pyroptosis biology have provided new directions for kidney cancer treatment. Therefore, there is an urgent need to identify potential targets and prognostic biomarkers for the combination of immunotherapy and pyroptosis-targeted therapy. METHODS: The expression of immune-pyroptosis-related differentially expressed genes (IPR-DEGs) between KIRC and healthy tissues was examined using the Gene Expression Omnibus datasets. The GSE168845 dataset was selected for subsequent analyses. Data of 1793 human immune-related genes were downloaded from the ImmPort database (https://www.immport.org./home), while those of 33 pyroptosis-related genes were extracted from previous reviews. The independent prognostic value of IPR-DEGs was determined using differential expression, prognostic, and univariate and multivariate Cox regression analyses. The GSE53757 dataset was used to further verify the GSDMB and PYCARD levels. In our cohorts, the association among DEGs and clinicopathological features and overall survival was analyzed. The least absolute shrinkage and selection operator Cox regression model was established to evaluate the correlation of IPR-DEGs with the immune score, immune checkpoint gene expression, and one-class logistic regression (OCLR) score. KIRC cells and clinical tissue samples were subjected to quantitative real-time polymerase chain reaction to examine the GSDMB and PYCARD mRNA levels. The GSDMB and PYCARD levels in a healthy kidney cell line (HK-2 cells) and two KIRC cell lines (786-O and Caki-1 cells) were verified. The tissue levels of GSDMB and PYCARD were evaluated using immunohistochemical analysis. GSDMB and PYCARD were knocked down in 786-O cells using short-interfering RNA. Cell proliferation was examined using the cell counting kit-8 assay. Cell migration was measured by transwell migration assays RESULTS: GSDMB and PYCARD were determined to be IPR-DEGs with independent prognostic values. A risk prognostic model based on GSDMB and PYCARD was successfully established. In the GSE53757 dataset, the GSDMB and PYCARD levels in KIRC tissues were significantly higher than those in healthy tissues. The GSDMB and PYCARD expression was related to T stage and OS in our cohort. The GSDMB and PYCARD levels were significantly correlated with the immune score, immune checkpoint gene expression, and OCLR score. The results of experimental studies were consistent with those of bioinformatics analysis. The GSDMB and PYCARD levels in KIRC cells were significantly upregulated when compared with those in healthy kidney cells. Consistently, GSDMB and PYCARD in KIRC tissues were significantly upregulated when compared with those in adjacent healthy kidney tissues. GSDMB and PYCARD knockdown significantly decreased 786-O cell proliferation (p < 0.05). Transwell migration result reflects that silencing GSDMB and PYCARD inhibited 786-O cell migration (p < 0.05) . CONCLUSIONS: GSDMB and PYCARD are potential targets and effective prognostic biomarkers for the combination of immunotherapy and pyroptosis-targeted therapy in KIRC.

In situ tumor-triggered subcellular precise delivery of multi-drugs for enhanced chemo-photothermal-starvation combination antitumor therapy.

Rationale: Drug combination therapy for cancer treatment exerts a more potent antitumor effect. The targeted delivery and release of multiple drugs in a patient's body thus presents a more effective treatment approach, warranting further research. Methods: Two antitumor drugs (ICG: indocyanine green and THP: pirarubicin) were successfully screened to sequentially trigger self-assembling peptides (P60) to produce bacteria-sized particles (500-1000 nm, P60-ICG-THP). First, after mixing equal amount of P60 and ICG, trace amount of water (the mass ratio between P60 and water: 100:1) was used to trigger their assembly into P60-ICG. Subsequently, the assembly of P60-ICG and THP was further triggered by ultrasound treatment to produce P60-ICG-THP. Results: P60-ICG-THP constituted a cluster of several nanoparticles (50-100 nm) and possessed a negative charge. Owing to its size and charge characteristics, P60-ICG-THP could remain outside the cell membrane, avoiding the phagocytic clearance of blood and normal tissue cells in vivo. However, after localizing in the tumor, the size and charge switches of P60-ICG-THP, rapidly triggered by the low pH of the tumor microenvironment, caused P60-ICG-THP to segregate into two parts: (i) positively charged nanoparticles with a size of approximately 50 nm, and (ii) negatively charged particles of an uneven size. The former, mainly carrying THP (chemotherapeutic agent), could immediately cross the cell membrane and deliver pirarubicin into the nucleus of tumor cells. The latter, carrying ICG (used for photothermal therapy), could also enter the cell via the endocytosis pathway or accumulate in tumor blood vessels to selectively block the supply of nutrients and oxygen (cancer starvation). Both these particles could avoid the rapid excretion of ICG in the liver and were conducive to accumulation in the tumor tissue for photothermal therapy. Conclusion: Our drug delivery system not only achieves the precise subcellular delivery of two anticancer drugs due to their size and charge switches in the tumor site, but also provides a new strategy to combine chemotherapy, photothermal therapy, and cancer starvation therapy for the development of a highly efficient antitumor therapeutic regimen.

Self-assembled peptide nanoparticles responsive to multiple tumor microenvironment triggers provide highly efficient targeted delivery and release of antitumor drug.

Stimuli-responsive drug delivery systems based on tumor microenvironment conditions show tremendous promise to enhance tumor-targeted delivery and drug release. Herein, a multifunctional peptide (P51) was developed for programmed delivery of the hydrophobic chemotherapeutic agent pirarubicin. P51 was prepared with a ligand-specific targeting for the cancer biomarker Arg-Gly-Asp (RGD), and three tumor microenvironment-sensitive release triggers, acid environment, reducing agent, and a specific enzyme. The peptides Cys-s-s-Cys (disulfide linkage) and Pro-Val-Gly-Leu-Ile-Gly correspond to the cleavage sites of a reducing agent (DTT) and an enzyme (MMP-2). The peptides act as a junction between Ser-Glu-Glu-Asp-Pro (a negatively charged sequence) and a 41-residue peptide containing an α-helix that has the capacity to encapsulate pirarubicin via electrostatic and hydrophobic interactions. These interactions can be disrupted by the acidic tumor microenvironment. Self-assembly of P51 and pirarubicin (P51-THP NPs) results into stable spherical nanoparticles in a single step. We have demonstrated that the acid environment, DTT, and MMP-2 stimulate the release of pirarubicin from P51-THP NPs and, more importantly, the efficiency of drug release is markedly increased when all three release triggers are present. In addition, more effective tumor targeting, antitumor effect, and reduced systemic toxicity of P51-THP NPs have been confirmed by in vitro and in vivo results.

Gold Biomineralization on Bacterial Biofilms for Leaching of Au3+ Damages Eukaryotic Cells.

Heavy metals not only pollute the environment but also are health and environmental hazard. Bacteria constitute inexpensive and eco-friendly material to eliminate and recycle heavy metals via biomineralization and biosorption. However, the effect of metal biomineralization in bacterial biofilms on the ecological balance of bacteria and infectious diseases is unclear. This study aimed to explore the interaction between a eukaryotic cell line HEK293T and mineralized Escherichia coli, using a model of gold biomineralization on E. coli biofilms (E. coli-Au). In our present model, bacterial activity was not disrupted and bacterial adhesion and invasion were enhanced. E. coli-Au invaded the cytoplasm and nuclei of HEK293T cells and damaged them via intracellular growth and multiplication. The present findings indicate that metal biomineralization in bacterial biofilms for leaching of heavy metal ions is hazardous to eukaryotic cells and even human health.

The relationship between the percent of euploid embryo and the tolerance of embryo biopsy in preimplantation genetic screening: A systematic review and meta-analysis of randomized controlled trials.

The aim of this study was to analyze the relationship between the percent of euploid embryo and the tolerance of embryo biopsy in preimplantation genetic screening (PGS).PubMed and trial registers were searched for clinical studies that patients were randomized to the PGS group or the control group from 1995 to October 2017. The patients of advanced maternal age, repeated implantation failure, and good prognosis with or without PGS in randomized controlled trials (RCTs) were collected.Original data from 9 RCT studies comparing in-vitro fertilization with and without PGS including 1642 patients were obtained and they were divided into 3 subgroups according to the percent of euploid embryo. PGS significantly increased live birth babies per embryo transferred (risk ratio: 2.98, 95% confidence interval: 1.54-5.75) in ≤30% of euploid embryo subgroups and but in other 2 groups, PGS has no effect. Significant negative correlation was found between the percent of euploid embryo and the tolerance of embryo biopsy in PGS (r = 0.80, P = 0.010)The tolerance of embryo biopsy in PGS was associated negatively with the percent of euploid embryo. There was a beneficial effect when PGS was used in the patients with the lowest percent of euploid embryo.

A facile strategy to fabricate a pH-responsive mesoporous silica nanoparticle end-capped with amphiphilic peptides by self-assembly.

A facile strategy with no modification processes was demonstrated to fabricate a pH-responsive end-capped mesoporous silica nanoparticle (MSN)-based drug delivery system (DDS). The simple but smart nanovalve systems were constructed by the self-assembly behavior of unbonded peptide-based amphiphile (P45) in the presence of Doxorubicin hydrochloride (Dox). A series of characterizations confirmed that the DDSs had been successfully fabricated. Dox molecules were entrapped by nanovalves in the pores of MSNs, and an in vitro release experiment demonstrated that the P45/Dox@MSNs exhibited "zero premature release" in the physical environment. However, an accelerated release was triggered by an acidic atmosphere in cellular cytosol. Moreover, detailed investigations confirmed that the enhanced cellular uptake of P45/Dox@MSNs due to the RGD motif of the nanovalves, exhibiting an obvious toxicity to cancer cells. Therefore, the DDSs constructed here can serve as a promising platform to realize targeted drug delivery and controlled drug release by using diversified bioactive native amphiphilic peptide.

A DNA-modified hydrogel for simultaneous purification, concentration and detection of targeted cfDNA in human serum.

Cell-free DNA (cfDNA) in blood, which stems from the fetus of pregnant women and tumor in cancer patients, has gained attention in molecular diagnosis. However, cfDNA is less stable, and its amount in the serum is extremely low; these are critical barriers for the utilization of this resource. In this study, a DNA-modified polyacrylamide hydrogel (DNA-Gel) was prepared, and a specialized device was designed to simultaneously catch, purify, concentrate, and detect targeted cfDNA by electrophoresis. We demonstrated that 20-1000 bp ssDNA and dsDNA could be caught and released by the DNA-Gel-based device with high specificity and sensitivity. Upon increasing the number of cycles and electrophoresis time, higher DNA purity and density were achieved, and the separation of serum proteins, untargeted cfDNA, and other charged molecules was promoted. As low as 10 pg µL-1 of DNA could be detected using the DNA-Gel after four cycles of concentration. We also detected 1 fg µL-1 of DNA in the serum with 16 cycles of concentration, followed by 25 PCR cycles. We also designed a device to obtain DNA from the DNA-Gel. We found that the DNA loss rate was around 50%, and A260/A280 was close to 1.7. Thus, we have designed a cost-effective and highly economical device to purify DNA at low concentrations with high specificity and selectivity.

[The rs4705342 gene mutation in the promoter region of the miR-143/ 145 cluster associated with the risk of prostate cancer in the Chinese Han population].

OBJECTIVE: To explore the correlation between the rs4705342 gene mutation in the promoter region of the miR-143/ miR-145 cluster and the risk of PCa in the Chinese Han population. METHODS: We enrolled in this study 156 cases of PCa and 188 cancer-free controls. Using TaqMan SNP genotyping assay, we detected the polymorphism of rs4705342 in the promoter region of the miR-143/ miR-145 gene, and performed a stratified analysis on the family history of cancer, body mass index (BMI), age, smoking status, and alcohol consumption of the subjects. RESULTS: The proportion of family history of cancer was significantly higher in the case group than in the control (P = 0.01). No statistically significant differences, however, were found between the two groups in age (P = 0.32), BMI (P = 0.79), smoking status (P = 0.47), and alcohol consumption (P = 0.34), nor in the distribution of the TT, CT and CC genotypes (P = 0.07) except in that of the CC and TT/CT combined genotypes (P = 0.01). There were statistically significant differences in the distribution of CC and TT/CT combined genotypes between the non-smoking subgroups, (P = 0.02) and alcohol drinking subgroups (P = 0.03), but not between the clinical stages of PCa (P = 0.81) or the levels of PSA (P = 0.39). CONCLUSIONS: The rs4705342 gene mutation in the promoter region of the miR-143/ miR-145 cluster is significantly associated with the pathogenesis of PCa in the Chinese Han population.

Virtual Network Embedding Based on Graph Entropy.

For embedding virtual networks into a large scale substrate network, a massive amount of time is needed to search the resource space even if the scale of the virtual network is small. The complexity of searching the candidate resource will be reduced if candidates in substrate network can be located in a group of particularly matched areas, in which the resource distribution and communication structure of the substrate network exhibit a maximal similarity with the objective virtual network. This work proposes to discover the optimally suitable resource in a substrate network corresponding to the objective virtual network through comparison of their graph entropies. Aiming for this, the substrate network is divided into substructures referring to the importance of nodes in it, and the entropies of these substructures are calculated. The virtual network will be embedded preferentially into the substructure with the closest entropy if the substrate resource satisfies the demand of the virtual network. The experimental results validate that the efficiency of virtual network embedding can be improved through our proposal. Simultaneously, the quality of embedding has been guaranteed without significant degradation.