Identification of SOCS3 and PTGS2 as new biomarkers for the diagnosis of gout by cross-species comprehensive analysis.

Background: Gout is the most common inflammatory arthritis in adults. Gout is an arthritic disease caused by the deposition of monosodium urate crystal (MSU) in the joints, which can lead to acute inflammation and damage adjacent tissue. Hyperuricemia is the main risk factor for MSU crystal deposition and gout. With the increasing burden of gout disease, the identification of potential biomarkers and novel targets for diagnosis is urgently needed. Methods: For the analysis of this subject paper, we downloaded the human gout data set GSE160170 and the gout mouse model data set GSE190138 from the GEO database. To obtain the differentially expressed genes (DEGs), we intersected the two data sets. Using the cytohubba algorithm, we identified the key genes and enriched them through GO and KEGG. The gene expression trends of three subgroups (normal control group, intermittent gout group and acute gout attack group) were analyzed by Series Test of Cluster (STC) analysis, and the key genes were screened out, and the diagnostic effect was verified by ROC curve. The expression of key genes in dorsal root nerve and spinal cord of gout mice was analyzed. Finally, the clinical samples of normal control group, hyperuricemia group, intermittent gout group and acute gout attack group were collected, and the expression of key genes at protein level was verified by ELISA. Result: We obtained 59 co-upregulated and 28 co-downregulated genes by comparing the DEGs between gout mouse model data set and human gout data set. 7 hub DEGs(IL1B, IL10, NLRP3, SOCS3, PTGS2) were screened out via Cytohubba algorithm. The results of both GO and KEGG enrichment analyses indicate that 7 hub genes play a significant role in regulating the inflammatory response, cytokine production in immune response, and the TNF signaling pathway. The most representative hub genes SOCS3 and PTGS2 were screened out by Series Test of Cluster, and ROC analysis results showed the AUC values were both up to 1.000. In addition, we found that PTGS2 expression was significantly elevated in the dorsal root ganglia and spinal cord in monosodium urate(MSU)-induced gout mouse model. The ELISA results revealed that the expression of SOCS3 and PTGS2 was notably higher in the acute gout attack and intermittent gout groups compared to the normal control group. This difference was statistically significant, indicating a clear distinction between the groups. Conclusion: Through cross-species comprehensive analysis and experimental verification, SOCS3 and PTGS2 were proved to be new biomarkers for diagnosing gout and predicting disease progression.

Effect of Precursors and Their Regulators on the Biosynthesis of Antibiotics in Actinomycetes.

During the life activities of microorganisms, a variety of secondary metabolites are produced, including antimicrobials and antitumor drugs, which are widely used in clinical practice. In addition to exploring new antibiotics, this makes it one of the research priorities of Actinomycetes to effectively increase the yield of antibiotics in production strains by various means. Most antibiotic-producing strains have a variety of functional regulatory factors that regulate their growth, development, and secondary metabolite biosynthesis processes. Through the study of precursor substances in antibiotic biosynthesis, researchers have revealed the precursor biosynthesis process and the mechanism by which precursor synthesis regulators affect the biosynthesis of secondary metabolites, which can be used to obtain engineered strains with high antibiotic production. This paper summarizes the supply of antibiotic biosynthesis precursors and the progress of research on the role of regulators in the process of precursors in biosynthesis. This lays the foundation for the establishment of effective breeding methods to improve antibiotic yields through the manipulation of precursor synthesis genes and related regulators.

Cr(VI)-bioremediation mechanism of a novel strain Bacillus paramycoides Cr6 with the powerful ability to remove Cr(VI) from contaminated water.

This research provided an excellent novel hexavalent chromium (Cr(VI))-removal bacterium, Bacillus paramycoides Cr6, and investigated its removal mechanism from the perspective of molecular biology. Cr6 could resist up to 2500 mg/L Cr(VI), and the removal rate of 2000 mg/L Cr(VI) reached 67.3% under the optimal culture conditions of 220 r/min, pH 8 and 31 â„ƒ. When the initial concentration of Cr(VI) was 200 mg/L, Cr6 had a removal rate of 100% within 18 h. The differential transcriptome analysis identified two key structural genes named bcr005 and bcb765 of Cr6, which were upregulated by Cr(VI). Their functions were predicted and further confirmed by bioinformatic analyses and in vitro experiments. bcr005 encodes Cr(VI)-reductase BCR005, and bcb765 encodes Cr(VI)-binding protein BCB765. Real-time fluorescent quantitative PCRs were performed, and the data illustrated a parallel pathway (one is Cr(VI) reduction, and the other is Cr(VI) immobilisation) of Cr6 to remove Cr(VI), which relies on the synergistic expression of the genes bcr005 and bcb765 induced by different concentrations of Cr(VI). In summary, a deeper molecular mechanism of Cr(VI) microorganism removal was elaborated; Bacillus paramycoides Cr6 was an exceptional novel Cr(VI)-removed bacterial resource, while BCR005 and BCB765 were two new-found efficient enzymes that have potential practical applications for sustainable microbial remediation of Cr-contaminated water.

Radiation prevents tumor progression by inhibiting the miR­93­5p/EphA4/NF­κB pathway in triple­negative breast cancer.

Breast cancer (BC) is the most common type of cancer in women. Triple­negative BC (TNBC) constitutes 10­15% of all BC cases and is associated with a poor prognosis. It has previously been reported that microRNA (miR)­93­5p is dysregulated in plasma exosomes from patients with BC and that miR­93­5p improves radiosensitivity in BC cells. The present study identified EphA4 as a potential target gene of miR­93­5p and investigated the pathway related to miR­93­5p in TNBC. Cell transfection and nude mouse experiments were performed to verify the role of the miR­93­5p/EphA4/NF­κB pathway. Moreover, miR­93­5p, EphA4 and NF­κB were detected in clinical patients. The results revealed that EphA4 and NF­κB were downregulated in the miR­93­5p overexpression group. By contrast, EphA4 and NF­κB expression levels were not significantly altered in the miR­93­5p overexpression + radiation group compared with those in the radiation group. Furthermore, overexpression of miR­93­5p with concomitant radiation therapy significantly decreased the growth of TNBC tumors in vivo. In conclusion, the present study revealed that miR­93­5p targeted EphA4 in TNBC through the NF­κB pathway. However, radiation therapy prevented tumor progression by inhibiting the miR­93­5p/EphA4/NF­κB pathway. Therefore, it would be interesting to elucidate the role of miR­93­5p in clinical research.

Mechanism and Role of Endoplasmic Reticulum Stress in Osteosarcoma.

Osteosarcoma is the most common malignant bone tumor, often occurring in children and adolescents. The etiology of most patients is unclear, and the current conventional treatment methods are chemotherapy, radiotherapy, and surgical resection. However, the sensitivity of osteosarcoma to radiotherapy and chemotherapy is low, and the prognosis is poor. The development of new and useful treatment strategies for improving patient survival is an urgent need. It has been found that endoplasmic reticulum (ER) stress (ERS) affects tumor angiogenesis, invasion, etc. By summarizing the literature related to osteosarcoma and ERS, we found that the unfolded protein response (UPR) pathway activated by ERS has a regulatory role in osteosarcoma proliferation, apoptosis, and chemoresistance. In osteosarcoma, the UPR pathway plays an important role by crosstalk with autophagy, oxidative stress, and other pathways. Overall, this article focuses on the relationship between ERS and osteosarcoma and reviews the potential of drugs or gene targets associated with ERS for the treatment of osteosarcoma.

[Application of MRI Water-Fat Separation Technology in Patients with Multiple Myeloma].

OBJECTIVE: To explore the clinical significance of magnetic resonance imaging water-fat separation (Dixon) technique in patients with multiple myeloma. METHODS: A total of 41 newly diagnosed patients with multiple myeloma who underwent Dixon in The Affiliated Hospital of Qingdao University from April 2019 to April 2021 were included in this study. Patients were divided into observation group and control group according to whether Dixon performance was normal or not. The differences of clinical data and fat fraction (FF) between the two groups were compared. The correlation between FF and clinical data, disease stages and differences before and after treatment were also compared. The receiver operator characteristic curve of patients was drawn to analyze the diagnostic value of FF combined with serum alkaline phosphatase for bone destruction in patients with multiple myeloma. RESULTS: Among the 41 patients, there were 12 cases in the control group and 29 cases in the observation group. There was no significant difference in age and sex between the two groups. In the observation group, ß2-microglobulin concentration and M protein were significantly higher than those in the control group, while serum alkaline phosphatase and FF were lower (P<0.05). In all 41 patients included in the study, there was a significant negative correlation between FF value and ß2-microglobulin concentration (r=-0.57), and a significant positive correlation between FF value and serum alkaline phosphatase (r=0.31). After treatment, FF value increased, while myeloma cell percentage, ß2-microglobulin concentration and M protein decreased in 11 patients who completed 4 cycles of chemotherapy, and the differences before and after treatment were statistically significant (P<0.05). The value of serum alkaline phosphatase combined with FF value in predicting bone destruction is higher than that of FF value or serum alkaline phosphatase alone. CONCLUSION: Dixon's different imaging manifestations can reflect the severity of the disease. FF value is correlated with clinical examination results and R-ISS staging, and there is a significant difference before and after treatment. Serum alkaline phosphatase combined with FF value is better than two indicators alone in predicting bone destruction.

Investigation of miR-93-5p and its effect on the radiosensitivity of breast cancer.

Accumulating evidence suggests that intrinsic resistance to radiotherapy reduces the survival of patients with cancer. The present study investigated whether miR-93-5p affects proliferation, migration, apoptosis, and radiosensitivity of breast cancer (BC) cells. MDA-MB-468, MCF-7, and MDA-MB-231 BC cells were incubated with hsa-miR-93-5p mimics, hsa-miR-93-5p inhibitor, and negative control RNA with or without exposure to ionizing radiation to determine cell proliferation, migration, and apoptosis using the Cell Counting Kit-8 assay, wound healing assay and apoptotic assay, respectively. Overexpression of miR-93-5p inhibited the migratory abilities (P = 0.001) and decreased the cell proliferation (P = 0.049) of MCF-7 cells. In MCF-7 cells, a significant increase in apoptosis was detected after treatment with miR-93-5p compared with the negative control (P = 0.001) and miR-93-5p inhibitor (P = 0.004). In MDA-MB-468 cells, the proportion of apoptotic cells increased following exposure to ionizing radiation (P = 0.001). The percentage of apoptotic MDA-MB-231 cells in the miR-93-5p group was significantly increase compared with that determined in the negative control (P = 0.044) and hsa-miR-93-5p inhibitor (P = 0.046) groups. In conclusion, our findings showed that miR-93-5p reduces BC cell proliferation and migratory capacity, and increases the ratio of apoptotic cells. Overexpression of miR-93-5p could increase radiosensitivity in BC cells by increasing apoptosis. This evidence provides new insight into the treatment of BC and identifies miR-93-5p as a potential therapeutic target.

Pretreatment Albumin-to-Fibrinogen Ratio Independently Predicts Chemotherapy Response and Prognosis in Patients with Locally Advanced Rectal Cancer Undergoing Total Mesorectal Excision After Neoadjuvant Chemoradiotherapy.

BACKGROUND: Neoadjuvant chemoradiotherapy (nCRT) followed by surgery of total mesorectal excision (TME) is currently accepted as the standard treatment for locally advanced rectal cancer (LARC). This study aimed to investigate the potential prognostic factors, including the albumin-to-fibrinogen ratio (AFR) for LARC patients. METHODS: We retrospectively recruited LARC patients (cT3-4 and/or cN1-2) who underwent nCRT followed by TME between January 2011 and January 2015. The cut-off value of pretreatment AFR for overall survival (OS) was determined by the receiver operating characteristic (ROC) curve. The potential predictive factors for prognosis in the LARC patients were assessed by the univariate and multivariate Cox's proportional hazard regression and Kaplan-Meier curve analyses. RESULTS: AFR was a significant predictor for OS with a cut-off value of 8.65 and an AUC of 0.882 (P<0.001). The pretreatment AFR level was the only independent risk factor for pathologic response to nCRT (HR: 2.44, 95% CI: 1.43-4.17, P=0.003), 5-year OS (HR: 3.31, 95% CI: 1.51-6.77, P=0.005) and disease-free survival (DFS) (HR: 2.73, 95% CI: 1.34-5.47, P=0.007) in LARC patients. A low pretreatment AFR level was significantly associated with a poor 5-year OS and DFS by the Log rank test (P=0.003 and 0.006, respectively). CONCLUSION: Pretreatment AFR level was an independent prognostic factor in LARC patients undergoing TME after nCRT.

3-Nitroacridine derivatives arrest cell cycle at G0/G1 phase and induce apoptosis in human breast cancer cells may act as DNA-target anticancer agents.

DNA is considered to be one of the most promising targets for anticancer agents. Acridine analogues have anticancer activity based on DNA binding and topoisomerases inhibition. However, due to the side effects, resistance and low bioavailability, a few have entered into clinical usage and the mechanisms of action are not fully understood. Novel acridine derivatives are needed for effective cancer therapy. A series of novel 3-nitroacridine-based derivatives were synthesized, their DNA binding and anticancer activities were evaluated. The chemical modifications at position 9 of the 3-nitroacridine were crucial for DNA affinity, thus optimizing anticancer activity. UV-Vis and circular dichroism (CD) spectroscopy indicated interaction of compounds with DNA, and the binding modes were intercalation and groove binding. MTT assay and clonogenic assay showed that compounds 1, 2 and 3 had obvious cell growth inhibition effect. They induced cell apoptosis in human breast cancer cells in a dose-dependent manner, and exhibited anticancer effect via DNA damage as well as cell cycle arrest at G0/G1 phage. Using confocal fluorescent microscope, the apoptotic features were observed. The results suggested that compounds 1-3 with high DNA binding affinity and good inhibitory effect of cancer cell proliferation can be developed as prime candidates for further chemical optimization.

Multi-Repeated Projection Lithography for High-Precision Linear Scale Based on Average Homogenization Effect.

A multi-repeated photolithography method for manufacturing an incremental linear scale using projection lithography is presented. The method is based on the average homogenization effect that periodically superposes the light intensity of different locations of pitches in the mask to make a consistent energy distribution at a specific wavelength, from which the accuracy of a linear scale can be improved precisely using the average pitch with different step distances. The method's theoretical error is within 0.01 µm for a periodic mask with a 2-µm sine-wave error. The intensity error models in the focal plane include the rectangular grating error on the mask, static positioning error, and lithography lens focal plane alignment error, which affect pitch uniformity less than in the common linear scale projection lithography splicing process. It was analyzed and confirmed that increasing the repeat exposure number of a single stripe could improve accuracy, as could adjusting the exposure spacing to achieve a set proportion of black and white stripes. According to the experimental results, the effectiveness of the multi-repeated photolithography method is confirmed to easily realize a pitch accuracy of 43 nm in any 10 locations of 1 m, and the whole length accuracy of the linear scale is less than 1 µm/m.

Biosynthesis of 3″-demethyl-gentamicin C components by genN disruption strain of Micromonospora echinospora and test their antimicrobial activities in vitro.

Gentamicin consists primarily of four components, which have different patterns of methylation at C-6' position. The methyl groups have a significant impact on gentamicin antimicrobial activity. Sequence analysis predicted that GenN was a methyltransferase in the gentamicin biosynthetic pathway. To study the function of genN, it was disrupted in Micromonospora echinospora. The genN disruption strains produced 3″-N-demethyl-gentamicin C complex instead of the gentamicin C complex. In this study, 3″-N-demethyl gentamicin C1a was purified from the broth of disruption strain, and its structure was elucidated using MS and NMR. Besides 3″-N-demethyl products corresponding to gentamicin C1a, C2, and C2a, two 3″-N-demethyl products corresponding to gentamicin C1 were detected, which were concluded as C-6' epimers originating from decreased substrate specificity of 6'-N methyltransferase. To explore the effects of 3″-N-methyl on gentamicin antimicrobial activity, antimicrobial activity of these demethyl gentamicin analogues were tested in vitro. 3″-N-Demethyl gentamicin components have identical activity with corresponding components of gentamicin. The results of bioassays showed that the 3″-N-methyl group has little impact on gentamicin activity. However, these highly bioactive compounds afforded a unique opportunity for creating new and high potent aminoglycoside antibiotics.

Assembly of a novel biosynthetic pathway for gentamicin B production in Micromonospora echinospora.

BACKGROUND: Isepamicin is a weakly toxic but highly active aminoglycoside antibiotic derivative of gentamicin B. Gentamicin B is a naturally occurring minor component isolated from Micromonospora echinospora. 2'-NH2-containing gentamicin C complex is a dominant compound produced by wild-type M. echinospora; by contrast, 2'-OH-containing gentamicin B is produced as a minor component. However, the biosynthetic pathway of gentamicin B remains unclear. Considering that gentamicin B shares a unique C2' hydroxyl group with kanamycin A, we aimed to design a new biosynthetic pathway of gentamicin B by combining twelve steps of gentamicin biosynthesis and two steps of kanamycin biosynthesis. RESULTS: We blocked the biosynthetic pathway of byproducts and generated a strain overproducing JI-20A, which was used as a precursor of gentamicin B biosynthesis, by disrupting genK and genP. The amount of JI-20A produced in M. echinospora ∆K∆P reached 911 µg/ml, which was 14-fold higher than that of M. echinospora ∆P. The enzymes KanJ and KanK necessary to convert 2'-NH2 into 2'-OH from the kanamycin biosynthetic pathway were heterologously expressed in M. echinospora ΔKΔP to transform JI-20A into gentamicin B. The strain with kanJK under PermE* could produce 80 µg/ml of gentamicin B, which was tenfold higher than that of the wild-type strain. To enhance gentamicin B production, we employed different promoters and gene integration combinations. When a PhrdB promoter was used and kanJ and kanK were integrated in the genome through gene replacement, gentamicin B was generated as the major product with a maximum yield of 880 µg/ml. CONCLUSION: We constructed a new biosynthetic pathway of high-level gentamicin B production; in this pathway, most byproducts were removed. This method also provided novel insights into the biosynthesis of secondary metabolites via the combinatorial biosynthesis.

Biosynthesis of Epimers C2 and C2a in the Gentamicin C Complex.

Gentamicin is a broad-spectrum aminoglycoside antibiotic widely used to treat life-threatening bacterial infections. The gentamicin C complex consists of gentamicin C1, gentamicin C1a, and epimers gentamicin C2 and gentamicin C2a. At present there is a generally accepted pathway of gentamicin biosynthesis, except for detailed understanding of the epimerization process involving gentamicins C2 and C2a. Here we have investigated the biosynthesis of these epimers. JI-20B-an intermediate in the gentamicin biosynthetic pathway-and its epimer JI-20Ba were generated by in-frame deletion within genP, which encodes a phosphotransferase that catalyzes the first step of 3',4'-bisdehydroxylation in gentamicin biosynthesis. GenB1 and GenB2 are aminotransferases with different substrate specificities and enantioselectivities. JI-20Ba, containing a 6'S chiral amine, a precursor of gentamicin C2a, was synthesized from G418 by GenQ/GenB1 through sequential oxidation/transamination at C-6'. GenQ/GenB2 catalyzed the synthesis of JI-20B, containing a 6'R chiral amine, a precursor of gentamicin C2, from G418. GenB2 catalyzed the epimerization of JI-20Ba/JI-20B and of gentamicins C2a/C2.