Biotechnology & Genetic Engineering Reviews最新文献

We investigate the impact of bovine pulmonary surfactant (PS) on LPS-induced ALI in vitro and in vivo to improve recognition and prevent mortality in sepsis-induced ALI. Primary alveolar type II (AT2) cells were treated with LPS alone or in combination with PS. Cell morphology observation, CCK-8 proliferation assay, flow cytometry apoptosis assay, and ELISA for inflammatory cytokine levels were performed at different time points after treatment. An LPS-induced ALI rat model was established and treated with vehicle or PS. Lung wet/dry weight ratio, histopathological changes, lung function parameters, and serum inflammatory cytokine levels were examined 6 h after PS treatment. Survival analysis by Kaplan-Meier method. RNA sequencing was conducted to identify LPS-induced differentially expressed genes in rat lungs. Proapoptotic gene expression in rat lungs was determined by Western blot. LPS significantly inhibited cell proliferation while promoting apoptosis of AT2 cells starting 2 h after treatment, accompanied by a significant increase in inflammatory cytokine production; PS reversed these effects. PS decreased the lung wet/dry ratio in septic rats, histological abnormalities, alterations in lung function parameters, and inflammatory cytokines production; while improving the overall survival of rats. LPS-induced differentially expressed genes were closely associated with apoptosis. PS attenuated LPS-induced upregulation of proapoptotic gene expression starting 2 h after treatment in AT2 cells while restoring lung ATPase activity in vivo. Bovine PS alleviates LPS-induced ALI in the early phase, possibly by suppressing inflammation and AT2 cell apoptosis, as a preemptive therapeutic agent for managing sepsis-induced ALI.

The aim of this study was to investigate the causal relationship between autoimmune disorders and celiac disease (CeD) through Mendelian randomization (MR). Single nucleotide polymorphisms (SNPs) significantly associated with 13 autoimmune diseases were extracted from the summary statistics of European genome-wide association studies (GWAS), and their effects were examined by Inverse variance-weighted (IVW) in a large European GWAS on CeD. Finally, reverse MR was performed to investigate the causal effects of CeD on autoimmune traits. Following the application of Bonferroni correction for multiple testing, genetically determined seven autoimmune diseases are causally associated with CeD: Crohn's disease (CD) (OR [95%CI] = 1.156 [1.106 ± 1.208], P = 1.27E-10), primary biliary cholangitis (PBC) (1.229 [1.143 ± 1.321], P = 2.53E-08), primary sclerosing cholangitis (PSC) (1.688 [1.466 ± 1.944], P = 3.56E-13), rheumatoid arthritis (RA) (1.231 [1.154 ± 1.313], P = 2.74E-10), systemic lupus erythematosus (SLE) (1.127 [1.081 ± 1.176], P = 2.59E-08), type 1 diabetes (T1D) (1.41 [1.238 ± 1.606], P = 2.24E-07), and asthma (1.414 [1.137 ± 1.758], P = 1.86E-03). The IVW analysis indicated that CeD increased the risk for seven diseases: CD (1.078 [1.044 ± 1.113], P = 3.71E-06), Graves' disease (GD) (1.251 [1.127 ± 1.387], P = 2.34E-05), PSC (1.304 [1.227 ± 1.386], P = 8.56E-18), psoriasis (PsO) (1.12 [1.062 ± 1.182], P = 3.38E-05), SLE (1.301[1.22 ± 1.388], P = 1.25E-15), T1D (1.3[1.228 ± 1.376], P = 1.57E-19), and asthma (1.045 [1.024 ± 1.067], P = 1.82E-05). The sensitivity analyses deemed the results reliable without pleiotropy. There are positive genetic correlations between various autoimmune diseases and CeD, and the latter also affects the predisposition to multiple autoimmune disorders in the European population.

Develop the ic-ELISA rapid detection method of Enrofloxacin (ENR). Corresponding antibodies are obtained by animal immunity to identify their titer and specificity. The optimal coating time was obtained by indirect competition ELISA, and the antigen coating time, suitable coating concentration, primary antibody dilution factor, blocking solution blocking time, primary antibody reaction time and secondary antibody reaction time were optimized, and the specificity and accuracy of the method were evaluated. The ic-ELISA rapid detection method of ENR, IC₅₀ was 9.13 ng/mL, and the linear detection range (IC₂₀-IC₈₀) was 4.16-20.03 ng/mL. The LOD limit is 2.11 ng/mL. The cross-reactivity rate of 9 fluoroquinolones was above 10%, and the average recovery rate was above 80%. The reason why the heterologous coating is more sensitive may be due to the fact that the piperazine group of ofloxacin is one less carbon atom than enrofloxacin, and ofloxacin is connected to the main ring by N and O hybridization, while enrofloxacin is connected to the main ring through a ternary ring, these two reasons may cause the charge density of extracyclic oxygen at the ofloxacin binding site to be higher than that of enrofloxacin, and the binding ability to antibodies is stronger. Therefore, when heterologous coating, the competitive inhibition rate against enrofloxacin is higher and the effect is better. The conclusion obtained through this experiment is that the detection method has strong broad spectrum and good sensitivity, and can quickly detect the total amount of enrofloxacin and its seven common fluoroquinolones in fish and eggs.

The leaves of Rumex vescarius L. are used locally to treat diabetes, a chronic illness. A flavonoid called Luteolin from R. vesicarius was chosen to explore for the antidiabetic potential through the in vivo antidiabetic test against male albino Wistar rats that had been induced with diabetes due to alloxan. Additionally, docking screening was carried out with the aid of autodock software to identify probable moiety that might be in charge of its anti-diabetic effect. Given at a dose of 100 mg/kg body weight, luteolin from R. vesicarius leaves had a significant (p < 0.05) hypoglycaemic impact after just one week. The blood glucose level significantly decreased during the third week (p < 0.05). All provided doses of luteolin from R. vesicarius leaves resulted in a reduction, however on all study days, the highest concentration (400 mg/kg body weight) produced the biggest reduction. The results of luteolin's molecular docking and dynamic modelling studies with a variety of targets revealed significant binding interactions at the active site binding pocket, with the target α-glucosidase having the highest binding affinity (-9.35 kcal/mol). In conclusion, the plant and the flavonoid luteolin it contains have potent anti-diabetic properties, possibly through an interaction with the enzyme α-glucosidase.

Pulmonary vascular remodeling (PVR) is the main factor of pulmonary hypertension (PH). The pathological characteristics of PVR are vascular smooth muscle hyperplasia, hypertrophy, and extensive damage. In vivo experiments, the expression of FTO in PH rat lung tissues of different rat models of hypoxia PH was observed by immunohistochemical method. mRNA microarray analysis was used to analyze the differential expressed genes in rat lung tissues. In vitro experiments, we developed models of overexpression and knockdown of FTO to study the effect of FTO protein expression on cell apoptotic, cell cycle, and the abundance of m⁶A. The expression of FTO was increased in PH rats. FTO knockdown can inhibit the proliferation of PASMCs, thereby regulating the cell cycle and reducing the expression of Cyclin D1 and the abundance of m⁶A, while overexpression of FTO leads to increased expression of Cyclin D1 and the abundance of m⁶A. FTO destroys the stability of Cyclin D1 by regulating the abundance of Cyclin D1 m⁶A, causing cell cycle arrest and inducing cell proliferation, thus inducing the occurrence and development of PVR in PH.

Cervical cancer (CC) is a frequent disease in women whose development is related with miRNA disorder. MiR-377-5p plays a negative role in the development of some tumors, while few studies have revealed its role in CC. In this study, the functions of miR-377-5p in CC were investigated by bioinformatics. Briefly, the expression and survival curve of miR-377-5p in CC was analyzed with the Cancer Genome Atlas (TCGA) database, and the abundance of miR-377-5p in clinical samples and CC cell lines were measured by qRT-PCR. Moreover, the MicroRNA Data Integration Portal (miRDIP) database was used to predict targets of miR-377-5p, and the Database for Annotation Visualization and Integrated Discovery (David) was used for enrichment analysis of the functions of the miR-377-5p. The Search Tool for the Retrieval of Interacting Genes (STRING) database was used to screen the hub targets of miR-377-5p. Moreover, the Gene Expression Profiling Interactive Analysis (GEPIA) database was used to analyze the abundance of the genes in CC. Results showed that decreased miR-377-5p was found in the CC tissues and cell lines, and low miR-377-5p was connected with poor prognosis of patients. Besides, the targets of miR-377-5p were enriched in the PI3K/AKT, MAPK and RAS signaling pathways. Moreover, CDC42, FLT1, TPM3 and CAV1 were screened as hub nodes in the targets of miR-377-5p, and increased CDC42, FLT1, TPM3 and CAV1 also indicated the poor survival rates of the patients in the long term. In conclusion, this study suggests that miR-377-5p downregulation is a biomarker event for CC progression.

This work sought to determine how lipopolysaccharide (LPS)-induced pro-inflammatory factor production in BV2 microglia was influenced by myeloid cell 2 (TREM2) expressions. LPS (0.1, 1, and 10 µg/mL) induced inflammation in BV2 cells, MTT and QPCR were used to detect the occurrence of inflammation; TREM2 activation and inhibition vectors were used to activate and inhibit TREM2; Cell Proliferation was detected using CCK-8 and cell cloning experiments. LY294002 was used to inhibit the activity of PI3K/AKT signal pathway; Western blot and ELISA were used to detect cell polarization and signal pathway changes. CCK-8 and cell clone experiments found that the activation of TERM2 can promote the proliferation of BV2 cells; and the activation of TERM2 can promote the expression of IL6, IL1β, TNFα and the expression of M2 cell phenotype molecules Arg-1 and CD206. The effect of adding LY294002 signaling pathway by TERM2 activation was inhibited, indicating that TERM2 can affect the occurrence of inflammation by regulating the activity of PI3K/AKT signaling pathway. Finally, Western blotting and ELISA showed that activation of TERM2 can promote the expression of Arg-1 and CD206 in BV2 cells, and promote the transformation of BV2 cells to M2 polarization. TERM2 can affect the inflammatory response in microglia through the PI3K/AKT signaling pathway, suggesting that TERM2 may be a target for the treatment of inflammatory response in glial cells. This study provides a treatment plan for alleviating the impact of inflammation on central nervous system.

Tuberculous uveitis can be a manifestation of extrapulmonary tuberculosis or an allergic reaction to tuberculosis infection. The clinical signs and symptoms of other uveitis causes are generic, making a false diagnosis simple. We present a brief introduction to theoretical modelling and simulation in systems biology and explore the consequences of TB uveitis if left untreated. Patients were admitted to our hospital with recurrent fever. They had a previous definitive diagnosis of binocular uveitis and a positive interferon gamma release assay (IGRA) test result. At the time, there was no antituberculosis medicine available, and immunosuppressive and glucocorticoid therapy did not work. After the admission, their pleural fluid tested positive for Mycobacterium tuberculosis. No other causes to explain the fever were found. A diagnosis of tuberculosis was made, and their body temperature normalized after antituberculosis treatment and closed chest drainage. Vigilance should be exercised to rule out tuberculous uveitis in cases of unexplained uveitis; this calls for IGRA screening, tuberculin skin testing, and cyst imaging. For patients with latent tuberculosis infections, it is recommended to administer antituberculosis treatment, after excluding other possible causes, and to avoid using glucocorticoids in isolation.

Hepatocellular carcinoma (HCC) is an intractable malignant disease with high incidence rate annually. LincRNA PRNCR1 has been confirmed as a tumor supporter, while its functions in HCC remain unclear. This study aims to explore the mechanism of LincRNA PRNCR1 in hepatocellular carcinoma. The qRT-PCR was applied to the quantification of non-coding RNAs. Cell counting Kit-8 (CCK-8), Transwell assay and flow cytometry assay were applied to reflect the change in the phenotype of HCC cells. Moreover, the databases including Targetscan and Starbase and dual-luciferase reporter assay were applied to investigate the interaction of the genes. The western blot was applied to detect the abundance of proteins and the activity of the related pathways. Elevated LincRNA PRNCR1 was dramatically upregulated in HCC pathological samples and cell lines. MiR-411-3p served as a target of LincRNA PRNCR1, and decreased miR-411-3p was found in the clinical samples and cell lines. LincRNA PRNCR1 downregulation could induce the expression of miR-411-3p, and LincRNA PRNCR1 silence could impede the malignant behaviors via increasing the abundance of miR-411-3p. Zinc finger E-box binding homeobox 1 (ZEB1) was confirmed as a target of miR-411-3p, which remarkably upregulated in HCC cells, and ZEB1 upregulation could significantly rescue the effect of miR-411-3p on malignant behaviors of HCC cells. Moreover, LincRNA PRNCR1 was confirmed to involve the Wnt/β-catenin pathway via regulating miR-411-3p/ZEB1 axis. This study suggested that LincRNA PRNCR1 could drive the malignant progression of HCC via regulating miR-411-3p/ZEB1 axis.

We aimed to explore the correlations of C-X-C motif chemokine receptor 2 (CXCR2) and chemokine (C-X-C motif) ligand 4 (CXCL4) gene polymorphisms with thoracic aortic aneurysm. A total of 50 patients with thoracic aortic aneurysm (disease group) and 50 healthy people in the physical examination center (control group) in our hospital were selected as the subjects. The CXCR2 and CXCL4 gene polymorphisms were detected by means of blood drawing, DNA extraction, PCR and sequencing. Moreover, the levels of serum CXCR2 and CXCL4 were measured using ELISA, and the levels of C-reactive protein (CRP) and low-density lipoprotein (LDL) were determined. The study found significant differences in the distribution of genotypes and alleles of CXCR2 and CXCL4 gene polymorphisms between the disease group and control group. The frequencies of certain genotypes (AA of rs3890158, CC of rs2230054, AT of rs352008, and CT of rs1801572) were higher in the disease group, as were the frequencies of certain alleles (C of rs2230054 and rs1801572). The distribution of recessive models of rs2230054 was also different, with a lower frequency of CC+CT in the disease group. The haplotype distributions of both gene polymorphisms differed between the groups. CXCR2 rs3890158 and CXCL4 rs352008 were correlated with lower serum levels of their respective proteins, while CXCL4 rs1801572 was associated with CRP levels and CXCR2 rs2230054 with LDL levels in patients (P < 0.05). The gene polymorphisms of CXCR2 and CXCL4 probably have apparent correlations with the susceptibility to thoracic aortic aneurysm.