Biotechnology & Genetic Engineering Reviews最新文献

Metabolic engineering principles have long been applied to explore the metabolic networks of complex microbial cell factories under a variety of environmental constraints for effective deployment of the microorganisms in the optimal production of biochemicals like biofuels, polymers, amino acids, recombinant proteins. One of the methodologies used for analyzing microbial metabolic networks is the Flux Balance Analysis (FBA), which employs applications of optimization techniques for forecasting biomass growth and metabolic flux distribution of industrially important products under specified environmental conditions. The in silico flux simulations are instrumental for designing the production-specific microbial cell factories. However, FBA has some inherent limitations. The present review emphasizes how the incorporation of additional kinetic, thermodynamic, expression and regulatory constraints and integration of omics data into the classical FBA platform improve the prediction accuracy of FBA. A programmed comparison of the simulated data with the experimental observations is presented for supporting the claim. The review further accounts for the successful implementation of classical FBA in biotechnological applications and identifies areas in which classical FBA fails to make correct predictions. The analysis of the predictive capabilities of the different FBA strategies presented here is expected to help researchers in finding new avenues in engineering highly efficient microbial metabolic pathways and identify the key metabolic bottlenecks during the process. Based on the appropriate metabolic network design, fermentation engineers will be able to effectively design the bioreactors and optimize large-scale biochemical production through suitable pathway modifications.

Multidrug resistance (MDR) is considered as a major obstacle in achieving an effective treatment of breast cancer. Paclitaxel has been used to treat cancers of the cervical, breast, ovarian and brain but MDR limits its therapeutic potential. Phytochemicals have received much interest in recent decades especially in combination approaches to tackle MDR due to their negligible harm to healthy cells and synergistic potential. Considering this notion, the present study aimed at investigating the synergistic activity of 4-MTBITC and PTX against a panel of breast cancer cells. Our results revealed that the combination had a significant antiproliferative activity against T-47D cells. Mechanistic studies revealed that 4-MTBITC and PTX also promoted the production of reactive oxygen species (ROS) and reduced mitochondrial membrane potential. In the presence of 4-MTBITC- PTX, T-47D cells were found to be arrested in the G₂/M phase which also confirmed the enhancement of late apoptotic cell population in the flow cytometer analysis. In western blot experiment, the combination had a significant decrease in Bcl-xl protein level, whereas a higher level of p53, cleaved caspase-3, and cleaved caspase-9 proteins compared to individual treatment in T-47D cells. The RT-qPCR analysis also showed that the combination had significant upregulation in the gene expression of p53, cytochrome-c, Apaf-1 and downregulation in the expression of Bcl-2 gene in T-47D cells. Hence, all the results showed that a combination of 4-MTBITC-PTX significantly enhanced the apoptosis pathway in the T-47D cell line which indicates its clinical application for the treatment of breast cancer.Abbreviations: Apaf-1: Apoptotic protease activating factor 1; AO/EB: Acridine orange/ethidium bromide; Bcl-2: B-cell lymphoma 2; CI: Combination Index; Cyt-c: Cytochrome c; CO₂: Carbon dioxide; DCFH-DA 2,7-Dichloroflourescein diacetate; DMEM: Dulbecco's modified Eagle's medium; ELISA: Enzyme-linked immunosorbent assay; EA: Early apoptosis; EDTA: Ethylenediaminetetraacetic acid; L929: Normal mouse fibroblast cells; LA: Late apoptosis; L: Live; 4-MTBITC: 4-methylthiobutyl isothiocyanate; MCF-7: Human breast cancer cells; MDA-MB-231: Human triple negative breast cancer cells; MMP: Mitochondria membrane potential; MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide; NCCS: National Centre for Cell Science; N: Necrotic; PTX Paclitaxel; PVDF: Polyvinylidene fluoride; PAGE: Polyacrylamide gel electrophoresis; PBS: Phosphate-buffered saline; RPMI-1640: Roswell Park Memorial Institute Medium- 1640; RT-qPCR: Quantitative real-time polymerase chain reaction; ROS: Reactive oxygen species; Rh-123: Rhodamine123; g Relative centrifugal force; SDS: Sodium dodecyl sulphate; SEM: Scanning electron microscopy; T-47D: Human estrogen positive breast cancer cells; WB: Western blotting.

Liver cancer, one of the most common malignant tumors, has seriously threatened people's physical and mental health. The paper was conducted to investigate the functions of two different anesthesia methods on Tumor necrosis factor-α (TNF-α), C-reactive protein (CRP) and T lymphocyte subsets in patients undergoing liver cancer resection, and to explore the liver function impact of patients undergoing liver cancer surgery. 80 liver cancer patients were split into propofol intravenous anesthesia group and sevoflurane inhalation anesthesia group. The patient's operation-related indicators and changes in TNF-α, CRP and T lymphocyte subsets before anesthesia, after anesthesia, 1 day after anesthesia, and 3 days after anesthesia were calculated. The levels of TNF-α and CRP in the two groups after operation were clearly higher than those before anesthesia, but there was no obvious difference between the two groups. After operation, the TNF-α of the propofol intravenous anesthesia group was lower than the sevoflurane inhalation anesthesia group. The CD3+, CD4+ and CD4+/CD8+ levels in both groups recovered to the pre-anaesthesia level 3 days after the operation. The levels of ALT, AST, ALB and TBIL returned to normal at d4 in both propofol intravenous anesthesia and sevoflurane inhalation anesthesia groups. Compared with evoflurane inhalation anesthesia for patients with liver cancer, intravenous anesthesia with propofol can reduce the body's inflammatory response to a certain extent, has weak inhibitory effect on T lymphocyte immune function, and a smaller effect on liver function.

Myocarditis in children is more common in clinical practice, which can cause different degrees of cardiac function damage. We investigated the effects of creatine phosphate in the treatment of myocarditis in children. Children in the control group were treated with sodium fructose diphosphate, and children in the observation group were treated with creatine phosphate on the basis of the control group. After treatment, the myocardial enzyme profile and cardiac function of children in the observation group were better than the control group. The total effective rate of treatment for children in the observation group was higher than that in the control group. In conclusion, creatine phosphate could significantly improve myocardial function, improve myocardial enzyme profile and reduce myocardial damage in children with pediatric myocarditis and had a high safety of use, which was worthy of clinical promotion.

To investigate the value of serum eotaxin-2, neutrophil/lymphocyte ratio (NLR), and platelet/lymphocyte ratio (PLR) in the diagnosis and prognosis of colorectal cancer (CRC). The association between different clinicopathological characteristics and eotaxin-2, NLR, and PLR in different patient groups was evaluated. The combined detection indicator and the combined detection's predictive effect on distant metastasis were examined. The receiver operating characteristic (ROC) curve was drawn to evaluate the efficacy of combined detection. The association between eotaxin-2, inflammation markers, and postoperative complications was assessed. Multivariate analysis to investigate the factors affecting the prognosis of patients with CRC. We detected a marked positive correlation between NLR and PLR (p < 0.05, r= 0.209). The AUC of serum eotaxin-2 combined with inflammation markers was 0.889, which was higher than single diagnosis. Compared with the single eotaxin-2 test, the combined detection of eotaxin-2 and inflammation markers might improve the specificity of CRC assessment. In univariate analysis, age, surgical method, high eotaxin-2, and high NLR were associated with postoperative complications. In multivariate analysis, age (≥ 60 years), high eotaxin-2, and high NLR were independent risk elements influencing postoperative complications of CRC. The distant metastasis, TNM staging -Ⅳ stage, NLR ≥ 3.18, and PLR ≥ 193 were independent factors affecting the prognosis of patients with CRC. The combined detection of eotaxin-2 and inflammatory markers has a particular value in improving the diagnosis of CRC, predicting distant metastasis, and guiding the frequency of reexamination after radical resection of CRC.

Hip fractures (VOHF) are a type of bone fracture that occurs in the hip bone, typically affecting older adults. Recovery from hip fractures in older adults can be challenging due to factors such as decreased mobility, weakened bones, and other health issues. This paper intends to determine the effect of mini-nutrition nursing plus Baduanjin rehabilitation exercise on postoperative rehabilitation of elderly patients with vertebral or hip fractures (VOHF). 106 senile VOHF patients were chosen for the study between May 2020 and May 2022. The research group (RG) consisted of 60 patients who received mini-nutrition nursing + Baduanjin rehabilitation exercise, while the control group (CG) had 46 patients who received mini-nutrition nursing + routine rehabilitation training. The study compared the fracture healing effect, MAMC, CC, and MNA-SF scores. The rehabilitation status of patients was evaluated using FMA, BI, BBS, and VAS. Adverse reactions during hospitalization and SF-36 scores were also recorded. Better fracture healing efficacy and higher MAMC, CC, and MNA-SF scores were determined in RG versus CG (P < 0.05). Elevated FMA, BI and BBS scores were observed in both groups after rehabilitation training, with more evident elevations in RG; while the VAS score reduced markedly and was lower in RG compared with CG (P < 0.05). In addition, RG was observed with fewer adverse reactions and higher SF-36 scores (P < 0.05). Mini-nutrition nursing + Baduanjin rehabilitation exercise can effectively accelerate the postoperative rehabilitation of elderly patients with VOHF, improve mobility, and enhance their quality of life.

Melanoma is a highly malignant cancer with a high differentiation potential and metastatic capacity. Sodium butyrate, known for its anti-cancer activity, is used in various types of solid tumors. This study aimed to investigate the effects of sodium butyrate on B16 melanoma cells using in vitro and in vivo mouse models. The study utilized MTT assay, flow cytometry, and immunoblot analysis. Mice were treated with normal saline (control) or 1 mM, 2 mM, 3 mM, or 5 mM sodium butyrate. Results showed that cell viabilities were significantly reduced in 2 mM, 3 mM, and 5 mM sodium butyrate groups after 24 to 48 hours (p < 0.01 for all). Moreover, sodium butyrate exhibited a tumor suppression effect that was time-dependent and lasted for 30 days (p < 0.01 for all). A significant tumor suppression effect was observed in the case of 5 mM sodium butyrate after 30 days (p < 0.001 for all). As compared to control (no sodium butyrate), tumor-associated macrophages were decreased in 2 mM, 3 mM, and 5 mM sodium butyrate groups (<0.01 for all). The maximum reduction was observed in 5 mM sodium butyrate groups. Sodium borate decreased the release of interleukin-10, vascular endothelial growth factor, transforming growth factor beta, and β-actin (<0.01 for all). A significant reduction was observed in the case of 5 mM concentration. Overall, these findings suggest that sodium butyrate is effective in the treatment of melanoma and may offer a promising new avenue for melanoma therapy.

To analyze the changes of immune function-related indicators with newly diagnosed glioblastoma before and after radiotherapy and their clinical significance. Clinical data of 104 patients were analyzed. The independent samples t-test or chi-square test was used to compare changes in immune function indicators and to ascertain the differences between groups with different doses or volumes. The grading of the lowest lymphocyte count during radiotherapy was compared. The log-rank (Mantel - Cox) test of the Kaplan - Meier method was used to compare the survival rate, and the relationship of radiotherapy-related parameters, with the survival rate was evaluated by using the Spearman correlation coefficient. A Cox regression model was used to determine the relationship between various immune function indicators and prognosis. The percentages of total T lymphocytes and CD4+ T cells, the CD4-to-CD8 subset ratio, and the percentages of B cells and NKT cells showed an overall decreasing trend, whereas the percentages of CD8+ T cells and NK cells displayed an overall increasing trend. The lower CD4+ T cell percentage and CD4/CD8 ratio after radiotherapy were independent risk factors for OS. Short OS was observed in patients with grade 3 or 4 lymphopenia or with low levels of hemoglobin and serum albumin before radiotherapy. The percentage of CD4+ T cells and the CD4/CD8 ratio were higher in patients with the low tumor-irradiated volume and irradiated volume and dose of the OAR, than in patients from the corresponding high indicator group. Different irradiation dose or volume can differentially alter various immune function indicators.

To explore the mechanism of Tongmai Zhuke decoction for promoting blood circulation by taking carotid artery atherosclerosis (CAA) as an example, two sets of in-depth transcriptomic data as well as two sets of single-cell RNA sequencing data related to the macrophages in CAA were included. STAR and DCC software were used to process in-depth transcriptomic data in order to measure the expression level of LncRNAs as well as mRNA according to FPKM analysis. Single-cell RNA sequencing data from Illumina NovaSeq 6000 were further analyzed by CellRanger channel, CellRanger count, Seurat R package, DoubletFinder package, CCA algorithm, LogNormalize, principal-component analysis, t-SNE and ToppGene online tools. Based On unsupervised clustering, a total of four diverse cell populations with distinct transcriptional features were found in human carotid atherosclerotic plaques. The macrophages were further annotated as the "effector cell" in the pathologic process of CAA, based on the expression of CD68+/CD440-. A total of 84 up-regulated genes and 58 down-regulated linc-RNAs were identified in samples with carotid atherosclerotic plaques. Thereinto, lincRNA-Cox2 is the most down-regulated LincRNA. For the macrophages in carotid atherosclerotic plaques, expression level of Il6, Ccl3, Ccl4 Il10 and Tnfa were significantly up-regulated, while Timp1 significantly down-regulated comparing with healthy carotid sample. The expression level of lincRNA-Cox2 was significantly increased in macrophages after treated by Tongmai Zhuke decoction, while Cxcl10, Ccl3, Ccl4, Cxcl2, Ccl5, and Ccl19 were significantly decreased. Collectively, Tongmai Zhuke decoction could restrain the inflammatory reaction of macrophages for carotid artery atherosclerosis by up-regulating lincRNA-Cox2.

Ventricular remodeling (VR) after acute ST-elevation myocardial infarction (STEMI) is an important predictor for medium- and long-term prognosis. This study focuses on the relevant indexes of VR in patients with AMI, in which, the intervention effects of sacubitril/valsartan and enalapril were compared, guiding the clinical treatment. 58 patients with acute STEMI treated with PCI were divided into research group and control group. UCG was performed at 1 week, 1 month and 3 months after MI, and the patients' indexes were collected to compare VR and adverse reactions in the two groups. The test results showed that there was no statistical difference in the baseline data of patients in the two groups, which were comparable. In the blood biochemical index examination, no statistical difference was found in cTnI and NT-proBNP between the two groups. At 1 week after operation, the levels of cTnI and NT-proBNP in research group were lower than those in the control group. In ECG examination, there was no statistical significance in the levels of LVEF, LVEDD and LVESD at admission between the two groups. After 1 week, the results of LVEF, LVEDD, LVESD in the research group were higher than those in the control group. The results of this study show that sacubitril/valsartan can be used in patients with AMI instead of enalapril. Sacubitril/valsartan improves cardiac function in patients with emergency percutaneous coronary intervention (PCI) for AMI, inhibits ventricular remodeling, and has a low incidence of adverse cardiac events and adverse drug reactions.