Tuning transcription factor DegU for developing extracellular protease overproducer in Bacillus pumilus.

BACKGROUND: Global transcription machinery engineering (gTME) is an effective approach employed in strain engineering to rewire gene expression and reshape cellular metabolic fluxes at the transcriptional level. RESULTS: In this study, we utilized gTME to engineer the positive transcription factor, DegU, in the regulation network of major alkaline protease, AprE, in Bacillus pumilus. To validate its functionality when incorporated into the chromosome, we performed several experiments. First, three negative transcription factors, SinR, Hpr, and AbrB, were deleted to promote AprE synthesis. Second, several hyper-active DegU mutants, designated as DegU(hy), were selected using the fluorescence colorimetric method with the host of the Bacillus subtilis ΔdegSU mutant. Third, we integrated a screened degU(L113F) sequence into the chromosome of the Δhpr mutant of B. pumilus SCU11 to replace the original degU gene using a CRISPR/Cas9 system. Finally, based on transcriptomic and molecular dynamic analysis, we interpreted the possible mechanism of high-yielding and found that the strain produced alkaline proteases 2.7 times higher than that of the control strain (B. pumilus SCU11) in LB medium. CONCLUSION: Our findings serve as a proof-of-concept that tuning the global regulator is feasible and crucial for improving the production performance of B. pumilus. Additionally, our study established a paradigm for gene function research in strains that are difficult to handle.

Transcriptome profiling analysis reveals metabolic changes across various growth phases in Bacillus pumilus BA06.

BACKGROUND: Bacillus pumilus can secret abundant extracellular enzymes, and may be used as a potential host for the industrial production of enzymes. It is necessary to understand the metabolic processes during cellular growth. Here, an RNA-seq based transcriptome analysis was applied to examine B. pumilus BA06 across various growth stages to reveal metabolic changes under two conditions. RESULTS: Based on the gene expression levels, changes to metabolism pathways that were specific to various growth phases were enriched by KEGG analysis. Upon entry into the transition from the exponential growth phase, striking changes were revealed that included down-regulation of the tricarboxylic acid cycle, oxidative phosphorylation, flagellar assembly, and chemotaxis signaling. In contrast, the expression of stress-responding genes was induced when entering the transition phase, suggesting that the cell may suffer from stress during this growth stage. As expected, up-regulation of sporulation-related genes was continuous during the stationary growth phase, which was consistent with the observed sporulation. However, the expression pattern of the various extracellular proteases was different, suggesting that the regulatory mechanism may be distinct for various proteases. In addition, two protein secretion pathways were enriched with genes responsive to the observed protein secretion in B. pumilus. However, the expression of some genes that encode sporulation-related proteins and extracellular proteases was delayed by the addition of gelatin to the minimal medium. CONCLUSIONS: The transcriptome data depict global alterations in the genome-wide transcriptome across the various growth phases, which will enable an understanding of the physiology and phenotype of B. pumilus through gene expression.

Analysis of the Rehabilitation Efficacy and Nutritional Status of Patients After Endoscopic Radical Thyroidectomy by Fast Track Surgery Based on Nutritional Support.

Objective: To investigate and analyze the effect of fast track surgery (FTS) based on nutritional support on the improvement of rehabilitation efficacy and nutritional status of patients after radical lumpectomy for thyroid cancer. Methods: Eighty-six patients admitted to our hospital for radical lumpectomy for thyroid cancer between April 2018 and April 2021 were selected, of which 40 patients admitted between April 2018 and April 2019 were included in the control group with conventional perioperative care. Forty-six patients admitted between May 2019 and April 2021 were included in the trial group with FTS care based on nutritional support. The two groups of patients were compared in terms of postoperative feeding time, length of stay, time out of bed, VAS scores, albumin (ALB), total protein (TP) and prealbumin (PA) levels, negative emotions [Mental Health Test Questionnaire (DCL-90)], quality of life [General Quality of Life Inventory (GQOLI-74)] and complication rates. Results: The patients in the trial group had shorter feeding time, hospitalization time and time out of bed than the control group (P < 0.05). After the intervention, ALB, TP and PA levels were higher in the trial group than in the control group vs. preoperatively (P < 0.05); VAS scores in the trial group were lower than VAS scores in the control group during the same period (P < 0.05). The postoperative DCL-90 scores of the trial group were lower than those of the control group (P < 0.05); the GQOLI-74 scores and total scores of the trial group were higher than those of the control group at the 3-month postoperative follow-up (P < 0.05). The overall incidence of complications such as hoarseness, choking on water, hand and foot numbness, wound infection, and hypocalemia was lower in the trial group than in the control group (P < 0.05). Conclusion: The implementation of FTS care based on nutritional support for patients after endoscopic radical thyroidectomyr can effectively improve the postoperative recovery and reduce their pain level, as well as help improve their nutritional status, negative emotions and improve their quality of life, which is worth promoting.

The Effect of Teach-Back Combined with King Interactive Standard Mode on the Life of COPD Patients.

Background: COPD is a common clinical chronic airway inflammatory disease that occurs mostly in middle-aged and older adults over the age of 40. The incidence of COPD is increasing year by year and the onset of age is gradually becoming younger. Objective: To observe the effect of teach-back combined with king interaction on the life of patients with chronic obstructive pulmonary disease (COPD). Methods: A total of 100 COPD patients admitted to our hospital from Jan 2021 to Jan 2022 were retrospectively selected to be divided into 50 cases in the control group and 50 cases in the observation group according to the nursing methods. The control group was treated with routine nursing intervention, while the observation group was treated with teach-back combined with king interactive standard mode intervention. The differences in Self-Care Ability Assessment Scale (ESCA) score, St. George's Respiratory Questionnaire (SGRQ) score, Mental State Assessment Scale (MSSNS) score, 6-minute walking distance (6MWD), and pulmonary function indexes were compared between the two groups before and after the intervention. The success rate and patient compliance of each index in the groups were also recorded. Results: After 3 months and 6 months of intervention, the total SGRO score and its factor scores of self-care skills, self-care responsibility, self-concept, health knowledge level in them were all higher than those before the intervention, while the total SGRO score and its factor scores of respiratory symptoms, activity limitation, disease influence, and so on were all decreased compared with those before the intervention. The ESCA score of the observation group was significantly higher than that of the control one after 3 months and 6 months of intervention, while the SGRQ score was significantly lower than that of the control one, with statistical significance (P < 0.05). After 3 months of intervention, the total score of MSSNS and the scores of anxiety, depression, loneliness, and other factors in both groups were decreased compared with those before intervention. After 6 months of intervention, the total score of MSSNS and scores of each factor in both groups were decreased compared with those before intervention, and the MSSNS scores in the observation group were significantly lower than those in the control group after the intervention, which was statistically significant (P < 0.05). After 3 months and 6 months of intervention, 6MWD, forced vital capacity (FVC), forced expiratory value in 1 second (FEV1), and FVC/FEV1 in them were all higher than those before intervention, and 6MWD and pulmonary function were significantly higher in the observation group than in the control group after 3 and 6 months of intervention, which was statistically significant (P < 0.05). The ESCA score, SGRQ score, MSSNS score, pulmonary function compliance rate, and compliance rate in the observation group were significantly higher than those in the control group, which was statistically significant (P < 0.05). Conclusion: The use of teach-back combined with king interactive standard mode in COPD patients can improve the patient's self-care ability, reduce psychological negative emotions, and improve the quality of life.

Effects of Disease-Related Knowledge on Illness Perception and Psychological Status of Patients With COVID-19 in Hunan, China.

OBJECTIVE: The aim of this study was to assess the current status of disease-related knowledge and to analyze the relationship among the general condition, illness perception, and psychological status of patients with coronavirus disease 2019 (COVID-19). METHODS: A hospital-based cross-sectional study was conducted on 118 patients using convenience sampling. The general questionnaire, disease-related knowledge questionnaire of COVID-19, Illness Perception Questionnaire (IPQ), and Profile of Mood States (POMS) were used to measure the current status of participants. RESULTS: The overall average score of the disease-related knowledge of patients with COVID-19 was (79.19 ± 14.25), the self-care situation was positively correlated with knowledge of prevention and control (r = 0.265; P = 0.004) and total score of disease-related knowledge (r = 0.206; P = 0.025); the degree of anxiety was negatively correlated with the knowledge of diagnosis and treatment (r = -0.182; P = 0.049). The score of disease-related knowledge was negatively correlated with negative cognition (volatility, consequences, emotional statements) and negative emotions (tension, fatigue, depression) (P < 0.05); positively correlated with positive cognition (disease coherence) and positive emotion (self-esteem) (P < 0.05). CONCLUSIONS: It was recommended that we should pay more attention to the elderly and low-income groups, and increase the knowledge about diagnosis and treatment of COVID-19 and self-care in the future health education for patients.

Humanistic care interventions in patients with lower extremity arteriosclerosis obliterans.

OBJECTIVE: This study was designed to investigate the application value of humanistic care interventions in patients with lower extremity arteriosclerosis obliterans (LEASO). METHODS: We enrolled 98 LEASO patients undergoing interventional therapy in our hospital into this study, among whom 47 cases received conventional care interventions (the regular group) while the other 51 received humanistic care interventions based on conventional care interventions (the research group). The two groups were compared in negative emotions, complications, and quality of life of patients. RESULTS: Scores of negative emotions were lower in the research group than in the regular group after care (P < 0.05). The pain intensity decreased in both groups after care, with a sharper decrease in the research group (P < 0.05). The incidence of complications was lower in the research group than in the regular group after care (P < 0.05). The pain-free walking distance (PFWD) increased remarkably in both groups after care, with a longer PFWD in the research group than in the regular group (P < 0.05). Scores of the 36-Item Short-Form Health Survey (SF-36) increased after care in both groups (P < 0.05), with higher SF-36 scores in the research group than in the regular group (P < 0.05). SF-36 reflects the quality of life of patients from four aspects including vitality, emotional state, social function, and role-emotional. The care compliance and satisfaction level with care of patients were higher in the research group than in the regular group (P < 0.05). The care compliance rate and patient satisfaction level with care were higher in the research group than in the regular group (P < 0.05). CONCLUSION: Humanistic care interventions can reduce postoperative pain intensity and improve the quality of life in patients with LEASO.

Post-traumatic Growth and Related Influencing Factors in Discharged COVID-19 Patients: A Cross-Sectional Study.

The purpose of this study is to investigate the current state of post-traumatic growth (PTG) and identify its influencing factors in discharged COVID-19 patients. PTG refers to individual experiences of significant positive change arising from the struggle with a major life crisis. This descriptive cross-sectional study used the convenient sampling method to recruit 140 discharged COVID-19 patients in Hunan, China. The results show that the PTG of the discharged COVID-19 patients was positively correlated with self-esteem, post-traumatic stress disorder, coping style tendency, and social support, but negatively correlated with the time from onset to diagnosis. Our findings could provide guidance on improving the psychological state and well-being of discharged COVID-19 patients.

Research on an Mg-Zn alloy as a degradable biomaterial.

In this study a binary Mg-Zn magnesium alloy was researched as a degradable biomedical material. An Mg-Zn alloy fabricated with high-purity raw materials and using a clean melting process had very low levels of impurities. After solid solution treatment and hot working the grain size of the Mg-Zn alloy was finer and a uniform single phase was gained. The mechanical properties of this Mg-Zn alloy were suitable for implant applications, i.e. the tensile strength and elongation achieved were approximately 279.5MPa and 18.8%, respectively. The results of in vitro degradation experiments including electrochemical measurements and immersion tests revealed that the zinc could elevate the corrosion potential of Mg in simulated body fluid (SBF) and reduce the degradation rate. The corrosion products on the surface of Mg-Zn were hydroxyapatite (HA) and other Mg/Ca phosphates in SBF. In addition, the influence caused by in vitro degradation on mechanical properties was studied, and the results showed that the bending strength of Mg-Zn alloy dropped sharply in the earlier stage of degradation, while smoothly during the later period. The in vitro cytotoxicity of Mg-Zn was examined. The result 0-1 grade revealed that the Mg-Zn alloy was harmless to L-929 cells. For in vivo experiments, Mg-Zn rods were implanted into the femoral shaft of rabbits. The radiographs illustrated that the magnesium alloy could be gradually absorbed in vivo at about 2.32mm/yr degradation rate obtained by weight loss method. Hematoxylin and eosin (HE) stained section around Mg-Zn rods suggested that there were newly formed bone surrounding the implant. HE stained tissue (containing heart, liver, kidney and spleen tissues) and the biochemical measurements, including serum magnesium, serum creatinine (CREA), blood urea nitrogen (BUN), glutamic-pyruvic transaminase (GPT) and creatine kinase (CK) proved that the in vivo degradation of Mg-Zn did not harm the important organs. Moreover, no adverse effects of hydrogen generated by degradation had been observed and also no negative effects caused by the release of zinc were detected. These results suggested that the novel Mg-Zn binary alloy had good biocompatibility in vivo.