In recent years, the bacteriophage Φ29 (Phi29) DNA polymerase has garnered increasing attention due to its high-fidelity amplification capacity at constant temperatures. To advance the industrial application of this type of isothermal polymerases, this study mined and characterized new enzymes from the microbial metagenome based on the known Phi29 DNA polymerase sequence. The results revealed that a new enzyme, Php29 DNA polymerase, was identified in the microbial metagenome with plants as the hosts. This enzyme exhibited higher strand displacement activity, with a 59.5% similarity to bacteriophage Φ29. Experimental validation demonstrated that the enzyme had 3'→5' exonuclease activity, and its amplification products can serve as substrates for further catalytic reactions. The discovery and validation of Php29 DNA polymerase gives insights into the future industrial application of isothermal polymerases.
DNA-Directed DNA Polymerase/metabolism* ; Bacillus Phages/genetics* ; Metagenome

Environmental Microbiology, as a compulsory foundational course for the major of environmental and ecological engineering. Its knowledge system not only lays a profound theoretical foundation but also evinces a close-knit nexus to practical applications in environmental protection and ecological restoration. This course thus serves as an outstanding platform facilitating both teaching reforms and practical innovations of professional courses. With Environmental Microbiology as a paradigmatic illustration, this paper concentrates on addressing issues like archaic course contents and enervating teaching approaches, with the aspiration of augmenting students' professional acuities, practical dexterities, and innovation and entrepreneurial aptitudes. In light of the aforesaid aim, we initiated an all-encompassing probe into the teaching reform and practicable paradigms from the perspective of amalgamating professional education with innovation-steered stratagems. Across multiple fronts, comprehensive reformative undertakings were implemented, subsuming the optimization and reconfiguration of course contents, the exploration of sundry teaching methodologies, and the scientific erection of an evaluation system. Ultimately, a nascent teaching model dubbed "three-dimensional, six-loop and five-collaborative" was formed. This series of comprehensive and innovative reform measures has not only substantially augmented the profundity and extensiveness of professional knowledge, permeating into the core of disciplinary essence, but also remarkably bolstered students' practical operational capabilities and innovative thinking patterns, thereby catalyzing their transformation into well-rounded individuals with a competitive edge. By virtue of these efforts, a solid foundation has been firmly laid for the teaching reform of Environmental Microbiology and the cultivation of high-caliber environmental professionals in the contemporary era, heralding a new chapter in the development of environmental education.
Environmental Microbiology ; Teaching ; Education, Professional/methods* ; Curriculum

Objective: To explore the effects of group sports game intervention on social ability and quality of life of children with austism spectrum disorders (ASD),so as to provide reference for rehabilitation intervention of social and quality of life of children with ASD. Methods: From September 2021 to January 2022, 72 children with ASD aged 4-6 in the children s rehabilitation department of Xiangyang Central Hospital were selected to participate in the study, and were randomly divided into experimental group ( n =36) and control group ( n =36). The control group received routine rehabilitation training (including individual sports game training), and the experimental group replaced individual sports game training with group sports game training on the basis of routine rehabilitation.The course content mainly included three parts: warm up before class, group sports games and relaxation after class. The course combined social skills with sports games, and was carried out in a group form (divided into 12 groups with 3 people in each group), and was trained five times a week for 60 minutes, for a total of 12 weeks. The scores of Childhood Autism Rating Scale (CARS), Autism Treatment Evaluation Checklist (ATEC), Social Responsiveness Scale (SRS) and Pediatric Quality of Life Inventory Version 4.0 Generic Core Scales (PedsQL 4.0) were observed before and after treatment. t-test and χ 2 test were used for statistical analysis. Results: There was no significant difference in SRS scores between the experimental group and the control group before intervention ( t =-0.63, P >0.05). After the intervention, the total response rate in the experimental group was 83.33 %, higher than 41.67% in the control group χ 2=13.33, P <0.05),and the SRS scores decreased in the experimental group and control group ( t =17.75,8.71, P <0.05). The SRS scale score of the experimental group Social perception (17.67±4.12) , social cognition (30.33±4.99) , social communication (50.33±9.39) , social motivation (24.25±6.78) scores and total scores ( 152.67± 25.82) were lower than those of the control group(22.17±5.34，36.00±4.13，62.58±11.07，34.42±7.13，186.33±29.03)( t = -4.88，-2.03，-2.13，-3.58，-3.01， P ＜0.05).After the intervention, the scores of social function (53.33±18.01) and total score (283.83±51.83) on PedsQL 4.0 scale in experimental group were higher than those in control group(23.33±15.13，218.00±39.01) ( t =4.42，3.52， P ＜0.05). After the intervention, Autism Treatment Evaluation Scale (ATEC) scores of experimental groups(44.33±14.72) was lower than that in control group ( 59.33±16.95)( t =-2.32, P ＜0.05). Conclusion The intervention of group sports game has a significant effect on improving social ability and life quality of children with ASD.

Objective:To explore the application effects of application of rehabilitation care decision-making scheme based on case management model in severe burn patients.Methods:The study was a non-randomized historical control study. Thirty patients who met the inclusion criteria and received routine rehabilitation nursing in the First Affiliated Hospital of Army Medical University (the Third Military Medical University, hereinafter referred to as the hospital) from April 2021 to March 2022 were included in routine rehabilitation nursing group (26 males and 4 females, aged 48.50 (31.75, 56.25) years), and 30 patients who met the inclusion criteria and received case management rehabilitation nursing in the hospital from April 2022 to March 2023 were included in case management rehabilitation nursing group (22 males and 8 females, aged 46.00 (36.75, 55.25) years). The length of intensive care unit (ICU) stay, total hospitalization day, and total hospitalization cost of the patients in two groups were recorded. At admission, convalescence, discharge, and 6 months after injury, the patients' life quality was evaluated by the concise burn specific health scale, the sleep quality was evaluated by the Pittsburgh sleep quality index, and the functional independence was evaluated by the functional independence rating scale. At convalescence, discharge, and 6 months after injury, the patients' scar status was evaluated by the Vancouver scar scale. At 6 months after injury, a third-party satisfaction questionnaire was used to investigate the efficacy satisfaction of patients.Results:The length of ICU stay and total hospitalization day of patients in case management rehabilitation nursing group were both significantly shorter than those in routine rehabilitation nursing group (with Z values of -1.97 and -1.99, respectively, P<0.05), and the total hospitalization cost was less than that in routine rehabilitation nursing group ( Z=-1.99, P<0.05). At discharge and 6 months after injury, the life quality scores of patients in case management rehabilitation nursing group were significantly higher than those in routine rehabilitation nursing group (with t values of -3.19 and -4.43, respectively, P<0.05), while the sleep quality scores were significantly lower than those in routine rehabilitation nursing group (with Z values of -2.18 and -3.33, respectively, P<0.05). There were no statistically significant differences in cognitive function scores of functional independence of patients between the 2 groups at admission, convalescence, discharge, and 6 months after injury ( P>0.05). The exercise function scores and total scores of functional independence of patients in case management rehabilitation nursing group at convalescence, discharge, and 6 months after injury were significantly higher than those in routine rehabilitation nursing group (with Z values of -4.37, -2.73, -4.10, -4.37, -2.64, and -4.06, respectively, P<0.05). The scar pigmentation scores of patients in case management rehabilitation nursing group at 6 months after injury were significantly lower than those in routine rehabilitation nursing group ( Z=-2.05, P<0.05), and the scar vascularity scores of patients in case management rehabilitation nursing group at discharge and 6 months after injury in case management rehabilitation nursing group were significantly lower than those in routine rehabilitation nursing group (with Z values of -3.16 and -2.07, respectively, P<0.05). The scar pliability scores (with Z values of -3.16, -2.45, and -4.38, respectively, P<0.05), thickness scores (with Z values of -2.56, -2.35, and -4.70, respectively, P<0.05), and total scores (with Z values of -3.77, -3.04, and -3.13, respectively, P<0.05) of patients in case management rehabilitation nursing group at convalescence, discharge, and 6 months after injury were significantly lower than those in routine rehabilitation nursing group. At 6 months after injury, the efficacy satisfaction scores of patients in case management rehabilitation nursing group were 4.00 (3.00, 4.25), which were significantly higher than 3.00 (2.00, 4.00) in routine rehabilitation nursing group ( Z=-2.72, P<0.05). Conclusions:The implementation of rehabilitation care decision-making scheme based on case management model can optimize the cost efficiency, improve the effectiveness of clinical treatment, and enhance the life quality and satisfaction of the curative effect of severe burn patients.

Objective:To conduct a scope review on the application of ICU diaries in critically ill patients, laying the foundation for further exploration and construction of ICU diary patterns and frameworks that were in line with the national conditions and tailored to different regions and cultural backgrounds.Methods:The Joanna Briggs Institute Reviewer′s Manual was used as the methodological framework, and a computer search was conducted in nine domestic and international databases, including China National Knowledge Infrastructure, Wanfang Database, China Biomedical Literature Database, Cochrane Library, PubMed, and Embase, etc. The search period was from the inception of the databases until March 13, 2023. The included literature was screened, summarized, and analyzed.Results:A total of 19 articles were included. ICU diaries were commonly recorded using a combination of text and visuals, with the involvement of both healthcare professionals and family members. Most patients received ICU diaries approximately one month after their transfer from the ICU. Out of the 15 studies, ICU diaries were found to be effective, while 4 studies indicated no significant improvement in patients′ psychological issues. However, ICU diaries were still considered acceptable by patients and their families.Conclusions:The application of ICU diaries has shown positive significance in critically ill patients, but further research and exploration are needed to investigate its impact on issues such as post-traumatic stress disorder, anxiety, depression, and quality of life. In the future, a combination of multiple forms and high-quality research designs with large samples, long periods, and structured approaches should be employed to explore its application effects and long-term outcomes on psychological problems.

Objective To understand the factors that affect the pulmonary rehabilitation behavior of elderly patients with lung cancer during the post-operative transitional period,and provide reference bases for formulating the pulmonary rehabilitation plan of elderly patients with lung cancer during the transitional period and improving the compliance of pulmonary rehabilitation.Methods Based on the capability,opportunity,and motivation-behavior model(COM-B model),12 elderly lung cancer patients who underwent surgery at a tertiary A tumor hospital in Tianjin from March to July 2022 were selected for semi-structured interviews using descriptive research methods and purposive sampling method.The data were analyzed using directed content analysis.Results The ability factors(physical strength and postoperative discomfort symptoms of the elderly limit lung rehabilitation exercise,lack of pulmonary rehabilitation knowledge and awareness of the importance of exercise in lung rehabilitation knowledge),opportunity factors(lack of grassroots medical and health services,limiting lung rehabilitation exercise;family and friends are important resources for promoting lung rehabilitation exercise;medical support is a guarantee for promoting lung rehabilitation exercise),motivational factors(high self-efficacy in exercise is the foundation for lung rehabilitation exercise,perception of lung rehabilitation effects increases motivation for lung rehabilitation exercise)and 7 sub themes were extracted.Conclusion There are certain obstacles and promoting factors in the transitional lung rehabilitation process for elderly lung cancer patients after surgery.It is necessary to improve their postoperative discomfort symptoms,increase their knowledge of lung rehabilitation,enhance their awareness of lung rehabilitation,face the social support role of family,friends,and medical staff,improve the infrastructure and lung rehabilitation system,increase community lung rehabilitation support,provide multi-channel high-quality lung rehabilitation resources,and promote the development of smart medical services,constructing a transitional lung rehabilitation strategy for elderly patients with lung cancer.

Objective To investigate the clinical effect of sedation hypnotic drugs combined with sleep health education in comorbid insomnia of adult type 2 diabetes mellitus.Methods The data of general condi-tions,examination indicators,Insomnia Severity Index Scale(ISI),Self-rating Anxiety Scale(SAS),Self-rat-ing Depression Scale(SDS)scores in the patients with type 2 diabetes mellitus receiving short-term(2 weeks)insulin intensive therapy in the neurology and endocrinology department of this hospital from January to De-cember 2023 were collected.Sixty-five patients receiving the sedation hypnotic drugs combined with sleep health education served as the observation group,among them,39 cases were complicated with mild anxiety(group A1),22 cases were complicated with mild depression(group A2);fifty-four patients without receiving sedative and hypnotic drugs combined with sleep health education served as the control group,among them,33 cases were complicated with mild anxiety(group B1),22 cases were complicated with mild depression(group B2).All patients received the routine diabetes diet,exercise instruction and intensive insulin therapy.The gly-cemic control and improvement of insomnia,anxiety and depression after 1,2 weeks of treatment were com-pared among various groups.Results The reaching standard rates of fasting blood glucose(FPG),2 h post-prandial glucose(2 hPG)and time in range(TIR)in 1 week after treatment showed no statistical difference between the observation group and control group(P＞0.05);FPG and 2 hPG after 2-week treatment in the observation group were were significantly lower than those in the control group(P＜0.05),and the TIR reac-hing standard rate in the observation group was significantly higher than that in the control group(P＜0.05).The scores of ISI,SAS and SDS after 1 week treatment had no statistical difference between the obser-vation and control groups(P＞0.05);the ISI and SAS scores after 2-week treatment in the observation group were significantly lower than those in the control group(P＜0.05),The SDS score had no statistical differ-ence between the observed group and the control group(P＞0.05);the remission rate of insomnia and anxiety symptoms in 2 weeks after treatment in the observation group was significantly increased compared with that in 1 week after treatment(P＜0.05);the remission rate of insomnia,anxiety and depression symptoms in the control group had no statistic difference between in 2 weeks after treatment and 1 week after treatment(P＞0.05).Conclusion Sedation hypnotic drugs combined with sleep health education for treating comorbid in-somnia in adult patients with type 2 diabetes mellitus can not only effectively increase the reaching standard rate of blood glucose control,but also improve the symptoms of insomnia and anxiety.

Glaucoma is a neurodegenerative disease characterized by progressive and irreversible necrosis and apopto-sis of retinal ganglion cells.The abnormal activation of microglia,astrocytes and Müller cells in the central nervous system of the retina and macrophages,T/B lymphocytes and regulatory T cells in the peripheral immune system is involved in the pathogenesis of glaucoma,and the secretion of inflammatory factors such as interleukin 6 and interleukin 8 is closely relat-ed to the development of glaucoma.In this paper,the role of immune cells of the central nervous system and peripheral im-mune system of the retina and the inflammatory factors they secreted in the development of glaucoma is reviewed,in order to understand the pathogenesis of glaucoma at the cellular and molecular levels and to provide ideas for exploring new treatment methods.

Objective:To study the clinical value of serum microRNA-34a (miR-34a) and aryl hydrocarbon receptor nuclear transcript-like protein 1 (BMAL1) in cervical cancer and their relationship with high-risk human papillomavirus (HR-HPV) infection.Methods:The clinical data of 76 patients with cervical cancer and 50 patients with benign cervical diseases in the Affiliated Hospital of Jining Medical University from January to December 2022 were retrospectively analyzed. The expression levels of serum miR-34a and BMAL1 were detected by real-time fluorescence quantitative polymerase chain reaction, and HR-HPV infection was detected by flow fluorescent hybridization. The patients were followed up until December 2023, and the death and poor prognosis (death, tumor recurrence and progression and severe complications at 1-year of follow-up) were recorded. The receiver operating characteristic (ROC) curve was used to analyze the efficacy of miR-34a, BMAL1 and related indexes in evaluating the poor prognosis in patients with cervical cancer at 1-year. Multivariate Cox regression analysis was used to analyze the independent risk factors for death in patients with cervical cancer. The Kaplan-Meier survival curve was used to analyze the relationship between miR-34a, BMAL1 expression and survival period, and the log-rank test was used for comparison.Results:The expression level of serum miR-34a in patients with cervical cancer was significantly lower than that in patients with benign cervical lesions (0.46 ± 0.08 vs. 0.67 ± 0.11), the expression level of serum BMAL1 was significantly higher than that in patients with benign cervical lesions (0.58 ± 0.07 vs. 0.41 ± 0.07), and there were statistical differences ( t= 12.40 and 13.34, P<0.01). The expression levels of serum miR-34a and BMAL1 in patients with cervical cancer were associated with tumor differentiation, myometrial invasion depth, lymph node metastasis, distant metastasis and International Federation of Gynecology and Obstetrics (FIGO) stage, and there were statistical differences ( P<0.05 or<0.01); they were not associated with age, menopause and pathological type, and there were no statistical differences ( P>0.05). In patients with cervical cancer, the expression level of miR-34a in patients with HR-HPV positive infection (60 cases) was significantly lower than that in patients with HR-HPV negative infection (16 cases): 0.41 ± 0.07 vs. 0.49 ± 0.08, the expression level of BMAL1 was significantly higher than that in patients with HR-HPV negative infection: 0.65 ± 0.09 vs. 0.53 ± 0.06, and there were statistical differences ( t = 3.68 and 4.24, P<0.05 or<0.01). In patients with benign cervical diseases, there were no statistical differences in the expression levels of miR-34a and BMAL1 between patients with HR-HPV positive infection (7 cases) and patients with HR-HPV negative infection (43 cases) ( P>0.05). ROC curve analysis result showed that miR-34a combined with BMAL1 had the highest sensitivity (90.4%), specificity (89.9%) and area under curve (0.911) in assessing the 1-year poor prognosis in patients with cervical cancer ( P<0.01), and the optimal cutoff values of miR-34a and BMAL1 expression level were ≤0.39 and ≥0.64. Multivariate Cox regression analysis result showed that poor differentiation, myometrial invasion depth ≥1/2, lymph node metastasis, distant metastasis, FIGO stage Ⅲ+Ⅳ, miR-34a expression level ≤0.46 and BMAL1 expression level ≥0.58 were independent risk factors for death in patients with cervical cancer ( OR = 1.857, 2.125, 2.337, 2.751, 2.457, 3.885 and 3.666; 95% CI 0.845 to 5.788, 0.726 to 5.924, 0.709 to 5.631, 0.693 to 5.727, 0.801 to 5.936, 1.244 to 6.423 and 1.031 to 5.612; P<0.01). Kaplan-Meier survival curve analysis result showed that the median survival time in cervical cancer patients with miR-34a expression level ≤0.39 and BMAL1 expression level ≥0.64 (21 cases) was significantly lower than that in the other cervical cancer patients (miR-34a expression level>0.39 or BMAL1 expression level<0.64, 55 cases): (26.4 ± 4.2) months vs. (34.2 ± 5.6) months, log-rank χ2 = 17.12, P<0.05. Conclusions:The expression level of serum miR-34a in patients with cervical cancer is significantly reduced and the expression level of BMAL1 is significantly increased, which is related to the condition, prognosis and HR-HPV infection. It can be used as a marker for the assessment of the condition and prognosis of cervical cancer. The combined detection of the two can significantly improve the sensitivity and specificity in predicting poor prognosis of cervical cancer.

Objective:This paper explores the predictive value of indicators of cardiopulmonary exercise test in the degree of coronary artery disease in patients with coronary heart disease.It provides an objective basis for predicting the degree of coronary artery disease by non-invasive detection methods in patients with coronary heart disease,and provides objective guidance for formulating exercise prescriptions for patients with coronary heart disease. Method:Fifty-three patients with coronary heart disease who were treated in the Affiliated Cardiovascular Hospi-tal of Kunming Medical University from March 2020 to May 2022 were selected to complete coronary angiogra-phy and cardiopulmonary exercise test.According to the coronary angiography results and the SYNTAX score,the patients were divided into low group(SYNTAX≤22 points)with 30 cases and medium-high group(SYN-TAX＞22 points)with 23 cases.The general information of the patients such as gender,age,BMI and comor-bidities were collected,and the indicators of cardiopulmonary exercise test were collected. Result:Compared with the medium-high group,maximum oxygen uptake(VO2max),maximum oxygen uptake as a percentage of the predicted value(VO2max％P),maximum metabolic equivalent(METSmax),maximum heart rate(HRmax)and maximum heart rate as a percentage of predicted value(HR％P)of the low group were high-er,and the difference between the two groups was statistically significant with p-value less than 0.05.Com-pared with the medium-high group,the heart rate reserve(HRR)in the low group was lower,and the differ-ence between the two groups was statistically significant with p-value less than 0.05.The SYNTAX score was negatively correlated with VO2max,VO2max％P,maximum oxygen uptake in kilograms(VO2max/kg),METSmax,HRmax and HR％P,and positively correlated with HRR with P-value less than 0.05.HRR has the highest sensitivity in predicting coronary heart disease. Conclusion:The low group with low SYNTAX scores and mild coronary lesions has higher VO2max,VO2max％P,VO2max/kg,METSmax,HRmax and HR％P values,suggesting that the patients in this group have better cardiopulmo-nary reserve function and better exercise capacity.The high group with severe coronary artery disease has high-er HRR,and the gap between the maximum heart rate and the target heart rate during exercise is larger,which means it is more difficult for patients in the high group to reach the target heart rate.