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

Background: Epimedii Folium, first recorded in the Shennong’s Classic of Materia Medica (Shen Nong Ben Cao Jing), is a traditional Chinese medicine (TCM) known for its effects of “benefiting Qi and strengthening the heart.” Icariside II (ICS II) is one of the main active components of Epimedii Folium, possessing cardiovascular protective and anti-inflammatory properties. However, the potential mechanisms of ICS II on myocardial ischemia (MI) remain unclear. Objective: The aim of the study was to investigate the effects and preliminary molecular mechanisms of ICS II in treating isoproterenolinduced MI in rats. Methods: A rat model of MI was established by subcutaneous injection of isoproterenol. Electrocardiography, echocardiography, myocardial enzymes analysis, heart weight index, triphenyltetrazolium chloride staining, histopathology, TUNEL staining, RT-qPCR, and Western blot were employed to evaluate the effects and preliminary molecular mechanisms of ICS II on MI rats. Results: Pharmacodynamic studies suggested that ICS II inhibited ST-segment elevation in electrocardiograms, improved cardiac function, reduced heart weight index and myocardial enzyme levels, decreased myocardial infarct size, alleviated cardiac histological damage, and inhibited apoptosis, thereby exerting cardioprotective effects in MI rats. Further studies revealed that ICS II may partially inhibit the expression of NLRP3/Caspase-1 axis-related targets at both protein and mRNA levels. Conclusions: Our findings indicate that ICS II exerts anti-MI effects, and its preliminary molecular mechanisms may be related to inhibiting the activation of the NLRP3/Caspase-1 axis to alleviate inflammatory responses.

Background: Epimedii Folium, first recorded in the Shennong’s Classic of Materia Medica (Shen Nong Ben Cao Jing), is a traditional Chinese medicine (TCM) known for its effects of “benefiting Qi and strengthening the heart.” Icariside II (ICS II) is one of the main active components of Epimedii Folium, possessing cardiovascular protective and anti-inflammatory properties. However, the potential mechanisms of ICS II on myocardial ischemia (MI) remain unclear. Objective: The aim of the study was to investigate the effects and preliminary molecular mechanisms of ICS II in treating isoproterenolinduced MI in rats. Methods: A rat model of MI was established by subcutaneous injection of isoproterenol. Electrocardiography, echocardiography, myocardial enzymes analysis, heart weight index, triphenyltetrazolium chloride staining, histopathology, TUNEL staining, RT-qPCR, and Western blot were employed to evaluate the effects and preliminary molecular mechanisms of ICS II on MI rats. Results: Pharmacodynamic studies suggested that ICS II inhibited ST-segment elevation in electrocardiograms, improved cardiac function, reduced heart weight index and myocardial enzyme levels, decreased myocardial infarct size, alleviated cardiac histological damage, and inhibited apoptosis, thereby exerting cardioprotective effects in MI rats. Further studies revealed that ICS II may partially inhibit the expression of NLRP3/Caspase-1 axis-related targets at both protein and mRNA levels. Conclusions: Our findings indicate that ICS II exerts anti-MI effects, and its preliminary molecular mechanisms may be related to inhibiting the activation of the NLRP3/Caspase-1 axis to alleviate inflammatory responses.

Background: Epimedii Folium, first recorded in the Shennong’s Classic of Materia Medica (Shen Nong Ben Cao Jing), is a traditional Chinese medicine (TCM) known for its effects of “benefiting Qi and strengthening the heart.” Icariside II (ICS II) is one of the main active components of Epimedii Folium, possessing cardiovascular protective and anti-inflammatory properties. However, the potential mechanisms of ICS II on myocardial ischemia (MI) remain unclear. Objective: The aim of the study was to investigate the effects and preliminary molecular mechanisms of ICS II in treating isoproterenolinduced MI in rats. Methods: A rat model of MI was established by subcutaneous injection of isoproterenol. Electrocardiography, echocardiography, myocardial enzymes analysis, heart weight index, triphenyltetrazolium chloride staining, histopathology, TUNEL staining, RT-qPCR, and Western blot were employed to evaluate the effects and preliminary molecular mechanisms of ICS II on MI rats. Results: Pharmacodynamic studies suggested that ICS II inhibited ST-segment elevation in electrocardiograms, improved cardiac function, reduced heart weight index and myocardial enzyme levels, decreased myocardial infarct size, alleviated cardiac histological damage, and inhibited apoptosis, thereby exerting cardioprotective effects in MI rats. Further studies revealed that ICS II may partially inhibit the expression of NLRP3/Caspase-1 axis-related targets at both protein and mRNA levels. Conclusions: Our findings indicate that ICS II exerts anti-MI effects, and its preliminary molecular mechanisms may be related to inhibiting the activation of the NLRP3/Caspase-1 axis to alleviate inflammatory responses.

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 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 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.

Objective:To summary the clinical phenotype and genotype characteristics of 2 cases of autosomal dominant mental retardation (MRD) caused by TRIO gene variation. Methods:Retrospective study of the clinical data of 2 cases of autosomal dominant MRD caused by TRIO gene mutations diagnosed at the Department of Neurology, Children′s Hospital Affiliated to Zhengzhou University in April 2019 and January 2023 was conducted. The clinical features were summarized and gene analysis and follow-up were carried out. Results:The 2 patients were 6 years and 5 months old and 5 months old males, respectively. Clinical manifestations included seizures, cognitive and motor disorders, low intelligent development; case 1 had microcephaly, attention deficit disorder, ataxia, and aggressive behavior, and case 2 had macrocephaly. Brain magnetic resonance imaging revealed cerebellar atrophy in case 1, and non-specific dilation of the subarachnoid space and hypoplasia of the corpus callosum in case 2. Analysis of chromosome karyotype and chromosome copy number variation in 2 children showed no abnormalities. Whole exome sequencing revealed novel missense mutations in the TRIO gene in both patients [NM_007118:c.4289C>A(p.Thr1430Lys), c.4111C>A(p.His1371Asn), respectively]. The application of rehabilitation function training and a variety of anti-seizure medications can not fully and effectively control the seizure. Conclusion:TRIO gene c.4289C>A(p.Thr1430Lys), c.4111C>A(p.His1371Asn) de novo missense variants were the genetic etiology of the 2 probands,causing rare autosomal dominant MRD type 44 and 63.