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

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.

Aspirin,a non-steroidal anti-inflammatory drug,is widely used in clinics with definite therapeutic effects and a few adverse drug reactions.Recent studies demonstrated that other effects besides antipyretic,analgesic,and anti-inflammatory effects have been discovered,such as anti-cancer,anti-cardiovascular diseases,anti-skeletal muscle diseases,and anti-nervous system diseases.This paper reviewed the therapeutic effects and possible mechanisms of aspirin on cancer,cardiovascular disease,muscle-skeletal disease,and nervous system disease based on the related research status at home and abroad.It will provide clues and a theoretical basis for clinical rational drug use and discover novel pharmacological effects of aspirin in the future.

Insufficient catalytic efficiency of flavonoid 6-hydroxylases in the fermentative production of scutellarin leads to the formation of at least about 18% of by-products. Here, the catalytic mechanisms of two flavonoid 6-hydroxylases, CYP82D4 and CYP706X, were investigated by molecular dynamics simulations and quantum chemical calculations. Our results show that CYP82D4 and CYP706X have almost identical energy barriers at the rate-determining step and thus similar reaction rates, while the relatively low substrate binding energy of CYP82D4 may facilitate product release, which is directly responsible for its higher catalytic efficiency. Based on the study of substrate entry and release processes, the catalytic efficiency of the L540A mutation of CYP82D4 increased by 1.37-fold, demonstrating the feasibility of theoretical calculations-guided engineering of flavonoid 6-hydroxylase. Overall, this study reveals the catalytic mechanism of flavonoid 6-hydroxylases, which may facilitate the modification and optimization of flavonoid 6-hydroxylases for efficient fermentative production of scutellarin.
Cytochrome P-450 Enzyme System/metabolism* ; Apigenin ; Glucuronates

Objective:To explore the intervention effect of different modes of ankle pump exercise on femoral vein peripherally inserted central catheter (PICC) catheter-related thrombosis in patients with lung cancer.Methods:Using the convenient sampling method, 96 cases of lung cancer patients with femoral vein PICC catheterization admitted to Xiangya Hospital of Central South University from January 2019 to October 2020 were set as the control group and 97 cases of lung cancer patients with femoral vein PICC cannula from October 2020 to October 2021 were set as the observation group. They adopted the comfort angle ankle pump exercise and the maximum angle ankle pump exercise mode, respectively. The differences in the incidence of catheter-related thrombosis, the incidence of unplanned extubation, fatigue score and catheter indwelling time were compared between the two groups.Results:The incidence of catheter-related thrombosis and unplanned extubation in the observation group were lower than those in the control group, and the differences were statistically significant ( P<0.05) . The fatigue score and catheter indwelling time of the observation group were higher than those of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The maximum angle ankle pump motion mode can reduce the incidence of catheter-related thrombosis and unplanned extubation of femoral vein PICC catheterization in patients with lung cancer and improve the catheter indwelling time, but it is easy to cause patient fatigue, so clinical promotion and use should consider the patients' tolerance.

Currently, there is still no effective curative treatment for the development of late-stage liver fibrosis. Here, we have illustrated that TB001, a dual glucagon-like peptide-1 receptor/glucagon receptor (GLP-1R/GCGR) agonist with higher affinity towards GCGR, could retard the progression of liver fibrosis in various rodent models, with remarkable potency, selectivity, extended half-life and low toxicity. Four types of liver fibrosis animal models which were induced by CCl₄, α-naphthyl-isothiocyanate (ANIT), bile duct ligation (BDL) and Schistosoma japonicum were used in our study. We found that TB001 treatment dose-dependently significantly attenuated liver injury and collagen accumulation in these animal models. In addition to decreased levels of extracellular matrix (ECM) accumulation during hepatic injury, activation of hepatic stellate cells was also inhibited via suppression of TGF-β expression as well as downstream Smad signaling pathways particularly in CCl₄-and S. japonicum-induced liver fibrosis. Moreover, TB001 attenuated liver fibrosis through blocking downstream activation of pro-inflammatory nuclear factor kappa B/NF-kappa-B inhibitor alpha (NFκB/IKBα) pathways as well as c-Jun N-terminal kinase (JNK)-dependent induction of hepatocyte apoptosis. Furthermore, GLP-1R and/or GCGR knock-down results represented GCGR played an important role in ameliorating CCl₄-induced hepatic fibrosis. Therefore, TB001 can be used as a promising therapeutic candidate for the treatment of multiple causes of hepatic fibrosis demonstrated by our extensive pre-clinical evaluation of TB001.

Objective:To investigate the present situation of unintended pregnancy within two years postpartum and its influencing factors in China.Methods:Participants who delivered a live birth at 60 hospitals in 15 provinces in the eastern, central and western regions of China during July 2015 to June 2016 were interviewed by using structured questionnaire. Information on occurrence of unintended pregnancy within 2 years after delivery, postpartum contraceptive use, sexual resumption, breastfeeding, and women′s socio-demographic characteristics, and so on, were collected. Life-table analysis, cluster log-rank tests and a 2-level Cox regression model were used for data analysis.Results:A total of 18 045 postpartum women were investigated. The cumulative 1- and 2-year unintended pregnancy rates after delivery were 5.3% (95% CI: 4.5%－6.1%) and 13.1% (95% CI: 11.3%－14.8%), respectively. Cox regression model analysis showed that the risk of unintended pregnancy within 2 years postpartum were increased in younger women, ethnic minorities, women with abortion history, and those who had a vaginal delivery with short lactation time and late postpartum contraceptive initiation (all P<0.01). The risk of postpartum unintended pregnancy was not associated with geographic regions and hospitals where women gave a birth (all P>0.05). Conclusions:In China, the risk of unintended pregnancy within 2 years after delivery is relatively high. Service institutions and service providers should improve the quality of postpartum family planning services, promote the use of high effect contraceptive methods, and educate women to use a method at the time of their sexual resumption or even before.

OBJECTIVES: To explore the expression of autophagy related genes 5 (ATG5) and cyclin E in coronary heart disease (CHD) and its clinical significance. METHODS: From April 2018 to August 2018, 80 patients diagnosed with CHD in the Second Xiangya Hospital, Central South University were selected as an observation group, and another 80 healthy subjects were selected as a control group. The expression of ATG5 and cyclin E mRNA in nucleate cells and the plasma protein in the 2 groups were detected and analyzed. The model of macrophage-derived foam cells induced by oxidized low density lipoprotein (ox-LDL) was used to simulate atherosclerosis. The proliferation of macrophage- derived foam cells and the protein levels of ATG5 and cyclin E induced by ox-LDL at different concentrations were examined. RESULTS: Compared with the control group, the levels of ATG5 mRNA and protein in the blood in the observation group were decreased, and the cyclin E mRNA and protein levels were increased, there were statistically difference (both <0.05). Receiver operating characteristic (ROC) curve showed that the area under curve (AUC) of ATG5 mRNA, cyclin E mRNA, ATG5 protein and cyclin E protein were 0.739, 0.780, 0.671 and 0.807, respectively. Pearson analysis showed that the ATG5 mRNA was negatively correlated with the cyclin E mRNA (=-0.734, <0.05),while the plasma ATG5 protein was negatively correlated with the plasma cyclin E protein (=-0.746, <0.05). Macrophage-derived foam cell model induced by ox-LDL showed that the proliferation of foam cells and the expression levels of cyclin E protein were increased in a concentration and time-dependent manner, and the expression levels of ATG5 protein were decreased in a concentration-dependent manner. CONCLUSIONS The levels of ATG5 mRNA and protein are lowly expressed while the levels of cyclin E mRNA and protein are highly expressed in the patients with CHD.The ATG5 protein levels are lowly expressed in ox-LDL-treated macrophage-derived foam cells while the cyclin E protein levels are highly expressed in ox-LDL-treated macrophage-derived foam cells. Based on these observations, we conclude that ATG5 inhibits the degradation of the cyclin E and promotes the proliferation of macrophages, involving in the occurrence and development of CHD.
Autophagy ; Autophagy-Related Protein 5 ; Coronary Disease ; Cyclin E ; Foam Cells ; Humans ; Lipoproteins, LDL

Objective To investigate the effects of reflection diary training on nurse's clinical decision-making capability.Methods A total of 489 nurses in our hospital were enrolled in the study for tests of their clinical decision capability between July 2013 and July 2015.Two hundred forty-eight of them were set as the experiment group and two hundred forty-one of them were set as the control group at random.The reflection diary training was applied in the experiment group for one year.Result After intervention,the scores on clinical decision-making ability,clinical thinking,knowledge structure,ability in nurse-patient communication were all significantly higher than those of control group (P＜0.01).Conclusion The reflection diary training can improve nurse's clinical decision-making capability and inspire their clinical thinking about the nursing problems.

Objective:The paper aims at developing a method of defining and visualizing the scope of the basic medical insurance pharmacy service, and provides a new way of thinking for the designated pharmacy planning. Methods:Collecting the basic data and information on administrative divisions in the planning area taking equity and efficiency as the guidance, using ArcGIS and its function modules to define and visualize the scope of the medical in-surance pharmacy service. The procedure of issue focus, method improvement, data simulation, expert consultation, methodology perfecting were followed to define and visualize the scope. Results:Forming a whole set of operative pro-cedures of defining and visualizing the scope of the medical insurance pharmacy service based on medical resources allocation standard, and the operation commands and procedures in ArcGIS were clarified. Conclusion:Operating ac-cording to the appropriate method steps, the following can be achieved:(1) The adjacent scope of medical insurance pharmacy service are adjacent to each other but do not overlap or cross;(2) Spatial relations can be clearly and ef-fectively expressed;(3) The shape is flat and regular;(4) The data collected at different times can be comparable in space, providing good prerequisites for medical insurance designated pharmacy planning.