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 evaluate the clinical value of transarterial catheterization C-arm CT perfusion scanning technique during prostatic artery embolization(PAE)for benign prostatic hyperplasia(BPH).Methods The clinical data of 46 patients with BPH received PAE were analyzed retrospectively.All patients underwent prostatic artery(PA)digital subtraction angiography(DSA)and C-arm CT perfusion scanning to identify PA and prevent non-target organ embolization.The final recognization of PA was consulted by three senior doctors.After C-arm CT confirmation,PA was embolized with 100-300 μm polyvinyl alcohol(PVA)particles or microspheres under fluoroscopy.The postoperative complications and 3-month clinical efficacy were observed.Results A total of 106 vessels were angioraphed in 46 patients,with 83 PA vessels and 23 non-PA vessels.PA was identified by DSA and C-arm CT with sensitivity of 81.9%(68/83)and 100%(83/83),respectively,which showed significance(χ2=22.3,P<0.01).Non-PA was identified by DSA and C-arm CT with specificity of 73.9%(17/23)and 100%(23/23),which showed significance(χ2=9.2,P=0.02).No serious complications were observed and 3-month clincial efficacy was 91.3%.Conclusion Transarterial catheterization C-arm CT perfusion scanning technique can accurately identify PA,reduce PA leakage and prevent non-target organ embolization.

Objective To evaluate the efficacy of microwave ablation(MWA)synchronously with biopsy for pulmonary nodules.Methods The data of 64 patients with MWA combined with biopsy were analyzed retrospectively.Thirty-one patients(non-synchronous group)were treated with ablation following biopsy in turn to identify malignant tumors,and 33 patients(synchronous group)were treated by ablation and biopsy synchronously.The technical success rate,operation time,complications,hospitalization time and expenses were compared between non-synchronous group and synchronous group.Results The technical success rate,pneumothorax,and pleural effusion rate showed no significance between the two groups(P>0.05).There were all significant differences in operation time(42.00 min vs 54.26 min),hospitalization time(5.09 days vs 9.26 days),hospitalization expenses(26 840.61 yuan vs 32 527.26 yuan),lung hemorrhage(27.27%vs 87.10%)and hemoptysis(3.03%vs 19.35%)between synchronous group and non-synchronous group,respectively(P<0.05).Conclusion MWA synchronously with biopsy for pulmonary nodules is safe and feasible,which can reduce intraoperative bleeding,shorten treatment period and reduce hospitalization expenses.

Objective To explore the efficiency and safety of trans-sheath intraluminal forceps biopsy under digital subtraction angiography(DSA)guidance for assisting diagnosis of pulmonary artery obstructive diseases.Methods Data of 16 patients who underwent trans-sheath intraluminal forceps biopsy for pulmonary artery obstructive diseases were retrospectively analyzed,and the clinical manifestations were recorded.The technical success of biopsy was defined as tissue obtained met the needs of pathology diagnosis.For patients with malignant pathology results,the final diagnosis was malignant,for those with benign pathology results after biopsy and no obvious changes after 6-month or longer follow-up,or benign pathology results after surgical resection,the final diagnosis was benign,otherwise was no clear diagnosis.The operation time,technical success rate,diagnostic efficiency,complications and changes of pulmonary artery pressure before and after the biopsy were observed.Results Among 16 patients,9 complained of intermittent chest tightness,4 complained of chest pain with chest tightness,2 complained of chest pain but 1 denied any symptoms.The lesions located in the left lung in 10 cases and in the right lung in 6 cases,all with enhanced CT showed filling defects of the involved branch of pulmonary artery.Totally 16 trans-sheath intraluminal forceps biopsies were performed in 16 patients,with an average operation time of(31.02±6.02)min and technical success rate of 100%.Malignant tumors were finally diagnosed in 10 cases,including 1 case of lung cancer with false-negative biopsy result,while biopsy correctly diagnosed benign lesions in the other 6 cases.Transient worsening chest pain with chest tightness occurred in 2 cases and relieved after symptomatic treatments.No statistically significant difference of pulmonary artery pressure was found before([53.38±14.28]mmHg)and after([53.69±14.15]mmHg)biopsy(P＞0.05).Conclusion DSA-guided trans-sheath intraluminal forceps biopsy was relatively safe and valuable for assisting diagnosis of pulmonary artery obstructive diseases.

Objective:To investigate the status and influencing factors of different types of cognitive frailty in aged inpatients with hypertension.Methods:Totally 300 aged patients with hypertension admitted to the Cardiology, Geriatrics, and Endocrinology Departments of Xuanwu Hospital, Capital Medical University, from April to October 2022 were selected by convenience sampling. Data were collected using a general information questionnaire, the Fried Frailty Phenotype (FP), the Mini-Mental State Examination (MMSE), the 9-item Subjective Cognitive Decline Questionnaire-9 (SCD-Q9), the Morisky Medication Adherence Scale (MMAS), the Pittsburgh Sleep Quality Index (PSQI), the Hospital Anxiety and Depression Scale (HADS), the Social Support Rating Scale (SSRS), and the Mini Nutritional Assessment (MNA) .Results:A total of 300 questionnaires were distributed and returned, yielding a 100.00% response rate. Among the 300 aged hypertensive patients, 74 were diagnosed with reversible cognitive frailty (RCF) and 98 with potentially reversible cognitive frailty (PRCF). Logistic vegression analysis showed that, factors influencing RCF included daily exercise habits and the use of thiazide diuretics ( P<0.05) ; factors influencing PRCF included age, pre-retirement occupation, daily exercise habits, sleep quality, Barthel Index score, and serum albumin levels ( P<0.05) . Conclusions:The status of both RCF and PRCF is relatively high among aged inpatients with hypertension. The influencing factors for different types of cognitive frailty vary. Targeted interventions based on both common and specific influencing factors should be developed to reverse or slow the progression of cognitive frailty in aged hypertensive patients.

Objective:To observe the proteomic changes in vitreous fluid samples from patients with rhegmatogenous retinal detachment combined with choroidal detachment (RRDCD).Methods:A prospective cross-sectional clinical study. Vitreous fluid samples were collected from 35 patients with RRDCD (RRDCD group) and 40 patients with rhegmatogenous retinal detachment (RRD group) who were diagnosed at Wuhan Aier Eye Hospital between November 2021 and December 2023. Prior to vitrectomy, 0.3-0.5 ml of vitreous fluid was collected from the affected eyes. Differentially expressed proteins were analyzed using Data-Independent Acquisition (DIA). Three of these proteins were randomly selected for validation using enzyme-linked immunosorbent assay (ELISA). Bioinformatics analyses, including gene ontology functional enrichment and kyoto encyclopedia of genes and genomes pathway enrichment, were performed to explore the functions of the differentially expressed proteins.Results:Significant differences were observed between the RRDCD and RRD groups in intraocular pressure ( t=-12.795), the number of retinal tears ( t=4.601), the extent of retinal detachment ( χ2=39.642), axial length ( t=0.840), postoperative proliferative vitreoretinopathy incidence ( χ2=4.730), single-surgery reattachment rate ( χ2=7.717), and best-corrected visual acuity ( t=7.033) at 6 months postoperatively ( P<0.05). A total of 237 differentially expressed proteins were identified between the RRDCD and RRD groups, with 63 upregulated and 174 downregulated. These proteins were involved in pathways such as extracellular matrix-receptor interaction, complement activation, coagulation, and lysosomal pathways. ELISA validation results showed that the expression trends of the three selected proteins in the RRDCD and RRD groups were consistent with the DIA proteomic analysis. Compared to the RRD group, proteins such as fibrin, coagulation factors, cathepsins, and trypsin inhibitors were significantly upregulated in the RRDCD group. Conclusions:The protein expression profile in vitreous fluid samples from RRDCD patients show significant alterations compared to the RRD group. These differential changes suggest that RRDCD is closely associated with complement and coagulation cascade activation, lysosomal pathways, and extracellular matrix remodeling.

Objective:To observe the diurnal changes of serum hypoxia inducible factor-1α (HIF-1α) and its relationship with polysomnography (PSG) parameters in obese patients with obstructive sleep apnea (OSA).Methods:In this study, 64 obese subjects with OSA (experimental group) and 24 normal subjects (control group) were included. PSG monitoring was performed in all subjects, and peripheral blood samples were collected at night and next early morning to detect the expression level of HIF-1 α mRNA and protein concentration in serum. The data were analyzed by SPSS25.0 software. The counting data was represented by n(%), and t-tests were used for inter group comparisons. The measurement data of normal distribution adopted mean ± standard deviation ( ± s), and the inter group comparison adopted Chi-square. Serum HIF-1α mRNA levels and protein concentrations in the two groups were compared and analyzed by Two-way ANOVA, and Spearman rank correlation was used to analyze the correlation. Results:The ratio of male to female in the control group was 15: 9 and the average age was (39.25 ±10.58) years old, while that in the experimental group was 41∶24 with an average age of (41.31 ±15.20) years. The body mass index (BMI) of experimental group was higher, while all the relevant parameters recorded by PSG were significantly different between two groups ( P<0.001). The results showed that there was no significant difference in the expression of HIF-1α mRNA between the control group and the experimental group ( P=0.914), or between night and morning ( P=0.167). The concentration of HIF-1α protein in the control group was significantly lower than that in the experimental group ( P<0.001), but there was no significant difference in the concentration of HIF-1α protein in the same group at night and in the morning ( P=0.798). The results of correlation analysis showed that there was a significant correlation between PSG related indexes(AHI, minimum blood oxygen, mean blood oxygen and SIT90), night and morning mRNA expression and protein concentration ( P<0.05). Conclusions:Compared with normal people, the concentration of serum HIF-1 α protein in obese patients with OSA is higher, but there is no difference between night and morning. The process of HIF-1 α induced by CIH may be involved in post-translational regulation rather than transcription, and other compensatory mechanisms may also be involved.

Objective To investigate the independent influences on the occurrence of ICU Acquired Weakness(ICU-AW)in elderly patients undergoing third-and fourth-level surgeries,to construct a prediction model and draw a column-line diagram,and to internally validate the model.Methods A convenience sampling method was used to select 186 elderly patients undergoing tertiary and quaternary surgeries who were hospitalized in 3 tertiary A hospitals in Beijing from May to December 2022 as the study subjects.Single-factor and multifactor logistic regression were used to analyze the risk factors for ICU-AW in elderly patients undergoing third-and fourth-degree surgeries.A risk prediction model was established and the model was visualized by drawing a column-line diagram,and the receiver operator characteristic curve(ROC)and the Hosmer-Lemeshow tests were applied to verify the predictive effect of the model.Results ICU-AW occurred in 40 of 186 cases in the modeling group,with an incidence rate of 21.5％.The results of univariate analysis showed that the 2 groups of preoperative physiology score and surgical severity score included in the physiology and surgical severity scoring system,age,presence of cardiac disease,hemoglobin(within 24 h of admission to the ICU),blood urea nitrogen(within 24 h of admission to the ICU),blood creatinine(within 24 h of admission to the ICU),presence of braking,mode of establishment of mechanical ventilation,presence of nutritional therapy,number of sedative or analgesic drugs used,whether vasoactive drugs were used,whether diuretics were used,and the level of hemoglobin,blood urea nitrogen and blood creatinine within 24 h after admission to ICU the difference is statistically significant(P＜0.05).The results of multifactorial logistic regression analysis showed that preoperative physiology scores included in the physiology and surgical severity scoring system,the presence of cardiac disease,the presence of braking,the presence of nutritional therapy,and the number of sedative or analgesic medications used were the predictors of the occurrence of ICU-AW in elderly patients undergoing third-and fourth-degree surgeries(OR were 1.364,2.344,5.568,5.823,1.109,all P＜0.05).The above 5 factors were plotted as independent variables in a column-line graph,and the area under the ROC curve of the model was 0.859(95％CI 0.793～0.924),with an optimal critical value of 0.156,a sensitivity of 0.875,a specificity of 0.705,and a Hosmer-Lemeshow goodness-of-fit test of x2=3.906,P=0.865,Brier score of 0.109,and a decision analysis curve indicating that patients could benefit.Conclusion The predictive effect of the constructed model is good,and it can be used as a reference for early and rapid identification of the risk of ICU-AW in elderly patients undergoing third-and fourth-degree surgeries by clinical staff,and timely provision of preventive intervention programs.

Objective:To carry out high-risk foot examination and grading combined with two examination methods for inpatients with type 2 diabetes, and explore the influencing factors of the occurrence and development of high-risk foot, and investigate their foot care behavior status.Methods:From July 2021 to January 2022, 409 patients with diabetes who were admitted to the Department of Endocrinology, Department of Geriatrics, Department of Cardiology, Urology Surgery and Department of Ophthalmology of Xuanwu Hospital of Capital Medical University were selected as research subjects by convenience sampling. The General Information Questionnaire, Standardized Process of At-risk Foot Screening and Stratification for Diabetic Patients, InIow's Screening for the High-Risk Diabetic Foot: A 60-Second Tool, and Foot Care Behavior Questionnaire for Diabetic Patients. Single factor analysis and Logistic regression analysis were used to explore the factors influencing the occurrence and development of high-risk foot in type 2 diabetes patients. A total of 409 questionnaires were distributed, and 392 valid questionnaires were collected, with an effective response rate of 95.8% (392/409) .Results:Among 392 patients, the detection rate of high-risk diabetic foot (HRDF) was 76.3% (299/392), and the proportion of high-risk foot grade 2 was the largest (193). Age ( OR=1.042, P<0.01) and years of hypertension ( OR=1.030, P<0.05) were independent influencing factors for the occurrence of HRDF, with statistically significant differences. Taking the high risk foot grade 3 as a reference, cerebrovascular disease [ OR=16.408, 95% CI (1.323, 203.417) ], diabetes course [ OR=1.066, 95% CI (1.008, 1.128) ], education level in middle school [ OR=0.180, 95% CI (0.056, 0.581) ], education level in primary school and below [ OR=0.126, 95% CI (0.019, 0.841) ] were independent influencing factors for the progress of high risk foot. The foot care behavior of high-risk foot patients with high risk levels was not superior to that of patients with low risk levels. Conclusions:The combination of the two screening methods can meet the examination needs of clinical and nursing medical staff for inpatients with type 2 diabetes. Emphasizing the influencing factors of HRDF occurrence and development can provide reference for early identification of high-risk foot.

Objective:To explore the correlation between frailty and foot care behavior in elderly patients with high-risk diabetic foot.Methods:From January to June 2022, 220 patients with high-risk diabetic foot who were admitted to the Department of Endocrinology and Department of Geriatrics of Xuanwu Hospital of Capital Medical University were selected by convenience sampling as the research object. The patients were investigated with the General Information Questionnaire, Gavin's Weighted Scale for Diabetic Foot Risk Factors for Progression to Ulceration, the Chinese version of the Frail Scale and the Foot Care Behavior Questionnaire for Diabetic Patients. Spearman correlation analysis was used to explore the correlation between frailty and foot care behavior in elderly patients with high-risk diabetic foot. Multiple linear regression was used to analyze the influencing factors of foot care behavior in elderly patients with high-risk diabetic foot. A total of 220 questionnaires were distributed, and 210 valid questionnaires were collected, with an effective response rate of 95.45% (210/220) .Results:The standardized score of the Foot Care Behavior Questionnaire for Diabetic Patients among 210 elderly patients with high-risk diabetic foot was (56.65±11.27), which was in the middle to low level. Among them, 126 patients (60.00%) were at a low level, and 80 patients (38.10%) were at a middle level. The incidence of frailty in 210 elderly patients with high-risk diabetic foot was 27.14% (57/210). The results of correlation analysis showed that the frailty score of elderly patients with high-risk diabetic foot were negatively correlated with the scores of the foot and footwear examination, foot cleaning and maintenance, footwear selection, and the total score of Foot Care Behavior Questionnaire for Diabetic Patients ( P<0.05). The results of multiple linear regression analysis showed that gender, frailty, foot risk classification and living conditions were the influencing factors of foot care behavior in elderly patients with high-risk diabetic foot ( P<0.05) . Conclusions:The foot care behavior of elderly patients with high-risk diabetic foot needs to be improved. The higher the degree of frailty, the lower the level of foot care behavior. Medical and nursing staff should formulate targeted intervention measures according to the characteristics of patients to improve or delay the progression of patients' frailty, thereby improving their foot care behavior and preventing the occurrence of diabetic foot.