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

ObjectiveBased on non-targeted metabolomics， to analyze the regulation of endogenous differential metabolites in serum of type 2 diabetes mellitus（T2DM） rats by Fuling Yunhua granules， and to clarify the metabolic pathways through which this granules exerted its effect on improving T2DM. MethodSeventy SD rats， half male and half female， were randomly divided into the control group， model group， and high， medium， low dose groups of Fuling Yunhua granules（20.70， 10.35， 5.18 g·kg^-1 in raw drug amount） and the positive drug group（pioglitazone hydrochloride tablets， 8.1 mg·kg^-1）. Except for the control group， other groups were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin（STZ） to establish a T2DM rat model. After successful modeling， the treatment groups were administered the corresponding drugs by gavage， and the control group and model group were treated with an equal volume of saline by gavage， once/d， for 28 d. Fasting blood glucose（FBG） and glycosylated hemoglobin A1c（GHbA1c） levels were measured in all groups of rats during the administration period， and hematoxylin-eosin（HE） staining was used to observe the pathomorphological changes in the pancreatic tissues of rats at the end of the administration period. The endogenous metabolite levels in rat serum were detected by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-LTQ-Orbitrap MS）， and the data were processed using principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）. Differential metabolites were identified by the Human Metabolome Database（HMDB） and the Kyoto Encyclopedia of Genes and Genomes（KEGG）， and screened for differential metabolites with variable importance in the projection（VIP） value>1， P<0.05， and fold change（FC）<0.6 or FC>1. And the metabolic pathway enrichment analysis of the screened differential metabolites was performed by MetaboAnalyst 5.0， then the screened differential metabolites were diagnosed and evaluated by the receiver operating characteristic（ROC） curves. ResultCompared with the control group， the FBG level of rats in the model group increased significantly（P<0.01）， the GHbA1c content tended to increase， but the difference was not statistically significant， and the pancreatic tissue of rats was obviously damaged， the number of pancreatic islets decreased， and the pancreatic β-cells were obviously reduced， atrophied and enlarged. Compared with the model group， the FBG levels of rats in the high dose group of Fuling Yunhua granules and the positive drug group were significantly reduced after 2 weeks of administration（P<0.05， P<0.01）， the GHbA1c content of rats in the high dose group of Fuling Yunhua granules was significantly reduced（P<0.05）， and the pancreatic tissue lesions of rats in the different dose groups of Fuling Yunhua granules were reduced. The results of non-targeted metabolomics showed that 46 differential metabolites were significantly changed in the model group compared with the blank group. Pathway enrichment analysis found that T2DM mainly affected biological processes including biosynthesis of primary bile acid， D-amino acid metabolism， steroid hormone biosynthesis， and glycerophospholipid metabolism in rats. Compared with the model group， the levels of 8 differential metabolites in the high dose group of Fuling Yunhua granules were significantly adjusted， and the pathway enrichment analysis found that D-amino acid metabolism， retinol metabolism， glycine， serine and threonine metabolism， tryptophan metabolism and other metabolic pathways were mainly involved. ROC curves further analysis revealed that the four characteristic differential markers of 11-cis-retinol， D-piperidinic acid， D-serine， and p-cresol sulfate had high diagnostic value for the treatment of T2DM with Fuling Yunhua granules. ConclusionFuling Yunhua granules can improve the symptoms of T2DM rats by regulating the amino acid metabolic and retinol metabolic pathways through the modulation of endogenous differential metabolites.

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with unclear etiology and limited treatment options. The median survival time for IPF patients is approximately 2-3 years and there is no effective intervention to treat IPF other than lung transplantation. As important components of lung tissue, endothelial cells (ECs) are associated with pulmonary diseases. However, the role of endothelial dysfunction in pulmonary fibrosis (PF) is incompletely understood. Sphingosine-1-phosphate receptor 1 (S1PR1) is a G protein-coupled receptor highly expressed in lung ECs. Its expression is markedly reduced in patients with IPF. Herein, we generated an endothelial-conditional S1pr1 knockout mouse model which exhibited inflammation and fibrosis with or without bleomycin (BLM) challenge. Selective activation of S1PR1 with an S1PR1 agonist, IMMH002, exerted a potent therapeutic effect in mice with bleomycin-induced fibrosis by protecting the integrity of the endothelial barrier. These results suggest that S1PR1 might be a promising drug target for IPF therapy.

BACKGROUND: In recent years, immunotherapy represented by programmed cell death 1 (PD-1)/programmed cell death ligand 1 (PD-L1) immunosuppressants has greatly changed the status of non-small cell lung cancer (NSCLC) treatment. PD-L1 has become an important biomarker for screening NSCLC immunotherapy beneficiaries, but how to easily and accurately detect whether PD-L1 is expressed in NSCLC patients is a difficult problem for clinicians. The aim of this study was to construct a Nomogram prediction model of PD-L1 expression in NSCLC patients based on 18F-fluorodeoxy glucose (18F-FDG) positron emission tomography/conputed tomography (PET/CT) metabolic parameters and to evaluate its predictive value. METHODS: Retrospective collection of 18F-FDG PET/CT metabolic parameters, clinicopathological information and PD-L1 test results of 155 NSCLC patients from Inner Mongolia People's Hospital between September 2016 and July 2021. The patients were divided into the training group (n=117) and the internal validation group (n=38), and another 51 cases of NSCLC patients in our hospital between August 2021 and July 2022 were collected as the external validation group according to the same criteria. Then all of them were categorized according to the results of PD-L1 assay into PD-L1+ group and PD-L1- group. The metabolic parameters and clinicopathological information of patients in the training group were analyzed by univariate and binary Logistic regression, and a Nomogram prediction model was constructed based on the screened independent influencing factors. The effect of the model was evaluated by receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis (DCA) in both the training group and the internal and external validation groups. RESULTS: Binary Logistic regression analysis showed that metabolic tumor volume (MTV), gender and tumor diameter were independent influences on PD-L1 expression. Then a Nomogram prediction model was constructed based on the above independent influences. The ROC curve for the model in the training group shows an area under the curve (AUC) of 0.769 (95%CI: 0.683-0.856) with an optimal cutoff value of 0.538. The AUC was 0.775 (95%CI: 0.614-0.936) in the internal validation group and 0.752 (95%CI: 0.612-0.893) in the external validation group. The calibration curves were tested by the Hosmer-Lemeshow test and showed that the training group (χ2=0.040, P=0.979), the internal validation group (χ2=2.605, P=0.271), and the external validation group (χ2=0.396, P=0.820) were well calibrated. The DCA curves show that the model provides clinical benefit to patients over a wide range of thresholds (training group: 0.00-0.72, internal validation group: 0.00-0.87, external validation group: 0.00-0.66). CONCLUSIONS The Nomogram prediction model constructed on the basis of 18F-FDG PET/CT metabolic parameters has greater application value in predicting PD-L1 expression in NSCLC patients.
Humans ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Positron Emission Tomography Computed Tomography ; Lung Neoplasms/drug therapy* ; Fluorodeoxyglucose F18/therapeutic use* ; Nomograms ; Retrospective Studies ; B7-H1 Antigen/metabolism* ; Glucose/therapeutic use* ; Positron-Emission Tomography/methods*

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.

Humans ; Blood Pressure/physiology* ; Cohort Studies ; Patient Discharge ; Ischemic Stroke ; Stroke ; Brain Ischemia ; Diabetes Mellitus ; Hypertension

Objective:To explore the effect of the combined effect of noise and heat on occupational hearing loss of workers by using Meta-analysis method.Methods:In August 2020, the Chinese and English literature on the relationship between exposure to noise and heat and occupational hearing loss published from January 2005 to August 2020 by CNKI, China Biomedical Literature Service System, Wanfang Data Knowledge Service Platform, VIP Official Database, Medline and PubMed Databases were searched, using noise, heat or hyperthermia, hearing as keywords. The selected data were analyzed by Stata 12.0 software, and the combined OR (95% CI) value included in the literature was calculated. Sensitivity analysis was used to explore the source of heterogeneity and analyze publication bias. Results:A total of 14 literatures (14 in Chinese, 0 in English) were included in the analysis, and 38654 subjects were included, including 6411 workers in the noise and heat combined effect group and 32243 workers in the noise alone group. The probability of hearing loss in the noise and heat combined effect group was 1.39 times higher than that in the noise alone group (95% CI: 1.14-1.69). The effect size OR was stable after sensitivity analysis, and there was no publication bias in the included literatures tested by Egger's and Begg's Method ( z=0.38, P=0.702, t=-0.74, P=0.476) . Conclusion:Simultaneous exposure to noise and heat may increase the risk of hearing loss for workers in noisy workplaces.

Objective:To explore the feasibility of using the sigma metrics calculated with the data of internal quality control for the comparison of the analytical performance between different biochemical analyzers.Methods:The internal quality control results of twenty-five biochemical assays in the biochemical analyzers of the department of clinical laboratory in Cancer Hospital from February 1, 2021 to July 31, 2021 were collected. The formula sigma =( TEa- Bias)/ CV was used to calculate the sigma metrics of two different levels of the biochemical assays including albumin, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, calcium, cholesterol, creatine kinase, chlorine, creatinine, γ- glutamyltranspeptidase, blood glucose, high density lipoprotein cholesterol, immunoglobulin A, immunoglobulin G, immunoglobulin M, potassium, lactate dehydrogenase, low density lipoprotein cholesterol, sodium, inorganic phosphorus, total bilirubin, triglyceride, total protein, urea, uric acid. The imprecision was obtained by the coefficient of variation of internal quality control. The bias was calculated by the deviation between the mean of internal quality control of the comparison instrument and the target instrument. The allowable total error ( TEa) was based on People's Republic of China Health Industry Standard (WS/T403-2012) or EQA standard of National Center for Clinical Laboratories (NCCL). Compared the sigma values of the comparison instrument relative to the target instrument with the average percentage bias obtained by the traditional comparison method. Quality goal index was used to analyze the causes of poor performance and judge the results of comparison. Results:Compared with the target instrument Beckman AU5800-3, the comparison instrument Beckman AU5800-1 had 10 assays with σ>6, accounting for 40%, 23 assays with σ>3, accounting for 92%, and only albumin and blood glucose showed σ<3. Through statostical analysis, the comparisons of all assays were passed. The comparison instrument Beckman AU5800-2 had 8 assays with σ>6, accounting for 32%, 20 assays with σ>3, accounting for 80%. Only alkaline phosphatase, calcium, lactate dehydrogenase, total protein and urea showed σ<3. Through statostical analysis, the comparisons of GGT and IgM failed. For the traditional comparison method, the percentage bias between the comparison instruments and the target instrument were all within the range of the evaluation standard. But there was no significant correlation between the σ value and the average bias of the traditional comparison method, and the biases were correlated.Conclusions:Using the sigma metrics calculated with the data of internal quality control for the comparison of different detection systems is a convenient and operable method. It can monitor the comparability between different detection systems in the laboratory at any time and be the supplement of the traditional comparison method.

Hepatocellular carcinoma (HCC) is an aggressive human cancer with increasing incidence worldwide. Multiple efforts have been made to explore pharmaceutical therapies to treat HCC, such as targeted tyrosine kinase inhibitors, immune based therapies and combination of chemotherapy. However, limitations exist in current strategies including chemoresistance for instance. Tumor initiation and progression is driven by reprogramming of metabolism, in particular during HCC development. Recently, metabolic associated fatty liver disease (MAFLD), a reappraisal of new nomenclature for non-alcoholic fatty liver disease (NAFLD), indicates growing appreciation of metabolism in the pathogenesis of liver disease, including HCC, thereby suggesting new strategies by targeting abnormal metabolism for HCC treatment. In this review, we introduce directions by highlighting the metabolic targets in glucose, fatty acid, amino acid and glutamine metabolism, which are suitable for HCC pharmaceutical intervention. We also summarize and discuss current pharmaceutical agents and studies targeting deregulated metabolism during HCC treatment. Furthermore, opportunities and challenges in the discovery and development of HCC therapy targeting metabolism are discussed.

Objective:To explore the effect of the combined effect of noise and heat on occupational hearing loss of workers by using Meta-analysis method.Methods:In August 2020, the Chinese and English literature on the relationship between exposure to noise and heat and occupational hearing loss published from January 2005 to August 2020 by CNKI, China Biomedical Literature Service System, Wanfang Data Knowledge Service Platform, VIP Official Database, Medline and PubMed Databases were searched, using noise, heat or hyperthermia, hearing as keywords. The selected data were analyzed by Stata 12.0 software, and the combined OR (95% CI) value included in the literature was calculated. Sensitivity analysis was used to explore the source of heterogeneity and analyze publication bias. Results:A total of 14 literatures (14 in Chinese, 0 in English) were included in the analysis, and 38654 subjects were included, including 6411 workers in the noise and heat combined effect group and 32243 workers in the noise alone group. The probability of hearing loss in the noise and heat combined effect group was 1.39 times higher than that in the noise alone group (95% CI: 1.14-1.69). The effect size OR was stable after sensitivity analysis, and there was no publication bias in the included literatures tested by Egger's and Begg's Method ( z=0.38, P=0.702, t=-0.74, P=0.476) . Conclusion:Simultaneous exposure to noise and heat may increase the risk of hearing loss for workers in noisy workplaces.