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

Bacterial therapy has attracted increasing attention due to its special mechanism and abundant applications. With the flourishing development of synthetic biology, therapeutic genes have been introduced into engineering bacteria to improve their antitumor efficacy. However, it is difficult to spatiotemporally control the expression of these therapeutic genes at the tumor site in vivo, thereby considerably limiting the application of engineered bacteria in tumor treatment. To resolve this problem, we constructed a temperature-responsive bacterial strain capable of triggering the expression of exogenous genes in a laser-controllable way. Noxa, a pro-apoptotic protein, is chosen to test the expression of exogenous protein and its anti-tumor effect in engineered bacteria upon laser irradiation. Firstly, Noxa was fused to the C-terminus of the bacterial outer membrane protein cytolysin A (ClyA), and then the recombinant gene fragment ClyA-Noxa was inserted into the temperature-sensitive plasmid pBV220 and the recombinant plasmid was transformed into non-pathogenic Escherichia coli MG1655. Thus, we constructed the engineering strain (TRB@Noxa) that could express Noxa on the bacterial surface. TRB@Noxa could target and colonize the tumor tissue without causing notable host toxicity. The bacterial infection triggered thrombosis in the tumor tissue, resulting in the darkness of tumor sites. In a xenograft mouse tumor model, our strategy demonstrated precise tumor targeting and strong tumor inhibition. In conclusion, we successfully constructed a new engineering bacterial strain TRB@Noxa. TRB@Noxa combined with photothermal therapy could arrest tumor growth in the absence of photosensitizers, which represents an appealing method for antitumor therapy in the future.
Escherichia coli/radiation effects* ; Animals ; Humans ; Lasers ; Mice ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Neoplasms/therapy* ; Genetic Engineering ; Cell Line, Tumor ; Escherichia coli Proteins/genetics*

OBJECTIVE Analyze the nasal nitric oxide(NNO)of CRS children,and explore the clinical value of NNO in the diagnosis and treatment of CRS in children.METHODS CRS children diagnosed in the outpatient clinic were selected,and were divided into CRS with and without AR according to their allergen results.VAS score and NNO test were performed for them.Healthy children during the same period were selected as the control group.Finally their results were compared and analyzed.RESULTS The NNO of CRS children with and without AR were(193±62)ppb and(138±49)ppb,both lower than the control group's[(243±51)ppb];There were negative correlations between NNO and VAS scores in CRS children without AR before and after treatment;The NNO of CRS children with and without AR were significantly increased after treatment(P<0.05);NNO has high predictive value for diagnosing CRS children without AR(P<0.01).CONCLUSION The levels of NNO in different types of CRS were lower than normal,and CRS children without AR was lower than those with AR.NNO could assist in the diagnosis of CRS,dynamically reflect the severity of nasal inflammation,and help to distinguish the allergic status of CRS.

Objective:To investigate associations of serum nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), apoptosis-associated speck like protein containing a caspase recruitment domain (ASC), caspase-1 with the outcome in patients with hypertensive intracerebral hemorrhage (HICH) after minimally invasive neuroendoscopic surgery.Methods:Patients with HICH underwent minimally invasive neuroendoscopic surgery at the Affiliated Hospital of Noncommissioned Officer School, Army Medical University between June 2022 and June 2024 were included prospectively. According to the Glasgow Outcome Scale score at 3 months after surgery, the patients were divided into a good outcome group (4-5) and a poor outcome group (1-3). The clinical data and peripheral blood levels of NLRP3, ASC, caspase-1, interleukin (IL)-18, and IL-1β between the two groups were compared. Multivariate logistic regression analysis was used to determine assocations of serum NLRP3, ASC, caspase-1 with the postoperative outcome. Results:A total of 121 patients with HICH were enrolled, including 71 males (58.68), aged 56.11±4.96 years. At 3 months after surgery, 70 patients (57.9%) had good outcome, 51 (42.1%) had poor outcome, and 3 died. Onset to admission time, onset to first CT scan time, onset to surgery time, baseline serum NLRP3, ASC, caspase-1, baseline hematoma volume, and the proportion of patients with hematoma rupture into the ventricles and midline shift in the poor outcome group were significantly higher than those in the good outcome group, while baseline Glasgow Coma Scale (GCS) score and hematoma clearance rate were significantly lower than those in the good outcome group ( P<0.05). Multivariate logistic regression analysis showed that after adjusting for other factors such as onset to surgery time, baseline GCS score, hematoma rupture into the ventricles, and hematoma clearance rate, baseline serum NLRP3 (odds ratio [ OR] 2.018, 95% confidence interval [ CI] 1.502-2.711; P<0.001), ASC [ OR 1.764, 95% CI 1.418-2.195; P<0.001], caspase-1 [ OR 1.901, 95% CI 1.476-2.449; P<0.001]) were significantly independently associated with the poor outcome. Conclusion:The serum levels of NLRP3, ASC, and caspase-1 are significantly higher in HICH patients with poor outcome, and are independently associated with the poor outcome after minimally invasive neuroendoscopic surgery.

Objective:To investigate the predictive value of D-dimer (DD)/platelet count (PLT) ratio (DPR) and fibrinogen (Fg)/C-reactive protein (CRP) ratio for lower extremity deep venous thrombosis (LEDVT) in patients with spontaneous intracerebral hemorrhage (ICH).Methods:Consecutive patients with ICH admitted to the Department of Neurosurgery, the Affiliated Hospital of Noncommissioned Officer School, Army Medical University from February 2023 to November 2024 were included retrospectively. The baseline clinical data and laboratory test results between the LEDVT group and the non-LEDVT group were compared. Multivariate logistic regression analysis was used to evaluate the independent influencing factors of LEDVT. Receiver operating characteristic (ROC) curves were used to evaluate the predictive efficacy of influencing factors for patients with ICH complicated with LEDVT. Results:A total of 156 patients with ICH were enrolled, including 67 males (42.9%), aged 61.54±7.91 years; 47 patients (30.1%) experienced LEDVT during hospitalization. Univariate analysis showed that DD, PLT, DPR, Fg, and CRP in the LEDVT group were significantly higher than those in the non-LEDVT group, while the Fg/CRP ratio was significantly lower than that in the non-LEDVT group (all P<0.05). Multivariate logistic regression analysis showed that DD (odds ratio [ OR] 5.499, 95% confidence interval [ CI] 2.909-10.395; P<0.001), PLT ( OR 1.044, 95% CI 1.026-1.062; P<0.001), Fg ( OR 2.119, 95% CI 1.482-3.031; P<0.001), DPR ( OR 5.924, 95% CI 3.058-11.475; P<0.001), and Fg/CRP ratio ( OR 0.614, 95% CI 0.505-0.746; P<0.001) were the independent influencing factor for the occurrence of LEDVT in patients with ICH. ROC curve analysis showed that the areas under the curves of DD, PLT, Fg, DPR and the Fg/CRP ratio for predicting LEDVT in patients with ICH were 0.784 (95% CI 0.711-0.846), 0.772 (95% CI 0.699-0.836), 0.711 (95% CI 0.633-0.781), 0.782 (95% CI 0.709-0.844), and 0.778 (95% CI 0.705-0.841), respectively. The area under the curve for the combined prediction of DD+PLT+Fg was 0.878 (95% CI 0.816-0.924), and the area under the curve for the combined prediction of DPR+Fg/CRP ratio was 0.921 (95% CI 0.867-0.958). The latter showed a higher predictive value. Conclusion:The combined detection of DPR and Fg/CRP ratio has higher predictive value for LEDVT in patients with ICH.

Objective:To study and analyze radiation doses to the eye of the lens dose in interventional radiology staff of Guandong medical institutions from 2019 to 2023.Methods:Employing a convenience sampling method, the lens dose and chest dose was monitored with TLD dosimeters in 34 level A tertiary medical institutions. The monitoring data were analyzed on the basis of a summary, and the relationship between the two was explored.Results:A total of 1 033 interventional radiology staff were monitored. The ocular lens doses were in the range of (

Objective:To investigate the inflammatory indicator levels and influencing factors on middle-aged and elderly male residents at different residential distances from a uranium mine in Guangdong, so as to provide scientific data for the health risk assessment of the residents therein.Methods:With stratified sampling method, two groups of the permanent middle-aged and elderly male residents were randomly sampled within < 10 km and 10-20 km of the uranium mine, along with the basic demographic characteristics and lifestyle information collected through face-to-face questionnaire survey. Both height and weight of the respondents were measured using standard method and their body mass indexes (BMI) were calculated. Through collection of the venous blood, the levels of a wide range of inflammatory indicators were measured, such as C-reactive protein (CRP), complement C3, complement C4, leukocytes, lymphocytes, neutrophils, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and others. Multiple linear regression was used to analyze the influencing factors on the levels of inflammation indicators.Results:A total of 867 middle-aged and elderly male residents were included in this study, including one group of 431 residents within the 10-20 km at an average age of 51.54±5.37, and a second group of 436 residents within <10 km at an average age of 52.05±5.24. The result of multiple linear regression showed a positive correlation of the distance groups with complement C4 level ( β=0.014, 95% CI: 0.001-0.027) and lymphocyte number ( β=0.086, 95% CI: 0.003-0.168) ( t=2.07, 2.04, P<0.05). The ages of middle-aged and elderly male residents around the uranium mine was positively correlated ( t=2.50, P<0.05) with the levels of complement C4 ( β=0.018, 95% CI: 0.004-0.031), and negatively correlated ( t=-3.25, -2.97, P<0.05) with the levels of TNF-α ( β=-63.022, 95% CI: -101.114 to -24.929) and IL-6 ( β=-14.694, 95% CI: -24.396 to -4.992). Smoking was positively correlated ( t=4.29, 4.81, 3.19, P<0.05) with leukocytes number ( β=0.630, 95% CI: 0.341-0.918), lymphocytes number ( β=0.226, 95% CI: 0.134-0.319) and neutrophils number ( β=0.372, 95% CI: 0.143-0.601), and negatively correlated ( t=-2.07, -1.98, P<0.05) with the levels of TNF-α ( β=-43.551, 95% CI: -84.778 to -2.324) and IL-6 ( β=-10.603, 95% CI: -21.103 to -0.102). BMI was positively correlated ( t=8.60, 3.62, 3.18, 4.01, 2.10, P<0.05) with complement C3 level ( β=0.108, 95% CI: 0.084-0.133), complement C4 level ( β=0.026, 95% CI: 0.012-0.039), leukocytes number ( β=0.433, 95% CI: 0.166-0.699), lymphocyte number ( β=0.175, 95% CI: 0.089-0.261), and neutrophil number ( β=0.226, 95% CI: 0.014-0.438). Fruit consumption ( β=0.017, 95% CI: 0.001-0.034) was positively correlated with complement C4 levels ( t=2.10, P<0.05). Conclusions:This study showed no significant correlation between the inflammatory index levels of middle-aged and elderly male residents around the uranium mine and uranium mining. Age, smoking, BMI and fruit consumption were the influencing factors on the levels of inflammatory indicators of middle-aged and elderly male residents.

Objective:To investigate differential gene expression, enriched biological processes, and pathway differences between residents of a high-background radiation area (HBRA), Yangjiang, and a control area—Enping.Methods:Seven residents were selected from the HBRA (the HBRA group) and seven from the control area (the control group) using the two-stage random sampling method. The cumulative radiation dose for each individual was calculated based on the ambient gamma exposure levels. Peripheral blood samples were collected and analyzed via high-throughput sequencing to identify differentially expressed genes. Subsequent analyses included gene ontology (GO) for biological process (BP), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and gene set enrichment analysis (GSEA).Results:The median ages of the HBRA and control groups had no statistically significant difference ( P = 0.370). The cumulative external doses for the HBRA and control groups were (99.59±20.07) and (33.82±10.61) mSv, respectively. This difference was statistically significant ( t = -5.88, P = 0.001). High-throughput sequencing identified 1 224 differentially expressed genes in the HBRA group, including 32 senescence-related genes, compared to the control group. The result of GO-BP analysis showed that these genes were predominantly enriched in cell signaling, biosynthesis, localization, cell cycle regulation, and cellular stress responses. KEGG analysis revealed significant enrichment in the chemokine and MAPK signaling pathways, as well as in pathways related to the cell cycle, autophagy, and mitophagy. Furthermore, GSEA analysis confirmed that the differentially expressed genes were mainly related to cell cycle regulation and mitochondrial functions. Conclusions:Differentially expressed mRNAs are found in the peripheral blood of residents in the HBRA. These mRNAs are predominantly associated with key biological processes and pathways, including cell cycle regulation, chemokine and MAPK signaling pathways, and mitophagy.

Vaccination is an effective measure to control infectious diseases and provides lifelong health protection for the public. In recent years, vaccination for adults, especially the elderly, has gradually attracted attention in China, but its vaccination rate is significantly lower than that in some developed countries. To improve adult vaccine coverage, an immunization service model featuring adult vaccine prescriptions has been explored in some provinces of China. This study describes the implementation and characteristics of an adult vaccine prescription pilot program in Shandong Province and discusses considerations related to the concept, compliance, advantages, and difficulties of the vaccine prescription model. Suggestions for developing the vaccine prescription system in China are also provided.

From July 23, 2018, to December 31, 2019, a total of 299 219 children in Shandong Province received the fifth dose of the diphtheria, tetanus, and acellular pertussis combined vaccine (DTaP). Among these recipients, the distribution by age was as follows: 20 children under 2 years old (0.01%), 273 996 children aged 2 years (91.57%), 20 242 children aged 3 years (6.76%), 3 932 children aged 4 years (1.31%), 963 children aged 5 years (0.32%), and 66 children aged 6 years and above (0.02%). In total, 1 972 cases of adverse events following immunization (AEFI) were reported after the administration of the fifth dose of DTaP, resulting in an incidence rate of 659.05 per 100 000 doses. Among these, 1 718 cases were classified as common vaccine reactions, with an incidence rate of 574.16 per 100 000 doses, while 247 cases were identified as rare reactions, yielding an incidence rate of 82.55 per 100 000 doses. The incidence of AEFIs, as well as the rates of common and rare reactions, exhibited a significant increasing trend with the number of doses administered (all P<0.001). Among the rare reactions, there were 10 cases classified as severe, resulting in a reported incidence of 3.34 per 100 000 doses.