Objective To investigate the knowledge and attitude of clinical nurses on Panvascular Disease(PVD)care in Zhejiang Province,and to analyze the influencing factors,so as to provide references for constructing a systematic PVD training program.Methods The convenient sampling method was used to investigate 454 nurses from 36 general hospitals in 11 prefectures and cities in Zhejiang Province in June 2024 with a self-designed questionnaire on general information and a questionnaire on PVD nursing knowledge and attitude questionnaire,and multiple stepwise linear regression was used to analyze the impact factors.Results 452 valid questionnaires were collected and the valid recovery rate was 99.56％.Nurses scored(8.57±2.02)in knowledge dimension,(60.82±6.45)in attitude dimension.The results of multiple stepwise linear regression analysis showed that hospital grade was the factor influencing the score of nurses'knowledge towards PVD nursing in Zhejiang Province(P＜0.05).Hospital grade,whether they had received systematic knowledge training related to PVD were the factors influencing the score of nurses'attitude towards PVD nursing(P＜0.05).Conclusion Nurses in Zhejiang Province are positive about PVD nursing,but their knowledge level needs to be improved.Nursing managers should pay full attention to the training intention of clinical nurses,establish a PVD knowledge training system,and strengthen the theoretical and practical training of clinical nurses on PVD nursing,so as to promote the development of PVD nursing work.

Objective To analyze various scanning parameters and contrast agent injection schemes,and explore the feasibility of three-low scanning technology characterized by lower radiation dose,slower injection rate and reduced contrast agent dosage in coronary angiography with wide-detector CT.Methods A total of 210 patients who underwent coronary angiography were recruited and randomly divided into group A(routine group,n=105)and group B(three-low group,n=105),and all of them were examined using the Revolution CT from the American GE company.Group A and group B adopted 120 and 100 kV tube voltages,respectively.The contrast injection rate and contrast agent dosage were calculated using formulas based on body mass index and beat per minute.Image quality was objectively evaluated using CT number,signal-to-noise ratio and contrast-to-noise ratio,and subjectively scored using Likert 5-point method.Results No statistical differences were found in the basic data between two groups(P>0.05).The CT numbers of the aortic sinus orifice in coronary artery images obtained from group A and group B were(442.70±58.26)and(454.11±62.36)HU,respectively.The differences between two groups in CT number,image score,signal-to-noise ratio,and contrast-to-noise ratio were trivial(P>0.05),while statistically significant differences were noted in injection rate,contrast agent dosage,tube current,and effective radiation dose(P<0.05).Group B reduced the effective radiation dose by 33.89%as compared with group A.Conclusion Based on the patient's body mass index and beat per minute,a personalized contrast agent injection scheme can be developed.Both routine group and three-low group can obtain image quality that meets imaging diagnosis requirements,and the latter has better image quality,demonstrating that the three-low scheme benefits the patients more for it can reduce the patient's radiation dose and contrast agent dosage while achieving higher image quality.

Thyroid cancer is the only non reproductive tumor that mainly occurs in women.The morbidity rate of female thyroid cancer is three times higher than that of male thyroid cancer.However,the ex-isting studies have not systematically summarized the factors that contribute to the high incidence of thyroid cancer in women.This paper reviews all the literatures that mention the high incidence of thyroid cancer in women,and sorts out the main reasons,such as estrogen,obesity,reproductive factors,oxidation antioxidant system imbalance,X chromosome,etc.,which are the main reasons for the current research finding that the in-cidence rate of thyroid cancer in women is higher than that in men.This paper makes a vivid graphic analysis and summary of the reasons,providing a research direction for future researchers to research on gender duality of thyroid cancer,and providing a certain reference for the clinical diagnosis and treatment of gender differences in thyroid cancer.

Objective To review the state of research on medullary thyroid carcinoma(MTC)over the past two decades,so as to offer a quick reference for understanding the current state and future trends in this field.Methods A computer was employed to search the Web of Science Core Collection database for MTC-re-lated literature(types;articles and reviews)published between 2000 and 2022.VOS viewer 1.6.18 software was utilized to create collaboration network diagrams,heat distribution diagrams for high-yielding countries(with publication volume ≥ 90),research institutions(with publication volume ≥ 60),and authors(with publication volume ≥30),and high-frequency keyword clustering network diagrams.Cite Space 5.7.R5 soft-ware was employed to conduct cluster analysis and explosive keyword detection on co-cited literature.Results A total of 4866 MTC related literature(3688 articles and 918 reviews)were obtained.Although the annual publication volume fluctuated,the overall trend showed a gradual increase.The 4866 articles are from 87 coun-tries,involving 4250 research institutions,21 016 authors,and 963 journals.The United States(1436 arti-cles),MD Anderson Cancer Center at the University of Texas(139 articles),and Dralle H(Department of General Surgery at Halle Wittenberg University Hospital,61 articles)were the countries,research institu-tions,and authors with the highest number of publications,respectively.High frequency keywords form four clusters:RET gene expression,prognostic factors,MTC clinical management,and MTC treatment.The key-words emerging explosively in recent years were management,survival,guideline,association,etc.The key areas of focus in highly cited literature were RET proto oncogene,targeted therapy,RET gene mutation,and so on.Conclusions In the past 20 years,MTC related research has shown a gradual upward trend,with de-veloped countries led by the United States taking a leading position in this field.The current research hotspots are the treatment,diagnosis,application of inhibitors,and exploration of the pathogenesis of MTC.

OBJECTIVE: To explore the clinical phenotype, pregnancy outcome and follow-up of fetuses with 15q11.2BP1-BP2 microdeletions in order to provide a basis for prenatal and reproductive consultation. METHODS: From March 2019 to December 2023, 20 fetuses who were diagnosed with 15q11.2BP1-BP2 microdeletion syndrome at the Prenatal Diagnosis Center of General Hospital of Ningxia Medical University were selected as the study subjects. Results of genetic testing and ultrasound examination, outcome of pregnancy, and postnatal follow-up were retrospectively analyzed. This study has been approved by the Ethics Committee of General Hospital of Ningxia Medical University ([2020]0520B). RESULTS: None of the 20 fetuses was found to have chromosomal abnormality, whilst all were found to harbor a 15q11.2 BP1-BP2 microdeletion by low-depth whole genome sequencing (CNV-seq). The range of deletions was determined as 0.26 ~ 0.87 Mb, and all were rated as pathogenic CNVs. Three fetuses had abnormal ultrasound findings, including 1 with widened renal pelvis, 1 with agenesis of corpus callosum, and 1 with nuchal fold thickening. Parental verification in 10 couples verified that two fetal deletions were de novo, whilst the remaining eight were inherited from a phenotypically normal parent. Following genetic counseling, three couples had opted to terminate the pregnancy, whilst the remaining 17 had continued with the pregnancy until delivery. The 17 liveborns were followed up for 2 months to 5 years, with no obvious abnormality in growth and development noted. CONCLUSION CNV-seq plays an important role in the prenatal diagnosis of 15q11.2 BP1-BP2 microdeletions. Such deletions may not always lead to disease phenotypes. Individualized consultation and long-term follow-up, in combination with intrauterine ultrasound and parental verification are necessary.
Humans ; Female ; Pregnancy ; Chromosomes, Human, Pair 15/genetics* ; Genetic Counseling ; Prenatal Diagnosis ; Chromosome Deletion ; Adult ; Fetus/abnormalities* ; Retrospective Studies ; Pregnancy Outcome ; Ultrasonography, Prenatal ; Genetic Testing ; DiGeorge Syndrome/diagnosis* ; Male

Objective To understand the driver gene mutation status in patients with non-small cell lung cancer (NSCLC) in Xi'an City, and to analyze the association with environmental exposure factors. Methods A total of 305 NSCLC patients admitted to the First Affiliated Hospital of the Air Force Medical University from January 2019 to December 2023 were included. The driver gene mutation status was observed, and the relationship with environmental exposure factors was analyzed. Results The driver gene mutation rate of 305 patients was 46.89%, with EGFR gene mutation accounting for the highest proportion, and 4 cases of gene co-mutations were detected. There was a difference in gender among patients with different single drive gene mutations (P<0.05), and the proportion of EGFR in women was significantly higher (P<0.05). Univariate analysis showed that there were statistical differences in family history, smoking history, long-term cooking history, and fried smoked food intake between patients with driver gene mutation and patients without driver gene mutation (P<0.05). Logistic regression analysis suggested that long-term cooking history (OR=2.392), and fried smoked food intake (OR=2.849) were the environmental exposure factors affecting EGFR gene mutation (P<0.05), and smoking history (OR=1.377) was an environmental exposure factor of KRAS gene mutation (P<0.05). Conclusion EGFR gene mutation accounts for the highest proportion of NSCLC patients in Xi'an City, and is mainly female. Long-term cooking history, and fried smoked food intake are related to EGFR gene mutation. There is a certain association between smoking history and KRAS gene mutation.

ObjectiveTo investigate the change in the activity of hepatitis B virus （HBV）-specific CD8⁺ T cells after pegylated interferon-α-2b （PEG-IFN-α-2b） therapy in HBeAg-negative patients with chronic HBV infection. MethodsA total of 53 HBeAg-negative patients with chronic HBV infection who attended The First Affiliated Hospital of Xinxiang Medical University and Tangdu Hospital of Air Force Mdical University from April 2020 to June 2022 were enrolled and treated with PEG-IFN-α-2b （180 μg/week， subcutaneous injection） antiviral therapy. The study endpoint was HBsAg clearance （course of treatment<48 weeks） or 48 weeks （course of treatment≥48 weeks）. Peripheral blood mononuclear cells were isolated at baseline and study endpoint， and peripheral blood T cell counts were measured. Enzyme-linked immunospot assay was used to measure the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ. A total of 17 HLA-A^*02-restricted patients were selected， and CD8⁺ T cells were purified to establish direct- and indirect-contact co-culture systems for HBV-specific CD8⁺ T cells and HepG2.2.15 cells. The level of lactate dehydrogenase in supernatant was measured to calculate the mortality rate of HepG2.2.15 cells， and the levels of HBV DNA， cytotoxic molecules， and cytokines in supernatant were also measured. Flow cytometry was used to measure the expression of apoptosis ligands， and the cytotoxicity of HBV-specific CD8⁺ T cells was evaluated. The independent samples t-test or the paired t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test or the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsThe HBsAg clearance rate at study endpoint was 30.19% （16/53）. There were no significant differences in peripheral blood T cell counts （CD3⁺， CD4⁺， and CD8⁺ T cells） between baseline and study endpoint （P>0.05）. At study endpoint， there was a significant increase in the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ （U=177.50， t=11.90， U=186.50， all P<0.001）， and the patients with HBsAg clearance had a significantly higher frequency of such HBV-specific CD8⁺ T cells than those without HBsAg clearance （U=120.50， t=2.73， U=121.50， all P<0.01）. In the direct- and indirect-contact co-culture systems at study endpoint， HBV-specific CD8⁺ T cells induced a significant reduction in HBV DNA in the supernatant of HepG2.2.15 cells （all P<0.001） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （all P<0.05）； in the direct-contact co-culture system， HBV-specific CD8⁺ T cells induced significant increases in the mortality rate of HepG2.2.15 cells （13.62%±3.27% vs 11.39%±2.40%， t=2.27， P=0.030） and the secretion of perforin and granzyme B （t=72.50， U=52.50， both P<0.05）. In the direct- and indirect-contact co-culture systems， compared with HBV-specific CD8⁺ T cells from the patients without HBsAg clearance， the HBV-specific CD8⁺ T cells from patients with HBsAg clearance had a significantly greater reduction in HBV DNA （P<0.05） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （P<0.05）. ConclusionPEG-IFN-‍α‍-2b therapy can help to achieve a relatively high HBsAg clearance rate in HBeAg-negative patients with chronic HBV infection， and the activity of HBV-specific CD8⁺ T cells is significantly enhanced， which is closely associated with HBsAg clearance.

L-theanine is an important natural non-protein amino acid that is widely used in food and medicine. Although in previous studies, a microbial fermentation method for L-theanine without the addition of ethylamine has been developed, the conversion rate of this process needs to be further improved. In this study, we constructed a de novo synthesis pathway of L-theanine with glucose as the substrate. First, an in vitro transformation pathway containing ω-transaminase (TA) and γ-glutamylmethylamide synthetase (GMAS) was designed, optimized, and introduced into the chassis strain Escherichia coli K12 W3110 to achieve de novo synthesis of L-theanine. To improve the synthesis efficiency through metabolic engineering, we increased the copies of the GMAS gene gams and the TA gene spuC and enhanced the expression of the aldehyde dehydrogenase gene eutE to provide sufficient acetaldehyde substrate, knocked out the lactate dehydrogenase gene ldhA and the pyruvate formate lyase gene pflB to block bypass metabolism, and introduced the alanine dehydrogenase gene alD to recycle alanine. Furthermore, we over-expressed the phosphoenolpyruvate carboxylase gene ppc to enhance the carbon flux of the TCA cycle, knocked out the succinyl-CoA synthase gene sucCD to reduce the loss of downstream flux of TCA, and integrated the glutamate dehydrogenase gene gdh to enhance the supply of L-glutamate. Finally, the polyphosphate kinase gene ppk was introduced to the ATP cycle, which enhanced the energy supply in L-theanine production. The recombinant strain Tea11 produced 22.60 g/L L-theanine in a 5 L fermenter in 28 h, with a conversion rate of 41.71%. This synthetic pathway in this study balanced the relationship between the supply of ethylamine and the production of theanine, providing a new idea for metabolic engineering of microorganisms to produce L-theanine.
Glutamates/biosynthesis* ; Metabolic Engineering/methods* ; Escherichia coli/genetics* ; Fermentation ; Transaminases/metabolism* ; Amide Synthases/metabolism* ; Glucose/metabolism*

L-tyrosine is one of the 20 amino acids that make up proteins and is an essential amino acid for mammals, often used as a nutritional supplement. The conventional methods for synthesizing L-tyrosine have some problems such as the production of many by-products, high requirements for production conditions, and environmental pollution. In this study, we designed and constructed a multi-enzyme cascade for the synthesis of L-tyrosine with alanine, glutamate, ammonium chloride, and phenol as substrates. Initially, the sources of glutamate oxidase, alanine aminotransferase, and tyrosine phenol lyase were screened and analyzed, which was followed by the identification of the rate-limiting enzyme in the reaction process. A colorimetric screening method was established, and the rate-limiting enzyme DbAlaA was engineered to enhance its activity by 40.0%. Subsequently, the reaction conditions, including temperature, pH, cell concentration, and surfactant and coenzyme dosages, were optimized. After optimization, the yield of L-tyrosine reached 9.93 g/L, with a alanine conversion rate of 54.90%. Finally, a feed-batch fermentation strategy was adopted, and the yield of L-tyrosine reached 56.07 g/L after 24 h, with a alanine conversion rate of 65.22%. This study provides a reference for the whole-cell catalytic synthesis of L-tyrosine and its industrialization.
Tyrosine/biosynthesis* ; Escherichia coli/metabolism* ; Tyrosine Phenol-Lyase/genetics* ; Multienzyme Complexes/metabolism* ; Fermentation

OBJECTIVE To investigate the current status and risk factors of carbapenem-resistant gram-negative or-ganisms(CRO)infections in neurosurgical ICU patients,and to construct and validate their prediction models.METHODS Clinical data and active screening microbiological results of 113 patients admitted to the Neurosurgical Intensive Care Unit(ICU)of The First Affiliated Hospital of Soochow University between Jul.2023 and Jan.2024 were retrospectively collected,and the patients were divided into a CRO-infected group(n=28)and a non-CRO-infected group(n=85).Predictive variables were screened using LASSO regression and logistic regression.Risk prediction models were constructed using random forest(RF)and logistic regression,the performance of the mod-el was evaluated by analyzing the area under the receiver operating characteristic(ROC)curve(AUC),calibration curves,and decision curves,and internal validation was performed using the bootstrap resampling method.RESULTS Among 113 neurosurgical ICU patients,28 cases developed CRO infections,with an infection rate of 24.78％.The highest infection rate was observed in lower respiratory tract infection,with 17 cases(15.04％).The predominant CRO pathogens were carbapenem-resistant Klebsiella pneumoniae(CRKP)and carbapenem-re-sistant Acinetobacter baumannii(CRAB),accounting for 50.00％and 42.86％of cases respectively.The AUC values for the RF prediction model and nomogram prediction modeling groups were 0.881 and 0.787 respectively,with Brier scores of 0.114 and 0.146,and threshold probabilities of net benefit ranging from 10％to 97％and 12％to 62％respectively.The RF prediction model exhibited superior discrimination,calibration,and clinical u-tility.The RF prediction model demonstrated that days of combined use of meropenem and vancomycin,GCS score,intestinal colonization,and hospitalization history were important predictors for CRO infections.CONCLUSION The prediction model for CRO infections in neurosurgical ICU patients established based on random forest algorithm has good predictive performance,and can be intervened with preventive and control measures for important predictive factors.