Objective To evaluate the performance of two molecular point-of-care testing(POCT)prod-ucts in the diagnosis of influenza A virus(Flu A)and influenza B virus(Flu B)of clinical samples,and pre-liminarily evaluate the clinical diagnostic value of the changes of infection-related indicators in peripheral blood.Methods A total of 491 oropharyngeal swabs from patients with influenza-like symptoms who were treated in the hospital were recruited into this study from November 1,2019 to June 30,2023.These swabs were collected using reverse transcription real-time quantitative fluorescent polymerase chain reaction(RT-qPCR),and two POCT molecular products,XpertTM Xpress Flu/RSV and EasyNAT? Flu Assay,respectively.The diagnostic performance of two POCT molecular products was analyzed using RT-qPCR reaction as a standard.According to the results of RT-qPCR method,the subjects were divided into Flu A positive group,Flu B positive group and negative group(both Flu A and Flu B were negative).The levels of indicators in pe-ripheral blood of the three groups were compared to evaluate the value of these indicators in the clinical diag-nosis of Flu A and Flu B.Results Among the 491 patient specimens,the XpertTM Xpress Flu/RSV assay showed the sensitivity for Flu A was 96.88%,and the specificity was 99.75%,and the sensitivity for Flu B was 100.00%,and the specificity was 100.00%.EasyNAT? Flu Assay assay showed the sensitivity for Flu A was 94.79%,and the specificity was 96.81%,and the sensitivity for Flu B was 100.00%,and the specificity was 100.00%.And two POCT molecular methods performed well consistency(Kappa value was 0.974).There was no significant difference in the levels of C-reactive protein and serum amyloid A among the negative group,Flu A positive group,and Flu B positive group(P>0.05).But the levels of white blood cell count in the negative group were higher than those in the Flu A positive group and Flu B positive group(P<0.01).Conclusion In this paper,two typical molecular POCT products are studied.Their sensitivity and specificity are highly consistent with the results of RT-qPCR.Molecular POCT products have the advantages of flexibil-ity and rapidity,which are of great value for the improvement of clinical diagnosis and treatment.Molecular detection combined with peripheral blood infection related indicators is helpful for the early diagnosis of influ-enza virus infectious diseases.

Objective:To investigate the impact of daily step count on glycemic outcomes in community residents with impaired glucose tolerance (IGT).Methods:This was a prospective cohort study, in October 2018, 204 residents who met the criteria of IGT were recruited in the Shijingshan District in Beijing. The subjects were tested for fasting blood glucose, oral glucose tolerance test 2-hour blood glucose (2hBG), glycated hemoglobin A 1c (HbA 1c), lipid profile, liver and kidney function, as well as measurements of height, weight and waist circumference. A dedicated mobile application was used to deliver prediabetes health knowledge monthly. Online guidance was provided to answer questions and daily step count was collected using the application. Three years later, a follow-up was conducted to assess the participants′ glycemic outcomes and other indexes, and a total of 142 participants completed the follow-up review. According to daily step count, the subjects were categorized into high step count group (42 cases,>7 000 steps daily), moderate step count group (54 cases, 5 000-7 000 steps daily), and low step count group (46 cases,<5 000 steps daily). Subjects were categorized into diabetes group (30 cases), prediabetes group (77 cases) and normal glucose tolerance group (35 cases) with glycemic outcomes. Independent sample t test was used to compare the differences in blood glucose, blood lipids, and step counts between the two groups. Kruskal-Wallis H test or one-way ANOVA was used to compare the differences in blood glucose, blood lipids, and step counts between multiple groups. The χ2 test was used to compare the differences in glycemic outcomes between multiple groups. Multivariate logistic regression analysis was used to assess the impact of daily step counts and body mass index on glycemic outcomes. Linear regression analysis was used to evaluate the relationship between daily step counts and 2 h BG. Results:A total of 142 participants completed the 3-year follow-up, including 43 males and 99 females, with a mean age of (60.15±5.67) years. At baseline, males had significantly higher body mass index, waist circumference, and fasting blood glucose when compared to those in females [(26.97±2.43) vs (24.89±2.93) kg/m 2, (92.68±7.75) vs (83.83±8.60) cm, (5.83±0.61) vs (5.62±0.52) mmol/L], the total cholesterol and HDL-C were also significantly lower in males than those in females [(5.10±1.16) vs (5.55±0.95) mmol/L, (1.35±0.34) vs (1.56±0.35) mmol/L] (all P<0.05). After 3-year follow-up, 21.1% (30/142) of IGT participants progressed to diabetes, with an annual conversion rate of approximately 7%. The normal glucose tolerance group showed significantly higher daily step counts when compared with the prediabetes and diabetes groups [(7 886±2 867) vs (5 981±2 655) vs (4 117±2 674) steps] ( H=31.778, P<0.001). Individuals with higher daily step counts exhibited lower body mass index, 2 h BG, and HbA 1c level when compared with those in the ones with moderate and low step counts [(24.26±3.09) vs (25.44±3.38) vs (26.26±3.59) kg/m 2, (7.50±1.71) vs (9.15±3.30) vs (11.19±3.84) mmol/L, 5.97%±0.46% vs 6.14%±0.99% vs 6.40%±0.96%] (all P<0.05). Higher step count was positively correlated with the reversal of prediabetes to normal blood glucose levels (moderate step count, OR=0.297, 95% CI: 0.109-0.804; low step count, OR=0.055, 95% CI: 0.010-0.287), lower daily step count correlated positively with prediabetes progressing to diabetes ( OR=4.857, 95% CI: 1.140-20.689) (all P<0.05). For every additional 1 000 steps per day, the 2 h BG decreased by 0.5 mmol/L. Conclusion:As daily step count increases, the glucose metabolism improves in IGT community residents. Higher daily step count is associated with reversal of IGT to normal glucose tolerance, while lower daily step count may be associated with the progression of IGT to diabetes.

γ-aminobutyric acid can be produced by a one-step enzymatic reaction catalyzed by glutamic acid decarboxylase. The reaction system is simple and environmentally friendly. However, the majority of GAD enzymes catalyze the reaction under acidic pH at a relatively narrow range. Thus, inorganic salts are usually needed to maintain the optimal catalytic environment, which adds additional components to the reaction system. In addition, the pH of solution will gradually rise along with the production of γ-aminobutyric acid, which is not conducive for GAD to function continuously. In this study, we cloned the glutamate decarboxylase LpGAD from a Lactobacillus plantarum capable of efficiently producing γ-aminobutyric acid, and rationally engineered the catalytic pH range of LpGAD based on surface charge. A triple point mutant LpGAD^{S24R/D88R/Y309K} was obtained from different combinations of 9 point mutations. The enzyme activity at pH 6.0 was 1.68 times of that of the wild type, suggesting the catalytic pH range of the mutant was widened, and the possible mechanism underpinning this increase was discussed through kinetic simulation. Furthermore, we overexpressed the Lpgad and Lpgad^{S24R/D88R/Y309K} genes in Corynebacterium glutamicum E01 and optimized the transformation conditions. An optimized whole cell transformation process was conducted under 40 ℃, cell mass (OD₆₀₀) 20, 100 g/L l-glutamic acid substrate and 100 μmol/L pyridoxal 5-phosphate. The γ-aminobutyric acid titer of the recombinant strain reached 402.8 g/L in a fed-batch reaction carried out in a 5 L fermenter without adjusting pH, which was 1.63 times higher than that of the control. This study expanded the catalytic pH range of and increased the enzyme activity of LpGAD. The improved production efficiency of γ-aminobutyric acid may facilitate its large-scale production.
Glutamate Decarboxylase/genetics* ; Lactobacillus plantarum/genetics* ; Catalysis ; gamma-Aminobutyric Acid ; Hydrogen-Ion Concentration ; Glutamic Acid

Objective:To investigate the role and diagnostic value of miRNA-205 in chronic kidney disease (CKD) patients with vascular calcification.Methods:It was divided into in vitro cell experiment and retrospective cohort study. In vitro experiments were conducted by using rat thoracic aortic smooth muscle cells. Alizarin red staining and calcium content detection were used to detect the calcification of vascular smooth muscle cells (VSMCs). Alkaline phosphatase (ALP) test kit was used to measure ALP activity. Western blotting was used to detect the protein expression levels of osteogenic transcription factors runt-related transcription factor 2 (Runx2), α smooth muscle actin (α-SMA) and smooth muscle-22α (SM-22α) in VSMCs. qRT-PCR was used to detect miRNA-205 and Runx2 expression levels. The double luciferase reporter gene assay was used to verify the targeted relationship between miRNA-205 and Runx2. The non-dialysis patients with CKD 3-5 stage from June 2020 to January 2021 in the Department of Nephrology of Fourth Hospital, Hebei Medical University were selected. According to coronary artery calcium score (CACs), the patients were divided into non-calcification group (CACs=0), mild-moderate calcification group (0 400). Spearman correlation analysis was used to analyze the correlation between miRNA-205 and Runx2 and vascular calcification. Logistic regression model and receiver operating characteristic (ROC) curve analysis were used to analyze the ability of miRNA-205 to predict the vascular calcification in patients with CKD. Results:(1)Compared with the control group, calcium nodules were more, and the calcium content, ALP activity and Runx2 protein level were higher, and the expression levels of miRNA-205, α-SMA and SM-22α were significantly lower in high phosphorus group (all P<0.05). Overexpression of miRNA-205 significantly reduced the calcification of VSMCs and Runx2 protein level, and increased the protein levels of α-SMA and SM-22α (all P<0.05). miRNA-205-5p reduced the activity of luciferase in the wild-type Runx2-3'-end non-coding region plasmid. (2) Eighty CKD patients were enrolled, with age of (57.50±14.93) years old and 49 males (61.3%). The results of comparison of miRNA-205 and Runx2 expression levels in non-calcification group ( n=26), mild- moderate calcification group ( n=30) and severe calcification group ( n=24) showed that, the higher degree of calcification, the lower miRNA-205 expression level and the higher Runx2 mRNA expression level (all P<0.05). miRNA-205 was negatively correlated with CACs ( r=-0.50, P<0.01) and Runx2 was positively correlated with CACs ( r=0.55, P<0.01). Multivariate logistic regression analysis results suggested that miRNA-205 ( OR=0.451, 95% CI 0.122-0.873) was an independent influencing factor of vascular calcification in CKD patients. The area under the ROC curve of miRNA-205 and miRNA-205 combined with Runx2 for predicting vascular calcification were 0.796 (95% CI 0.697-0.859) and 0.924 (95% CI 0.866-0.982), respectively. Conclusions:miRNA-205 inhibits vascular calcification by targeting Runx2 to negatively regulate osteogenetic phenotype transformation of VSMCs and is expected to be an early diagnostic marker of vascular calcification in CKD patients.

Objective:To investigate the role and mechanism of N 6-methyladenosine (m 6A) methyltransferase-like 3 (METTL3) in vascular calcification (VC) of chronic kidney disease (CKD) through apoptosis-associated protein. Methods:(1) Real-time fluorescence quantitative PCR was used to test METTL3 mRNA in serum of maintenance hemodialysis (MHD) patients. (2) Western blotting was used to detect the expression of METTL3 protein in high-phosphorus stimulated vascular smooth muscle cells (VSMCs), and immunofluorescence double lable was used to observe the distribution of METTL3 and Runt-related transcription factor 2 (Runx2). The METTL3 overexpressed and knockdown plasmids were constructed and transfected into VSMCs. Alizarin red staining was used to detect calcification degree. Western blotting was used to detect the expressions of osteogenic markers [Runx2, bone morphogenetic protein-2(BMP-2), collagen Ⅰ] and apoptosis- related proteins Bax and Bcl-2. (3) SD rats were randomly divided into control group, CKD-VC group and S-adenosylhomocysteine (SAH) intervention group. The calcification of thoracic aorta was evaluated by von Kossa staining, and the protein expressions of Runx2, Bax and Bcl-2 were detected by immunohistochemistry and Western blotting.Results:(1) METTL3 mRNA expression in MHD patients with VC was significantly lower than that in non-VC patients ( P<0.05), and was negatively correlated with coronary artery calcium score ( r=-0.65, P<0.001). (2) The expression of METTL3 in VSMCs stimulated by high phosphorus was decreased and showed a time dependence. Immunofluorescence double label showed that METTL3 and Runx2 were co-expressed in the nucleus. METTL3 was overexpressed in high-phosphorus induced VSMCs, and the expressions of Runx2, collagen I and BMP-2 were significantly decreased, accompanied by the decrease of calcified nodules and Bax/Bcl-2 ratio (all P<0.05). Conversely, METTL3 knockdown aggravated VSMCs calcification by inducing apoptosis. (3) Furthermore, METTL3 inhibitor SAH was administered in vivo, and it was found that inhibition of METTL3 expression significantly increased the calcification of rat thoracic aorta, and the Bax/Bcl-2 ratio and Runx2 expression were up-regulated. Conclusions:Serum METTL3 level is reduced in MHD patients with VC. In vivo and in vitro studies demonstrate that METTL3 inhibits VC in CKD by mediating the apoptosis-related protein Bax/Bcl-2.

ObjectiveTo investigate the status quo of emergency rescue for public health emergencies by medical staff under regular pandemic prevention and control， analyze its influencing factors， and provide evidence for formulating intervention measures. MethodsAn online questionnaire was conducted among the medical staff of Jinshan District public medical institutions， and the possible influencing factors were analyzed by multiple linear stepwise regression analysis. The questionnaire included general information and knowledge， attitude， and behavior of emergency rescue. ResultsA total of 1，614 valid questionnaires were collected. The total average score of emergency rescue performance was （4.01±0.83）， which was at a high level. The average scores of emergency rescue knowledge and attitude were （4.07±0.80） and （4.21±0.70） respectively， which were at a high level， and the average scores of emergency rescue behavior were （3.25±1.23）， which was at a medium level. Regression analysis， age， department， occupation， hospital level， whether to participate in emergency training， whether to participate in emergency drills were the main influencing factors of emergency rescue knowledge and practice （P<0.05）. ConclusionUnder the situation of major public health emergencies， the knowledge， attitude and practice of emergency rescue of medical personnel are generally at a good level， and the behavior of emergency rescue is at a medium level. Emergency rescue training and drills should be strengthened for medical personnel of different ages， occupations， departments， levels of hospitals， and sections.

Under the background of efficiency medical reform, the authors introduced the nursing efficiency management system of a large public hospital, which was divided into two dimensions: nursing resource allocation efficiency and nursing service efficiency. The specific four measures included the establishment of multi-campus unified management organization structure based on nursing management committee, nursing performance reform based on structured big data information platform, the construction of nurse-led patient whole process management model and the accelerated rehabilitation nursing practice from surgery to the whole hospital, so as to provide reference for the nursing efficiency management of large general public hospitals under the medical reform.

【Objective】 To investigate the airway parameters of adult-onset eosinophilic asthma （EA） and analyze the correlation between airway remodeling and lung function by quantitative CT. 【Methods】 From March 2015 to November 2016， totally 94 subjects from the “FACT-Digital Lung” Multi-research Center were divided into three groups： 30 normal subjects， 33 EA patients and 31 non-eosinophilic asthma （NEA） patients. We measured and recorded the bronchial parameters of RB1， LB1+2， RB10， and LB10， and small airway disease parameters. The indicators for quantitative evaluation of bronchial parameters include lumen area （LA）， wall thickness （WT）， wall area （WA）， and wall area percentage （WA%）. The parameters for the quantitative assessment of small airway disease included the percentage of inspiratory voxels below -950HU （IN-950）， the mean lung density （MLDin）， and the whole volume of the lung in inspiration （Vin）. Pearson or Spearman rank correlation analysis was used to evaluate the correlation between these parameters and lung function. 【Results】 The differences in LA/BSA， WT/√BSA， and WA/BSA between the three groups were statistically significant （P<0.05）. FEV1% had a significant correlation with IN-950 and MLDin （P<0.01）. FEV1/FVC had a significant correlation with Vin， IN-950， and MLDin （P<0.01）. EOS counts were positively related to IN-950 （r=0.343， P=0.011）， while EOS had a negative correlation with FEV1% （r=－0.343， P=0.015）. 【Conclusion】 With the increase of eosinophils counts in peripheral blood， the airway's stenosis in asthma patients gradually increased， and the extent of airflow limitation steadily increased. The IN-950 may be a sensitive imaging biomarker for evaluating the small airway disease in adult-onset eosinophilic asthma patients.

Objective:To analyze the effect and safety of levonorgestrel-releasing intrauterine system(LNG-IUS) for the premenopausal patients with breast cancer who took tamoxifen as adjuvant therapy.Methods:From June 2014 to June 2016, 84 patients with breast cancer who met the inclusion criteria in the First People′s Hospital of Xiaoshan District were randomly divided into two groups according to the digital table.The treatment group (39 cases) underwent LNG-IUS insertion, while the control group (45 cases) received no LNG-IUS insertion.The general condition of patients before the use of tamoxifen and LNG-IUS was evaluated.Transvaginal ultrasound was used to measure the thickness of endometrium, hysteroscope was used for pathological examination of endometrium and the measurement of ER/PR expression, and blood lipid level was also detected.All above was done before the treatment of tamoxifen and LNG-IUS, 1 year after treatment and 2 years after treatment.Results:Before the therapy, there were no statistically significant differences between the two groups in general condition and uterine cavity condition(all P>0.05). After 1 year, the incidences of endometrial polyp, endometrial hyperplasia/secretion, benign lesion and endometrial atrophy in the treatment group were 2.6%, 5.1%, 15.4%, 76.9%, respectively, which in the control group were 6.7%, 20.0%, 17.8%, 55.6%, respectively.the differences between the two groups were statistically significant(χ 2=4.06, 4.22, all P<0.05). After 2 years, the incidences of endometrial polyp, endometrial hyperplasia/secretion, benign lesion and endometrial atrophy in the treatment group were 0.0%, 2.6%, 84.6%, respectively, which in the control group were 11.1%, 15.6%, 60.0%, respectively, the differences between the two groups were statistically significant(χ 2=4.608, 4.092, 6.203, all P<0.05). Conclusion:LNG-IUS can prevent the benign endometrial lesions of breast cancer patients caused by tamoxifen therapy after surgery, and can decrease the incidence of endometrial polyp and endometrial hyperplasia/secretion, while increase the incidence of endometrial atrophy, without increasing the recurrence risk of breast cancer.

Objective To establish and validate a voxel-based method for the quantitative detection of air trapping (AT),and to explore its diagnostic value by preliminarily apply this method in chronic obstructive pulmonary disease (COPD) patients.Methods From March 2015 to February 2016,fifty healthy young volunteers and eighteen COPD patients who underwent both end-inspiratory and end-expiratory CT were included from the Digital Lung Multi-center Study.The quantitative parameters of AT and emphysema were measured by both the voxel-based quantitative method and the conventional threshold method,respectively.All subjects underwent pulmonary function examination within 3 days after CT examination.For healthy volunteers,paired sample rank-sum test was used to compare the difference of quantitative parameters between voxel-based method and threshold method,Spearman rank correlation analysis was used to investigate the correlation between quantitative parameters of the two methods and pulmonary function.For COPD patients,the distribution and extent of AT and emphysema in patients with similar degree of pulmonary function (PFT) injury were observed.Results There were varying degrees of AT in the asymptomatic youth,with a median value of 5.70％ for the voxel-based method and with a median value of 7.96％ for the conventional threshold method,there was significant difference(Z=-4.015,P＜0.001).The correlation between AT and emphysema parameters of the voxel-based method and PFT parameters (r=-0.399 and-0.494,-0.335 and-0.439 separately,P＜0.05) were higher than that of the conventional threshold method,respectively (r=-0.357 and-0.453,-0.284 and-0.391,respectively;all P＜0.05).Furthermore,the voxel-based method can classify COPD patients with similar degree of pulmonary function injury into three subtypes:AT-dominant,emphysema-dominant,and mixed.Conclusions The voxel-based AT quantitative measurement method not only has high sensitivity and accuracy,but also provides imaging phenotype for the diagnosis of COPD and provides assistant decision-making for clinical management.