Objective　To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Listeria monocytogenes, and analyze the structure and homology of CRISPR loci. Methods Totally 34 strains of Listeria monocytogenes isolated in our laboratory were identified, PCR amplified and sequenced. The repeat sequence structure and spacer sequence homology in CRISPR loci were analyzed by bioinformatics software. Results A total of 7 CRISPR loci were detected in 34 strains. The mutation rate of the first 2 and last 2 bases of the Repeat sequence of CRISPR loci was higher, while the mutation rate of the middle part was lower. Seven CRISPR sites form eight CRISPR structural types, among which the Repeat sequences of CRISPR1 and CRISPR2 are relatively conserved, while the Repeat sequences of CRISPR1 and CRISPR5 can form dumbbell shaped secondary structures. The number of Spacer sequences contained in each CRISPR site ranges from 2 to 15, with an average of 2.43. The 136 Spacer sequences detected were not only homologous to Listeria plasmids and bacteriophages, but also homologous to uncultured virus sequences, staphylococcal bacteriophages, and Listeria innocua. The same CRISPR genotype did not show large-scale clustering, but some strains in the same year were in the same evolutionary cluster with close genetic relationships. Conclusion The CRISPR structure of Listeria monocytogenes in this study exhibits high specificity, and its homology with bacteriophages provides a theoretical basis for the application of bacteriophages in the control and prevention of Listeria monocytogenes.

Objective To evaluate the change of energy metabolism during cisplatin-induced acute kidney injury.Methods Adult CD-1 male mice were intraperitoneally injected with a single dose of cisplatin (20 mg/kg), and renal function and renal tissue pathology were tested;gene expression was analyzed and signaling pathways were en-riched in cisplatin-treated renal tubular epithelial cells using transcriptome; the contents of renal glycolysis and a-mino acid metabolites were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) .Re-sults Serum urea nitrogen and blood creatinine significantly increased in cisplatin-treated mice.Pathological his-tology observed swelling and shedding of renal tubular epithelial cells.Transcriptome analysis revealed that 2632 genes were upregulated and 2799 genes were downregulated in cisplatin-treated HK-2 cells.GO and KEGG analy-sis showed that differential genes were enriched in energy metabolism.The GSEA analysis results showed that cispl-atin caused an upregulation of the oxidative phosphorylation pathway and a downregulation of the glycolysis pathway in renal tubular epithelial cells, further KEGG analysis demonstrated that cisplatin caused changes in the expression of amino acid genes in renal cells.Metabolomics showed that the contents of glycolytic intermediates and several a-mino acids were altered in the kidney of cisplatin-treated mice.Conclusion Cisplatin-induced acute renal injury is accompanied by modification in renal tubular cell glycolysis and amino acid metabolism.

With the increasing proportion of human papilloma virus(HPV)infection in the pathogenic factors of oro-pharyngeal cancer,a series of changes have occurred in the surgical treatment.While the treatment mode has been im-proved,there are still many problems,including the inconsistency between diagnosis and treatment modes,the lack of popularization of reconstruction technology,the imperfect post-treatment rehabilitation system,and the lack of effective preventive measures.Especially in terms of treatment mode for early oropharyngeal cancer,there is no unified conclu-sion whether it is surgery alone or radiotherapy alone,and whether robotic minimally invasive surgery has better func-tional protection than radiotherapy.For advanced oropharyngeal cancer,there is greater controversy over the treatment mode.It is still unclear whether to adopt a non-surgical treatment mode of synchronous chemoradiotherapy or induction chemotherapy combined with synchronous chemoradiotherapy,or a treatment mode of surgery combined with postopera-tive chemoradiotherapy.In order to standardize the surgical treatment of oropharyngeal cancer in China and clarify the indications for surgical treatment of oropharyngeal cancer,this expert consensus,based on the characteristics and treat-ment status of oropharyngeal cancer in China and combined with the international latest theories and practices,forms consensus opinions in multiple aspects of preoperative evaluation,surgical indication determination,primary tumor re-section,neck lymph node dissection,postoperative defect repair,postoperative complication management prognosis and follow-up of oropharyngeal cancer patients.The key points include:① Before the treatment of oropharyngeal cancer,the expression of P16 protein should be detected to clarify HPV status;② Perform enhanced magnetic resonance imaging of the maxillofacial region before surgery to evaluate the invasion of oropharyngeal cancer and guide precise surgical resec-tion of oropharyngeal cancer.Evaluating mouth opening and airway status is crucial for surgical approach decisions and postoperative risk prediction;③ For oropharyngeal cancer patients who have to undergo major surgery and cannot eat for one to two months,it is recommended to undergo percutaneous endoscopic gastrostomy before surgery to effectively improve their nutritional intake during treatment;④ Early-stage oropharyngeal cancer patients may opt for either sur-gery alone or radiation therapy alone.For intermediate and advanced stages,HPV-related oropharyngeal cancer general-ly prioritizes radiation therapy,with concurrent chemotherapy considered based on tumor staging.Surgical treatment is recommended as the first choice for HPV unrelated oropharyngeal squamous cell carcinoma(including primary and re-current)and recurrent HPV related oropharyngeal squamous cell carcinoma after radiotherapy and chemotherapy;⑤ For primary exogenous T1-2 oropharyngeal cancer,direct surgery through the oral approach or da Vinci robotic sur-gery is preferred.For T3-4 patients with advanced oropharyngeal cancer,it is recommended to use temporary mandibu-lectomy approach and lateral pharyngotomy approach for surgery as appropriate;⑥ For cT1-2N0 oropharyngeal cancer patients with tumor invasion depth＞3 mm and cT3-4N0 HPV unrelated oropharyngeal cancer patients,selective neck dissection of levels ⅠB to Ⅳ is recommended.For cN+HPV unrelated oropharyngeal cancer patients,therapeutic neck dissection in regions Ⅰ-Ⅴ is advised;⑦ If PET-CT scan at 12 or more weeks after completion of radiation shows intense FDG uptake in any node,or imaging suggests continuous enlargement of lymph nodes,the patient should undergo neck dissection;⑧ For patients with suspected extracapsular invasion preoperatively,lymph node dissection should include removal of surrounding muscle and adipose connective tissue;⑨ The reconstruction of oropharyngeal cancer defects should follow the principle of reconstruction steps,with priority given to adjacent flaps,followed by distal pedicled flaps,and finally free flaps.The anterolateral thigh flap with abundant tissue can be used as the preferred flap for large-scale postoperative defects.

Objective:To investigate the diagnostic value of CT-guided puncture biopsy combined with serum gamma-glutamyltransferase (GGT) and abnormal prothrombin (PIVKA-Ⅱ) in serum alpha-fetoprotein(AFP) negative primary liver cancer (PHC).Methods:Eighty patients with AFP negative PHC treatment in Fuyang Women and Children′s Hospital from January 2018 to March 2021 were selected as AFP negative PHC group, and another 85 patients diagnosed with benign liver tumor during the same period were selected as the control group retrospectively. The patients of the two groups underwent CT-guided biopsy and the levels of GGT and PIVKA-Ⅱ were detected. The single diagnostic value and combined diagnostic value of AFP negative PHC were analyzed by receiver operating characteristic (ROC) curve.Results:Seventy-five of the 80 patients in the AFP negative PHC group were confirmed as liver malignant lesions by biopsy, with a coincidence of 93.75%, and 84 of the 85 patients in the control group were confirmed as liver benign lesions by biopsy, with a coincidence of 98.82%. The levels of AFP, GGT and PIVKA-Ⅱ in AFP negative PHC group were significantly higher than those in the control group: (175.67 ± 39.58) μg/L vs. (18.74 ± 7.42) μg/L, (1 245.37 ± 255.41) U/L vs. (486.63 ± 89.05) U/L, (385.49 ± 30.27) AU/L vs. (25.84 ± 7.66) AU/L, there were statistical differences ( P<0.05). Spearman correlation analysis showed that serum AFP was positively correlated with GGT and PIVKA-Ⅱ ( r = 0.858 and 0.429, P<0.05). The results of ROC curve showed that the area under curve of CT-guided biopsy combined with GGT and PIVKA-Ⅱ in the diagnosis of AFP negative PHC was 0.877, the sensitivity was 91.19%, the specificity was 87.34%. Conclusions:CT-guided biopsy combined with GGT and PIVKA-Ⅱ detection of AFP negative PHC can effectively improve the diagnostic value.

This paper summarized the current status of the commonly used quality of life assessment scales of bronchiectasis （BE） at home and abroad， and compared the characteristics and differences of each scale from the basic framework， domain， item pool， application status and quality assessment， so as to provide a basis for the development and application of scales based on the mode of combination of disease and syndrome. There are currently 12 assessment tools for evaluating the quality of life in BE with good reliability， validity， and responsiveness， but only quality of life - bronchiectasis （QOL-B） and bronchiectasis health questionnaire （BHQ） are specifically developed for BE patients. The development process of these scales is mostly rooted in western culture and modern medicine， making it difficult to highlight the characteristics and advantages of traditional Chinese medicine （TCM）. The content of the scales lacks the outcome indicators that BE patients are most concerned about， and cannot fully reflect the subjective feelings of patients. The development and assessment of scales lacks clinical research data and methodological support. Under the guidance of TCM syndrome differentiation and treatment， the development of scales based on the mode of combination of disease and syndrome can emerge the characteristics of TCM， reflect the true feelings of patients， and make up for the limitations of the existing scales of BE. Therefore， based on domestic and international scales， factors of China's national conditions and cultural characteristics of TCM should be considered to develop the scale suitable for Chinese people， which requires the construction of a theoretical model for the combination of disease and syndrome， by taking patients as the center and evaluating the performance and quality of the scale.

H 1-antihistamines are widely used in the treatment of various allergic diseases, but there are still many challenges in the safe and rational use of H 1-antihistamines in pediatrics, and there is a lack of guidance on the prescription review of H 1-antihistamines for children.In this paper, suggestions are put forward from the indications, dosage, route of administration, pathophysiological characteristics of children with individual difference and drug interactions, so as to provide reference for clinicians and pharmacists.

AlM: The Chinese medicinal herb Hanfangji is dried roots of Stephania tetrandra S. Moore (Family, Menispermaceae). Tetrandrine and fangchinoline are two major constituents of Hanfangji and these bisbenzyltetrahydroisoquinoline alkaloids possess anti - cancer and other pharmacological activities. To facilitate further pharmacodynamic investigation of these compounds, a pharmacokinetic investigation was performed in rats and in vitro. METHODS: Pharmacokinetics of tetrandrine and fangchinoline were characterized in rats p.o. or i.v. dosing an aqueous extract of Hanfangji or the individual compound. Unbound levels of systemic exposure to these two alkaloids were assessed using in vitro studies of plasma protein binding, blood-plasma partition, and lysosomal trapping. All the study samples were analyzed by liquid chromatography/mass spectrometry.RESULTS: We found two pharmacokinetic features of tetrandrine and fangchinoline. First, the two compounds had blood levels of systemic exposure substantially higher than the respective plasma levels of systemic exposure. Second, the two compounds exhibited significantly higher systemic exposure levels after p.o. dosing an aqueous extract of Hanfangji than the respective exposure levels after p.o. dosing the individual compound, at the same compound dose levels and under the same conditions for analytical measurement and the same conditions for animal study. Unbound fractions of tetrandrine and fangchinoline in rat plasma were 2%-5% and the concentrations of the alkaloids in rat erythrocytes were 5-times higher than those in rat plasma. Lysosomal inhibitor could block their trapping in lysosomes and significantly reduce their concentrations in HEK-293 cells. CONCLUSlON: The following pharmacokinetic aspects should be noted in pharmacodynamic investigation of tetrandrine and fangchinoline: extensive binding with plasma proteins, extensive binding with erythrocytes, and trapping by lysosomes of tissue cells substantially reduce the levels of unbound tetrandrine and fangchinoline in the systemic circulation.

Objective:To discuss the influence of mindfulness-based stress reduction plus micro-class education on the complications and knowledge mastery rate in surgical patients with diabetes mellitus.Methods:A total of 105 patients, diagnosed as diabetes mellitus complicated with appendicitis in the Affiliated Hospital of Chengde Medical College were selected from January 2019 to January 2020. They were hospitalized for laparoscopic appendectomy and were randomly divided into the control group ( n = 52) and the study group ( n = 53) in accordance with the random number table. The patients in the control group were given routine nursing care, and the patients in the study group were given mindfulness decompression therapy combined with micro-classroom education. The blood glucose control and psychological emotion of the two groups before and after operation, and the postoperative complications and the mastery rate of disease knowledge of the two groups were compared. Results:There was no significant difference in blood glucose indexes between the two groups at baseline ( P>0.05); FBG, HbA1c and other indicators in the two groups were improved during operation and 24h after operation, but FBG (7.38±0.54) mmol/L, HbA1c (6.39±0.21)% and FBG (6.90±0.52) mmol/L and HbA1c (6.10±0.39)% in the study group were lower than those in the control group [(8.16±1.21) mmol/L, (7.53±1.05)%, (7.60±0.57) mmol/L, (6.50±0.41)%], the difference was statistically significant ( t value was 6.789-13.264, P < 0.05); there was no significant difference in SAS and SDS scores between the two groups at baseline ( P > 0.05), until discharge, the SAS and SDS scores of the treatment group were 35.81±5.49 and 42.08±4.91 respectively, which were significantly lower than those of the control group (42.21±5.53, 6.51±4.72) respectively, compared with the corresponding scale scores of the control group, the difference between the two groups after treatment was statistically significant ( t value was 5.386, 4.265, P < 0.05). Compared with 17.31% (9/52) of the control group, the incidence of complications in the study group decreased to 5.66% (3/53), there was significant difference ( χ2 value was 6.789, P < 0.05). The qualified rate of disease knowledge mastery in the study group (98.11%,52/53) was significantly higher than that in the control group (86.53%, 45/52), and the difference was significant ( χ2 value was 5.062, P < 0.05). Conclusion:The mindfulness-based stress reduction plus micro-class education can effectively control the laparoscopic appendectomy patients blood glucose, stabilize the mental emotions, increase the illness knowledge mastery degree, keep in good mood, reduce the postoperative complications and promote the fast recovery.

BACKGROUND: Many studies have investigated heavy metal exposure could increase the occurrence of congenital heart defects (CHDs). However, there are limited data regarding the relationship between cobalt exposure and CHD occurrence in offspring. The aim of this study was to analyze the association between cobalt exposure in mothers and the risk of CHDs in offspring. MATERIALS AND METHODS: In order to explore the association between cobalt exposure and occurrence of congenital heart defect (CHD), a case-control study with 490 controls and 399 cases with CHDs in China were developed. The concentrations of cobalt in hair of pregnant woman and fetal placental tissue were measured and processed by a logistic regression analysis to explore the relationship between cobalt exposure and risk of CHDs. RESULTS: The median concentration of hair cobalt in the control and case group was 0.023 ng/mg and 0.033 ng/mg (aOR, 1.837; 95% CI, 1.468-2.299; P < 0.001), respectively. And the median (5-95% range) fetal placental cobalt concentrations were 19.350 ng/g and 42.500 ng/g (aOR, 2.924; 95% CI, 2.211-3.868; P < 0.001) in the control and case groups, respectively. Significant differences in the middle level of cobalt in hair were found in the different CHD subtypes, including septal defects, conotruncal defects, right ventricular outflow tract obstruction, and left ventricular outflow tract obstruction (P < 0.001). Dramatically, different cobalt concentrations in fetal placental tissue were found in all subtypes of cases with CHDs (P < 0.01). CONCLUSIONS The finding suggested that the occurrence of CHDs may be associated with cobalt exposure.
Adolescent ; Adult ; Case-Control Studies ; China ; Cobalt ; adverse effects ; Female ; Hair ; chemistry ; Heart Defects, Congenital ; chemically induced ; Humans ; Maternal Exposure ; adverse effects ; Placenta ; chemistry ; Pregnancy ; Prenatal Exposure Delayed Effects ; chemically induced ; Risk Factors ; Young Adult

Objective To analyze the characteristics and applicability of the quality of life (QOL) evaluation scale for patients with idiopathic pulmonary fibrosis (IPF). Methods The IPF-QOL evaluation scales at home and abroad were retrieved and analyzed comprehensively and deeply in terms of fields, number of items, scoring methods, applicability and correlation. Results In the literature area of IPF-QOL evaluation scale, the contents mainly included symptoms,daily activity ability, psychological activity, social activity, economic status, curative effect, etc, and the IPF health-related quality of life, dyspnea, cough and other symptoms were evaluated; the number of related items was between 5-74, and Likert 5 grade scoring method was mostly adopted,with scoring results of 0-100 points; the classical scoring theory such as reliability, validity, responsiveness, feasibility, etc were mainly used to analyze the scale assessment, lacking the application of modern measurement theory; mostly, the studies were concentrated in foreign countries,and relatively few in China, especially lack of the development of the evaluation scale for the combination of disease and symptoms based on Chinese culture. Conclusion The development of IPF-QOL evaluation scale has become a research hotspot, and in clinical application, it is necessary to base on the different characteristics of the scale to improve the accuracy of the evaluation results; in the scale development process, the standardization, comprehensiveness of assessment, and the rationality of scoring results should be strengthened in order to obtain an effective tool which can be provided and suitable for IPF-QOL evaluation.