Beta-amylases (BAMs), key enzymes in starch hydrolysis, play an important role in plant growth, development, and resistance to abiotic stress. To mine the saline-alkali tolerance-related BAM genes in alfalfa (Medicago sativa L.), we identified MsBAM genes in the whole genome. The physicochemical properties, phylogeny, gene structures, conserved motifs, secondary structures, promoter cis-acting elements, chromosome localization, and gene replication relationships of BAM gene family members were analyzed. RNA-seq and quantitative real-time PCR (qRT-PCR) were employed to analyze the expression patterns of BAM family members under saline-alkali stress. The results showed that 54 BAM genes were identified in the genome, which were classified into 8 subgroups according to the phylogenetic tree. The members of the same subgroup had similar gene structures except that those of subgroups 1 and 7 had large differences. Conserved motif analysis showed that all MsBAM proteins had a typical glycohydrolysis domain. The chromosome localization analysis showed that MsBAM gene family members were unevenly distributed on 27 chromosomes. The duplication of gene segments led to the increase in BAM gene number in alfalfa. The promoters of BAM genes contained a large number of elements in response to plant hormones and stress. Transcriptome data and qRT-PCR results showed that the expression levels of most MsBAM genes were up-regulated in response to saline-alkali stress. Under the saline-alkali stress, the expression levels of 28 genes, including MsBAM6, were up-regulated on days 1 and 7, and those of 5 genes, including MsBAM9, were up-regulated by over 2 folds. In addition, under salt-alkali stress, BAM activity and soluble sugar content were significantly increased. These results indicate that BAM genes play a key role in alfalfa in response to saline-alkali stress, laying a foundation for further research in this field.
Medicago sativa/physiology* ; beta-Amylase/metabolism* ; Phylogeny ; Gene Expression Regulation, Plant ; Stress, Physiological/genetics* ; Multigene Family ; Alkalies ; Plant Proteins/genetics*

On-site supervision is a risk-based regulatory system that requires the scientific development of supervision plans for quality risks and hidden dangers in pharmaceutical enterprises， the rational allocation of supervision resources based on their risk levels， and the implementation of classified supervision measures. In this study， the quality risk monitoring business support system is set up for pharmaceutical enterprises by establishing the quality risk expert database and quality risk monitoring index system for pharmaceutical enterprises based on the difficulty analysis of on-site drug supervision. Based on this support system， the quality risk classification method， the differentiated spot check strategy and business auxiliary visualization system are established. This support system is used to learn the risk level of pharmaceutical enterprises， so as to innovate supervision methods and optimize monitoring strategies. Taking Jiangxi Province as an example， it is verified that the support system can guide the risk assessment of sample enterprises， can improve the targeting of on-site drug supervision in the process of technical review， scheme editing， on-site implementation and comprehensive evaluation， and can effectively improve the quality and efficiency of supervision.

OBJECTIVE To explore the causal relationship of circulating cytokines and sensorineural deafness by Mendelian randomization.METHODS Large-scale genome-wide association studies(GWAS)data sets based on UK Biobank were used to identify single nucleotide polymorphisms(SNP)associated with exposure and outcome.The SNP(F>10)significantly associated with exposure data(P<5e-6)and strongly associated with exposure in the F statistic was selected as the instrumental variable for a two-sample Mendelian randomization analysis.The causal effect was estimated using MR-Egger,regression and inverse variance weighting(IVW).Sensitivity analysis was performed to assess the robustness of the results.RESULTS This study confirmed that VEGF(OR=1.04,95%CI:1.01-1.07,P=0.00),SDF-1α(OR=1.10,95%CI:1.02-1.18,P=0.01)and IL-13(OR=1.04,95%CI:1.01-1.06,P=0.01),IL-10(OR=1.06,95%CI:1.02-1.11,P=0.01),IL-7(OR=1.05,95%CI:1.01-1.08,P=0.00)increased the risk of sensorineural deafness.The backward analysis did not find that sensorineural deafness significantly affected the levels of these cytokines.No significant heterogeneity or horizontal pleiotropy was observed among the instrumental variables.CONCLUSION This study reveals a positive correlation between elevated levels of VEGF,SDF-1α,IL-13,IL-10,and IL-7 with sensorineural hearing loss,providing novel targets for further development of biomarkers for the prediction,treatment,and prevention of sensorineural hearing loss.

Objective:To explore the predictive value and prognosis effect of calprotectin on acute kidney injury (AKI) in patients with sepsis.Methods:A prospective observational study was conducted. From December 2018 to November 2020, patients with sepsis admitted to the Emergency Department of China Rehabilitation Research Center were enrolled. General clinical data of patients were collected continuously, and the acute physiology and chronic health evaluationⅡ (APACHEⅡ) score, sequential organ failure assessment (SOFA) score and calprotectin were evaluated in 24 h after admission. The patients were divided into the AKI group and non-AKI group according to the occurrence of AKI within 7 days after admission. Calprotectin level and other clinical data were compared between the two groups. Logistic regression was used to analyze the risk factors for AKI in patients with sepsis, and receiver operating characteristic (ROC) curve was plotted to evaluate the predictive value of calprotectin for AKI in patients with sepsis. The patients with AKI were further divided into the survival group and death group according to the 28-day outcome, and the calprotectin levels between the two groups were compared.Results:A total of 207 patients with sepsis were enrolled, and the incidence of AKI was 68.12% (141/207). The level of calprotectin in patients with AKI was higher than that in patients without AKI [4.65 (3.25, 5.61) μg/mL vs. 3.42 (2.29, 4.09) μg/mL, P < 0.001]. Multivariable Logistic regression analysis showed that APACHEⅡ score ( OR=1.090, 95% CI: 1.043-1.139), C-reactive protein ( OR=1.004, 95% CI: 1.001-1.008) and calprotectin ( OR=1.590, 95% CI: 1.269-1.991) were independent risk factors for AKI in patients with sepsis. The area under ROC curve (AUC) of calprotectin for predicting AKI was 0.716 (95% CI: 0.643-0.788). The cutoff value of prediction was 4.63 μg/mL with the Yoden index of 0.405, which yielded a sensitivity of 0.511 and a specificity of 0.894. When calprotectin was combined with APACHE II score and SOFA score respectively, the predictive ability was significantly improved with the AUC of 0.768 (95% CI: 0.701-0.834) and 0.769 (95% CI: 0.701-0.837), respectively. We further divided patients with AKI into the survival group and non-survival group according to the 28-day outcome and there was no significant difference in calprotectin between the two groups [4.80 (3.40, 5.76) μg/mL vs. 4.19 (2.89, 5.29) μg/mL, P < 0.05]. Conclusions:The level of calprotectin in the AKI group is higher than that in the non-AKI group. Calprotectin can be regarded as an effective predictor of AKI in patients with sepsis, and the combination with APACHEⅡ score or SOFA score will improve its predictive efficacy. However, there is no significant difference in the concentration of calprotectin for patients with sepsis associated AKI with different prognosis.

Objective:To explore the values of albumin-bilirubin (ALBI) score, carcinoembryonic antigen (CEA) and combination of the two in the prognostic assessment of colorectal cancer patients with postoperative liver metastasis.Methods:The clinicopathological data of 98 colorectal cancer patients with postoperative liver metastasis who were admitted to Lianyungang Oriental Hospital and receiving adjuvant chemotherapy from January 2016 to March 2020 were retrospective analyzed. The data of serum protein, bilirubin, and CEA before chemotherapy were obtained, the relationship between serum protein and bilirubin was analyzed, and the ALBI score was calculated. The ALBI-CEA score was judged according to the ALBI score and the CEA level. ALBI score > -2.60 points was categorized as high ALBI group, and ALBI score ≤ -2.60 points was categorized as low ALBI group; CEA >5 ng/ml was categorized as high CEA group, and CEA ≤5 ng/ml was categorized as low CEA group; patients were categorized into 0, 1, and 2 points groups based on ALBI-CEA score. Overall survival (OS) and progression-free survival (PFS) of ALBI score, CEA and ALBI-CEA score subgroups were analyzed by Kaplan-Meier method; with the actual survival and progress status of the patients as the gold standard, receiver operating characteristic (ROC) curve was used to analyze the effect of 3 indicators to assess patients' OS and PFS, and area under the curve (AUC) was compared; Cox proportional hazards model was used to analyze the influencing factors of OS and PFS.Results:The median albumin and bilirubin levels of the 98 patients were 34.4 g/L (26.8-42.8 g/L) and 16.6 μmol/L (7.6-44.6 μmol/L), and the result of Pearson correlation analysis showed a negative correlation between the levels of albumin and bilirubin ( r = -0.282, P < 0.001). The 3-year OS and PFS rates in the high ALBI group were lower than those in the low ALBI group (OS rate: 9.2% vs. 33.3%, PFS rate: 7.7% vs. 18.2%), and the differences in OS and PFS between the two groups were statistically significant ( χ2 values were 27.64, 23.30, both P < 0.001). The 3-year OS and PFS rates in the high CEA group were lower than those in the low CEA group (OS rate: 7.1% vs. 42.9%, PFS rate: 7.1% vs. 21.4%), and the differences in OS and PFS between the two groups were statistically significant ( χ2 values were 23.71, 17.14, both P < 0.001). The 3-year OS rates in the ALBI-CEA score 0, 1 and 2 points groups were 77.8%, 20.9% and 2.2%, and the 3-year PFS rates were 44.4%, 9.3% and 6.5%, and there were statistical differences in OS and PFS among the three groups ( χ2 values were 102.36, 76.55, both P < 0.001). The ROC curve analysis showed that the AUC of ALBI score, CEA and ALBI-CEA score for assessing OS were 0.688 (95% CI 0.544-0.832), 0.754 (95% CI 0.618-0.890) and 0.828 (95% CI 0.723-0.933) (all P < 0.05), and the AUC for assessing PFS were 0.618 (95% CI 0.436-0.799), 0.646 (95% CI 0.464-0.829) and 0.682 (95% CI 0.494-0.870) (all P > 0.05). Multivariate Cox regression analysis showed that ALBI-CEA score was an independent influencing factor for OS (2 points vs. 0 point: HR = 17.254, 95% CI 8.385-35.504, P < 0.001) and PFS (2 points vs. 0 point: HR = 6.144, 95% CI 3.725-10.134, P < 0.001) of patients. Conclusions:The colorectal cancer patients with liver metastasis and high ALBI-CEA score are at high risk of death and disease progression and have a poor prognosis, and they are recommended to receive intensive treatment.

Objective: To analyze the latent category characteristics of lifestyle habits of adolescents aged 12-18 years, and to explore its relationship with obesity. Methods: Data of 3 252 adolescents aged 12-18 years in the established China Nutrition and Health Survey were used for this study. The contents of the survey include basic situation, living habits, and body mass index. Latent category analysis was performed on 11 lifestyle habits on eating and exercise. Chi square test was used to explore the relationship between the potential categories of lifestyle habits and obesity. Results: All the participants were classified into 3 categories:mixed ( n =1 092, 35.58%), passive ( n =765, 23.52%), and healthy ( n =1 395, 42.90%) groups. Body mass index showed significant differences across three categories ( χ 2=19.66, P ＜0.01). In the BMI classification of adolescents, the healthy type has the highest proportion in the light type, overweight type and obesity type(41.61%,47.87%,50.00%). Conclusion The lifestyle habits of adolescents aged 12-18 years show obvious classification characteristics, which have different correlations with obesity. Targeted interventions should be carried out according to the characteristics of lifestyle habits under different body mass index.

Objective: To examine the effects of obesity and central obesity on hypertension, so as to provide insights into the prevention and control measures of hypertension. Methods: From September to December 2018, residents at ages of 35 to 75 years were sampled using the multi-stage random sampling method in Baiyin District, Baiyin City, Gansu Province, and subjected to questionnaire surveys and physical examinations. The interaction between obesity/central obesity and hypertension was evaluated using logistic regression analysis. The synergy index ( SI ), relative excess risk due to interaction ( RERI ) and attributable proportion due to interaction ( AP ) were calculated using Excel compiled by Andersson et al. Results: A total of 6 246 questionnaires were allocated and 6 169 valid questionnaires were recovered, with an effective recovery rate of 98.77%. The respondents included 3 038 men ( 49.25% ) and 3 131 women (50.75%), with a mean age of ( 52.05±8.78 ) years. There were 832 respondents with obesity ( 13.49% ) and 2 278 with central obesity ( 36.93% ). The crude and standardized prevalence rates of hypertension were 35.89% and 33.05%, respectively. Multivariable logistic regression analysis showed that obesity ( OR=2.020, 95%CI: 1.705-2.393 ) and central obesity ( OR=1.622, 95%CI: 1.433-1.836 ) were statistically associated with hypertension. There was no multiplicative interaction between obesity or central obesity and hypertension ( OR=1.011, 95%CI: 0.655-1.560 ), and no additive interaction was detected between obesity or central obesityand hypertension ( SI=1.405, 95%CI: 0.815-2.424; RERI=0.658, 95%CI: -0.298 to 1.614; AP=0.201, 95%CI: -0.075 to 0.476 ). Conclusions Obesity and central obesity increase the risk of hypertension; however, no interaction is detected between obesity or central obesity and hypertension.

Objective:To explore the effect of continuing care based on timing theory in the postoperative rehabilitation of patients with ectopic pregnancy.Methods:Using the convenient sampling method, a total of 86 patients with ectopic pregnancy who were admitted to Department of Obstetrics and Gynecology of the Anhui No.2 Provincial People 's Hospital were selected as the research objects from November 2018 to November 2020. According to the random number table method, they were divided into the observation group and the control group, with 43 cases in each group. The patients in the control group were given routine care and follow-up, and patients in the observation group implemented the continuing care based on timing theory on the basis of the control group. The Exercise of Self-Care Agency Scale (ESCA) , Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS) scores, nursing satisfaction and the incidence of complications were compared between the two groups at the time of admission and one month after discharge. Results:The scores of all dimensions in the ESCA of the observation group were higher than those of the control group, and the differences were statistically significant (all P<0.05) . The SAS and SDS scores of the observation group were lower than those of the control group, and the differences were statistically significant (all P<0.05) . The incidence of complications in the observation group was 4.65% (2/43) , which was lower than 18.60% (8/43) in the control group, and the difference was statistically significant ( P<0.05) . The satisfaction of nursing care of patients in the observation group was 95.35% (41/43) , which was higher than 81.40% (35/43) of the control group, and the difference was statistically significant ( P<0.05) . Conclusions:The implementation of transitional care under the guidance of timing theory for patients after ectopic pregnancy can achieve good rehabilitation effects and improve the self-care ability of patients, which is worthy of clinical promotion.

The aim of the study was to develop a simple, rapid and accurate LC-MS/MS method for the determination of digoxin.Digoxin-d3 was taken as the internal standard (IS), and sample preparation was achieved by liquid-liquid extraction.Chromatographic separation was performed on a Kinetex C18 column (2.1 mm × 50 mm, 2.6 μm; Phenomenex) using an isocratic elution with merely 2 min for each sample.The mobile phase consisted of water and acetonitrile solutions, both containing 1 mmol/L ammonium acetate and 1 mmol/L formic acid (55∶45).The detection was conducted on a TripleQuadTM 4500MD mass spectrometer coupled with electrospray ionization interface under positive-ion multiple reaction monitoring mode.The transitions were m/z 798.5 → 651.3 and m/z 801.6 → 654.4 for digoxin and digoxin-d3, respectively.Results showed that the method was linear over the range of 0.100-20.0 ng/mL.The selectivity, accuracy and precision, recovery and stability of the method were all within the acceptable limits with no matrix effect.This method was successfully applied to a girl treated with digoxin with substantial improvement of therapeutic effect and elimination of toxic reaction, so it can provide valuable fuidance and reference for individualized medication in clinical practice.

With the rapid development of biopharmaceuticals, therapeutic monoclonal antibodies (mAbs) are widely accepted due to their low side-effects and high therapeutic efficacy. Individual differences in the response to mAb drugs put forward new requirements for therapeutic drug monitoring of antibody drugs.Therefore, the need for accurate and robust bioanalytical methods is increasing.Recently, LC-MS/MS has been gaining increasing interest in the field of large molecules.In this article, the recent advances in this emerging field are reviewed, along with common issues and analytical approaches.Thus, this review article is helpful for better understanding the advance of LC-MS/MS technique in the field of therapeutic drug monitoring for mAbs.