In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.

Objective:To investigate the expressions of tissue inhibitor of matrix metalloproteinase-1 (TIMP1) and fibronectin 1 (FN1) in pregnancy associated breast cancer (PABC) and their correlations with expression of E-cadherin (E-cad).Methods:The clinicopathological data of 55 PABC patients in Binzhou People's Hospital Affiliated to Shandong First Medical University from January 2011 to December 2020 were retrospectively analyzed. Immunohistochemistry was used to detect expressions of TIMP1, FN1 and E-cad in cancer tissues and corresponding paracancerous tissues (>3 cm from the edge of the tumor foci). The expressions of TIMP1 and FN1 proteins in fresh intraoperative frozen cancer tissues and paracancerous tissues of 10 PABC patients were detected by Western blotting. The correlations of TIMP1 and FN1 expressions with clinicopathological characteristics of patients were analyzed by χ2 test, the correlation of TIMP1 and FN1 expressions with E-cad expression was analyzed by Spearman method, and the correlation of TIMP1 and FN1 expressions with survival was analyzed by Kaplan-Meier method. Results:The positive rates of TIMP1 and FN1 in PABC tissues were 72.7% (40/55) and 58.2% (32/55), and 25.5% (14/55) and 18.2% (10/55) in paracancerous tissues, and the differences were statistically significant ( χ2 values were 24.59 and 18.64, both P < 0.001). The results of Western blotting showed that the relative expressions of TIMP1 and FN1 proteins in the fresh cancer tissues of 10 PABC patients was higher than those in the corresponding paracancerous tissues (1.60±0.76 vs. 0.62±0.29, 1.31±0.62 vs. 0.44±0.15), and the differences were statistically significant ( t values were 5.92 and 4.86, both P < 0.001). The expressions of TIMP1 and FN1 in PABC tissues were correlated with estrogen receptor expression, Ki-67 positivity index, TNM stage and lymph node metastasis (all P < 0.05). The expressions of TIMP1 and FN1 were negatively correlated with expression of E-cad in PABC ( r values were -0.471 and -0.432, both P < 0.001). Five cases were lost to follow-up, and the remaining 50 cases had a median follow-up time of 43 months (12-90 months). Among the 50 cases, 36 cases were TMP1-positive and 29 cases were FN1-positive. The overall survival of TIMP1-negative group and FN1-negative group were better than those of the corresponding positive group ( χ2 values were 4.49 and 6.06, both P < 0.05); the median overall survival time of TIMP1-positive group and FN1-positive group were 51 months (95% CI 37-65 months) and 43 months (95% CI 32-53 months), while that of TIMP1-negative group and FN1-negative group were 89 months (95% CI 84-93 months) and 87 months (95% CI 85-92 months). Conclusions:TIMP1 and FN1 expressions are elevated in PABC tissues and negatively correlated with E-cad expression, TIMP1 and FN1 may be involved in PABC invasion through epithelial-mesenchymal transition and affect the prognosis of patients.

Objective:To explore the change of medical students' self-regulated learning from traditional medicine to clinical practice in emergency department, and to provide direction for optimizing the curriculum system and exploring new education and teaching methods.Methods:Through questionnaires and in-depth interviews, the self-regulated learning scale was applied, and the 6-level Likert scale was used for scoring. A total of 118 medical students, including 8-year undergraduates majoring in clinical medicine of Peking Union Medical College, were evaluated in the traditional medicine course stage (traditional group), emergency clinical practice stage (clinical group), and emergency clinical practice stage of the "4 + 4" reform pilot class ("4 + 4" pilot class group) of clinical medicine. The learning situation and related influencing factors were self-regulated. SPSS 23.0 was used for one-way analysis of variance.Results:Among the eight-year medical students of Peking Union Medical College, there were 48 males (40.7%) and 70 females (59.3%). The three groups of 8-year traditional medicine course stage, 8-year emergency clinical practice stage and "4 + 4" pilot class of emergency clinical practice stage were analyzed. The results showed that the total score of self-regulated learning ability in the clinical group was significantly lower than that in the traditional group [(326.2±31.9) vs. (347.7±40.2) points]. The subscales of self-regulated learning ability were analyzed respectively. In the learning motivation subscale, the score of external goal orientation of the clinical group is significantly lower than that of the traditional group [(8.9±2.3) vs. (10.0±2.9) points] and the score of the "4 + 4" pilot class group is significantly higher than that of the clinical group [(11.0±3.5) vs. (8.9±2.3) points]. In the learning strategy subscale, there was no significant difference among the three groups. In the resource management subscale, the scores of time and study environment [(6.5±1.1) vs. (7.5±1.9) points], learning management [(37.7±4.0) vs. (40.3±3.0) points] and help-seeking [(32.7±5.3) vs. (37.5±9.5) points] of the clinical group decreased significantly compared with those of the traditional group, while the scores of learning management [(40.2±7.3) vs. (37.7±4.0) points] and help-seeking [(38.7±7.6) vs. (32.7±5.3) points] of the "4+4" pilot class group increased significantly compared with those of the clinical group.Conclusion:The self-regulated learning of clinical medical students has changed significantly during the transition from traditional medical class to emergency clinical practice. The decrease of external goal orientation and resource management may be the important reason for the decline of the self-regulated learning ability of 8-year undergraduate medical students in the clinical environment during emergency practice.

ObjectiveTo screen and validate key enzyme genes affecting the polysaccharide content in different Polygonatum species and perform in-depth amino acid sequence analysis by transcriptomic analysis of P. zanlanscianense， P. kingianum， and P. cyrtonema rhizomes to enrich the transcriptome data of Polygonatum plants and provide references for polysaccharide biosynthesis mechanism and genetic improvement. MethodThe Polygonatum transcriptome was sequenced and analyzed using the Illumina NovaSeq high-throughput sequencing platform， and the differences in the transcriptomes of the three Polygonatum species were compared and according to the annotations of Nr， Gene Ontology （GO）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） databases. The key enzymes in the polysaccharide metabolism pathway were screened， and the expression of key enzyme genes was clustered and correlated with the polysaccharide content. Finally， Real-time polymerase chain reaction （Real-time PCR） was performed to validate the eight key enzyme genes， and the key genes of polysaccharide biosynthesis were further screened for homologous gene sequence analysis in combination with sequencing results， followed by constructing phylogenetic trees， predicting motifs， conserved structural domains， protein sequence isoelectric points， and molecular weights， and constructing 3D protein structures by using homology modeling method. ResultThe annotation of the Nr database revealed that three Polygonatum species had the highest gene homology with Asparagus officinalis. GO database annotation results showed that three Polygonatum species differed significantly in binding， catalytic activity， metabolic processes， and cellular components， while the KEGG pathway annotation results indicated that three Polygonatum species differed significantly in the starch and sucrose metabolic pathway and galactose metabolic pathway. According to clustering analysis， correlation analysis， Real-time PCR， expression profiles， and structural and functional predictions of amino acid sequences， the key enzyme significantly affecting the polysaccharide content in different Polygonatum species was inferred to be β-fructofuranosidase （sacA）. ConclusionSacA may be the main influencing factor for the difference in polysaccharide content of Polygonatum， and is also an important reason why Polygonatum polysaccharides are mainly fructans.

Objective:To compare clinical characteristics and results on parameters for muscle measurement in elderly sarcopenia patients of different age groups.Methods:This was a cross-sectional study, and 404 outpatients aged 60 years and over were enrolled and received a long-term follow-up.According to the World Health Organization classification of the elderly, the subjects were divided into a youngest-old group, a middle old group and an oldest-old group; and according to the diagnostic criteria of the Asian Working Group for Sarcopenia in 2019, elderly people of all ages were divided into a sarcopenia group and a non-sarcopenia group.General clinical data were recorded, and the upper arm circumference, leg circumference, skeletal muscle index, grip strength and walking speed, the Short Physical Performance Battery and timed up and go were conducted.At the same time, the body composition was measured by multifrequency bioelectrical impedance analysis.Results:The subjects had a mean age of(85.24 ± 8.23)years, including 90(22.28%)youngest-old, 165(40.84%)middle old and 149(36.88%)oldest-old.123 patients(30.45%)were diagnosed with sarcopenia, with a prevalence of 27.78%(25)in the youngest-old group, 28.48%(47)in the middle old group and 34.23%(51)in the oldest-old group.Compared with those without sarcopenia, the basal metabolic rate in sarcopenia patients of all age groups decreased significantly, the levels of hemoglobin and albumin in youngest-oldsarcopenia patients decreased, the waist circumference and body mass index in middle old and oldest-old sarcopenia patients decreased, and the body fat rate in middle old sarcopenia patients increased.In the subjects with sarcopenia, the proportion of men in the oldest-old group was higher than in the other two groups.Compared with the youngest-old group, the levels of alanine aminotransferase and prealbumin decreased, serum creatinine increased, and skeletal muscle index values did not change in the middle old and longest-old groups, but handgrip strength(27.7±5.9 vs.23.2±6.9 vs.21.4±5.8, F=5.81, P=0.004), gait speed(0.8±0.2 vs.0.7±0.2 vs.0.5±0.2, F=11.88, P=0.000)and calf circumference(33.4±3.3 vs.15.9± 8.4 vs.31.5±3.1 vs.30.9±3.3, F=3.58, P=0.031)significantly decreased, and time up and go values(10.0±2.1 vs.15.9±8.4 vs.20.8±12.8, F=6.98, P=0.001)increased in the middle old and longest-old groups.Partial correlation analysis showed that age had a significant negative correlation with handgrip( r=-0.374, P=0.001), daily gait speed( r=-0.441, P=0.000), and calf circumference( r=-0.223, P=0.017), but a significant positive correlation with timed up and go( r=0.319, P=0.009). Conclusions:Compared with youngest-old sarcopenia patients, middle old patients show significant decline in muscle function, muscle strength and muscle mass in the lower limb.With increasing age, the impact of changes in muscle function, muscle strength and regional muscle mass on adverse events should receive heightened attention.

Objective:To investigate the prevalence of sarcopenia and associated factors in hospitalized elderly patients.Methods:This was a cross-sectional study.A total of 578 patients admitted to the Geriatrics Department of our hospital were consecutively recruited according to the admission criteria.Patients were divided into the sarcopenia group(n=202, 34.95%)and non-sarcopenia group(n=376, 65.05%)based on the diagnostic criteria(2014)of the Asian Working Group for Sarcopenia.Their clinical data and laboratory parameters were collected.All patients underwent comprehensive geriatric assessment.Results:(1)The detection rate of sarcopenia in these hospitalized patients was 34.95%.(2)Age, free thyroxine(FT4), and the prevalences of diabetes and osteoporosis and the incidence of falls in the past year were higher while body mass index(BMI), calf circumference, hemoglobin(Hb), albumin(ALB), triglycerides(TG), low-density cholesterol(LDL), glycosylated hemoglobin(HbA1c)and free triiodothyronine(FT3)were lower in sarcopenia patients than in non-sarcopenia patients, with statistical significance.Scores on the mini-mental state examination(MMSE)and activities of daily living(ADL)were lower in sarcopenia patients than in non-sarcopenia patients while scores on nutritional risk screening 2002(NRS2002)and the FRAIL scale were higher, all with statistical significance.(3)Binary logistic regression analysis revealed that osteoporosis and FRAIL score were risk factors for sarcopenia( OR=9.083 and 2.505, P<0.001)and BMI, calf circumference and ADL score were protective factors for sarcopenia( OR=0.735, 0.774 and 0.967, P<0.05). (4)ROC analysis showed that the areas under the curve for FRAIL score, BMI, calf circumference and ADL score were 0.832, 0.805, 0.841 and 0.812, respectively, with threshold values at 2.5 points, 23.52 kg/m 2, 32.5 cm and 92.5 points.(5)The chi-square test for sarcopenia screening using various related factors found osteoporosis, calf circumference <32.5 cm and ADL <90 points had higher sensitivity(0.787, 0.807, 0.817)while FRAIL ≥ 3 points and BMI <23.5 had slightly lower sensitivity(0.683, 0.708), with each related factor having a high negative detection rate(0.833-0.888). Conclusions:The prevalence of sarcopenia in hospitalized elderly is more than 1/3.Osteoporosis and FRAIL score are risk factors for sarcopenia.BMI, calf circumference and ADL score are protective factors for sarcopenia.All relevant factors have practical clinical value and can be used for preliminary screening of sarcopenia.

Objective:To compare the in vitro susceptibility of fluconazole-resistant Candida albicans strains from superficial and deep infections to 8 antifungal drugs, and to compare drug resistance mutations in these strains. Methods:According to the Clinical and Laboratory Standards Institute (CLSI) protocol M27-A4, 26 deep infection-derived and 33 superficial infection-derived drug-resistant Candida albicans strains were tested for in vitro susceptibility to 8 antifungal drugs (fluconazole, voriconazole, itraconazole, posaconazole, amphotericin B, fluorocytosine, terbinafine, and micafungin) alone or in combination. DNA was extracted from all drug-resistant strains, and mutations in 3 drug resistance genes, including ERG3, ERG11 and FUR1, were detected by PCR. Normally distributed measurement data with homogeneous variance were compared between two groups by using two-independent-sample t test, non-normally distributed measurement data with non-homogeneous variance were compared using Mann-Whitney U test, and enumeration data were compared using chi-square test. Results:The minimum inhibitory concentrations (MICs) of fluconazole, itraconazole, voriconazole, posaconazole and fluorocytosine all significantly differed between the superficial infection group and deep infection group (all P < 0.05) , while there was no significant difference in the MIC of amphotericin B or micafungin between the two groups (both P > 0.05) . The MIC of terbinafine was >64 μg/ml in 96.6% of the above strains, so could not be compared between groups. As combination drug susceptibility testing revealed, the combination of terbinafine with azoles (fluconazole, voriconazole, itraconazole or posaconazole) showed synergistic inhibitory effects against 15 Candida albicans strains (7 strains from deep infections, 8 strains from superficial infections) , with fractional inhibitory concentration (FIC) indices being 0.033 to 0.187; no marked synergistic effect was observed in the combinations between fluorocytosine and azoles, between fluorocytosine and amphotericin B, or between amphotericin B and fluconazole, with the FIC indices being 0.56 to 1.125. The missense mutation V351A in the ERG3 gene was identified in all the 33 (100%) superficial infection-derived strains, as well as in 13 (50%) deep infection-derived strains, and the mutation A353T in the ERG3 gene was identified in 4 (15%) deep infection-derived strains; as for the ERG11 gene, missense mutations identified in the superficial infection-derived strains included I437V (32 strains, 97%) , Y132H (23 strains, 70%) , T123I (16 strains, 48%) , K128T (6 strains, 18%) , D116E (5 strains, 15%) , A114S (4 strains, 12%) , E266D (2 strains, 6%) , G448E (2 strains, 6%) , and G465S (2 strains, 6%) , while missense mutations identified in the deep infection-derived strains included I437V (23 strains, 88%) , E266D (13 strains, 50%) , E260G (5 strains, 19%) , and V488I (4 strains, 15%) ; the missense mutation R101C in the FUR1 gene was identified in 11 (33%) superficial infection-derived strains, but not identified in deep infection-derived strains. Conclusion:The drug susceptibility and drug resistance mutations differed to some extent between superficial infection- and deep infection-derived fluconazole-resistant Candida albicans strains.

Objective To understand and evaluate the knowledge, attitude and practice of female human papillomavirus and HPV vaccine in Chinese mainland. Methods We searched the relevant studies about the knowledge, attitude and behavior factors of HPV infection and HPV vaccine published in PubMed, MEDLINE, CNKI and Wanfang databases about Chinese mainland women from January 1, 1995 to January 31, 2021.Two researchers independently screened the literatures, extracted data and the literatures with quality score≥5 points.Chi square test was used to evaluate its heterogeneity.Begg' s test was used to evaluate publication bias. Results A total of 33 literatures were included, with a total number of 46013 people.The results showed that the total awareness rate of HPV in the population was 65.9%, the lowest rates were 16.0%, 41.2% and 14.4% in Northeast, rural areas and middle school students, respectively; while the highest rates of HPV were 77.4%, 56.3% and 71.0% in Central China, cities and towns and college students, respectively.The main source of HPV knowledge was the network/WeChat official account number (38.9%), and the lowest was family/friend (4.5%).The population's awareness rate of HPV vaccine related knowledge was 41.4%, the highest rates were 51.8% and 69.7% in East China and medical staff, respectively, while the lowest were 23.6% and 12.7% in Southwest and middle school students. Conclusion In order to achieve the total elimination of cervical cancer in China by 2030 as soon as possible, people should strengthen their awareness of HPV and HPV vaccine related knowledge.

Peking Union Medical College Hospital, as one of the most stressful medical institutions in China, is facing the problem of emergency department overcrowding. In order to effectively alleviate the emergency overcrowding, improve the medical quality and patients′ medical experience, the hospital firmly grasped the two incremental links of " throughput" and " output" factors, established a multidisciplinary and multi-department cooperation team, constructed a close medical alliance cooperation mode, and innovated and explored a harmonious emergency overcrowding relief mode with the goal of unblocking the " exit" of patients. The practice showed that the comprehensive measures could effectively alleviate the problem of emergency overcrowding, and improve the medical environment and medical quality.

Objective To investigate the health status of centenarian hospitalized patients and analyze the risk factors for in-hospital death in Nanjing district.Methods All centenarians hospitalized patients who were discharged from wards of 10 upper first-class general hospitals in Nanjing district during the past five years were retrieved from their hospital information systems.Then,a retrospective study was performed on centenarians' data of general information,laboratory test results,Charlson comorbidity index (CCI),neutrophil to lymphocyte ratio (NLR) and shock index(SI),etc.were calculated and collected.Relevant risk factors for in-hospital death were analyzed by multivariate logistic regression analysis.Results A total of 156 patients aged 100 years and over,with an average age of (101.0±2.1)years,were enrolled during the past 5 years.The top 3 admitting diagnosis for the patients were pulmonary infection(30.1％,47/156 cases),coronary heart disease(10.9％,17/156 cases)and cerebrovascular disease(7.1％,11/156 cases).Fifty patients died during hospitalization,with a mortality of 32.1％ (50/156).Pneumonia was the most common admitting diagnosis(40.0％,20/50 case).Among causes of death,the combined admitting diagnosis with dementia,chronic renal insufficiency,one or more basic disease were significantly associated with death.There were statistically significant differences between bad vs.good vs.indifferent prognosis in heart rate,shock index,leukocyte count,neutrophil count,NLR,hemoglobin,albumin,albumin/globulin,fasting blood glucose,blood urea nitrogen,serum creatinine,C-reactive protein(CRP)and CCI levels.Multivariate logistic regression analysis suggested that NLR≥13.18,fasting blood glucose ≥7.56 mmol/L,blood urea nitrogen ≥20.74 mmol/L,CRP≥65 mg/L and CCI≥3 might be predictors for in-hospital death in the cohort(OR =48.91、3.43、1.22、6.55、1.55,all P＜0.05).Conclusions Pulmonary infection is the most common reason for admission and the cause of death in centenarian inpatients.Comorbidities increase the risk of death.To lower in-hospital mortality,CCI and other assessment indicators should be used to strengthen the comprehensive assessment and chronic disease management of hospitalized centenarians.Infectious diseases should be prevented beforehand.