In order to investigate the effects of porin genes ompW,ompS and ompD on the biological properties and virulence of Salmonella typhimurium,the corresponding mutant strains were con-structed using the λ Red homologous recombination system,and the growth curves,motility,bio-chemical properties,in vitro genetic stability,biofilm-forming ability,drug resistance,and lethal dose at half capacity(LD50)between the standard strain and each mutant strain were detected by comparative assays for Salmonella typhimurium.The results showed that,compared with the standard strain,the ompD and ompW mutation had less effect on the growth rate and motility of the bacteria,while the ompS mutation significantly reduced the growth rate and motility;none of the three genetic mutation affected the biochemical characteristics of Salmonella typhimurium,nor the genetic stability,but affected its susceptibility to a variety of commonly used antibiotics to varying degrees and caused a highly significant decrease(P＜0.01)in the ability to form a biofilm,and the results showed that the three mutant strains had a significant reduction in the ability to form a biofilm.The result of LD50 virulence assay showed that all three genetic mutation led to a decrease in the virulence of Salmonella typhimurium,among which the ompS mutant strain showed the most obvious decrease in virulence,LD50 was 25 times that of the standard strain.In conclusion,mutations of the pore protein ompW,ompS,and ompD genes can affect some biological properties of Salmonella typhimurium.The results of this study laid an experimental foundation for further research on the biological functions of the pore protein ompW,ompS and ompD genes and Salmonella pathogenicity.

Background and purpose:For patients with human epidermal growth factor receptor 2(HER2)-positive metastatic breast cancer,trastuzumab treatment can prolong the overall survival and significantly improve the prognosis of patients.However,the reference original research trastuzumab(Herceptin?)is more expensive.Biosimilars have comparable efficacy and safety profiles while increasing patient access to treatment.This clinical trial aimed to evaluate the efficacy,pharmacokinetics,safety and immunogenicity of the trastuzumab biosimilar AK-HER2 compared to trastuzumab(Herceptin?)in patients with HER2-positive metastatic breast cancer.Methods:This multi-center,randomised,double-blind phase Ⅲ clinical trial was conducted in 43 subcenters in China.This study complied with the research protocol,the ethical principles stated in the Declaration of Helsinki and the quality management standards for drug clinical trials.It was approved by the hospital's medical ethics committee.The clinical trial registration agency is the State Food and Drug Administration(clinical trial approval number:2015L04224;clinical trial registration number:CTR20170516).Written informed consent was obtained from subjects before enrollment.Enrolled patients were randomly assigned to the AK-HER2 group and the control group,respectively receiving AK-HER2 or trastuzumab(initial loading dose 8 mg/kg,maintenance dose 6 mg/kg,every 3 weeks as a treatment cycle,total treatment time is 16 cycles)in combination with docetaxel(75 mg/m2,treatment duration is at least 9 cycles).The primary endpoint of this clinical trial was the objective response rate(ORR9)between the AK-HER2 group and the control group in the 9th cycle.Secondary efficacy endpoints included ORR16,disease control rate(DCR),clinical benefit rate(CBR),progression-free survival(PFS)and 1-year survival rate.In this study,100 subjects(AK-HER2 group to control group=1:1)were randomly selected for blood sample collection after the 6th cycle of medication,The collection time points were 45 minutes after infusion(the end of administration),4,8,24,72,120,168,336,and 504 hours after the end of administration.After collection,blood samples were analyzed by PK parameter set(PKPS).Other evaluation parameters included safety and immunogenicity assessment.Results:A total of 550 patients with HER2-positive metastatic breast cancer were enrolled in this clinical trial between Sep.2017 and Mar.2021.In the AK-HER2 group(n=237),129 subjects in the experimental group achieved complete response(CR)or partial response(PR),and the ORR9 was 54.4%.There were 134 subjects in the control group(n=241)who achieved CR or PR,and the ORR9 was 55.6%.The ORR9 ratio between the AK-HER2 group and the control group was 97.9%[90%confidence interval(CI):85.4%-112.2%,P=0.784],which was not statistically significant.In all secondary efficacy endpoints,no statistically significant differences were observed between the two groups.We conducted a mean ratio analysis of pharmacokinetics(PK)parameters between the AK-HER2 group and the control group,and the results suggested that the pharmacokinetic characteristics of the two drugs are similar.The incidence of treatment emergent adverse event(TEAE)leading to drug reduction or suspension during trastuzumab treatment was 3.6%(10 cases)in the AK-HER2 group and 8.1%(22 cases)in the control group.There was statistically significant difference between the two groups(P=0.027).The incidence rate was significantly lower in the AK-HER2 group than in the control group,and there was no statistically significant difference among the other groups.The differences in the positive rates of anti-drug antibodies(ADA)and neutralizing antibodies(NAB)between groups were of no statistical significance(P=0.385 and P=0.752).Conclusion:In patients with HER2-positive metastatic breast cancer,AK-HER2 was comparable to the trastuzumab(Herceptin?)in terms of drug efficacy,pharmacokinetics,safety and immunogenicity.

Objective:To explore the clinical and electrophysiological characteristics of peripheral neuropathy in prediabetic patients.Methods:Subjects aged 20-65 years with high-risk factors of impaired glycemia enrolled in Beijing Tiantan Hospital, Capital Medical University from 2019 to 2022 were recruited to conduct oral glucose tolerance test, after excluding other causes of neuropathy or radiculopathy. Patients with impaired fasting glucose or impaired glucose tolerance were defined by American Diabetes Association criteria. These patients were divided into clinical polyneuropathy (PN) and clinical non-PN groups, according to the 2010 Toronto consensus criteria and the presence of PN symptoms and signs or not. Nerve conduction studies (NCS), F wave, sympathetic skin response (SSR), R-R interval variation (RRIV) and current perception thresholds (CPT) were performed and the abnormal rate was compared between different electrodiagnostic methods and between clinical subgroups.Results:Among the 73 prediabetic patients ultimately enrolled, only 20 (27.4%) can be diagnosed as clinical PN according to the Toronto consensus criteria. The abnormal rate of CPT (68.5%, 50/73) was significantly higher than those of F wave (2.7%, 2/73), lower limb NCS (0, 0/73), upper limb NCS changes of carpal tunnel syndrome (26.0%, 19/73), SSR (6.8%, 5/73) and RRIV (5.5%, 4/73; McNemar test, all P<0.001). With sinusoid-waveform current stimuli at frequencies of 2 000 Hz, 250 Hz and 5 Hz, the CPT device was used to measure cutaneous sensory thresholds of large myelinated, small myelinated and small unmyelinated sensory fibers respectively. CPT revealed a 21.9% (16/73) abnormal rate of unmyelinated C fiber in the hands of prediabetic patients, significantly higher than that of large myelinated Aβ fibers [8.2% (6/73), χ2=5.352, P=0.021]. Both abnormal rates of small myelinated Aδ [42.5% (31/73)] and unmyelinated C fibers [39.7% (29/73)] in the feet of prediabetic patients were significantly higher than that of large myelinated Aβ fibers [11.0% (8/73), χ2=18.508, 15.965, both P<0.001]. Compared with the clinical non-PN group, the abnormal rates of CPT [90.0% (18/20) vs 60.4% (32/53), χ2=5.904, P=0.015] and SSR [20.0% (4/20) vs 1.9% (1/53), P=0.016) were significantly higher in the clinical PN group. Conclusions:Peripheral neuropathies in prediabetic patients are usually asymptomatic or subclinical, and predispose to affect unmyelinated and small myelinated sensory fibers. Selective electrodiagnostic measurements of small fibers help to detect prediabetic neuropathies in the earliest stages of the disease.

Objective:To validate the feasibility of the gamma analysis method in the study of prescription dose conversion between logistic nanodosimetry model (LNDM) and microdosimetric kinetic model (MKM) basing on the Chinese self-developed model LNDM by applying clinical experiences of National Institute of Radiological Science (NIRS).Methods:Physical dose distributions derived from the MKM- and LNDM-based carbon ion treatment plans were compared via the method of gamma analysis under the open-source treatment planning platform matRad. In this way, the prescribed dose conversion factor between the MKM- and LNDM-based treatment plans was obtained. Using water phantoms, the influence of geometric shape, size, depth of target volume (TV), prescribed dose and field setting on the conversion factor was investigated comprehensively. Moreover, preliminary verification of the acquired conversion factor was conducted on the C-shape model and a case of liver cancer patient.Results:The conversion factor depended on the field setting rather than the TV shape. Under the condition of single field, the conversion factor was positively correlated with the size and depth of TV, and the prescribed dose. Moreover, the conversion factor was successfully verified using the C-shape model and the patient with liver cancer, where the gamma passing rates (2%/2 mm) of the physical dose distribution generated by the MKM and LNDM treatment plans were 92.79% and 91.19%, respectively.Conclusions:The conversion factors (f=D LNDM/D MKM) obtained in this study might provide guidance for the prescribed dose setting during the carbon ion treatment planning based on the LNDM. Besides, the gamma analysis method could be used for the study of the prescribed dose conversion between different models.

Entinostat plus exemestane in hormone receptor-positive (HR+) advanced breast cancer (ABC) previously showed encouraging outcomes. This multicenter phase 3 trial evaluated the efficacy and safety of entinostat plus exemestane in Chinese patients with HR + ABC that relapsed/progressed after ≥1 endocrine therapy. Patients were randomized (2:1) to oral exemestane 25 mg/day plus entinostat (n = 235) or placebo (n = 119) 5 mg/week in 28-day cycles. The primary endpoint was the independent radiographic committee (IRC)-assessed progression-free survival (PFS). The median age was 52 (range, 28-75) years and 222 (62.7%) patients were postmenopausal. CDK4/6 inhibitors and fulvestrant were previously used in 23 (6.5%) and 92 (26.0%) patients, respectively. The baseline characteristics were comparable between the entinostat and placebo groups. The median PFS was 6.32 (95% CI, 5.30-9.11) and 3.72 (95% CI, 1.91-5.49) months in the entinostat and placebo groups (HR, 0.76; 95% CI, 0.58-0.98; P = 0.046), respectively. Grade ≥3 adverse events (AEs) occurred in 154 (65.5%) patients in the entinostat group versus 23 (19.3%) in the placebo group, and the most common grade ≥3 treatment-related AEs were neutropenia [103 (43.8%)], thrombocytopenia [20 (8.5%)], and leucopenia [15 (6.4%)]. Entinostat plus exemestane significantly improved PFS compared with exemestane, with generally manageable toxicities in HR + ABC (ClinicalTrials.gov #NCT03538171).

Objective:To explore the different types and characteristics of rehabilitation exercise compliance of patients with first stroke at 2 weeks of onset, and analyze its influencing factors, so as to provide reference for the formulation of targeted health education.Methods:A cross-sectional investigation was conducted in Neurology Department of the Second Affiliated Hospital of Zhejiang University School of Medicine from January to June, 2021. 276 patients with first-episode stroke were investigated by the Questionnaire of Exercise Adherence at 2 weeks of the onset. The potential profile analysis was conducted to explore characteristics classification of the rehabilitation exercise compliance. And the chi-square test was used to compare demographic differences among different categories and ordered multi classification Logistic regression was used to explore the influencing factors of rehabilitation exercise compliance.Results:The patients were divided into 109 cases with high rehabilitation exercise compliance (39.5%), 114 cases with moderate rehabilitation exercise compliance (41.3%), and 53 cases with low rehabilitation exercise compliance (19.2%). There were statistically significant differences in education level, consciousness level at admission, complications and limb muscle strength among the three types of patients ( χ2 values were 6.17-31.50, all P<0.05). Ordered multi classification Logistic regression showed that the patient′s education level, the patient′s consciousness level at admission, whether there were complications and limb muscle strength would affect the rehabilitation exercise compliance of stroke patients ( P<0.05). Conclusions:There are three potential categories of rehabilitation exercise compliance in patients with first stroke. Patients with illiteracy, drowsiness, complications and poor limb muscle strength have poor rehabilitation exercise compliance.

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 learn the characteristics of second-line drug resistance and related gene mutations of multidrug-resistant Mycobacterium tuberculosis ( MDR-TB ) Beijing genotype strains. Methods: The MDR-TB isolates in Hwa Mei Hospital from 2017 to 2019 were enrolled and detected using RD105 deletion-targeted multiplex polymerase chain reaction (PCR). The proportion method for drug susceptibility test was used to detect the drug-resistant profiles against kanamycin, amikacin, capreomycin, ofloxacin and levofloxacin. The gene sequencing of rrs, tlyA, eis, gidB, gyrA and gyrB was conducted by PCR compared with H37RV strain. The differences in the rates of drug resistance and mutation between Beijing and non-Beijing genotype strains were examined to understand the characteristics of Beijing genotype strains. Results: There were 106 Beijing genotype and 27 non-Beijing genotype strains in 133 MDR-TB isolates. The drug resistance rates of kanamycin, amikacin, capreomycin, ofloxacin and levofloxacin in Beijing genotype strains were 9.43%, 7.55%, 3.77%, 32.08% and 32.08%, respectively. The rates of quasi-extensive and extensive drug resistance in Beijing genotype strains were 30.19% and 7.55%. The gene mutation rates of rrs, tlyA, eis, gidB, gyrA and gyrB in Beijing genotype strains were 7.55%, 7.55%, 1.89%, 2.83%, 36.79% and 2.83%, respectively. There were no significantly differences between Beijing and Non-Beijing genotype strains in the factors above ( P>0.05 ). The gene rrs, tlyA, eis, gidB, gyrA and gyrB had 2, 1, 2, 2, 5 and 3 mutation types, respectively, with single base substitution as the main type. Conclusion Beijing genotype strains are dominant in MDR-TB, with high resistance to fluoroquinolones and mainly gyrA gene mutation.

Objective: This study aims to investigate the impact of individual and family factors on anxiety and loneliness of middle school students during quarantine in the COVID-19 pandemic and to provide reference for discussing the possible effective intervention paths in the stress process of public health emergencies. Methods: A total of 11 596 students from junior and high schools in Shanghai were selected to complete the questionnaire with the stratified random cluster sampling method. Generalized Anxiety Disorder(GAD-7) and UCLA Loneliness Scale, University of California at Los Angeles(ULS-6) were used for the survey. Results: The prevalence of anxiety and loneliness were 34.78% and 52.44% respectively. The rate of anxiety and loneliness in girls (36.13%, 54.62%) was higher than that of boys (33.44%, 50.28%)(χ 2=17.03, 21.82, P<0.05). Logistic regression analysis showed that graduates (OR=1.34, 95%CI=1.18-1.52), students ranking in the middle (OR=1.25, 95%CI=1.08-1.45) or in the bottom 25% (OR=1.76, 95%CI=1.50-2.05), non only children in the family (OR=1.25, 95%CI=1.09-1.43) and low socioeconomic status (OR=1.42, 95%CI=1.23-1.65) were associated with increased risk of moderate and severe level of anxiety(P<0.05). Girls(OR=1.20, 95%CI=1.12-1.30), graduates(OR=1.13, 95%CI=1.04-1.24), non residents(OR=1.20, 95%CI=1.09-1.33), lower level of grades(OR=1.72, 95%CI=1.55-1.91) and not living with parents(OR=1.32, 95%CI=1.18-1.48) were associated with increased risk of loneliness(P<0.05). Conclusion During the quarantine period caused by the COVID-19 pandemic, negative emotions such as anxiety and loneliness among adolescents need attention, especially females, graduates, only children and students with low rankings or low socioeconomic status.

Objective:To investigate the serum level of low density lipoprotein cholesterol (sdLDL-C) in elderly patients (age≥65 years) with type 2 diabetes mellitus (T2DM) and the its predictive value in evaluatingthe risk of cardiovascular and cerebrovascularevents in elderly patients with T2DM.Methods:In this retrospective study,386 elderly patients with T2DM were collected from December 2014 to December 2016, the averageage was (72.7±5.4) years old, including 269 males and 117 females; 92 of whom had cardiovascular events during follow-up, the averageage was (72.9 ± 5.2) years old, including 65 males and 27 females, and serum sdLDL-C level was detected. Meanwhile, biomarkers such as lipoprotein (a), apolipoprotein B, apolipoprotein AI and hypersensitive c-reactive protein were analyzedin all the patients.Univariate and multivariate factors were used to analyze the relationship between each variable and the occurrence of cardiovascular and cerebrovascular events. The correlation between LDL-C, sdLDL-C of all subjects with age and other lipid indexes were analyzed. ROC curve was used to determine the predictive value of sdLDL-C elevation for cardiovascular and cerebrovascular events in elderly patients with T2DM.Results:The levels of LDL-c, sdLDL-C, non-HDL-C, GLU, HbA1c and ApoB were significantly higher in theCardio-cerebral vascular event group ( t=3.26, 3.46, 2.91, 2.47, 4.03, 3.00, P<0.05). While the levels of apolipoprotein AI was significantly lower than those in theNon-cardio-cerebral vascular event group ( t=-2.39, P<0.05). Cox regression analysis showed that sdLDL-C per 10 mg/dl was independently correlated with the risk of cardiovascular and cerebrovascular events( HR 1.281, 95 %CI 1.225-16.032, P<0.01)after adjusted for age. SdLDL-C was positively correlated with TG, non-HDL-C and ApoB ( r=0.323, 0.588, 0.623, P<0.01) and was negativelycorrelated correlation with age, HDL-C and apolipoprotein AI ( r=-0.363, -0.301, P<0.01), ROC curve analysis showed that sdLDL-C had a strong ability to predict the occurrence of cardiovascular and cerebrovascular events (AUC=0.736, 95 %CI 0.554 9-0.918 2, P=0.003), while HbA1c also had a strong ability to predict the occurrence of cardiovascular and cerebrovascular events(AUC=0.725, 95 %CI 0.524 3-0.927 3, P=0.006). Combined sdLDL-C with HbA1had the strong ability to predict the occurrence of cardiovascular and cerebrovascular events (AUC=0.837, 95 %CI 0.711 4-0.973 5, P=0.001). Conclusions:The elevation of serum sdLDL-C in elderly patients with T2DM wasa significantly independentrisk factorof cardiovascular and cerebrovascular events. Serum sdLDL-C had a higher clinical value than LDL-C, which was expected to be the most effective predictor of lipid profile in riak assessment of cardiovascular and cerebrovascular events in elderly patients with T2DM.