Objective:To construct an evaluation system for clinical thinking ability of general practitioners in the treatment of multimorbidity.Methods:This was a cross-sectional study. The draft of evaluation indexes for clinical thinking ability of general practitioners in treatment of multimorbidity was preliminary developed through literature review, collation, analysis and discussion. Nineteen clinical and teaching experts of general practice were selected for consultation via anonymous convenient sampling. From January to June 2022, 2 rounds of expert consultation were conducted using the Delphi method. During the first round of consultation, according to the survey feedback, we modified and improved the evaluation system of general practitioners′ clinical thinking ability for multi-disease co-treatment. During the second round, experts were asked to assess the importance of each index, and to calculate the weight of each index accordingly. Questionnaires were sent to experts via letters. The content of the questionnaires encompasses the basic information of experts, evaluation for various indexes and relevant opinions. The mean value of importance assignment ≥3.5, coefficient of variation ≤0.25 and the full score frequency ≥30% were taken as the criteria. Indexes unsatisfying the criteria were removed, so that the final index system could be constructed.Results:The average age of 19 experts was 50.2 years old, 9 of them were male. A total of 2 rounds of expert consultation were conducted, 19 questionnaires were issued in each round, and 19 effective questionnaires were received afterwards. In the first round of consultation, 10 experts put forward revised opinions, and some indexes were adjusted according to the definition criteria and the discussion of the research group. In the second round of consultation, 3 experts put forward suggestions for modification. According to the definition criteria, no need to delete the indexes. After discussion by the research group, some indexes were adjusted, and finally an evaluation system of clinical thinking ability for multi-disease co-treatment of general practitioners was established, including 4 first-level indexes and 30 second-level indexes. The weights of the 4 first-level indexes in descending order were "overall thinking ability" (38.01%), "diagnostic thinking ability" (33.96%), "evidence-based thinking ability" (14.75%), and "critical thinking ability" (13.28%). Among the 30 secondary indexes, the top 5 were "ability to identify and handle priority emergency incidents" (5.04%), "risk assessment and critical illness identification ability" (4.63%), "emergency referral ability" (4.61%), "communication and expression ability" (4.57%), and "standardized diagnosis and treatment ability" (4.23%).Conclusion:This study successfully constructed an evaluation system for clinical thinking ability of general practitioners in the treatment of multimorbidity.

In situ and real-time monitoring of responsive drug release is critical for the assessment of pharmacodynamics in chemotherapy. In this study, a novel pH-responsive nanosystem is proposed for real-time monitoring of drug release and chemo-phototherapy by surface-enhanced Raman spectroscopy (SERS). The Fe₃O₄@Au@Ag nanoparticles (NPs) deposited graphene oxide (GO) nanocomposites with a high SERS activity and stability are synthesized and labeled with a Raman reporter 4-mercaptophenylboronic acid (4-MPBA) to form SERS probes (GO-Fe₃O₄@Au@Ag-MPBA). Furthermore, doxorubicin (DOX) is attached to SERS probes through a pH-responsive linker boronic ester (GO-Fe₃O₄@Au@Ag-MPBA-DOX), accompanying the 4-MPBA signal change in SERS. After the entry into tumor, the breakage of boronic ester in the acidic environment gives rise to the release of DOX and the recovery of 4-MPBA SERS signal. Thus, the DOX dynamic release can be monitored by the real-time changes of 4-MPBA SERS spectra. Additionally, the strong T₂ magnetic resonance (MR) signal and NIR photothermal transduction efficiency of the nanocomposites make it available for MR imaging and photothermal therapy (PTT). Altogether, this GO-Fe₃O₄@Au@Ag-MPBA-DOX can simultaneously fulfill the synergistic combination of cancer cell targeting, pH-sensitive drug release, SERS-traceable detection and MR imaging, endowing it great potential for SERS/MR imaging-guided efficient chemo-phototherapy on cancer treatment.

[This corrects the article DOI: 10.1016/j.apsb.2022.08.024.].

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Antibodies, Viral ; Humans ; Porcine Reproductive and Respiratory Syndrome/prevention & control* ; Porcine respiratory and reproductive syndrome virus/genetics* ; Swine

Objective To compare the tumor characteristics and survival between postoperative incidentally discovered gallbladder cancer (ID-GBC) and preoperatively suspected gallbladder cancer (PS-GBC).Methods The data of 276 GBC patients who underwent surgical resection with curative intent between January 2004 and December 2014 at the Eastern Hepatobiliary Surgery Hospital were retrospectively analyzed.Results The 1-,3-,and 5-year cumulative survival rates of the ID-GBC group (88.8％,52.2％,and 33.0％,respectively) were significantly better than those in the PS-GBC group (57.5％,25.7％,and 16.6％,P ＜ 0.05).In the ID-GBC group,multivariate analysis revealed that T staging,hepatic invasion and time interval from cholecystectomy to re-operation were independent prognostic factors.The overall survival (OS) in the group with the time interval within 2 weeks was significantly better than those in the other two groups (both P ＜ 0.05).However,there were no significant differences in OS between the groups with the time interval of 2 weeks to 1 month and more than 1 month (P ＞ 0.05).Conclusions Postoperative ID-GBC had significantly better survival outcomes than PS-GBC.Reoperation within two weeks in patients with ID-GBC is a good strategy.

Objective To clarify the characteristics and postoperative benefit of gallbladder carcinoma in elderly patients (≥ 65 years old).Methods Two hundred and seventy-three patients of gallbladder carcinoma were collected,who were treated intent resection from January 2004 to December 2012 in the Department of Hepatobiliary Surgery,Eastern Hepatobiliary Hospital,Second Military Medical University.More than 65 years old was defined as the elderly,else was defined as the younger.The clinical-pathological features and prognosis of 85 elderly patients(elderly group) and 188 younger patients (younger group) were retrospectively analyzed.The survival of patients were followed up by telephone or outpatient.The incidence of hypertension,incidence of diabetes,TNM staging,and median CA19-9 were compared between the two groups.Continuous variables using a two-sample t test or Mann-Whitney U test.Categorical variables were compared by the Chi-square test or Fisher probability method.The survival curve was drawn by the Kaplan-Meier method.The univariate analysis and multivariate analysis of prognosis were respectively done using the Log-rank test and COX regression model.Results The incidence of hypertension,incidence of diabetes,TNM stage ratio (Ⅲ + Ⅳ/Ⅰ + Ⅱ),and CA19-9 median in the elderly group were 30.6％,11.8％,27.6 and 69.3 U/ml,respectively.The differences in the younger group were 13.8％,4.8％,7.9 and 28.2 U/ml,respectively,with statistically significant difference between the two groups (all P ＜ 0.05).The incidence of complications was 54.1％ in the elderly group and 48.9％ in the younger group,with no significant difference between the two groups (P =0.302).The median survival of the elderly group was 28.01 months,and the median survival of the younger group was 36.20 months,with no statistical difference between the two groups (P =0.131).Cox analysis showed that independent prognostic risk factors for the elderly patients with gallbladder cancer included liver invasion (HR =2.386,95％ CI:1.379-4.127,P =0.002) and lymph node metastasis (HR =1.866,95 ％ CI:1.100-3.167,P =0.021).Conclusions Radical resection is safe and feasible for elderly patients with gallbladder carcinoma.Age is not a contraindication for surgery.Radical resection can get the same benefits as young people.Liver invasion and lymph node metastasis are independent risk factors affecting the prognosis of the elderly patients with gallbladder carcinoma.

Objective Reference standard of the RPOB(rifampin resistance)gene recommended by CLSI-MM18A(Interpretive Criteria for Identification of Bacteria and Fungi by DNA Target Sequencing) was used to evaluate the ability of MALDI-TOFMS techniques for the identification and classification of non-tuberculous Mycobacterium.Methods Fifty five clinicalstrains were collected from 2012 to 2016 with different sources.The RPOB gene was sequenced, and results were applied to phylogenetics analysis. MALDI-TOF MS technology was implemented to identify the strains, and cluster analysis was conducted based on protein fingerprint.The consistency of two methods for NTM identification and typing was evaluated.Results The RPOB gene method showed a good ability of identification(similarity>99.0%) and subtyping(to subspeciesof the complex level).The French BioMérieux MALDI-TOF MS identified 89.1% of 55 strains to genus level and 78.2% to species level.The phylogeneticsanalysis of protein fingerprint by SARAMS Premium software also showed good typing ability.Conclusions MALDI-TOF MS technology can identify and classify non-tuberculous Mycobacterium effectively,which is rapid and easy.It is complementary to RPOB gene method in laboratory application.

Objectives To identify the Francisella strain isolated from blood of a patient with drowning-associated pneumonia.Methods The whole genome of the strain,designated Wenzhou1,was sequenced using the high throughput sequencing technology by 2000/miSeq system of Illumina platform,and the obtained genome draft was assembled by MicrobeTrakr Plus software.The phylogenetic neighbors of Wenzhou1 were obtained by NCBI BLAST analysis from GenBank database for the gene sequences of 16S rRNA,malate dehydrogenase(mdh),DNA-directed RNA polymerase subunit beta (rpoB) and succinate dehydrogenase subunit alpha (sdhA).The average nucleotide identity(ANI) between Wenzhou1 and its phylogenetic neighbors was analyzed by the software OrthoANI using NCBI BLAST search under the Java Runtime Environment Version 8.Results The genome size of Wenzhou1 was 1.96 × 106 bp,containing 74 contigs.The genomic G + C mol％ of Wenzhou1 was 32.1％,which was similar to the other species of genus Francisella and Allofranicella.Based on the analysis of NCBI BLAST of GenBank for the similarities of 16S rRNA gene,mdh gene,rpoB gene and sdbA gene sequences,Wenzhou1 was most closely related to F.hispaniensis FSC454 and Francisella cf.novicida 3523.The ANI of Wenzhou1 was 97.8％ to F.hispaniensis FSC454,97.5％ to 97.6％ to Francisella cf.novicida 3523,but only 91.3％ to 91.5％ to the four subspecies of F.tularensis.Conclusion ANI analysis based on whole genome sequence should be an accurate,effective method for bacterial identification.Wenzhou1 could be identified as F.hispaniensis by ANI with high-throughput whole genome sequencing technology.

Objectives To identify and characterize 10 strains of Francisella philomiragia-like organisms isolated from blood samples and environmental water.Methods The 10 clinical and environmental isolates were identified by traditional morphological examination and biochemical characterization,matrix-assisted laser desorption/ionization time of flight(MALDI-TOF) mass spectrometry(MS) systems and sequencing based on 16S rRNA gene.The minimum inhibitory concentrations were tested by E-test methods.Results All the 10 isolates were gram-negative coccobacilli appearing tiny and faint counterstain of safranin,negative for urease,nitrate reduction and X and/or V factor requirement,but positive for oxidase and catalase.The isolates grew rapidly in sheep blood agar,chocolate agar and BCYE plate forming white opaque,colorless transparent or gray smooth colonies with about 2-mm diameters,but did not grow in M-H agar and MacConkey agar.The sequencing for 16S rRNA gene indicated that the 10 isolates shared more than 99.6％ similarity to Francisella philomiragia,and fell into the same clusters of Francisella philomiragia on phylogenetic tree.The MALDI-TOF MS analysis also showed the typical peaks with 6 153 m/z,5 180 m/z,7 757 m/z and 9 392 m/z which were similar to Francisella philomiragia ATCC 25015.However,they may be misidentified to be Sphingomonas paucimobilis by using Vitek 2 GN cards,Neisseria cinerea by using Vitek 2 NH cards,Myroides odoratimimus by using API 20NE strips and Haemophilus by using API NH cards.The results of antimicrobial susceptibility showed that they were all sensitive to chloramphenicol,doxycycline,tetracycline,gentamicin,ofloxacin and ciprofloxacin.Conclusion The 10 isolates could be identified as Francisella philomiragia,so we should pay more attention to the infrequent pathogen for its inactive biochemical reaction and the misidentification by commercial detection systems.

Objective To identify the biotype and analyze the epidemiological characteristics of twentyfour Brurella strains from the primary hospitals in Guangdong Province.Methods The twenty-four Brucella strains,collected from Oct.2009 to Oct.2015,were identified by routine biochemical methods,VITEK 2 COMPACT automatic microbial identification analyzer,16S rRNA gene sequencing and biology phenotype based on serological and bacteriophages lysis test.The etiology was analyzed based on clinical data,biotypes of the isolates and other clinical information.Results All of the twenty-four strains were Gram-negative coccobacilli,including two strains of Brucella suis biotype Ⅱ,four strains of Brucella melitensis biotype Ⅰ and eighteen strains of Brucella melitensis biotype Ⅲ.By GN card of VITEK 2 COMPACT automatic microbial identification analyzer,one strain was mistaken as Bordetella bronchiseptica and two strains were mistaken as Ochrobaetrum anthropi.The 16S rRNA gene sequencing showed they were high homology to Ochrobactrum intermedium and Ochrobaetrum anthropi,which completely excluded the possibility of Bordetella bronchiseptica.Tbe clinical data showed that all of the twenty-four patients were adults with an average age of 49.0 years old,men and women were twelve people respectively,with no significant gender differences and no occupational exposure,which presenting a wide and diverse range of nonspecific clinical signs and symptoms,but brucellosis was not aware of by the physician.Conclusion Brucella melitensis biotype Ⅲ is the main pathogen of brucellosis,with the characteristics of sporadic outbreak and occult infection in the primary hospitals in Guangdong Province.