Objective:To study the distribution of species type, biotype and genotype of human Brucella isolated and identified in Nanjing. Methods:A total of 89 strains of human Brucella were collected from microbiology laboratories of three sentinel hospitals in Nanjing from 2017 to 2022. The species type was identified using biological methods and Brucella nucleic acid detection (BCSP31-PCR and AMOS-PCR). Further biotyping of Brucella melitensis isolates was conducted by serological results of A and M factors. Meanwhile, genotype analysis was performed using multiple-locus variable number tandem repeat analysis (MLVA), multilocus sequence typing (MLST) and single nucleotide polymorphism (SNP). Results:From 2017 to 2022, 89 strains of Brucella isolated and identified in Nanjing were all Brucella melitensis. Among them, Brucella melitensis biotype 3 accounted for 82.02% (73/89), and biotype 1 accounted for 17.98% (16/89). MLVA typing showed that 89 strains of Brucella melitensis belong to the "Eastern Mediterranean" cluster and could be divided into 50 MLVA genotypes; among which panel 1 had 3 genotypes, namely Type 42 (84.27%, 75/89), Type 63 (8.99%, 8/89) and Type 43 (6.74%, 6/89). The results of MLST-9 and MLST-21 were both ST8, and the core genome multilocus sequence typing (cgMLST) classified 89 strains into 11 genotypes. SNP analysis revealed a total of 4 013 SNP loci, with SNPs ranging from 0 to 409 across different strains, involving 59 SNP genotypes. Conclusions:The human Brucella strains isolated and identified in Nanjing are all Brucella melitensis, mainly biotype 3. The MLVA cluster is the "Eastern Mediterranean" cluster. The traditional MLST-9 and MLST-21 typing results are all ST8 type, while cgMLST divides all the strains into 11 genotypes with higher resolution.

Objective:To identify human infection with Brucella suis, analyze its biological and molecular characteristics, and to provide basis for prevention and control of brucellosis. Methods:Brucella suis strains were isolated from the body of the first case of human Brucella suis infection in Jiangsu Province. Serum agglutination test was used for serotyping. The specific gene bcsp-31 of Brucella was detected by PCR. AMOS-PCR was used to identify IS-711. The species and biotypes were identified by multiplex PCR. The wboA gene products were sequenced and phylogenetic tree was constructed. Multilocus sequence analysis (MLSA) was used for molecular typing, and cluster analysis was performed with reference strains. Results:The strain was confirmed to be Brucella suis biotype 3 by serum agglutination test and PCR. After sequencing the wboA gene, cluster analysis of the reference sequence showed that the wboA gene was closest to the biotype 3 strain Brucella suis str. 686 (CP007719). MLSA was typed into ST17(1-6-4-1-5-3-5-2-4). Conclusions:Brucella suis biotype 3 is reported in Jiangsu Province for the first time. The MLSA type is ST17. In the future, the prevention and control of human brucellosis should be carried out. We should actively cooperate with the animal husbandry and veterinary department to increase the quarantine, immunization and other control measures.

Objective:To explore the feasibility of direct renin inhibitor aliskiren for the treatment of severe or critical coronavirus disease 2019 (COVID-19) patients with hypertension.Methods:The antihypertensive effects and safety of aliskiren was retrospectively analyzed in three severe and one critical COVID-19 patients with hypertension.Results:Four patients, two males and two females, with an average age of 78 years (66-87 years), were referred to hospital mainly because of respiratory symptoms. Three were diagnosed by positive novel coronavirus 2019 (2019-nCoV) nucleic acid or antibody, and the critical patient with cardiac insufficiency was clinically determined. Two patients were treated with calcium channel antagonist (CCB), one with angiotensin converting enzyme inhibitor (ACEI), and one with angiotensin Ⅱ receptor antagonist (ARB). After admission, ACEI and ARB were discontinued, one patient with heart failure was treated by aliskiren combined with diuretic.Three patients were treated with aliskiren combined with CCB among whom two withdrew CCB due to low blood pressure after 1 to 2 weeks. Based on comprehensive treatment including antiviral and oxygenation treatment, blood pressure was satisfactorily controlled by aliskiren after three to four weeks without serious adverse events. All patients were finally discharged.Conclusion:Our preliminary clinical data shows that antihypertensive effect of aliskiren is satisfactory and safe for severe COVID-19 patients complicated with hypertension.

Background: Zhongyi paste is a traditional Chinese medicine herbal paste that is externally applied to reduce inflammation and relieve pain. Methods: An acute foot swelling inflammation model in C57BL/6J mice was established by carrageenan-induced pathogenesis. Zhongyi paste raised the pain threshold and also reduced the degree of swelling in mice with carrageenan-induced foot swelling. Results: Analysis indicated that serum tumor necrosis factor-alpha, interleukin-1 beta, and prostaglandin E2 (PGE2) cytokine levels and PGE2levels in the paw tissue of the mice were decreased by Zhongyi paste treatment. The quantitative polymerase chain reaction and western blot results showed that Zhongyi paste downregulated the mRNA and protein expression of extracellular signal-regulated kinase 1/2 (ERK1/2), and cyclooxygenase-2 (COX-2), and also downregulated the mRNA expression of PGE2 . At the same time, the Zhongyi paste exerted a stronger effect as an external drug than that of indomethacin, which is an oral drug, and voltaren, which is an externally applied drug. Conclusions Our results indicated that Zhongyi paste is a very effective drug to reduce inflammatory swelling of the foot, and its mechanism of action is related to regulation of the ERK1/2–COX-2–PGE2 pathway.

COVID-19 is caused by the 2019 novel coronavirus (2019-nCoV). COVID-19 clinical cases are considered as the principal source of infection, however, asymptomatic cases may also play a role in the transmission. Significant gap exists in terms of the proportion or prevalence and transmissibility of asymptomatic cases. This study design plans to use data from areas with different epidemiological profiles to investigate the COVID-19 epidemic in China. In each selected region, both general community residents and key populations at high risk of COVID-19 infection, including recovered COVID-19 cases, close contacts of confirmed COVID-19 cases, medical professionals, investigators at CDCs, and visitors to fever clinics, will be recruited and examined for viral RNA of 2019-nCoV and serum antibodies. Prevalence and characterization of asymptomatic cases will be determined, stratified by varied demographics and exposure risk. During the follow-up, the change in the serum antibodies will be studied prospectively in the symptomatic and asymptomatic cases to address the scientific and public health concerns of infectivity and transmissibility of 2019-nCoV.

Criteria for diagnosis of Alzheimer's disease (AD) is not available in China.The international criteria is not a proper choice due to issues such as translation and lead to low diagnostic rate and high rate of missed diagnosis.The research group of Alzheimer's Disease Chinese (ADC) reviewed knowledge and techniques in neuropsychology,neuroimaging,molecular biology,and clinical neurology,and systematically studied the detection techniques such as memory,language,visuospatial,executive function,and medial temporal lobe visual scores on MRI,and their optimal threshold and diagnostic value for the diagnosis of AD.Through a systematic review and consensus meeting,a diagnostic framework for screeningAD in the Chinese population was established.Among these methods,an operational standard for clinical pathology models increased the diagnostic sensitivity by 15％.The sensitivity and specificity of screening memory impairment increased by 18.1％ and 11.6％,respectively.The sensitivity of screening medialtemporal lobe atrophy increased by 24.5％ and missed diagnosis was decreased by 34.5％.An operational standard for clinical biology models,incorporating the latest molecular imaging and molecular biology techniques,has enabled the early diagnosis of AD in China.The framework combines a principled diagnostic guideline with an operational screening protocol,which is applicable to all clinical settings and of great significance for the early detection,early diagnosis and early treatment of AD.

Objectives To evaluate the effectiveness and safety of peramivir trihydrate in patients with influenza.Methods This was a randomized,double-blind,double-dummy,placebo and positive control,multicenter clinical trial,comparing peramivir trihydrate with oseltamivir and placebo.The inclusive criteria were 15-70 years old,onset within 48 h,positive rapid influenza antigen test,and febrile(＞38℃) accompanied with at least two associated symptoms.The severe cases complicated with chronic pulmonary and cardiac diseases,malignancies,organ transplantation,hemodialysis,uncontrolled diabetes,immunocompromised status,pregnancy and coexistence of bacterium infections were excluded.All patients were randomized 2:2:1 to receive peramivir,oseltamivir and placebo respectively.The primary endpoint was the disease duration,the secondary endpoints included time to normal axillary temperature and normal living activities,viral response,and adverse effects.Results Following informed consent,133 patients were included in this study.Four patients were exclude due to missing medical records,not fitting inclusion or exclusion criteria and poor compliance.A total of 129 patients were finally analyzed,including 49 cases,54 cases and 26 cases in peramivir group,oseltamivir group and placebo group.The median disease duration were 96 (76,120)hours,105(90,124) hours,and 124 (104,172)hours in three groups respectively(P＞0.05).The time to normal axillary temperature,normal living activities and viral response were not significantly different in three groups(P＞0.05).Conclusion The value of antiviral therapy in patients with mild influenza needs to be further determined.

Objective To investigate the effects of the number of harvested lymph nodes in neoadjuvant chemoradiotherapy (nCRT) combined with surgery on prognosis of middle-low rectal cancer.Methods The retrospective case-control study was conducted.The clinicopathological data of 373 patients with middle-low rectal cancer who underwent nCRT combined with surgery in the Fujian Medical University Union Hospital from January 2009 to December 2013 were collected.There were 241 males and 132 females,aged from 26 to 81 years,with the age of (55 ± 11) years.Observation indicators:(1) treatment situations;(2) follow-up and survival;(3)influencing factors for the number of harvested lymph nodes;(4) prognostic analysis of the different number of harvested lymph nodes as cut-off for grouping;(5) stratified analysis.Follow-up using telephone interview and outpatient examination was performed to detect postoperative survival of patients once every three months within postoperative 2 years and once every 6 months during the postoperative third year up to March 2016.The endpoint of follow-up was tumor recurrence,retastasis or death.Measurement data with normal distribution were represented as Mean±SD,and comparison between groups was done using the independent sample t test.Measurement data with skewed distribution were represented as M (range),and comparison between groups was done using the Kruska1-Wallis H test.Count data was described as absolute numbers.Univariate and multivariate analyses were done by the multiple linear regression model.Survival rate was calculated by the Kaplan-Meier method,and Logrank test was used for survival analysis.Results (l) Treatment situations:373 patients underwent nCRT combined with surgery,including 329 combined with sphincter-sparing rectal resection and 44 combined with abdominoperineal rectal resection.The number of harvested lymph nodes was 12 ± 6 in 373 patients.There were 185 patients with the number of harvested lymph nodes ＜ 12 and 188 with the number of harvested lymph nodes ≥ 12.(2) Follow-up and survival:373 patients were followed up for 5-77 months,with a median follow-up time of 43 months.During the follow-up,the 1-,3-,5-year disease-free survival rates were respectively 90.4％,76.3％,and 67.5％ in the 373 patients.(3) Influencing factors for the number of harvested lymph nodes:univariate analysis showed that distance between the tumor and anal verge,tumor diameter,tumor pathological N staging,and regression grade of rectal cancer were associated factors for the number of harvested lymph nodes (t =3.156,2.992,x2=8.183,10.839,P＜0.05).Multivariate analysis showed that distance between the tumor and anal verge,regression grade of rectal cancer,and tumor pathological N staging were independent factors for the number of harvested lymph nodes (t=3.308,2.690,2.584,95％ confidence interval:0.808-3.180,0.446-2.873,0.332-2.448,P＜0.05).(4) Prognostic analysis of the different number of harvested lymph nodes as cut-off for grouping:with the number of harvested lymph nodes of 6,7,8,9,10,11,12,13,14,15,and 16 as cut-off for grouping,there was no significant difference in the 3-year disease-free survival rate,cumulative local recurrence rate,and cumulative distant metastasis rate between ＜6 group and ≥6 group,between ＜7 group and ≥7 group,between＜8 group and ≥8 group,between ＜9 group and ≥9 group,between ＜10 group and ≥ 10 group,between ＜11 group and ≥ll group,between ＜12 group and ≥12 group,between ＜13 group and ≥13 group,between ＜ 14 group and ≥ 14 group,between ＜ 15 group and ≥ 15 group,between ＜ 16 group and ≥ 16 group,respectively (P＞0.05).(5) Stratified analysis:with the number of harvested lymph nodes of 7,8,9,and 10 as cut-off for grouping in 45 of 373 patients with Ⅱ-Ⅲ regression grade of rectal cancer and negative lymph nodes (NO staging),there was no significant difference in the 3-year disease-free survival rate between ＜7 group and ≥ 7group,between ＜8 group and ≥8 group,between ＜9 group and ≥9 group,between＜10 group and ≥ 10 group,respetively (x2 =3.946,5.346,6.375,4.297,P＜0.05).Conclusions The number of lymph nodes as 12 is not the independent factor for prognosis of patients with middle-low rectal cancer after nCRT combined with surgery.The number of harvested lymph nodes as 7 to 10 is the important factor for evaluating the prognosis of middle-low rectal cancer patients with Ⅱ-Ⅲ regression grade of rectal cancer and negative lymph nodes after nCRT combined with surgery.

Objective To explore the etiology and treatment of acute intestinal obstruction.Methods Clinical data of patients who underwent operation for acute intestinal obstruction in Zhongshan Hospital from May 2012 to May 2017 were collected and retrospectively analyzed.Results 721 patients were included and the ratio of males to females was 1.55 ∶ 1.There were 48.8％ in old-aged group and 51.1％ in young-middle-aged group.The most common causes of ileus included tumor in 376 cases (51.5％),adhesion in 168 cases (23.3％),hernia in 70 cases (9.7％),intraluminal obstruction in 42 cases (5.8％) and others in 79 cases.There was a significant difference between incarcerated hernia in elderly group and middle-young-aged group (4.6％ vs.15.1％,x2 =22.4,P ＜ 0.01).The length of hospitalized days in patients with tumor and incarcerated hernia in elderly group were significantly longer than young-middle-aged patients [(15.3±8.6)d vs.(13.4±6.3)d,t =-2.5,P＜0.05;(10.1 ± 6.7) d vs.(6.4 ± 2.9) d,t =-2.2,P ＜ 0.05].The length of hospitalized days by limited operation in patients with tumor were significantly shorter than those by emergency operation [(16 ± 12)d vs.(18 ± 24) d,t =-0.3,P ＞ 0.05].Conclusion Tumor and adhesion are the main causes of acute intestinal obstruction.Neoplastic bowel obstruction from small intestine or proximal colon requires emergency surgery.

The accelerating growth of the public microbial genomic data imposes substantial bur-den on the research community that uses such resources. Building databases for non-redundant ref-erence sequences from massive microbial genomic data based on clustering analysis is essential. However, existing clustering algorithms perform poorly on long genomic sequences. In this article, we present Gclust, a parallel program for clustering complete or draft genomic sequences, where clustering is accelerated with a novel parallelization strategy and a fast sequence comparison algo-rithm using sparse suffix arrays (SSAs). Moreover, genome identity measures between two sequences are calculated based on their maximal exact matches (MEMs). In this paper, we demon-strate the high speed and clustering quality of Gclust by examining four genome sequence datasets. Gclust is freely available for non-commercial use at https://github.com/niu-lab/gclust. We also introduce a web server for clustering user-uploaded genomes at http://niulab.scgrid.cn/gclust.