OBJECTIVE: To investigate the molecular mechanism underlying inherent fosfomycin resistance of Klebsiella pneumoniae (K. pneumoniae). METHODS: The draft genomic sequences of 14 clinical hypervirulent/hypermucoviscous K. pneumoniae (HvKP/ HmKP) isolates were obtained using the next-generation sequencing technology. The genomic sequences were analyzed using the Resistance Gene Identifier (RGI) software for predicting the resistome based on homology and SNP models in the Comprehensive Antibiotic Resistance Database (CARD) and for identification of the presence of phosphomycin resistancerelated genes uhpt and fosA and their mutations in the bacterial genomes. The results were verified by analyzing a total of 521 full-length genomic sequences of K. pneumonia strains obtained from GenBank. RESULTS: All the 14 clinical isolates of HvKP/ HmKP carried hexose phosphate transporter (UhpT) gene mutation, in which the glutamic acid was mutated to glutamine at 350aa (UhpT^E350Q mutation); the presence of fosA6 gene was detected in 12 (85.71%) of the isolates and fosA5 gene was detected in the other 2 (14.29%) isolates. Analysis of the genomic sequences of 521 K. pneumonia strains from GenBank showed that 508 (97.50%) strains carried UhpT^E350Q mutation, 439 (84.26%) strains harbored fosA6, and 80 (15.36%) strains harbored fosA5; 507 (97.31%) strains were found to have both UhpT^E350Q mutation and fosA6/5 genes in the genome. Only 12 (2.30%) strains carried fosA6/5 genes without UhpT^E350Q mutation; 1 (0.19%) strain had only UhpT^E350Q mutation without fosA6/5 genes, and another strain contained neither UhpT^E350Q mutation nor fosA6/5 genes. CONCLUSION UhpT^E350Q mutation with the presence of fosA6/5 genes are ubiquitous in K. pneumonia genomes, indicating a possible intrinsic mechanism of fosfomycin resistance in the bacterium to limit the use of fosfomycin against infections caused by K. pneumoniae, especially the multi-resistant HvKP/HmKP strains.
Fosfomycin ; Klebsiella pneumoniae ; Mutation ; Databases, Factual ; High-Throughput Nucleotide Sequencing

Platelets are reprogrammed by cancer via a process called education, which favors cancer development. The transcriptional profile of tumor-educated platelets (TEPs) is skewed and therefore practicable for cancer detection. This intercontinental, hospital-based, diagnostic study included 761 treatment-naïve inpatients with histologically confirmed adnexal masses and 167 healthy controls from nine medical centers (China, n = 3; Netherlands, n = 5; Poland, n = 1) between September 2016 and May 2019. The main outcomes were the performance of TEPs and their combination with CA125 in two Chinese (VC1 and VC2) and the European (VC3) validation cohorts collectively and independently. Exploratory outcome was the value of TEPs in public pan-cancer platelet transcriptome datasets. The AUCs for TEPs in the combined validation cohort, VC1, VC2, and VC3 were 0.918 (95% CI 0.889-0.948), 0.923 (0.855-0.990), 0.918 (0.872-0.963), and 0.887 (0.813-0.960), respectively. Combination of TEPs and CA125 demonstrated an AUC of 0.922 (0.889-0.955) in the combined validation cohort; 0.955 (0.912-0.997) in VC1; 0.939 (0.901-0.977) in VC2; 0.917 (0.824-1.000) in VC3. For subgroup analysis, TEPs exhibited an AUC of 0.858, 0.859, and 0.920 to detect early-stage, borderline, non-epithelial diseases and 0.899 to discriminate ovarian cancer from endometriosis. TEPs had robustness, compatibility, and universality for preoperative diagnosis of ovarian cancer since it withstood validations in populations of different ethnicities, heterogeneous histological subtypes, and early-stage ovarian cancer. However, these observations warrant prospective validations in a larger population before clinical utilities.
Humans ; Female ; Blood Platelets/pathology* ; Biomarkers, Tumor/genetics* ; Ovarian Neoplasms/pathology* ; China

Professor GAO Wei-bin's clinical experience of electric eye acupuncture and stagnant-moving needling for ophthalmopathy was introduced. The indications of electric eye acupuncture and stagnant-moving needling include external ophtalmoplegia and visual impairment. Professor GAO has proposed new acupoints at the ocular muscles attachment of eyeball, and put forward five experience points: Shangming point, Neiming point, Xiaming point, Waiming point and Tijian point. The points are selected according to different pathological changes of ocular muscles. In the treatment of ophthalmopathy, the tendons and vessels are often regulated at the same time. Neiming point, Shangming point, Xiaming point and Qiuhou point are the main points, with Fengchi (GB 20) and Gongxue (Extra) as the matching points. In addition, attention is paid to the application of stagnant-moving needling and electroacupuncture (continuous dense wave, frequency of 50 Hz).
Acupuncture ; Acupuncture Points ; Acupuncture Therapy ; Electroacupuncture ; Eye Diseases ; Humans

Multidrug resistance (MDR) is the main cause of clinical treatment failure and poor prognosis in cancer. Targeting P-glycoprotein (P-gp) has been regarded as an effective strategy to overcome MDR. In this work, we reported our preclinical studies of the triazolo[1,5-a]pyrimidine-based compound WS-716 as a highly potent, specific, and orally active P-gp inhibitor. Through direct binding to P-gp, WS-716 inhibited efflux function of P-gp and specifically reversed P-gp-mediated MDR to paclitaxel (PTX) in multiple resistant cell lines, without changing its expression or subcellular localization. WS-716 and PTX synergistically inhibited formation of colony and 3D spheroid, induced apoptosis and cell cycle arrest at G2/M phase in resistant SW620/Ad300 cells. In addition, WS-716 displayed minimal effect on the drug-metabolizing enzyme cytochrome P4503A4 (CYP3A4). Importantly, WS-716 increased sensitivity of both pre-clinically and clinically derived MDR tumors to PTX in vivo with the T/C value of 29.7% in patient-derived xenograft (PDX) models. Relative to PTX treatment alone, combination of WS-716 and PTX caused no obvious adverse reactions. Taken together, our preclinical studies revealed therapeutic promise of WS-716 against MDR cancer, the promising data warrant its further development for cancer therapy.

【Objective】 To compare the volumes of brain regions and the white matter hyperintensities(WMH) in Alzheimer's disease(AD) and vascular dementia(VaD) patients, and discuss values of magnetic resonance imaging (MRI) in the differential diagnosis of AD and VaD. 【Methods】 The clinical data and MRI images of 35 patients with VaD and 74 patients with AD were retrospectively analyzed. Volumes of different brain regions and WMH were measured by using AccuBrain^TM system and visual rating scale was used to assess WMH. We then compared the volumes of brain regions and the WMH between AD and VaD. The principal component analysis(PCA) and logistic regression analysis were adopted for differential diagnosis. Receiver operating characteristic (ROC) analysis was performed to evaluate the diagnostic efficiency of the indexes. 【Results】 ①Compared with VaD patients, AD patients showed statistically smaller volumes of total intracranial volume, brain parenchyma, gray matter, white matter, hippocampus, amygdala, hypothalamus, frontal lobe(left, right), occipital lobe(left, right), temporal lobe(left, right), parietal lobe(left, right), hippocampus (left, right), amygdala(left, right), hypothalamus(left, right) (P<0.05). ②There was statistically significant difference in the visual scores for deep WMH(DWMH) between AD and VaD patients(P=0.015). The absolute and relative volumes of WMH in VaDpatients were statistically larger than those in AD patients(P<0.05). ③PCA revealed eight important parameters including brain parenchyma, hippocampus, amygdala, hypothalamus, frontal lobe(left), occipital lobe(left), temporal lobe(left) and parietal lobe(left) .The identification model consisting of amygdala, occipital lobe (left) and absolute volume of WMH was established based on logistic regression analysis. ④Among all the indexes, the identification model had the best diagnostic performance to differentiate AD from VaD and its sensitivity and specificity were 81.1% and 74.3%, respectively. 【Conclusions】 There are significant differences in both volumes of certain brain regions and severity of WMH between VaD and AD patients. The cranial MRI is of great value for the differential diagnosis of VaD and AD.

Objective:To investigate the quantitative changes of γδT cells in peripheral blood before and after anti-Brucella treatment in patients with chronic brucellosis.Methods:A prospective design was used to 88 patients with chronic brucellosis who were admitted to the Second People's Hospital of Tianjin from September 2012 to April 2018. The patients took anti-brucella drugs, And the changes in the number of γδT cell, CD3 +, CD4 +, CD8 +T lymphocytes and CD4/8 in peripheral blood before treatment, 6 weeks and 12 weeks after treatment were analyzed. Thirty volunteers were selected as the healthy control group from Tianjin Second People's Hospital employee health checkup in 2014. Results:After 6 weeks antibacterial therapy, the counts of CD3 +, CD4 + and CD8 +T lymphocytes were significantly lower than before treatment in patients with chronic brucellosis ( P<0.05) . After 12 weeks antibacterial therapy, the counts of γδT cell, CD3 +, CD4 + and CD8 +T lymphocytes were significantly lower than before treatment ( P<0.05) , but CD4/8 was higher than before treatment in patients with chronic brucellosis ( P<0.05) . Compared with healthy control group, after 6 weeks antibacterial treatment, the γδT cell count was still significantly higher, but the CD4 +T lymphocyte count was lower ( P<0.05) . After 12 weeks treatment, the γδT cell count was still significantly higher than that of the healthy control group ( P<0.01) . Conclusion:γδ T cells, CD4 +, CD8 + and CD3 +T lymphocytes may play a role in human body resistance to chronic Brucella infection.

Objective:To investigate the quantitative changes of γδT cells in peripheral blood before and after anti-Brucella treatment in patients with chronic brucellosis.Methods:A prospective design was used to 88 patients with chronic brucellosis who were admitted to the Second People's Hospital of Tianjin from September 2012 to April 2018. The patients took anti-brucella drugs, And the changes in the number of γδT cell, CD3 +, CD4 +, CD8 +T lymphocytes and CD4/8 in peripheral blood before treatment, 6 weeks and 12 weeks after treatment were analyzed. Thirty volunteers were selected as the healthy control group from Tianjin Second People's Hospital employee health checkup in 2014. Results:After 6 weeks antibacterial therapy, the counts of CD3 +, CD4 + and CD8 +T lymphocytes were significantly lower than before treatment in patients with chronic brucellosis ( P<0.05) . After 12 weeks antibacterial therapy, the counts of γδT cell, CD3 +, CD4 + and CD8 +T lymphocytes were significantly lower than before treatment ( P<0.05) , but CD4/8 was higher than before treatment in patients with chronic brucellosis ( P<0.05) . Compared with healthy control group, after 6 weeks antibacterial treatment, the γδT cell count was still significantly higher, but the CD4 +T lymphocyte count was lower ( P<0.05) . After 12 weeks treatment, the γδT cell count was still significantly higher than that of the healthy control group ( P<0.01) . Conclusion:γδ T cells, CD4 +, CD8 + and CD3 +T lymphocytes may play a role in human body resistance to chronic Brucella infection.

Objective To explore the effects of different skull-brain interfaces and mesh density of the cerebrospinal fluid (CSF) on dynamic responses of the brain. Methods The impact kinematics on cadaver head under rotation and translation impacts were reconstructed based on the 50th percentile adult head finite element model. The interfaces between skull and CSF, CSF and brain were modeled with different types of interfaces, which were set as sharing nodes, tied, frictionless sliding, so as to investigate the effect of different interface types on dynamic responses of the brain. Then, the interfaces between CSF, skull and brain were set as sharing nodes, while CSF was divided into single-layer and tri-layer of hexahedral element with the constant thickness of CSF, to study influences of CSF with different mesh density layers on dynamic responses of the brain. Results The intracranial pressure was highly sensitive to the interface types, while the brain response seemed to be relatively insensitive to the variation in CSF layers. Conclusions The research findings provide theoretical references for the construction of CSF and the selection of skull-brain contact interface of the head finite element model.

Background: The increased permeability of the blood-brain barrier (BBB) induced by ischemia/hypoxia is generally correlated with alteration of tight junctions (TJs). DL-3-n-butylphthalide (NBP) has been shown to exert neuroprotective effects after ischemic injury. However, few studies have assessed the correlation between NBP and TJs. This study aimed to investigate the potential effect of NBP on the TJ proteins claudin-5, zonula occludens-1 (ZO-1), and occludin during brain ischemia. Methods: A chronic cerebral hypoperfusion (CCH) Sprague-Dawley rat model was established, and NBP (20, 40, or 80 mg/kg, gavage, once a day) treatment was performed for 14 days. NBP (0.1 or 1.0 μmol/L) pre-treatment was applied to an in vitro hypoxia microvascular endothelial cell model (1% O₂, 24 h). BBB permeability was assessed by performing the Evans blue assay. The expressions and localization of claudin-5, ZO-1, occludin, phosphorylated/total protein kinase B (p-Akt/Akt), phosphorylated/total glycogen synthase kinase 3β (GSK-3β)/GSK-3β, and β-catenin/β-actin were evaluated by Western blotting or immunofluorescence. Reactive oxygen species (ROS) generation was measured by flow cytometry analysis. TJ ultrastructure was observed by transmission electron microscopy. Results: In CCH rats, treatment with 40 and 80 mg/kg NBP decreased the Evans blue content in brain tissue (9.0 ± 0.9 μg/g vs. 12.3 ± 1.9 μg/g, P = 0.005; 6.7 ± 0.6 μg/g vs. 12.3 ± 1.9 μg/g, P < 0.01), increased the expression of claudin-5 (0.79 ± 0.08 vs. 0.41 ± 0.06, P < 0.01; 0.97 ± 0.07 vs. 0.41 ± 0.06, P < 0.01), and elevated the ZO-1 protein level (P < 0.05) in brain microvascular segments in a dose-dependent manner in comparison with the corresponding values in the model group. There was no significant difference in occludin expression (P > 0.05). In the hypoxia cell model, NBP pre-treatment improved TJ ultrastructure, decreased intracellular ROS level, and increased the expression of claudin-5 (P < 0.01) and ZO-1 (P < 0.01) in comparison with the corresponding values in the hypoxia group. NBP treatment also elevated the relative expression levels of p-Akt/Akt, p-GSK-3β/GSK-3β, and β-catenin/β-actin in comparison with the corresponding values in the hypoxia group (all P < 0.05). Conclusion NBP improves the barrier function of BBB against ischemic injury by upregulating the expression of TJ proteins, possibly by reducing oxidative stress and activating the Akt/GSK-3β/β-catenin signaling pathway.

OBJECTIVE: To establish a compact and efficient hypergraph representation and a graph-similarity-based retrieval method of molecules to achieve effective and efficient medicine information retrieval. METHODS: Chemical structural formula (CSF) was a primary search target as a unique and precise identifier for each compound at the molecular level in the research field of medicine information retrieval. To retrieve medicine information effectively and efficiently, a complete workflow of the graph-based CSF retrieval system was introduced. This system accepted the photos taken from smartphones and the sketches drawn on tablet personal computers as CSF inputs, and formalized the CSFs with the corresponding graphs. Then this paper proposed a compact and efficient hypergraph representation for molecules on the basis of analyzing factors that directly affected the efficiency of graph matching. According to the characteristics of CSFs, a hierarchical collapsing method combining graph isomorphism and frequent subgraph mining was adopted. There was yet a fundamental challenge, subgraph overlapping during the collapsing procedure, which hindered the method from establishing the correct compact hypergraph of an original CSF graph. Therefore, a graph-isomorphism-based algorithm was proposed to select dominant acyclic subgraphs on the basis of overlapping analysis. Finally, the spatial similarity among graphical CSFs was evaluated by multi-dimensional measures of similarity. RESULTS: To evaluate the performance of the proposed method, the proposed system was firstly compared with Wikipedia Chemical Structure Explorer (WCSE), the state-of-the-art system that allowed CSF similarity searching within Wikipedia molecules dataset, on retrieval accuracy. The system achieved higher values on mean average precision, discounted cumulative gain, rank-biased precision, and expected reciprocal rank than WCSE from the top-2 to the top-10 retrieved results. Specifically, the system achieved 10%, 1.41, 6.42%, and 1.32% higher than WCSE on these metrics for top-10 retrieval results, respectively. Moreover, several retrieval cases were presented to intuitively compare with WCSE. The results of the above comparative study demonstrated that the proposed method outperformed the existing method with regard to accuracy and effectiveness. CONCLUSION This paper proposes a graph-similarity-based retrieval approach for medicine information. To obtain satisfactory retrieval results, an isomorphism-based algorithm is proposed for dominant subgraph selection based on the subgraph overlapping analysis, as well as an effective and efficient hypergraph representation of molecules. Experiment results demonstrate the effectiveness of the proposed approach.
Algorithms ; Databases, Chemical ; Information Storage and Retrieval ; Molecular Structure