In recent years,the clinical utility of circulating tumor cells (CTCs)analysis from peripheral blood has undoubtedly become a popular topic in the field of precision medicine.CTCs analysis is a noninvasive,easily obtained and highly repeatable testing approach,and could be carried out in a real-time manner,which has never failed to surprise us with great progression in the detection of early disease,assessing prognosis,treatment response monitoring and individualized therapeutic direction in breast cancer.

Objective To study the biocompatibility of carboxymethyl chitosan (CMCS) membrane with melanocytes from healthy human skin,and to investigate the feasibility to transport and carry melanocytes by using CMCS membrane.Methods CMCS membrane was prepared by a casting method combined with a glutaraldehydebased cross-linking method.Melanocytes were isolated from the foreskin of healthy men,and subjected to primary culture and subculture.The third-passage melanocytes were classified into two groups to be cultured on the CMCS membrane (test group) or traditional culture plates (control group).Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of melanocytes,and a sodium hydroxide-based lysis method to determine melanin content.HMB45 staining was conducted,and tyrosinase activity was estimated for melanocytes.Results Inverted microscopy showed that melanocytes were evenly distributed on the CMCS membrane with a normal shape.The melanocytes adherent to the CMCS membrane stained positive for anti-HMB45 monoclonal antibody.The growth curve of the melanocytes on the CMCS membrane,which was obtained from MTT assay,demonstrated that CMCS membrane could support the normal growth of melanocytes.No significant difference was observed between the test group and control group in melanin content (0.083 ± 0.015 vs.0.066 ± 0.008,t =2.38,P ＞ 0.01) or tyrosinase activity (0.234 ± 0.083 vs.0.241 ± 0.061,t =0.23,P ＞ 0.05).Conclusion CMCS membrane can maintain the normal biological activity of melanocytes and have good biocompatibility with skin melanocytes.

Objective To explore the relationship between operation scores and the expression of caveolin-1 ( Cav-1 ) protein in the cortex of mice.Methods Male Kunming mice were used and divided into two groups (excellent group and bad group) according to the training scores after 4 days' Y-maze learning training.The expression of cav-1 protein in cortex and hippocampus of the mice with different scores were measured through Western blot technology.Results Cav-1 protein expression of excellent group( Cav-1/β-actin was 5.71 + 1.11 ) in the cortex was significantly higher than that of the bad group ( Cav-1/β-actin was 1.69 + 0.20) (P ＜ 0.01 ).While there was no significant difference in the two groups in the expression of Cav-1 protein in hippocampus(P ＞ 0.05 ).Conclusion The operation scores have distinct correlation with the expression of Cav-1 protein in the cortex of mice.

Objective To express the novel fibronectin-binding protein Fba of group A streptococcus(GAS)and analyze its immunogenicity,so that to evaluate the immune responses to GAS infection.Methods fba gene was amplified by polymerase chain reaction(PCR)and confirmed by sequencing.Then it was cloned into pGEX4T-2 vector and Fba protein was expressed in E.coli BL21.The protein expression was identified by enzyme-linked immunosorbent assay(ELISA)and Westernblot.The sera from mice infected with GAS and anti-streptolysin-O positive patients were detected using microtiter plates coated with purified Fba protein as antigen.Afterward Balb/C mice were immunized with this purified protein and the sera were collected after the third immunization for the detection of IgG titer.Results It was confirmed by ELISA and Western blot that the recombinant Fba protein had a specific affinity with anti-Fba sera of rabbit.The anti-serum IgG titer of mice imrnunized with Fba protein was up to 1:4800.Conclusions GAS infection or Fba protein immunization are able to induce high serum titer of anti-Fba which could react specifically with the recombinant Fba protein.It indicates that Fba protein has good immunogenicity and antigenicity.So Fba protein could be a GAS candidate vaccine and an important tool to detect anti-GAS titer in GAS infected patients.

Objective To establish the HPLC fingerprint of Xuanbo Shuangsheng Granule for its quality control.Methods The components were separated on an Agilent TC-C18 column (150 mm ×4.6 mm,5 μm) at 25 ℃,with a gradient elution in 0-60 min at the flow rate of 1 mL/min using 0.2％ acetic acid aqueous solution and methanol as the mobile phase.The detection wavelength was 278 nm.By detecting 11 batches of Xuanbo Shuangsheng Granule,the HPLC fingerprint was established using Similarity Evaluation System for Chromatographic Fingerprint of TCMs (Version 2004A)and the common peaks were analyzed and identified in raw herbal material.Results The HPLC fingerprint of Xuanbo Shuangsheng Granule was obtained with similarity all over 0.9.Totally 27 common peaks were confirmed,and each common peak could be found in raw herbal medicines.Based on the reference substances,5 common peaks were identified,including phellodendrine (peak 11),liquiritin (peak 22),angoroside C (peak 25),cinnamic acid (peak 26) and harpagoside (peak 27).Conclusions This method is simple,accurate,repeatable and reliable,which could be applied in the quality control of Xuanbo Shuangsheng Granule.

Objective To compare the central foveal thickness (CFT) and subfoveal choroidal thickness (SFCT) in healthy pregnant women and patients with pre-eclampsia.Methods A prospective control study.Twenty normal subjects,20 healthy pregnant women and 20 patients with pre-eclampsia were included.The difference of gestational weeks between healthy pregnant women and patients with pre-eclampsia was not significant (x2=0.012,P=0.913).The differences of age and spherical equivalent among normal subjects,healthy pregnant women and patients with pre-eclampsia were not significant (x2=1.760,0.087;P=0.413,0.957).All eyes underwent optical coherence tomography examination to measure the CFT and SFCT.Results The mean CFT of normal subjects,healthy pregnant women and patients with pre-eclampsia were (194.40± 16.17),(201.2±17.33),(199.00±15.46) μm,there was no significant difference among the three groups (x2=0.888,P=0.641).The mean SFCT of normal subjects,healthy pregnant women and patients with pre-eclampsia were (263.45±69.66),(330.00±49.20),(373.40±52.00) μm,there was significant difference among the three groups (x2=22.818,P=0.000).The mean SFCT of healthy pregnant women was increased than that of normal subjects (Z=-2.991,P=0.002).The mean SFCT of patients with pre-eclampsia was increased than that of healthy pregnant women (Z=-2.638,P=0.007).Conclusion The mean SFCT of patients with pre-eclampsia is increased than healthy pregnant women.

Objective:To understand the characteristics of brucellosis in key population in rural areas of Hulunbuir City, Inner Mongolia Autonomous Region, and clarify the influencing factors of brucellosis infection, such as knowledge, belief and behavior, so as to provide scientific basis for the formulation of brucellosis prevention strategies and measures.Methods:A case-control study was used to study the confirmed cases of brucellosis among the key population of brucellosis in Hulunbuir rural areas in 2018. The patients without brucellosis were selected as control group. A questionnaire survey was used to investigated brucellosis related knowledge. The health-related behaviors (including raising livestock, cleaning houses, wells shared by human and livestock, immunizing livestock) were analyzed by single factor variation analysis and logistic regression analysis.Results:This study investigated 892 cases of population, 881 people responded effectively to the questionnaire, including 445 males, accounting for 50.51%, 436 females, accounting for 49.49%; the age range of 5-83 years old, the average age of (51.34 ± 11.87) years old; 153 cases of brucellosis antibody positive. The infection rate of brucellosis in this area was 17.37%, in which the male infection rate was 22.47%(100/445), and the female infection rate was 12.16% (53/436). There was a significant difference between male and female(χ 2=16.331, P < 0.05). Logistic regression analysis showed that raising cattle, cleaning houses, wells shared by human and livestock, using manure from cattle and sheep to fertilize, immunizing livestock were risk factors of brucellosis infection ( OR=1.905, 1.568, 1.320, 1.428, 1.488, P < 0.05). Conclusions:The incidence rate of brucellosis in Hulunbuir is higher than that in females. Close contact with infectious livestock and their excreta is a risk factor for brucellosis infection. According to the results of this survey. It is necessary to strengthen health education and behavior intervention of key population in order to effectively prevent and control brucellosis.

OBJECTIVE To explore mecha-nisms of imperatorin on regulating P-glycoprotein(P-gp)in blood-brain barrier(BBB)based on net-work pharmacology combined with in vitro experi-ment.METHODS Drug targets were predicted using the Pharmapper and Swiss targets data-bases;disease targets were obtained through the Genecards database;intersections between drugs and disease targets were screened by Cytoscape software;the obtained core targets were used to construct protein-protein interaction(PPI)network,gene ontology(GO)functions,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The effects of imperatorin(20,50,100 μ mol·L-1)on P-gp activity were monitored in hCMEC/D3in vitro BBB model,and the effects of imperatorin on the expression of target proteins were verified using Western blot method.RESULTS 55 drug targets and 3102 disease targets were obtained from the network pharmacology screening,and 37 core targets were obtained after the combination.Enrichment analysis showed that core targets were closely related to chemical synaptic trans-mission regulation,neurotransmitter receptor activity,proteinkinaseregulationactivity,G protein-coupled receptor signaling pathway,neural active ligand receptor interaction pathway,PI3K-Akt sig-naling pathway,VEGF signaling pathway,etc..In vitro experimental validation suggested that all tested concentration groups of imperatorin signifi-cantly reduced the activity and expression of P-gp,which were achieved by significantly downregu-lating the phosphorylation levels of PI3K and Akt,and repressing the expression of VEGFR2 pro-tein.CONCLUSION Network pharmacology was used to predict the core targets and signaling pathways of imperatorin on regulating P-gp in BBB and relevant validation was conducted through in vitro experiments,providing a refer-ence basis for further exploration of the mecha-nisms of imperatorin on regulating P-gp in BBB.

Objective: To compare the survival data of elderly advanced breast cancer (ABC) patients in China National Cancer Center with USA and summarize the therapeutic characteristics in elderly ABC patients via real world study. Methods: We summarized the clinicopathological characteristics, therapeutic regimens and survival outcome of 1 425 females with ABC who were initially hospitalized between January 2003 and December 2013 from Database in China National Cancer Center and compared with 21 185 ABC patients in the Surveillance, Epidemiology, and End Results: (SEER) database. Results The median overall survival (OS) of elderly patients was significantly shorter than that of the young group in China National Cancer Center (35.5 months vs. 43.9 months; χ²=8.747, P=0.003), which was similar to the survival feature in SEER database (24.0 months vs. 36.0 months; χ²=540.227, P<0.001). Compared with the young population, significantly more elderly patients suffered from the medical complications of hypertension [30.3% (67/221) vs. 9.5% (114/1 204); χ²=73.073, P<0.001], diabetes [14.5% (32/221) vs. 4.7% (57/1 204); χ²=30.220, P<0.001] and heart disease [6.3% (14/221) vs. 1.7% (20/1 204); χ²=17.638, P<0.001]. In estrogen receptor (ER) and/or progesterone receptor (PR)-positive patients, the percentage of receiving first-line endocrine therapy in elderly patients was significantly larger than that of the young population [26.9% (43/160) vs. 9.5% (80/841); χ²=37.599, P<0.001]. Moreover, in ER and/or PR-positive population, the elderly patients underwent first-line endocrine therapy resulted in better OS than those underwent first-line chemotherapy (49.9 months vs. 32.6 months; χ²=4.774, P=0.029), while no significant difference was observed between these two therapeutic modes in the young population (56.9 months vs. 48.8 months; χ²=1.103, P=0.294). Conclusion The proportion of elderly ABC patients with the medical complication of hypertension, diabetes and heart disease is significantly larger than that of the young population, which may lead to the difference in treatment decision making. In ER and/or PR-positive elderly ABC patients, receiving first-line endocrine therapy may result in better survival than first-line chemotherapy.