Objective To investigate the effects of miR-139-5p-modified human umbilical cord mensenchymal stem cell(UC-MSC)-derived exosomes on stemness and radiotherapy sensitivity of breast cancer cells.Methods MCF-7 cells were divided into the control,miR-NC Exo,and miR-139-5p Exo groups.Immunofluorescence staining was used to observe the cellular uptake of exosomes.A tumor sphere formation assay determined the number of tumor spheres formed.Western blotting detected NANOG,SOX2,and OCT4 protein expression levels.MCF-7 cells were divided into control,2 Gy X ray,miR-139-5p Exo,and miR-139-5p Exo+2 Gy X ray groups.The survival rate of MCF-7 cells was detected by the MTT method.The apoptosis rate of MCF-7 cells was determined using flow cytometry.Results Compared to the control and miR-NC Exo groups,the number of tumor spheres formed by MCF-7 cells in the miR-139-5p Exo group was significantly decreased,and the relative expression levels of Nanog,SOX2,and OCT4 proteins were significantly downregulated(P＜0.05).Compared to the control group,the survival rate of MCF-7 cells in the 2 Gy X ray and miR-139-5p Exo groups was significantly decreased,and the apoptosis rate was significantly increased(P＜0.05).Compared to the 2 Gy X ray group,the survival rate of MCF-7 cells in the miR-139-5p Exo+2 Gy X ray group was significantly decreased,and the apoptosis rate was significantly increased(P＜0.05).Conclusion UC-MSC-derived exosomes modified by miR-139-5p inhibit the stemness of MCF-7 cells and enhance the radiotherapy sen-sitivity of MCF-7 cells.

Objective To investigate the effects of miR-139-5p-modified human umbilical cord mensenchymal stem cell(UC-MSC)-derived exosomes on stemness and radiotherapy sensitivity of breast cancer cells.Methods MCF-7 cells were divided into the control,miR-NC Exo,and miR-139-5p Exo groups.Immunofluorescence staining was used to observe the cellular uptake of exosomes.A tumor sphere formation assay determined the number of tumor spheres formed.Western blotting detected NANOG,SOX2,and OCT4 protein expression levels.MCF-7 cells were divided into control,2 Gy X ray,miR-139-5p Exo,and miR-139-5p Exo+2 Gy X ray groups.The survival rate of MCF-7 cells was detected by the MTT method.The apoptosis rate of MCF-7 cells was determined using flow cytometry.Results Compared to the control and miR-NC Exo groups,the number of tumor spheres formed by MCF-7 cells in the miR-139-5p Exo group was significantly decreased,and the relative expression levels of Nanog,SOX2,and OCT4 proteins were significantly downregulated(P＜0.05).Compared to the control group,the survival rate of MCF-7 cells in the 2 Gy X ray and miR-139-5p Exo groups was significantly decreased,and the apoptosis rate was significantly increased(P＜0.05).Compared to the 2 Gy X ray group,the survival rate of MCF-7 cells in the miR-139-5p Exo+2 Gy X ray group was significantly decreased,and the apoptosis rate was significantly increased(P＜0.05).Conclusion UC-MSC-derived exosomes modified by miR-139-5p inhibit the stemness of MCF-7 cells and enhance the radiotherapy sen-sitivity of MCF-7 cells.

Objective To investigate the development and dynamic changes of cysts in the brain of mice following infection with different forms of Toxoplasma gondii, so as to provide insights into for toxoplasmosis prevention and control. Methods ICR mice at ages of 6 to 8 weeks, each weighing 20 to 25 g, were intraperitoneally injected with tachyzoites of the T. gondii PRU strain at a dose of 1 × 10⁵ tachyzoites per mouse, orally administered with cysts at a dose of 20 oocysts per mouse or oocysts at a dose of 200 oocysts per mouse for modeling chronic T. gondii infection in mice, and the clinical symptoms and survival of mice were observed post-infection. Mice were orally infected with T. gondii cysts at doses of 10 (low-dose group), 20 (medium-dose group), 40 cysts per mouse (high-dose group), and the effect of different doses of T. gondii infections on the number of cysts was examined in the mouse brain. Mice were orally administered with T. gondii cysts at a dose of 20 cysts per mouse, and grouped according to gender (female and male) and time points of infections (20, 30, 60, 90, 120, 150, 180 days post-infection), and the effects of gender and time points of infections on the number of cysts was examined in the mouse brain. In addition, mice were divided into the tachyzoite group (Group T), the first-generation cyst group (Group C1), the second-generation cyst group (Group C2), the third-generation cyst (Group C3) and the fourth-generation cyst group (Group C4). Mice in the Group T were intraperitoneally injected with T. gondii tachyzoites at a dose of 1 × 10⁵ tachyzoites per mouse, and the cysts were collected from the mouse brain tissues 30 days post-infection, while mice in the Group C1 were orally infected with the collected cysts at a dose of 30 cysts per mouse. Continuous passage was performed by oral administration with cysts produced by the previous generation in mice, and the effect of continuous passage on the number of cysts was examined in the mouse brain. Results Following infection with T. gondii tachyzoites, cysts and oocysts in mice, obvious clinical symptoms were observed on days 6 to 13 and mice frequently died on days 7 to 12. The survival rates of mice were 67.0%, 87.0% and 53.0%, and the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0) and (581.0 ± 183.1) in the mouse brain (F = 11.94, P < 0.01) on day 30 post-infection with T. gondii tachyzoites, cysts and oocysts, respectively, and the numbers of cysts in the brain tissues were significantly lower in mice infected with T. gondii tachyzoites and oocysts than in those infected with cysts (all P values < 0.01). The survival rates of mice were 87.0%, 87.0% and 60.0%, and the mean numbers of cysts were (953.0 ± 355.5), (1 084.0 ± 474.3) and (1 113.0 ± 546.0) in the mouse brain in the low-, medium- and high-dose groups on day 30 post-infection, respectively (F = 0.42, P > 0.05). The survival rates of male and female mice were 73.0% and 80.0%, and the mean numbers of cysts were (946.4 ± 411.4) and (932.1 ± 322.4) in the brain tissues of male and female mice, respectively (F = 1.63, P > 0.05). Following continuous passage, the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0), (896.8 ± 332.3), (782.5 ± 423.9) and (829.2 ± 306.0) in the brain tissues of mice in the T, C1, C2, C3 and C4 groups, respectively (F = 4.82, P < 0.01), and the number of cysts was higher in the mouse brain in Group 1 than in Group T (P < 0.01). Following oral administration of 20 T. gondii cysts in mice, cysts were found in the moues brain for the first time on day 20 post-infection, and the number of cysts gradually increased over time, peaked on days 30 and 90 post-infection and then gradually decreased; however, the cysts were still found in the mouse brain on day 180 post-infection. Conclusions There is a higher possibility of developing chronic T. gondii infection in mice following infection with cysts than with oocysts or tachyzoites and the most severe chronic infection is seen following infection with cysts. The number of cysts does not correlate with the severity of chronic T. gondii infection, and the number of cysts peaks in the mouse brain on days 30 and 90 post-infection.

The effect of radiotherapy on anti-tumor immunity is bidirectional, immunotherapy, especially the combination of immune checkpoint inhibitors (ICIs) and radiotherapy, can produce synergistic effects on anti-tumor immunity. Compared with conventional radiotherapy, stereotactic ablative body radiotherapy (SABR) can achieve high-precision and high-dose irradiation on target lesions, and has stronger anti-tumor immune activation effect. At the same time, due to the steep dose gradient, SABR can better protect the surrounding normal tissues, which is an effective means for the rapid control of local lesions in advanced non-small cell lung cancer (NSCLC). ICIs are an important component of standard treatment for advanced NSCLC. There is growing evidence that SABR in combination with ICIs can benefit patients with advanced NSCLC. This article reviews the biological basis and clinical research progress on the combination of these two therapies, aiming to provide reference for the domestic counterparts to better use this new treatment model.

Objective: To investigate the current status of malaria rapid diagnostic test (RDT) strips application and malaria laboratory technicians' evaluation about them at primary healthcare provider level in Jiangsu Province. Methods: From November to December 2016, 878 medical institutions and 118 CDCs of city, county and township/community level in Jiangsu Province were selected as study samples using stratified random sampling method. Self-designed questionnaire was distributed to investigate the institution's malaria work task, RDT strips application and evaluation status in 2015. We also investigated the socio-demographic information and collected the RDT strips evaluation score from the malaria laboratory technicians selected from the institutions investigated (one technician from each institution). Rank sum test was performed to compare the RDT strips evaluation scores between medical institutions and CDCs, and among different medical institutions and CDCs. Results: In 2015, 405 cases of malaria were reported, 362 200 person-time of malaria blood testing task was conducted, and 100 000 RDT strips were procured and provided for healthcare providers in Jiangsu province for free. Of the 996 healthcare institutions investigated, 628 used RDT strips in the year 2015 and the median (P₂₅, P₇₅) of RDT strips volume used in these institutions was 10 (2, 25). The volume of RDT strips used in CDCs (15 (5, 52)) was significantly higher than that in medical institutions (10 (2, 25), (Z=3.42, P=0.001)). The investigated CDCs gave higher score on RDT strips' testing time per operation (10 (8.5, 10)) than medical institutions (9(8, 10), (Z=-2.20, P=0.028)). The employers of 614 investigated malaria laboratory technicians used RDT strips in 2015. The median of the scores given by CDC malaria laboratory technicians for RDT strips in terms of testing time per operation, testing operation and results judgement difficulties were 10 (9, 10), 10 (9, 10) and 10 (9, 10), respectively, which were significantly higher than those from technicians of medical institutions (9 (8, 10), 9 (8, 10), 9 (8, 10), (Z values were -2.55, -2.97 and -2.96, respectively; P values were all less than 0.05)). Conclusion RDT strips had been widely performed in health institutions in Jiangsu Province. The amount of RDT strips used in CDCs was significantly higher than that in medical institutions. Primary-level institutions and malaria laboratory technicians generally recognized RDT strips' advantage for application in terms of testing time and operational procedure. CDCs and malaria laboratory technicians from them gave higher regards on RDT strips in terms of testing time per operation, testing operation and results judgement difficulties compared with that of medical institutions.

OBJECTIVETo study the population genetic variation, genetic diversity and phylogenesis of Anopheles sinensis in China.

METHODSAnopheles sinensis samples collected from Shandong, Anhui, Jiangsu, Guizhou, and Yunnan Provinces and Guangxi Zhuang Autonomous Region with different geographical conditions between 2010 and 2012 were analyzed by mitochondrial DNA cytochrome oxidase subunit I (mtDNA-COI) gene amplification and sequencing. Bioedit 7.0 and DnaSP 5.0 software was used to compare the gene sequences and analyze the population genetic structure, respectively. Arlequin 3.1 was used to calculate the genetic distance and parameters of population differentiation. The relationship between the geographic and genetic distances was analyzed using IBD Web Service. PHYLIP 3.6 was used to construct the phylogenetic tree.

RESULTSPCR amplification and sequencing was performed successfully for 6 Anopheles sinensis populations containing 123 female mosquitoes. The length of mtDNA-COI gene fragment was 841 bp with an average A+T content of 71.2% and G+C content of 28.8%. High nucleotide diversity and genetic differentiation were observed among the Anopheles sinensis populations based on mtDNA-COI gene. Analysis of the molecular variance revealed a greater variation between populations than that within populations with isolation by distance between the populations. The Anopheles sinensis populations appeared to have undergone expansion, but the Yunnan population constituted an isolated branch in the phylogenetic tree.

CONCLUSIONmtDNA-COI can serve as the molecular marker to analyze population genetic variation and phylogenesis of Anopheles sinensis. The Yunnan population shows a phylogenetic difference from the other populations analyzed in this study.

Animals ; Anopheles ; genetics ; China ; DNA, Mitochondrial ; genetics ; Electron Transport Complex IV ; genetics ; Female ; Genetic Variation ; Genetics, Population ; Phylogeny

Objective To study the population genetic variation, genetic diversity and phylogenesis of Anopheles sinensis in China. Methods Anopheles sinensis samples collected from Shandong, Anhui, Jiangsu, Guizhou, and Yunnan Provinces and Guangxi Zhuang Autonomous Region with different geographical conditions between 2010 and 2012 were analyzed by mitochondrial DNA cytochrome oxidase subunit I (mtDNA-COI) gene amplification and sequencing. Bioedit 7.0 and DnaSP 5.0 software was used to compare the gene sequences and analyze the population genetic structure, respectively. Arlequin 3.1 was used to calculate the genetic distance and parameters of population differentiation. The relationship between the geographic and genetic distances was analyzed using IBD Web Service. PHYLIP 3.6 was used to construct the phylogenetic tree. Results PCR amplification and sequencing was performed successfully for 6 Anopheles sinensis populations containing 123 female mosquitoes. The length of mtDNA-COI gene fragment was 841 bp with an average A+T content of 71.2% and G+C content of 28.8%. High nucleotide diversity and genetic differentiation were observed among the Anopheles sinensis populations based on mtDNA-COI gene. Analysis of the molecular variance revealed a greater variation between populations than that within populations with isolation by distance between the populations. The Anopheles sinensis populations appeared to have undergone expansion, but the Yunnan population constituted an isolated branch in the phylogenetic tree. Conclusion mtDNA-COI can serve as the molecular marker to analyze population genetic variation and phylogenesis of Anopheles sinensis. The Yunnan population shows a phylogenetic difference from the other populations analyzed in this study.

Objective To study the population genetic variation, genetic diversity and phylogenesis of Anopheles sinensis in China. Methods Anopheles sinensis samples collected from Shandong, Anhui, Jiangsu, Guizhou, and Yunnan Provinces and Guangxi Zhuang Autonomous Region with different geographical conditions between 2010 and 2012 were analyzed by mitochondrial DNA cytochrome oxidase subunit I (mtDNA-COI) gene amplification and sequencing. Bioedit 7.0 and DnaSP 5.0 software was used to compare the gene sequences and analyze the population genetic structure, respectively. Arlequin 3.1 was used to calculate the genetic distance and parameters of population differentiation. The relationship between the geographic and genetic distances was analyzed using IBD Web Service. PHYLIP 3.6 was used to construct the phylogenetic tree. Results PCR amplification and sequencing was performed successfully for 6 Anopheles sinensis populations containing 123 female mosquitoes. The length of mtDNA-COI gene fragment was 841 bp with an average A+T content of 71.2% and G+C content of 28.8%. High nucleotide diversity and genetic differentiation were observed among the Anopheles sinensis populations based on mtDNA-COI gene. Analysis of the molecular variance revealed a greater variation between populations than that within populations with isolation by distance between the populations. The Anopheles sinensis populations appeared to have undergone expansion, but the Yunnan population constituted an isolated branch in the phylogenetic tree. Conclusion mtDNA-COI can serve as the molecular marker to analyze population genetic variation and phylogenesis of Anopheles sinensis. The Yunnan population shows a phylogenetic difference from the other populations analyzed in this study.