ABSTRACT
Objective To explore the potential predictive value of plasma exosomal miR-422a in different time after ischaemic stroke (1, 2, 3, 4 day) .Methods Retrospective study.Forty patients diagnosed with ischaemic stroke ( IS ) and ten age and sex matched people who underwent a standard physical examination were recruited from the First Affiliated Hospital of Guangxi Medical University between May 2016 and December 2016.Plasma exosomes were extracted by use of related kit and the expression level of plasma exosomal miR-422a in both IS patients (1, 2, 3, 4 day) and healthy controls were examined via quantitative real-time polymerase chain reaction ( qRT-PCR ) .The areas under the curve ( AUC ) of the receiver operating characteristic curve were constructed to evaluate the diagnostic accuracy of the miR -422a in IS, and one-way analysis of variance followed by the Games-Howell post hoc test was used for difference analysis.Results The exosomes isolated from human plasma showed round or oval vesicles , the average particle size was 128.2 nm and with maximum peak distribution of 186.9 nm.Moreover , all of the isolated exosomes were positive for a marker , with the CD63-positive rate at 80.6％ and the CD81-positive rate at 91.7％.qRT-PCR confirmed that miR-422a was expressed in human plasma exosomes , and the expression levels of plasma exosomal miR-422a [median relative values, 1.221 (95％CI:0.640-1.802) for healthy group, 4.418 (95％CI:2.642-6.193) for group of 1 dayafter IS, 2.912 (95％CI:2.262-3.562) for 2 day, 2.744 (95％CI:2.000-3.487) for 3 day and 0.449 (95％CI:0.170-0.727) for 4 day] were significantly increased in the time after IS 1, 2, 3 day ( F=13.57, P<0.05, P<0.01, P<0.05, respectively;and the AUC values were 0.920, 0.945, 0.870, respectively ) .The expression of plasma exosomal miR-422a on the 4th day after IS were lower than those in other IS groups ( F=13.57,P<0.005, P<0.001, P<0.001, respectively), and no statistical significance was found between the expression of plasma exosomal miR-422a on the 4th day after IS than that in the control group [0.449 (95％CI:0.170-0.727)].Conclusion Plasma exosomal miR-422a showed high diagnostic value as blood-based biomarker for diagnosing IS in the early phase (1-3 d).
ABSTRACT
Objective To improve the success rate of cerebrospinal fluid (CSF) collection in rats using a modified suction device with negative pressure. Methods A flow regulator was added to the original apparatus to adjust the intensity of negative pressure and to limit the volume of CSF collected each time. A total of 36 male Sprague- Dawley (SD) rats were randomly divided into 3 groups, and the CSF was collected in 3 ways: drawing from the cerebellomedullary cistern via percutaneous puncture, using a suction device with negative pressure and using a modified device. Kruskal-Wallis test was used to evaluate the differences in result between these three methods. Results The modified suction device with negative pressure effectively reduced bleeding. The quality of CSF collected with it was 75％ of 12 rats, significantly higher than that obtained with the original device (33. 3％, P ＜ 0. 01) or drawn from the cerebellomedullary cistern via percutaneous puncture (0％, P ＜ 0. 001). The success rate of qualified sample collection was highly superior to those of previously reported CSF collection methods. Conclusions Our modified suction device for CSF collection is simple and user-friendly, with minimal damage and little bleeding to rats. It can obviously improve the success rate of CSF collection in rats.
ABSTRACT
Objective To explore the clinical distribution states of human papillomavirus genotypes in tissues of 691 women with vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province and genotyping clinical significance.Methods Polymerase chain reaction(PCR)and gene-chips technology were utilized for the detection of 23 kinds of HPV genotypes in tissue specimens from 619 women of vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province.And related materials of all subjects were analyzed.Results In 691 women of vulva condyloma acuminates,597 women of HPV infecton,total infection rate of HPV was 86.40％(597/691),including single genotype infection rate of HPV was 51.38％(355/691),11、6 and 16 genotypes are the most common in single genotypes,they are successively 51.55％(183/355)、41.97％(149/355)and 3.38％(12/355).multiple genotypes infection rate of HPV was 35.02％(242/691),6+11、11+18、6+16 and 11+16 genotypes are the most common in multiple genotypes,they are successively 9.92％(24/242)、9.09％(22/242)、4.96％(12/242)and 4.13％(10/242).Conclusion The low-risk HPV types are the main factors to cause the female vulva CA,a few high-risk HPV types may cause warts as well in tissues of women with vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province.The vulva examine of HPV types should be held to the vulva CA patients.This precaution will has extremely important meaning to the prevention and treatment of the female vulva CA and cervical lesion in our nation.
ABSTRACT
Objective: To investigate the effect of mitoxantrone (MIT) on calreticulin (CRT) expression in B16 cells, and to observe the immune effect of B16-membrane antigen vaccine highly expressing CRT on B16 tumor-bearing mice. Methods: The expression of CRT on membrane of B16 cells was detected by immunofluorescence after treatment with different concentrations of MIT. B16-implanted mouse model was established, and the growth of B16-implanted tumors and CRT expression in B16-implanted tumor tissues were observed after treatment with different concentrations of MIT. Membrane antigen vaccines from both normal B16 cells and MIT-treated B16 cells were prepared, and mice were immunized before B16 cell implantation. The infiltration of immune cells into B16 tumor tissues and the ratios of CD4~+ and CD8~+ T cells in the spleen of B16 tumor-bearing mice were examined by immunohistochemistry and flow cytometry, respectively. Results: Flow cytometry results showed that MIT dose-dependently increased CRT expression on B16 cell membrane, with CRT expression in control and high dosage MIT groups being (29.40±3.57)% and (72.20±2.94)% (P<0.05), respectively. MIT also increased CRT expression in B16 tumor tissues, with those in the control and high dosage MIT groups being 3.21±1.37 and 9.17±1.06 (P<0.05), respectively. MIT effectively inhibited the growth of B16 tumors (P<0.05). Compared with normal B16 cell membrane antigen vaccine, the vaccine highly expressing CRT increased the numbers of DCs and T cells in B16 tumors tissues and the ratios of CD4~+ and CD8(+) T cells in the spleen (P<0.05). Conclusion: MIT can increase CRT expression on membrane of B16 cells. B16-membrane antigen vaccine highly expressing CRT can enhance the infiltration of DCs and T cells in melanoma, thus improving the immune effect of B16-membrane antigen vaccine.