Objective To explore the effects of polydopamine (PDA) plus luteolin (LUT) with photothermal therapy on tumor killing by cytotoxic lymphocytes (CTL). Methods The synthesized PDA was characterized by using a scanning electron microscope (SEM) and a contact angle analyzer. RAW264.7 cells were divided into control group, LPS group (2 μg/L), and PDA' group (50 000 μg/L). After 48 h culture, cell viability was determined by cell counting kit-8. The effect of LUT on the differentiation of RAW264.7 cells was determined by flow cytometry. Female C57BL/6 mice were randomly divided into model group, PDA group, LUT group, and PDA+LUT group. B16F10 melanoma cells were subcutaneously injected into the back of mice in each group. On the 10th day after modeling, the PDA and PDA+LUT groups received injection of PDA into the tumor as well as photothermal therapy. The mice in the four groups received injection of PBS, PDA (2.5 μg/piece), LUT (500 μg/piece), and PDA (2.5 μg/piece) plus LUT (500 μg/piece) into the muscle of the right thigh, respectively. We observed the tumor growth and survival of the model mice. On days 3 and 7 after treatment, the spleen of each mouse was taken to prepare a single cell suspension for analysis of macrophage differentiation and T cell expression by flow cytometry. Results SEM showed that the synthesized PDA had good adhesion and affinity for water. CCK8 showed that the PDA did not affect cell viability (P>0.05). Significant differences were found in the mean fluorescence intensities of CD206 and iNOS between different groups of RAW264.7 cells (F=30.72,1 516.00,P<0.05). There were significant differences in tumor size and the expression levels of iNOS, CD206, IFN-γ+CD4+, and TNF-α+CD8+ of immune cells between mice with and without LUT injection, under the condition of either injecting or not injecting PDA (F=23.10-235.52,P<0.05). There were significant differences in tumor size and the expression levels of iNOS, CD206, IFN-γ+CD4+, and TNF-α+CD8+ of immune cells between mice with and without PDA injection, under the condition of either injecting or not injecting LUT (F=8.98-200.67,P<0.05). The survival rate of the PDA+LUT group was significantly higher than those of the other groups (χ2=9.70,P<0.01). Conclusion LUT could inhibit the differentiation of macrophages to M2 and promote the differentiation to M1 in mice. PDA combined with LUT could effectively inhibit tumor growth, improve survival rate, and enhance the tumor-kil-ling effect of CTL in mice.

目的:观察按不同时间方案注射的乙肝免疫球蛋白(HBIG)与乙肝疫苗对小鼠脾脏淋巴细胞亚群的影响.方法:饲养BALB/c小鼠120只(雌雄各60只),将小鼠按HBIG与第1针乙肝疫苗接种时间间隔分为三组(0周组、2周组、4周组),每组小鼠按接种的乙肝疫苗剂量分为2个亚组(1μg组、2μg组).每只小鼠于第0周注射50 IU HBIG.所有小鼠于第3针乙肝疫苗接种后4周分离脾脏淋巴细胞,使用流式细胞仪检测淋巴细胞亚群(CD3+、CD3+CD4+、CD3+CD8+、CD3?CD19+)比例.结果:0、2、4周组比较,两亚组(1μg、2μg)显示,与体液免疫相关的CD3?CD19+B淋巴细胞所占比例在雄、雌鼠4周组中均分别高于0、2周组,差异有统计学意义;与细胞免疫相关的T淋巴细胞及其亚群(CD3+、CD3+CD4+、CD3+CD8+)均未显示4周组高于其他两组.乙肝疫苗1μg组、2μg组比较,在雄、雌鼠中均显示4周组中2μg组CD3?CD19+B淋巴细胞所占比例高于1μg组(P<0.05).与细胞免疫相关的T淋巴细胞亚群未显示出2μg组高于1μg组的趋势.结论:小鼠注射HBIG的前提下,推迟接种乙肝疫苗及增加乙肝疫苗剂量有利于B淋巴细胞的增殖,推迟接种乙肝疫苗对T淋巴细胞亚群比例无明显影响.

目的通过预测分析严重急性呼吸综合征冠状病毒（SARS-CoV）与SARS-CoV-2结构蛋白中潜在的辅助性T细胞（Th细胞）表位,为新型冠状病毒疫苗的开发设计寻找合适的靶点。方法在美国国家生物技术信息中心的GenBank数据库中检索SARS-CoV与SARS-CoV-2病毒的参考序列,利用序列对比软件MEGA7和GeneDoc对2种病毒的刺突蛋白（S蛋白）、包膜蛋白（E蛋白）、膜蛋白（M蛋白）和核衣壳蛋白（N蛋白）的氨基酸序列进行比较分析。利用SYFPEITHI、IEDB和NetMHCIIpan在线生物信息学工具预测筛选SARS-CoV和SARS-CoV-2可能的Th细胞表位,并在此基础上寻找2种病毒同源的潜在表位。最后利用ProtScale对含同源表位的蛋白质进行疏水性分析比较。结果 2种病毒S蛋白同源性为75%,其他蛋白同源性分别为E蛋白94%、M蛋白90%及N蛋白90%。在线软件预测到SARS-CoV-2有22个潜在的Th细胞表位,SARS-CoV有25个潜在的Th细胞表位。进一步对比获得2对完全同源表位和6对高度同源表位,位于3对蛋白中,且ProtScale对比分析2种病毒相应蛋白的疏水性差异微小。结论对比分析得到8对完全同源或高度同源表位,且含差异氨基酸的对应肽段疏水性差异较小,同源性较高的Th细胞表位可考虑作为SARS-CoV-2疫苗设计的候选表位。

Objective:To summarize the diagnosis and treatment characteristics of multiple ovarian follicles growed after pituitary downregulation by gonadotropin-releasing hormone agonist (GnRH-a).Methods:This paper reported one patient with multiple ovarian follicles growed after pituitary downregulation using GnRH-a in our hospital, and the characteristics of 16 cases reported in Chinese and foreign literatures were retrospectively analyzed.Results:It is difficult to predict and avoid the development of multiple ovarian follicles after application of GnRH-a, and the "flare-up" effect after application of GnRH-a and the mutation of follicle-stimulating hormone (FSH) receptor gene may be the cause of the disease.Conclusion:The oocyte extraction operation is feasible for multiple ovarian follicles growed after pituitary downregulation. It is suggested to freeze the whole embryos in fresh cycle, and conducted embryo thawing transplantation at another time.

目的 利用生物信息学方法分析预测人巨细胞病毒(HCMV)膜表面糖蛋白gH以及磷酸化蛋白65(PP65)的B细胞、T细胞优势抗原表位,为HCMV亚单位疫苗的研发提供理论依据.方法 从美国国立生物技术信息中心(NCBI)数据库中获取HCMV的gH和PP65蛋白的氨基酸序列.通过ABCpred、BCPred和BepiPred软件预测gH和PP65蛋白的B细胞优势抗原表位;利用NetMHCⅡ2.3 Server和NetMHCⅡpan 4.0 Server软件预测gH和PP65蛋白的CD4+T细胞优势抗原表位;使用NetMHC 4.0 Server和IEDB软件预测gH和PP65蛋白的CD8+T细胞优势抗原表位;最后由VaxiJen v.2.0 Server软件对上述得到的所有抗原表位进行抗原性分析.结果 gH和PP65蛋白的二级结构中无规卷曲均占比最高,分别为40.51％、52.94％;综合所有软件的预测结果并进行抗原性分析后,获得g H蛋白CD8+T细胞优势抗原表位20个,CD4+T细胞优势抗原表位15个,B细胞优势抗原表位1个,获得PP65蛋白CD8+T细胞优势抗原表位8个,CD4+T细胞优势抗原表位8个,B细胞优势抗原表位3个.结论 HCMV的gH和PP65蛋白均含有丰富的B、T细胞优势抗原表位,可为HCMV亚单位疫苗的抗原肽选择提供理论依据.

Objective:To detect immunological effects of procyanidin ,gastrodin,baicalein and apigenin on antigen presenting cells of RAW264.7 and DC2.4 and search for new tumor vaccine adjuvant.Methods: After different concentrations of four kinds of Chinese herbal monomer and LPS were used to stimulate RAW 264.7 or DC2.4 cells for 48 hours,cells were obtained and stained with CD80,CD86,MHCⅠand MHCⅡ antibody.Then protein expression levels of CD80,CD86,MHCⅠ and MHCⅡ on RAW264.7 and DC2.4 cells were analyzed by flow cytometry.Results: Results showed that after 48 hours stimulated with different concentrations of four kinds of Chinese herbal monomer on RAW 264.7 or DC2.4 cells,the expression levels of CD80,CD86,MHCⅠ,MHCⅡ were all up-regulated in comparison with control , procyanidin and baicalein exhibited stronger immunological stimulating activity on a cellular level.Conclusion:Procyanidin and baicalein were found having potential application value in clinical treatment as tumor vaccine adju -vants.

目的 观察经角膜缘切口的无晶体眼行人工晶体悬吊术后角膜表面形态以及屈光状态变化.方法 无晶体眼病人19例(19只眼),采用英国RAYNER公司620H折叠式人工晶体对患眼行经角膜缘切口的折叠式人工晶体悬吊术,使用角膜地形图分析仪观察其术前、术后1周、术后1个月及术后2个月的角膜表面形态以及角膜表面非对称指数(SAI)、角膜表面散光值(CYL)、角膜表面规则指数(SRI)的改变.结果 术后1周及术后1个月CYL、SAI与术前相比均明显增大,差异有显著性(F=19.563、22.352,t=3.359～7.253,P＜0.05).SRI与术前相比差异无显著性(P＞0.05).三者在术后2个月与术前相比差异无显著性(P＞0.05).结论 经角膜缘切口行人工晶体悬吊术后SRI、SAI、CYL变化小、散光小、愈合快,可获得较满意的手术效果,该术式是治疗无后囊膜无晶体眼的一种较好的、可选择的手术方式.

Objective:To investigate whether women with high risks of ovarian hyperstimulation syndrome(OHSS) undergoing controlled ovarian hyperstimulation have the altered levels of the luteinizing hormone/choriogonadotropin receptor(LHCGR) mRNA in the lymphocytes.Methods:In this single center prospective study,108 IVF-ET patients were divided into 3 groups according ovarian response.In 42 cases with high OHSS risks,those 19 cases diagnosed OHSS were group A,the others 23 cases were group B.Those 62 cases with normal ovarian response were group C.The levels of LHCGR mRNA in lymphocytes were detected by PCR technology in the days of hCG injection and ovum pick-up(OPU day).Results:The level of basal serum FSH in group A was lower than that in group C(P=0.028).The serum concentration of estradiol(E2)in both group A and group B was higher than that in group C on the days of hCG injection and OPU(P=0.000).The median numbers of oocytes retrieved per patient in group A and group B were more than that in group C(P= 0.000).Group A had higher expression of LHCGR in lymphocytes mRNA on the hCG day and OPU day than group B and group C(P<0.05).The difference of the levels of LHCGR mRNA in lymphocytes between group B and group C was not significant(P >0.05).Conclusions:Expression of LHCGR mRNA in lymphocytes was related to ovarian response,which increased in OHSS patients.It could be used as a marker to predict OHSS.

Objective:To observe the effect of Leuprorelin on down-regulation of long protocol ovulation induction in in vitro fertilization and embryo transfer(IVF-ET).Methods:The clinical data of 148 patients who had received IVF-ET/intracytoplasmic sperm injection(ICSI) with long protocol for controlled ovarian hyperstimulation(COH) in reproductive medical center of the hospital from April 2010 to May 2011 were analyzed retrospectively.Thirty-two patients were treated with subcutaneous injection of 1.2 mg homemade Leuprorelin(Leuprorelin group) during mid-luteal phase for pituitary down-regulation,while 116 patients were treated with intramuscular injection of 1.2 mg Diphereline(Diphereline group) during mid-luteal phase for pituitary down-regulation.The effects of down-regulation and outcomes of IVF-ET in the two groups were analyzed and compared.Results:The patients in the two groups achieved the standard of pituitary down-regulation,there was no significant difference in the serum levels of estradiol,follicle stimulating hormone(FSH),and luteinizing hormone(LH) between the two groups(P>0.05).There was no significant difference in the number of retrieved oocytes,fertilization rate,the number of high quality embryos,implantation rate,and clinical pregnancy rate between the two groups(P>0.05).Conclusion:Subcutaneous injection of 1.2 mg homemade Leuprorelin during mid-luteal phase can achieve pituitary down-regulation effect of COH in IVF-ET long protocol ovulation induction,the clinical efficacy is the same as Diphereline.

<正>1984年Porter等[1]首次报道促性腺激素释放激素类似物(gonadotropin releasing hormone agonist,GnRH-a)用于体外受精-胚胎移植控制性促超排卵以来,GnRH-a与促性腺激素(Gn)联合促超排卵已广泛用于IVF-ET,尤以长方案应用最多。GnRH-a能有效地抑制内源性促黄体生成素(LH)峰出现,避免自发性排卵而获得较好的卵子质量与数量。但目前