Ethyl acetate fraction of Sargassum pallidum extract attenuates particulate matter-induced oxidative stress and inflammation in keratinocytes and zebrafish

Objective:To evaluate the effect of the ethyl acetate fraction derived from Sargassum pallidum extract against particulate matter(PM)-induced oxidative stress and inflammation in HaCaT cells and zebrafish.Methods:HaCaT cells and zebrafish were used to evaluate the protective effects of the ethyl acetate fraction of Sargassum pallidum extract against PM-induced oxidative stress and inflammation.The production of nitric oxide(NO),intracellular ROS,prostaglandin E_(2)(PGE_(2)),and pro-inflammatory cytokines,and the expression levels of COX-2,iNOS,and NF-κB were evaluated in PM-induced HaCaT cells.Furthermore,the levels of ROS,NO,and lipid peroxidation were assessed in the PM-exposed zebrafish model.Results:The ethyl acetate fraction of Sargassum pallidum extract significantly decreased the production of NO,intracellular ROS,and PGE_(2) in PM-induced HaCaT cells.In addition,the fraction markedly suppressed the levels of pro-inflammatory cytokines and inhibited the expression levels of COX-2,iNOS,and NF-κB.Furthermore,it displayed remarkable protective effects against PM-induced inflammatory response and oxidative stress,represented by the reduction of NO,ROS,and lipid peroxidation in zebrafish.Conclusions:The ethyl acetate fraction of Sargassum pallidum extract exhibits a protective effect against PM-induced oxidative stress and inflammation both in vitro and in vivo and has the potential as a candidate for the development of pharmaceutical and cosmeceutical products.

黄牛毛藓(Ditrichum pallidum)抑菌效果的生物测定

为了研究苔藓植物的抑菌效果,以山东省内分布较广泛、生物量较大的黄牛毛藓(Ditrichum pal-lidum)为试验材料,用分光光度法对黄牛毛藓水浸提液进行抑菌作用的初步研究。结果显示,黄牛毛藓对金黄色葡萄球菌、大肠杆菌、变形杆菌均有不同程度的抑制作用,抑制强度表现为大肠杆菌>金黄色葡萄球菌>变形杆菌。作为我国重要药用苔藓植物的一种,黄牛毛藓的抑菌效果对不同的供试细菌均表现出一定的抑制作用,在有害生物防治和疾病的治疗方面具有一定的应用价值。

Construction and Expression of Tp0453 Recombinant Protein of Treponema pallidum and Development of Indirect ELISA for Diagnosinq Syphilis

Objectives： To clone and express Tp0453 outer membrane protein of Treponema pallidum, and to evaluate its significance in the serodiagnosis of syphilis. Methods： The immuno-dominant epitope of Tp0453 gene was amplified from the complete genome of T.pallidum by polymerase chain reactions （PCR）, subcloned into the expression vector Pqe32 to generate recombinant plasmid Pqe32/Tp0453, and was then expressed in E. coli M15. The fusion protein was purified with Ni-NTA affinity purification. Indirect ELISA was developed to detect human serum IgG antibody to T. pallidum. Results： The recombinant Tp0453 protein was successfully expressed and purified. The recombinant protein had a molecular weight of approximately 32KDa.Indirect ELISA to the recombinant protein was developed.Sixty control sera were tested by ELISA and yielded a sensitivity of 100% （30/30） and a specificity of 100% （30/30）. While testing for T. pallidum in human sera, the sensitivity and specificity of ELISA were 96.8% and 100%, respectively, when compared with TPPA test results. The concordance of results between the ELISA test and the TPPA test was 98.2%. Conclusion： The recombinant Tp0453 outer membrane protein elicited a strong immunoreaction to anti-T.pallidum IgG antibody and has great potential use in ELISA for the serodiagnosis of syphilis.

Development and Clinical Application of a Single-tube Nested PCR Method to Amplify the DNA Polymerase Ⅰ Gene of Treponema Pallidum

Objective: To develop a sensitive, specific and simple method for detection of extremely low numbers of T. pallidum in clinical specimens, as a significant addition to the serologic tests for syphilis diagnosis. Methods: Double-tube nested PCR(DN-PCR) and single-tube nested PCR(SN-PCR) assays were performed to amplify specific fragments of the DNA poly-merase I gene(polA) of T. pallidum. Sensitivity and specificity of the two PCR assays were tested. Eighty-six whole blood specimens from persons with suspected syphilis were detected by the two nested PCR methods. The TPPA test was used as a comparison for detecting syphilis in sera from corresponding patients. Results: Only specific amplicons could be obtained during amplification of the T. pallidum polA gene and the detection limit was approximately 1 organism when analyzed on gel by the two PCR methods. Of 86 clinical specimens, 62 were positive by TPPA. Of these, 54 and 51 were positive by the DN-PCR and SN-PCR, respectively, which does not represent a statistically significant difference between the two PCR tests. Of 24 TPPA-negative specimens, 5 were positive by both DN-PCR assay and SN-PCR assay. Conclusion: The SN- polA PCR method is extremely sensitive, specific and easy to perform for detecting low numbers of T. pallidum in clinical blood specimens as a complementary to serology for syphilis diagnosis.

Treponema pallidum and Haemophilus ducreyi DNA detection by A Multi-Nested Polymerase Chain Reaction

Objectives: To develop a multi-nested polymerase chain reaction in an assay to detect early Treponema pallidum and Haemophilius ducreyi DNA in the swabs of genital ulcers. Methods: Four pairs of outer and inner primers, specific to the basic membrane protein gene of Treponema pallidum and to the 16s rRNA gene of H ducreyi were synthesized. The multi-nested PCR was developed and applied to detect Treponema pallidum and Haemophilus dicreyi in clinical swabs. Result: The two samples of standard strains of Haemophilus ducreyi and one Treponema pallidum were amplified and showed 309-bp rRNA gene of Haemophilus ducreyi and 506-bp DNA of Treponema palidum, respectively. Out of 51 samples of genital ulcer detected, 29 showed Treponema pallidum positive product and no Haemophilus ducreyi DNA was found. Conclusion: The multi-nested PCR for Treponema pallidum and Haemophilus ducreyi could be useful for early detection and distinguishing diagnosis between syphilis and chancroid.

CLINICAL EVALUATION OF FOUR RECOMBINANT TREPONEMA PALLIDUM ANTIGEN-BASED RAPID TESTS IN THE DIAGNOSIS OF SYPHILIS

Objective To assess the sensitivity, specificity, and feasibility of 4 recombinant Treponema pallidum antigenbased rapid tests in the diagnosis of syphilis. Methods A total of 970 outpatients were selected from the Sexually Transmitted Diseases Centre of Peking Union Medical College Hospital. Venous blood was collected and serum was extracted. T. paUidum antibodies in whole blood, anticoagulant whole blood, and serum were detected using 4 recombinant T. pallidum antigen-based rapid tests. T. pallidum haemagglutination test （TPHA） was considered as the gold standard for the detection of T. pallidum specific antibodies in serum. The sensitivities and specificities of four methods were analyzed. Results The sensitivities and specificities of Abbott Determine Syphilis TP test, SD-BIOLINE Syphilis 3.0 test, VISITECT-SYPHILIS test, and Syphicheck-WB test for serum specimens were 100% and 98. 9%, 95.7% and 98.0%, 94.6% and 98.2%, 68.1% and 98.9% ; for whole blood were 74. 1% and 99. 5%, 87.9% and 99.4% , 73.2% and 99.7%, 64. 7% and 99.7%. The observed sensitivities of the 4 rapid diagnosis tests were not significantly different with TPHA （ P 〉 0.05 ）. Conclusions The 4 rapid tests show good performance and characteristics in the diagnosis of syphilis. Furthermore, they are more sensitive for serum specimens than whole blood.

Construction of the Eukaryotic Expression Vector for Outer Membrane Protein Tp92 from Treponema pallidum and Its Preliminary Study on the Immune Responses in New Zealand Rabbits

To construct the recombinant plasmid of eukaryotic expression containing Tp92 gene from Treponema pallidum and study its immunogenicity in New Zealand white rabbits. Tp92 gene was amplified from the genomic DNA of T. pallidum by polymerase chain reaction (PCR) and subcloned into appropriate site of pcDNA3.1(+) vector. After identification by sequencing and restrictive enzyme digestion, the recombinant plasmid was transfected into HeLa cells using liposome, and the expressed protein was identified by immunocytochemistry and Western blotting. After verifying that the Tp92 antigen gene fragment could be expressed in HeLa cells, 100?μg of recombinant plasmids [pcDNA3.1(+)-Tp92], 100 μg of control plasmids [pcDNA3.1(+)] or 0.5 ml PBS buffer were administered in 3 groups of New Zealand white rabbits (6 rabbits/group), and the booster immunizations were employed at 2-week interval for 3 times. ELISA assay was used for the quantitative detection of the specific antibody in the sera of rabbits, and the proliferation response of spleen cells was detected by MTT assay. It was found that the target gene Tp92 segment about 2103 bp was obtained, and the DNA sequence of Tp92 gene constructed in pcDNA3.1 (+) vector was consistent with the published nucleotide sequence. The homologies of the nucleotide and putative amino acid sequences of Tp92 gene between T.pallidum subsp. pallidum Nichols and various pathogenic treponeme strains were 95.5%-100%. The analysis of immunocytochemistry and Western blotting showed that Tp92 gene segment constructed in pcDNA3.1(+) vector could express a fusion protein with a calculated molecular mass of 77 kDa in HeLa cells and the expressed protein could react with positive blood serum from syphilis patient. The specific antibody IgG titers were observed and the highest titer was 1∶1024 in rabbits after 3 times with pcDNA3.1(+)-Tp92. The proliferation response of spleen cells were significantly higher than that of rabbits injected with pcDNA3.1(+) ( P <0.05). The successful expression of the eukaryotic expression plasmid of Tp92 gene from T. pallidum was obtained in eukaryotic system and strong responses of humoral and cellular immunity was evoked by DNA vaccine of pcDNA3.1(+)-Tp92 in rabbits thus establishing a solid basis for the future studies in the biological activities and for the development of the syphilis DNA vaccine.

Expression and purification of the recombinant outermembrane protein Tp0453 of Treponema pallidum and its characterization of immuno-competence

To clone and express the recombinant outer membrane protein Tp0453 of Treponema pallidum and to analyze the immuno-reactivity and immunogenicity of the expressed protein, the immuno-dominant epitope of the Tp0453 was amplified from the complete genome of T.pallidum by PCR, subcloned into expression vector pQE32 to generate the recombinant plasmid pQE32/Tp0453, then expressed in E.coli M15 and analyzed by SDS/PAGE and Western blotting. The fusion protein expressed was purified with Ni-NTA affinity chromatography. Its immuno-reactivity was assayed by indirect ELISA, and the immunogenicity was determined by immunization with this fusion protein in New Zealand rabbits. In the present study, a fusion protein of molecular weight about 32 kDa was obtained. As demonstrated by Western blotting, the recombinant protein could react specifically with positive IgG sera of patients with syphilis, and the antibodies against T.pallidum in human sera were successfully detected by indirect ELISA. Both the sensitivity and specificity of ELISA based on the Tp0453 fusion protein as were 100% (30/30) when detected with control sera. In comparison with the results of IgG ELISA with those of TPPA. It was found that the sensitivity of ELISA was 96.8% and the specificity was 100%. The difference of ELISA and TPPA was not significant, and the concordance of results between ELISA and TPPA was 98.2%. In addition, specific humoral responses could be elicited by immunization with the recombinant fusion protein in New Zealand rabbits with a specific antibody titer of 1∶1280 after 3 successive doses of immunization. These results demonstrate that the expressed recombinant fusion protein shows excellent immuno-competence and provide foundation to develop a quick diagnostic kid applied to detect the presence of T.pallidum infections.

Eukaryotic expression of outer membrane protein Gpd from Treponema pallidum and preliminary studies on its immune response in rabbits

The Gpd gene was amplified from the genomic DNA of Treponema the appropriate site of pcDNA3.1 （ ＋ ） vector. The expression of pcDNA3. I was tested with Western blotting and technology of immunoeytochemisty. New munized with the eukaryotic expression recombinant pcDNA3, 1 （ ＋ ）-Gpd pallidum and cloned into （ ＋ ）-Gpd in Hel,a cells Zealand rabbits were imA fusion protein of C, pd with 4.1 kDa has been effectively expressed in HeLa cells, which were detected bv Western blotting and the immunocytochemistry techniques. The New Zealand rabbits were able to elicit the specific antibody after immunization with the nucleic acid vaccine. The antibody titer could reach as high as 1 ： 1024 after 2 weeks of the third injection; and the splenocytes proliferated evidently due to the Gpd protein stimulation. Both the antibody titer and the splenocytes proliferation were higher substantially than those of controls （ P 〈 0.01 ）. All above data will contribute to an experimental basis of further study of the biological function of Gpd protein as well as DNA vaccine for syphilis.

Cloning and Expression of the Tpp17 Gene of Treponema pallidum And Clinical Application

Objective: To obtain recombinant Treponema pallidumsubsp pallidum (TP 17KD) lipoprotein in large quantities byamplification and to further purify antigens for laboratorydiagnosis of syphilis and development of a syphilis vaccine. Method: The Tpp17 lipoprotein gene was amplified fromthe TP(strain Nichols), and then it was recombinated into aplasmid pMAL-2c and cloned within E coli l2-TB1. The hostbacteria containing recombinant plasmids were induced withIPTG. The Tpp 17KD lipoprotein gene was amplified by us-ing PCR and positive clones were screened with double diges-tion and PCR. Recombinant plasmids were transformed intoE. coli and the E coli carrying recombinant plasmids wereinduced. The expression of TP 17KD was detected by sodiumdedecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and immunoblot. Results:Gel staining with Coomassie blue G-250 showedthat the induced E coli carrying recombinant plasmid couldproduce 60KD fusion protein at high levels. Gel scanningshowed that 17KD protein expression in E coli accounted for10% of total cellular protein. The recombinant protein antigenreacted with the sera of syphilis patients. Conclusion: Our study lays a cornerstone for developingnew techniques of laboratory diagnosis for syphilis and newvaccines. Preliminary clinical application showed that thefusion protein could be used for the diagnosis of syphilis.

Detection of Treponema pallidum,Herpes Simplex Virus,and Haemophilus ducreyi from Genital Ulcers by Multiples Polymerase Chain Reaction

Objective: To evaluate the clinical application of multiplex PCR inthe detection of Treponema pallidum, Herpes simplex virus (HSV), andHaemophilus ducreyi. Method: Three standard strains were used to set up a multiplexPCR (MPCR) for detecting syphilis, herpes genitalis, and chancroidsimultaneously. Samples from 122 patients with genital ulcer disease(GUD) were subjected to MPCR and the results were compared withthose of dark-field microscopy and TP serology, HSV antigen ELISA,and H. ducreyi culture. Results: In the 122 patients with GUD, MPCR identified 34 casesof T. pallidum infection, 40 cases of HSV infection, and 2 cases of mixedinfection of T. pollidum and herpes. No positive results of H. ducreyiwere found. The sensitivity of MPCR to T. pallidum and herpes was100% and 93.3%, respectively. The sensitivities of dark-field mi-croscopy and TP serology, HSV antigen ELISA, and H. ducreyi cul-ture was 35.3%, 50% and 100%, respectively. Conclusion: MPCR showed a relatively higher sensitivity for T.pallidum as compared with the routine techniques. Although its sensi-tivity for HSV was not as good as that of antigen ELISA, it also yieldeda high detection rate. MPCR can detect more than one pathogen. It issimple, quick, sensitive, and suitable for clinical use or epidemiologicalinvestigation.

The Construction of the Eukaryotic Expression Vector of Glycerophosphodiester Phosphodiesterase Gene from Treponema pallidum and its Expression in Hela Cells

Objective: To construct the recombinant plasmid containing Glycerophosphodiester phosphodiesterase (Gpd) gene from Treponema pallidum and transfect it into Hela cells to express the encoded outer membrane protein. Methods: The Gpd gene was amplified from the genomic DNA of T.pallidum by polymerase chain reaction (PCR) and inserted into cloning vector pUCm-T. The inserted Gpd gene was subcloned into the appropriate site of pcDNA3.1(+) vector. After identification by sequencing and restrictive enzymes digestion, the recombinant plasmid was transfected into Hela cells using liposomes. The expressed protein was identified by immunocytochemistry and Western blot. Results: The target Gpd gene segment was approximately 1059bp. The DNA sequence of the Gpd gene contained in the pcDNA3.1(+) vector was consistent with the published nucleotide sequence. The homology of the nucleotide and putative amino acid sequences of the Gpd gene between T. pallidum subsp. pallidum Nichols and various pathogenic treponemal strains ranged from 98% to 100%. Immunocytochemistry and Western blot analysis showed that the constructed Gpd-pcDNA3.1(+) vector expressed a fusion protein with a calculated molecular mass of 41KDa in Hela cells and that the expressed protein reacted with the sera from syphilis patients. Conclusion: The successful construction and expression of the eukaryotic expression plasmid of the Gpd gene from T.pallidum provide a promising tool to further study the biological activity of T.pallidum and develop a DNA vaccine for syphilis.

酒依赖患者的腹侧苍白球改变及其与复饮的关系

目的:探讨酒依赖患者腹侧苍白球(VP)的体积变化及其与复饮的关系。方法:采用横断面与队列研究相结合的方法,选取符合美国精神障碍诊断与统计手册第4版(DSM-IV)酒精依赖综合征诊断标准的患者33例,在停酒14~28 d内进行头部磁共振成像检查,采用自制调查问卷、临床机构酒精戒断症状评定量表(CIWA)、酒精使用障碍筛查量表(AUDIT)进行评估,采用自制随访问卷评估半年内治疗结局。比较酒依赖组与对照组,复饮组与非复饮组间VP体积。结果:酒依赖组的VP体积小于正常对照组[(1749±492)mm^(3) vs.(2116±189)mm^(3),P<0.05],复饮组的VP体积较非复饮组大[(1956±452)mm^(3) vs.(1555±458)mm^(3),P<0.05]。结论:酒依赖患者的VP可能参与病理性奖赏环路功能,并与复饮有关。

梅毒实验室诊断的研究现状与展望

[背景]近年来,梅毒发病数呈逐年上升趋势,引起广泛关注,已成为全球性公共卫生问题.鉴于梅毒病程的隐匿性、复杂性和多样性,迫切需要高灵敏度的诊断方法和有效的疗效监测指标,以实现梅毒的早诊断和早治疗.[进展]本文对梅毒实验室检测方法及诊断程序的研究现状进行综述.病原学检查可以直接确诊感染,但易受取材部位及取材方法等因素影响,灵敏度较低.血清学检测是梅毒诊断的主要检测方法,但由于抗体至少在感染2周后才产生,难以筛查窗口期的患者.由于血清学诊断程序的行业标准尚未统一,各实验室对3种血清学诊断程序的选择也不尽相同.聚合酶链式反应(polymerase chain reaction,PCR)对于一期或二期梅毒患者皮损组织具有较高灵敏度和特异度,但对于较易获得的血液样本,其梅毒螺旋体(Treponema pallidum)DNA检出率低,国内至今尚无获得医疗器械注册证的梅毒PCR检测试剂,无法应用于临床检测.[展望]首先,研发梅毒螺旋体抗体和非特异性抗体联合检测试剂,避免单一试验假阴性造成的漏诊;其次,开展血液、脑脊液、尿液等标本的梅毒螺旋体抗原谱研究,缩短检测的窗口期;最后,引入质量管理体系,制定梅毒分子诊断的统一标准.

活动性梅毒患者梅毒血清特异性抗体水平与体内免疫球蛋白水平的相关性研究

目的:探究活动性梅毒患者梅毒血清特异性抗体水平(TPAb)与体内免疫球蛋白A/E/M/G(IgA/E/M/G)及IgG亚型的相关性,为临床梅毒的综合诊疗与预后评估提供依据。方法:选取2023年就诊于苏州大学附属第一医院的活动性梅毒患者,根据梅毒甲苯胺红不加热血清试验(TRUST)分为1∶4+与1∶16+两组,另选取健康对照组,均进行化学发光微粒子免疫检测(CMIA)测定TPAb,免疫比浊法测定IgA/E/M/G及IgG1/2/3/4含量。结果:本次共纳入活动性梅毒患者206例(男性125例,女性81例),年龄(46.05±19.58)岁。相对健康对照组,可见活动性梅毒组IgG2降低,TPAb、IgA/E/M/G和IgG1/3/4水平均升高,其中TPAb、IgE、IgG1/2/3水平差异具有统计学意义(P<0.05)。相比于健康对照组,TRUST 1∶4+组TPAb、IgE、IgG2差异具有统计学意义,TPAb与IgE、IgG2/4存在相关性;而TRUST 1∶16+组TPAb、IgA/E/M/G、IgG1/2/3水平差异均具有统计学意义,TPAb与IgE、IgG4存在相关性(P<0.05)。另外,TRUST阳性组之间的TPAb、IgG和IgG1/3水平差异具有统计学意义(P<0.05)。结论:不同TRUST滴度的活动性梅毒患者TPAb与Ig水平存在较大差异,TPAb联合Ig可用于监测梅毒活动性强弱,IgA/M/G及IgG1/3可能成为候选生物标志物。

抗-TP阳性献血者中抗病毒治疗药物使用情况分析

目的了解深圳市梅毒螺旋体(TP)抗体(抗-TP)阳性献血者中抗逆转录病毒治疗(ART)药物使用情况,评估人类免疫缺陷病毒(HIV)诊疗新趋势带来的血液安全风险。方法采用分层随机抽样方法选取2019年3月至2023年1月深圳地区血液筛查合格的重复献血者60例(阴性对照组),规律服用已知ART药物人群3例(阳性对照组),抗-TP阳性/抗-HIV阴性献血者353例(实验1组),抗-TP阳性/抗-HIV阳性献血者25例(实验2组),应用高效液相色谱-串联质谱(HPLC-MS/MS)方法检测各组血浆样本中8种ART药物浓度,分析ART药物使用情况。结果阳性对照组血浆采用1∶6稀释混样后ART药物仍可检出,实验1组和实验2组1∶6人份混合血浆阳性样本经拆分确证,实验2组检出1例ART药物阳性样本,该样本抗-HIV、蛋白免疫印迹、HIV RNA阳性,抗TP阳性献血者ART药物检出率为0.26%,实验1组ART药物检出率为0.00%,实验2组ART药物检出率为4.00%。结论深圳地区抗-TP阳性献血者中发现使用ART药物情况,合并HIV感染及高危性行为人群更可能使用ART药物。

2019—2023年江西省血液中心无偿献血者人类免疫缺陷病毒及梅毒螺旋体感染现状及特征分析

目的:调查2019—2023年江西省血液中心无偿献血者人类免疫缺陷病毒(HIV)及梅毒螺旋体(TP)感染现状,并分析其特征。方法:选取2019年1月—2023年12月江西省血液中心的408605名无偿献血者血液标本进行调查,对标本进行HIV、TP项目常规检测,HIV反应性标本送HIV确证实验室进行确证。结果:2019—2023年无偿献血者HIV初筛阳性率为0.062%,TP初筛阳性率为0.227%,总阳性率为0.289%。各年度初筛总阳性率比较,差异有统计学意义(P<0.05)。免疫印迹法确证检测结果显示,34名无偿献血者为HIV阳性,HIV阳性率为0.008%。2019—2023年的无偿献血者中,女性HIV及TP初筛阳性率(0.324%)高于男性(0.268%),45~<56岁的无偿献血者HIV及TP初筛阳性率(0.542%)较高,研究生HIV及TP初筛阳性率(0.160%)较低,初筛阳性率随学历降低而呈现升高的趋势,农民HIV及TP初筛阳性率(0.629%)较高,初次献血者HIV及TP初筛阳性率(0.417%)较高。2019—2023年34名HIV确证阳性的献血者中,女性占比(5.882%)低于男性(94.118%),18~<25岁者占比(50.000%)最高,学生占比(41.176%)最高。结论:不同特征的无偿献血者HIV及TP检测结果存在差异,可通过加强健康教育和“献血后回告”宣教、强化献血前征询与筛查、构建区域性的献血者监测系统和艾滋病疫情管理共享系统等方式推进我国血液安全。

2018-2022年北京市密云区医院孕产妇人类免疫缺陷病毒、梅毒螺旋体、乙型肝炎病毒检测结果分析

目的:分析2018-2022年北京市密云区医院孕产妇人类免疫缺陷病毒(HIV)、梅毒螺旋体、乙型肝炎病毒(HBV)检测结果。方法:选取2018年1月-2022年12月于密云区医院进行孕产期保健及分娩的孕产妇19 327例作为研究对象,收集并分析孕产妇一般资料及HIV、梅毒螺旋体和HBV初筛结果。结果:19 327例孕产妇中,HIV阳性0例,梅毒螺旋体阳性101例(0.52%),HBV阳性175例(0.91%)。2018-2022年梅毒螺旋体和HBV阳性呈小幅度波动,其中以2021年明显降低。孕产妇梅毒螺旋体、HBV阳性率及构成比均以20~29岁最高,40~45岁最低。结论:2018-2022年北京市密云区医院孕产妇HIV、梅毒螺旋体、HBV阳性率较低,以20~29岁最多,临床应针对此情况采取应对措施,消除艾滋病、梅毒、乙型肝炎的母婴传播。

探讨TRUST与TP-ELISA对梅毒患者的诊断价值

目的 探讨甲苯胺红不加热血清试验(toludine red unheated serum test, TRUST)、梅毒螺旋体-酶联免疫吸附试验(treponema pallidum-enzyme linked immunosorbent assay, TP-ELISA)检测方法在梅毒诊断中的诊断价值。方法 非随机选取2022年1月—2023年10月兴义市人民医院收治的86例疑似梅毒患者为研究对象,所有患者均完成TRUST与TP-ELISA检测,并将梅毒螺旋体明胶颗粒凝集试验(treponema palli-dum particle agglutination, TPPA)检测作为金标准,分析TRUST与TP-ELISA的诊断价值。结果 86例疑诊梅毒患者中,61例患者确诊为梅毒,患病率为70.93%(61/86)。TRUST检查的阳性率为59.30%(51/86),阴性率为40.70%(35/86);TP-ELISA检查的阳性率为67.44%(58/86),阳性率为32.56%(28/86)。TP-ELISA的敏感度为93.44%(57/61),高于TRUST检查的80.33%(49/61),差异有统计学意义(χ^(2)=4.604,P=0.032);且TP-ELISA准确度为94.19%(81/86),高于TRUST的83.72%(72/86),差异有统计学意义(χ^(2)=4.793,P=0.029)。结论 在梅毒诊断中,TP-ELISA的敏感度、准确度高于TRUST,但筛查过程仍存在假阴性风险,临床应综合考虑患者的危险因素筛查结果与其他临床指标。

响应面优化提取海蒿子多酚及其抗氧化和抗糖尿病活性分析

海蒿子是一种可药食两用的褐藻,具有多种功能活性。该研究采用响应面法优化了海蒿子多酚超声辅助提取的工艺,并测定了海蒿子多酚提取物及其萃取组分的总酚含量、抗氧化和抗糖尿病活性。结果表明,响应面优化超声辅助提取的最佳条件为乙醇体积分数8.5%,提取时间55 min,提取温度42℃,超声功率494 W,海蒿子多酚的含量为1.81 mg GAE/g DM。海蒿子粗提物正丁醇相的总酚含量最高,为24.45 mg GAE/g E。生物活性研究表明,二氯甲烷相的DPPH自由基清除能力(49.34μmol TE/g)和ABTS阳离子自由基清除能力(IC_(50)=249.22μg/mL)最强。乙酸乙酯和二氯甲烷相的α-葡萄糖苷酶抑制活性明显高于阿卡波糖,为其9.61和10.33倍。同时,乙酸乙酯相有最强的DPP-Ⅳ酶抑制活性(IC_(50)=25.83μg/mL),该溶剂能很好地富集海蒿子中的抗糖尿病成分。相关性分析表明,多酚是海蒿子中主要的抗氧化剂,α-葡萄糖苷酶和DPP-Ⅳ酶抑制活性呈高度相关。该研究发现海蒿子多酚作为多靶点抗糖尿病药物或健康食品的潜力很大,可促进海蒿子高附加值产品的开发。