细菌性阴道病肠道菌群与阴道菌群16S rDNA测序分析

目的通过对细菌性阴道病(BV)患者肠道菌群及阴道菌群16S rDNA扩增子测序,分析其结构、多样性、相关性以及BV对肠道菌群的影响,为今后治疗BV提供新的思路。方法选取符合纳入标准的BV患者11例(BV组),健康者9例(C组),留存阴道分泌物及新鲜粪便进行16S rDNA基因检测分析。结果C组阴道菌群以乳杆菌属为主,BV与加德纳菌属、普雷沃菌属、Sneathia、窄食单胞菌属(Stenotrophomonas)、阿托波菌属、Shuttleworthia、巨型球菌属密切相关。BV组肠道、阴道菌群丰富度均高于C组。Alpha多样性分析中C组和BV组肠道菌群、阴道菌群的Shannon指数组间比较,χ²值为29.137, P=0.000<0.05,两组阴道菌群Shannon指数组间比较差异具有统计学意义(P<0.05),BV组高于C组。物种多样性曲线反映本研究样本测序数据量的合理性,表明BV组的肠道菌群多样性、丰富度均高于C组,主坐标分析表明C组肠道与阴道的菌群结构差距较大,BV组肠道与阴道的菌群结构有相似之处,且两组肠道菌群结构接近。BV组阴道菌群中厚壁菌门丰度较C组低,放线菌门、拟杆菌门较C组高;BV组肠道菌群中拟杆菌门丰度较C组低;C组肠道中拟杆菌门明显高于阴道,厚壁菌门明显低于阴道; BV组阴道菌群中放线菌门丰度高于C组,差异均具有统计学意义(P<0.05)。结论BV阴道菌群与肠道菌群具有相关性,BV可能引起肠道菌群结构比例和多样性的改变。     

中国汉族育龄女性阴道内乳杆菌的鉴定与系统发育分析

目的比较基质辅助激光解吸电离飞行时间质谱(MALDITOF MS)与16S rRNA基因测序鉴定女性阴道乳杆菌方法,同时探究不同阴道微生态背景的汉族育龄女性阴道内乳杆菌的种类及其分布情况。方法对260例汉族育龄女性阴道拭子进行阴道微生态分析及乳杆菌的分离培养,根据阴道微生态鉴定结果分为正常组(173例)、中间型细菌性阴道病组(IBV,55例)、细菌性阴道病组(BV,9例)和外阴阴道炎假丝酵母菌病组(VVC,23例),分离乳杆菌并进行纯化培养,纯化的乳杆菌经MALDITOF MS和16S rRNA基因测序鉴定并进行聚类分析。结果2种方法鉴定的一致率为95.3%。260例标本中149例(57.3%)检出了乳杆菌,共发现17种乳杆菌,检出的乳杆菌分布于5个乳杆菌系统发育群,其中分离出较多的乳杆菌为卷曲乳杆菌(47例)、加氏乳杆菌(38例)、阴道乳杆菌(21例)。正常组116例(67.1%)标本分离出乳杆菌,IBV组19例(34.5%),BV组2例(22.2%),VVC组12例(52.2%)。正常育龄女性与患BV女性乳杆菌种类与数量差异有统计学意义(χ²=22.936 0,P<0.000 1),与患VVC女性差异无统计学意义(χ²=1.983 0,P=0.160 0)。与其他组相比,正常组育龄女性中分离出卷曲乳杆菌的比例显著高于BV组和VVC组(χ²=4.548 0,P=0.033 0;χ²=5.605 0,P=0.020 0),VVC组育龄女性分离出加氏乳杆菌的比例显著高于正常组(χ²=5.090 0,P=0.035 0)。结论对比16S rRNA基因测序与MALDITOF MS方法的乳杆菌鉴定结果,除了阴道乳杆菌和人阴道乳杆菌出现不一致的鉴定结果,其余15种乳杆菌鉴定结果一致性均为100%,但基于测序与基于质谱的方法产生的乳杆菌聚类分析结果不完全一致。卷曲乳杆菌可能是反映育龄女性正常阴道微生态的重要标志物,加氏乳杆菌可能与女性VVC的发生有关。     

宫颈癌患者阴道微生态变化及其与细胞免疫的相关性探讨

目的探讨宫颈癌患者阴道微生态变化及其与细胞免疫的相关性。方法回顾性分析2018年1月-2020年5月本院收治的且经病理确诊的73例宫颈癌患者(宫颈癌组)和同期65例体检筛查宫颈正常者(正常宫颈组)的临床病例资料。两组均采用直接涂片法、革兰染色法检测阴道分泌物中乳酸杆菌、滴虫、支原体、衣原体、细菌性阴道病、霉菌、淋菌、假丝酵母菌、人乳头瘤病毒(HPV)及阴道唾液酸甘酶活性、白细胞酯酶、过氧化氢等。另采用流式细胞仪检测外周血T淋巴细胞(CD3⁺、CD4⁺、CD8⁺、CD4⁺/CD8⁺)及自然杀伤细胞(CD16⁺CD56⁺)水平。观察各组阴道微生态变化情况及外周血CD3⁺、CD4⁺、CD8⁺、CD4⁺/CD8⁺、CD16⁺CD56⁺水平。另对比宫颈癌组中感染致病微生物者、阴道微生态失调者与未感染致病微生物者、阴道微环境正常者的CD3⁺、CD4⁺、CD8⁺、CD4⁺/CD8⁺、CD16⁺CD56⁺水平,采用Pearson相关性分析法分析宫颈癌组阴道乳酸杆菌定植密度与细胞免疫的相关性。结果宫颈癌组滴虫、支原体、衣原体、霉菌、细菌性阴道病、HPV感染检出率及阴道微生态失调率均高于正常宫颈组,宫颈癌组乳酸杆菌定植密度低于正常宫颈组,差异均有统计学意义(P<0.05),宫颈癌组淋菌、假丝酵母菌感染检出率与正常宫颈组比较差异均无统计学意义(P>0.05)；宫颈癌组CD3⁺、CD4⁺、CD4⁺/CD8⁺、CD16⁺CD56⁺水平均低于正常宫颈组,CD8⁺水平高于正常宫颈组,差异均有统计学意义(P<0.05)；宫颈癌组中滴虫、支原体、衣原体、霉菌、细菌性阴道病、HPV感染者及阴道微生态失调者CD3⁺、CD4⁺、CD4⁺/CD8⁺、CD16⁺CD56⁺均低于未感染者、阴道微环境正常者,CD8⁺高于未感染者、阴道微环境正常者,差异均有统计学意义(P<0.05)；宫颈癌患者乳酸杆菌定植密度与CD3⁺、CD4⁺、CD4⁺/CD8⁺、CD16⁺CD56⁺呈正相关性(P<0.05),与CD8⁺呈负相关性(P<0.05)。结论宫颈癌患者阴道致病微生物感染率较健康人群高,同时细胞免疫功能降低,增加罹患宫颈癌的风险。     

重组人干扰素α2b凝胶联合乳酸菌阴道胶囊对宫颈高危型HPV持续感染的治疗

目的观察重组人干扰素α2b凝胶联合乳酸菌阴道胶囊治疗宫颈高危型人乳头瘤病毒(highrisk human papillomavirus,HPV)持续感染的临床效果。方法将110例宫颈高危型HPV持续感染而液基细胞学检查阴性的患者随机分为两组,其中对照组(58例)予以重组人干扰素α2b凝胶治疗,观察组(52例)予以重组人干扰素α2b凝胶联合乳酸菌阴道胶囊治疗,两组患者均连续治疗3个月经周期。比较两组患者HPV转阴率及阴道微生态改善情况。结果观察组患者治疗总有效率(82.69%)高于对照组(63.79%),差异有统计学意义(χ²=4.937,P=0.026)。治疗后,两组患者阴道微生态失调率均有所下降,且观察组改善情况更为显著(均P<0.05)。治疗后观察组患者阴道菌群密集度及多样性显著低于治疗前,其微生物功能指标优于对照组,乳杆菌定植率高于对照组,病原菌检出率低于对照组(均P<0.05)。结论在重组人干扰素α2b凝胶治疗的基础上联合应用乳酸菌阴道胶囊能明显提高宫颈高危型HPV持续感染的转阴率,其中促进阴道微生态恢复平衡可能是其发挥作用的机制之一。     

不同干预时机下复方嗜酸乳杆菌片联合四联疗法在根除幽门螺杆菌失败补救治疗中的应用

目的探讨不同干预时机下复方嗜酸乳杆菌片联合四联疗法在首次根除幽门螺杆菌(H. pylori)失败患者补救治疗中的作用。方法选择西安市第一医院消化内科90例经标准四联疗法根除H. pylori失败的患者为研究对象,随机分为研究组A(A组)、研究组B(B组)与对照组(C组),各30例。A组患者使用复方嗜酸乳杆菌片联合补救四联疗法治疗。B组患者在补救四联疗法结束后序贯使用复方嗜酸乳杆菌片2周。C组患者单独采用补救四联疗法治疗。比较3组患者临床症状、不良反应发生率及H. pylori根除率。结果A组、B组、C组患者总有效率分别为93.33%(28/30),90.00%(27/30),66.67%(20/30),差异有统计学意义(χ²=9.120,P=0.010)。A组、B组、C组患者总不良反应发生率分别为13.33%(4/30)、16.67%(5/30)、66.67%(20/30),差异有统计学意义(χ²=24.522,P<0.001)。A组、B组、C组患者H. pylori根除率分别为60.00%(18/30)、63.33%(19/30)、33.33%(10/30),差异有统计学意义(χ² =6.502,P=0.039)。A组与B组患者H. pylori根除率相比差异无统计学意义(χ²=0.071,P=0.791)。结论复方嗜酸乳杆菌片联合四联疗法可提高首次四联疗法根除H. pylori失败患者H. pylori根除率,改善患者临床症状,降低总体不良反应发生率,但不同干预时机下复方嗜酸乳杆菌片对H. pylori根除率无明显影响。     

荧光染色法和传统KOH湿片法在甲真菌病诊断中的应用比较

目的探讨真菌荧光染色法与传统KOH湿片法在临床甲真菌镜检中的应用效果。方法选择2017年6月至2018年4月我院600例皮肤科门诊就诊疑似真菌感染患者指(趾)甲镜检标本,分别运用荧光染色法和KOH湿片法对这些标本进行检测,记录实验数据,并对检测结果进行比较。结果荧光染色法检测阳性为418例,阳性率为70%,其中指甲阳性130例,趾甲阳性288例。KOH湿片法检测阳性为354例,阳性率为59%,其中指甲97例,趾甲257例。指甲(χ²=8.816,P=0.003)、趾甲(χ²=7.884,P=0.005)及总阳性率(χ²=14.876,P<0.001)在两种方法中差异均有统计学意义。结论在对指(趾)甲真菌感染的诊断中,荧光染色法的阳性检出率明显高于KOH湿片法,是一种快速、准确的真菌镜检方法,值得临床推广使用。     

益生菌制剂对轮状病毒性肠炎患儿肠道菌群和免疫功能的影响

目的观察益生菌制剂辅助治疗儿童轮状病毒性肠炎的临床疗效及对患儿肠道菌群和免疫功能的影响。方法以160例轮状病毒性肠炎患儿为研究对象,随机分为对照组(n=80)和研究组(n=80)。对照组患儿给予消旋卡多曲颗粒联合蒙脱石散治疗,研究组患儿在此基础上联合双歧杆菌乳杆菌三联活菌片治疗。观察患儿治疗期间药物不良反应,患儿腹泻停止时间、发热消退时间和呕吐停止时间。分别于治疗前后检测患儿血清免疫球蛋白水平和肠道菌群状况,评价患儿治疗效果。结果研究组患儿腹泻停止时间[(2.41±0.70)d]、发热消退时间[(1.71±0.52)d]和呕吐停止时间[(2.09±0.62)d]均显著短于对照组(均P<0.05)。研究组患儿轮状病毒性肠炎总体治疗有效率显著高于对照组(95.00% vs 83.75%,χ²=5.331,P=0.021)。研究组患儿治疗后血清IgG和IgM水平[(5.83±0.59)g/L和(1.07±0.16)g/L]显著高于对照组(均P<0.05)。研究组患儿治疗后肠道菌群正常者比例显著高于对照组(92.50% vs 76.25%,χ²=8.012,P=0.005)。两组患儿恶心、便秘、皮疹和嗜睡等药物不良反应总体发生率差异无统计学意义(10.00% vs 8.75%,χ²=0.074,P=0.786)。结论益生菌制剂可有效缩短轮状病毒性肠炎患儿临床症状改善周期,提高疗效,改善患儿免疫功能,纠正肠道菌群失调状况,且不增加药物不良反应,值得临床推广使用。     

利用LightCycler研究食管鳞状细胞癌与MTHFR C677T多态性的关系

亚甲基四氢叶酸还原酶(MTHFR)在叶酸代谢中起重要作用.MTHFR基因第677位核苷酸的多态性（C→T）能影响其酶活性并与肿瘤易感性有关.为比较中国北方人群MTHFR C677T多态性与食管鳞状细胞癌（ESCC）易感性之间的关系, 通过高速实时聚合酶链反应（real-time PCR）和解链曲线（melting curve）方法分析了189名ESCC患者和141名健康对照的MTHFR C677T多态性位点的基因型.结果显示,健康对照组的MTHFR C/C（纯合野生）、C/T和T/T（纯合突变）基因型的频率分别为17.7%、38.3%和44.0%.ESCC患者的T/T基因型频率（42.3%）与健康对照组无显著差异（χ²=0.089, P＞0.05）,其C/T基因型频率（49.2%）仅略高于对照组（χ²＝3.890, P＜0.05）,而患者组的C/C基因型频率(8.5%)显著低于健康对照组(17.7%) (χ²=6.37, P=0.012).与C/T和T/T基因型相比,C/C基因型可显著降低ESCC的发病风险(相对风险度的比值比（OR）=0.43, 95%可信区间（CI）=0.22～0.84),在吸烟者和有上消化道肿瘤家族史的患者中这一倾向更加明显.研究提示,在中国北方人群中, MTHFR C677T纯合野生基因型对ESCC的发病起保护作用.     

香菇多糖对老年进展期肺癌患者同步放化疗效果及呼吸道菌群的影响

目的探讨香菇多糖对老年进展期肺癌患者同步放化疗治疗效果及呼吸道菌群的影响,为后续研究提供参考。方法选取2016年6月至2018年10月我院收治的86例老年进展期肺癌患者为研究对象,将其随机分为研究组与常规组,各43例。常规组患者给予基础治疗联合同步放化疗,研究组在常规组基础上给予香菇多糖治疗。比较两组患者治疗后细胞免疫功能指标水平变化,呼吸道菌群结构变化,患者疗效及治疗期间不良反应发生情况。结果治疗后研究组患者CD3⁺细胞、CD4⁺细胞、CD4⁺/CD8⁺水平均提高,常规组水平均降低(均P<0.05)。治疗后两组患者CD3⁺细胞、CD4⁺细胞、CD4⁺/CD8⁺水平差异有统计学意义(均P<0.05)。治疗后两组患者CD8⁺细胞水平均降低,且研究组低于对照组(均P<0.05)。治疗后两组患者呼吸道中优势菌属丰度差异有统计学意义(均P<0.05)。两组患者近期疗效分布差异有统计学意义(P<0.05),其中研究组患者临床有效率为58.14%,高于常规组的34.88%(P<0.05)。两组患者药物不良反应发生率差异无统计学意义(P>0.05)。结论香菇多糖可显著提高老年进展期肺癌同步放化疗患者免疫能力,改善患者呼吸道菌群结构,提高治疗效果且安全性较高。     

双歧杆菌三联活菌散联合非营养性吸吮对喂养不耐受早产儿血清胃肠激素水平的影响

目的探讨双歧杆菌三联活菌散联合非营养性吸吮对喂养不耐受早产儿血清胃肠激素水平的影响。方法选取2017年1月至2019年9月于台州医院住院治疗的76例喂养不耐受早产儿为研究对象,随机分为联合组和单用组各38例。单用组早产儿入院后即予以非营养性吸吮,联合组早产儿在单用组基础上加用双歧杆菌三联活菌散溶于母乳中口服或鼻饲给药,0.5 g/次,2次/d。两组对象连用3周。观察两组早产儿治疗前后血清胃肠激素水平的变化,并比较治疗后患儿恢复情况、治疗效果及不良反应。结果治疗3周后,两组早产儿血清胃动素(MTL)和胃泌素(GAS)水平较治疗前明显上升,且治疗后联合组上升幅度大于单用组(均P<0.05)。联合组早产儿呕吐消失时间、腹胀消失时间、恢复至出生体质量时间及达全胃肠喂养时间均明显短于单用组(均P<0.05)。联合组早产儿临床总有效率(94.7%)显著高于单用组(78.9%),差异有统计学意义(χ²=4.15,P=0.044 2)。单用组和联合组分别出现不良反应1例(2.63%)和3例(7.89%),症状均较轻,两组差异无统计学意义(χ²=0.26,P>0.05)。结论双歧杆菌三联活菌散联合非营养性吸吮治疗喂养不耐受早产儿的效果较确切,安全性较高,能加快早产儿的恢复,其作用机制可能与其能促进MTL和GAS的分泌相关。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

Chemokines and cytokines: axis and allies in asthma and allergy

Allergic asthma can be precipitated by many factors. For the atopic person, fungus, pollen, dust mites, cockroach antigens, and diesel exhaust are all agents that may trigger an allergic attack. Cytokines and chemokines are integral mediators of fungal asthma. From the earliest time points, they recruit and activate the cells required for the clearance of fungus as well as being critical factors involved in the immunopathology of this disease. In the final analysis, it is clear that these mediators can act to the benefit or the detriment of the host.     

人血管能抑素基因的克隆、表达及其表达产物的纯化和生物活性测定

以人胎盘脐带组织为材料,提取组织总RNA,用netRTPCR方法合成人血管能抑素cDNA基因,将该cDNA克隆进pSP72载体获得重组质粒pSP72C, DNA序列分析结果与预期序列一致。用BamHⅠ和NdeⅠ双酶切,切下pSP72C上的血管能抑素cDNA,插入pET3c载体的相应位点获得重组表达质粒pETC, 转化E. coli BL21(DE3), SDSPAGE分析显示：在IPTG诱导下,血管能抑素基因获得了高效表达,表达量约占菌体总蛋白的 27.9 %,主要以包涵体形式存在。包涵体经过洗涤、裂解、蛋白复性以及Sephadex G75凝胶过滤层析等步骤后,获得了纯度达91.4 %的人血管能抑素。CAM实验证明10 μg纯化蛋白就能显著抑制鸡胚新生血管生成。     

Oxidation and nitration of ribonuclease and lysozyme by peroxynitrite and myeloperoxidase

In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier.