高血压微血管反应性变化及粉防己碱的影响

目的观察肾血管性高血压大鼠（ＲＨＲ）微血管反应性变化及粉防己碱的影响。方法用冷热刺激诱导大鼠耳廓微血管舒缩,用血流量变化与恢复时间评价微血管舒缩反应能力。结果ＲＨＲ热刺激引起的微血管舒张反应降低,冷刺激导致的微血管收缩反应增强,去除刺激后血流的完全恢复延迟,伴血清ＡＣＥ活性升高和ＮＯ含量降低。粉防己碱可有效改善上述变化。结论ＲＨＲ微血管对缩舒血管刺激的反应性异常,粉防己碱具有良好的改善微血管反应性作用,其机制与维护内皮功能的完整性有关。     

不同剂量氟康唑治疗尿路念珠菌感染疗效比较

目的:比较不同剂量氟康唑治疗尿路念珠菌感染临床疗效.方法:选取2019年6月至2020年6月洛阳市第一人民医院收治的93例为尿路念珠菌感染患者研究对象,随机分为A、B、C三组各31例,均采用氟康唑治疗,A组使用剂量为400 mg?d-1,B组使用剂量为300 mg?d-1,C组使用剂量为200 mg?d-1进行治疗.治疗1周后比较三组临床疗效,对比治疗前后三组患者尿液标本培养念珠菌菌落计数,统计治疗1周内三组药物不良反应发生情况.结果:治疗1周后,A组总有效率明显高于C组(P<0.05);A组尿液标本培养念珠菌菌落计数明显低于B、C组(P<0.05);A组尿液氟康唑浓度明显高于B、C组(P<0.05);C组药物不良反应发生率明显低于A、B组(P<0.05).结论:400 mg?d-1氟康唑治疗尿路念珠菌感染临床疗效较高,但其药物安全性相对较低,需结合临床实际给药.     

粉防己碱诱导人红白血病细胞凋亡的研究

目的　探讨粉防己碱诱导人红白血病细胞 (K56 2 )凋亡的作用。　方法　采用光镜、电镜、免疫荧光观察K56 2 细胞形态的改变 ,以流式细胞仪分析细胞周期 ,以ABC法检测细胞中Bcl 2和p5 3蛋白的改变 ,以TUNEL法检测细胞凋亡。　结果　K56 2 细胞经粉防己碱诱导 4 8h后 ,出现细胞凋亡早期形态学改变 ;流式细胞仪显示细胞DNA合成受到抑制 ;ABC法检测出细胞中Bcl 2水平降低和p5 3水平升高 ;TUNEL法原位检测揭示有DNA断裂。结论　粉防己碱可抑制K56 2 细胞的生长 ,其作用与浓度相关 ,最终可导致细胞凋亡     

氟康唑序贯疗法治疗老年人肺部白色念珠菌感染的临床评价

目的评价氟康唑静脉滴注后改口服的序贯疗法治疗老年人肺部白色念珠菌感染的疗效及不良反应。方法用氟康唑序贯疗法治疗老年人肺部白色念珠菌感染60例,第1d 400mg静脉滴注,第2d改为200mg静脉滴注,每日1次,连续用7～10d后改口服,200mg／d,每天1次。疗程3～4周。结果总有效率为90．0％,白色念珠菌清除率为850％,不良反应发生率为8．3％。结论氟康唑序贯疗法疗效高,不良反应少,用药方便,尤适用于老年人。     

粉防己碱对心肌顿抑损伤的保护作用

用兔冠脉前降支结扎缺血再灌模型,观察粉防己碱（Ｔｅｔ）对心肌顿抑损伤的保护作用并分析其作用机理。结果示：①顿抑心肌收缩舒张功能明显降低,心电图ＳＴ段抬高,血清ＬＤＨ水平升高,左室湿重增大,Ｔｅｔ可减轻缺血及再灌后心肌功能的损害,降低血清ＬＤＨ水平与心肌湿重;②顿抑组血清ＮＯ水平略升高,Ｔｅｔ组血清ＮＯ无明显变化;③顿抑组心肌Ｎａ＾＋－ｋ＾＋－ＡＴＰ酶及Ｃａ＾２＋－ＡＴＰ酶活力均明显下降,Ｔｅｔ组Ｔ     

粉防己碱拮抗豚鼠水杨酸急性耳肾损伤的实验研究

目的 :探讨粉防己碱 (Tetrandrine ,Tet)在水杨酸钠 (主要成分Salicylateacid ,SA)急性耳肾损伤中的拮抗作用及其保护机制。方法 :将两耳阈值均≤ 5dB红色白目豚鼠分为SA组和Tet+SA组 ,于给药后 1h、2h、4h分别测定ABR阈值后留取尿标本和肾组织 ,测定尿N -乙酰 - β -D -氨基葡萄糖苷酶 (NAG)活性 ,用光镜、电镜观察肾组织学改变 ,用免疫组化方法测定肾组织肌动蛋白 (Actin)和转化生长因子 β1(TGFβ1)蛋白表达水平的变化 ,用原位末端标记法检测肾脏细胞凋亡情况。结果 :用药前SA +Tet组和SA组间ABR阈值差异无显著意义 (P >0 …     

粉防己碱对急性胰腺炎大鼠血清灌注的离体大鼠胃的保护作用及其机制

目的:研究急性胰腺炎(AP)大鼠血清对带血管灌流的离体大鼠胃内、外分泌功能的影响以及粉防己碱(Tet)的干预作用及其机制。方法:采用大鼠胆胰管逆行注射3%去氧胆酸钠复制AP模型,造模后5 h取动物血清。制备带血管灌流的离体大鼠胃, 经胃动脉输注AP血清, 并用Tet干预。从门静脉收集流出液,检测其中胃泌素、生长抑素、白细胞介素6(IL-6)及细胞因子诱导的中性粒细胞趋化因子1(CINC-1)水平的变化。同时从食管下段经贲门插管灌注生理盐水,从幽门收集流出液检测其中胃蛋白酶及胃酸分泌的变化。结果:AP大鼠胰腺组织破坏严重,胃壁出现多处糜烂、出血和溃疡,血清淀粉酶活性明显升高。用此AP血清灌流可引起离体胃胃泌素分泌增多,生长抑素分泌减少,IL-6和CINC-1分泌也增加,同时胃腔流出液中胃酸和胃蛋白酶含量增加。Tet可一定程度逆转AP血清刺激引起的胃泌素、胃酸和胃蛋白酶分泌的增多以及生长抑素分泌的抑制, 减少炎症介质的释放(P<0.05),减轻胃黏膜损伤。结论:急性胰腺炎相关的胃损害与炎症血清有害成分的刺激有关。Tet可干预AP血清刺激所致的离体大鼠胃内、外分泌功能的改变,从而发挥对胃的保护作用。     

粉防己碱对离体狗隐静脉的收缩作用

粉防己碱对狗隐静脉（ＤＳＶ）的去极化收缩有抑制作用。但在抑制后冲洗时却引起明显收缩。α２－肾上腺素能受体拮抗剂Ｒａｕｗｏｓｃｉｎｅ１０－７ｍｏｌ及ＫＣｌ４０ｍｍｏｌ／Ｌ能抑制冲洗波的出现；单独使用Ｔｅｔ也能诱发ＤＳＶ缓慢的张力增加，而ＫＣｌ２０ｍｍｏｌ／Ｌ既能加速Ｔｅｔ诱发收缩的出现也能增强α２－肾上腺素能受体激动剂Ｂ－ＨＴ９２０的缩血管效应。对ＢａｙＫ８６４４诱发的ＤＳＶ收缩。Ｔｅｔ既不能抑制也无冲洗波出现。提示在低Ｋ＋条件下Ｔｅｔ能激活α２－肾上腺素能受体而引起收缩，在高Ｋ＋存在时激活Ｎａ－ＫＡＴＰ酶导致去极化收缩的舒张。     

特比萘芬与氟康唑或伊曲康唑联合对氟康唑诱导耐药稳定白念珠菌的影响

目的探讨特比萘芬与氟康唑或伊曲康唑联合抗氟康唑诱导产生的耐药稳定白念珠菌的作用.方法采用多步诱导法,在YEPD培养基中,利用氟康唑诱导白念珠菌敏感株产生耐药稳定菌株.根据美国国家临床实验标准委员会（NCCLS）制定的标准,采用棋盘微量稀释法测定特比萘芬与氟康唑或伊曲康唑对耐药稳定菌株的联合药敏试验,并对诱导耐药稳定菌株ERG11基因的编码区序列进行DNA测序.结果临床敏感菌株和标准敏感菌株能被诱导形成耐药菌株,但大部分不稳定,诱导耐药稳定株ERG11基因的编码区DNA测序有突变点存在,特比萘芬与氟康唑或伊曲康唑联用对诱导耐药稳定株可产生协同作用.结论特比萘芬与氟康唑或伊曲康唑联合应用对基因突变产生的耐药株有协同作用,可阻止或延迟氟康唑诱导的耐药性白念珠菌菌株的产生.     

Candida tropicalis in human disease

Candida tropicalis is one of the more common Candida causing human disease in tropical countries; the frequency of invasive disease varies by geography causing 3–66% of candidaemia. C. tropicalis is taxonomically close to C. albicans and shares many pathogenic traits. C. tropicalis is particularly virulent in neutropenic hosts commonly with hematogenous seeding to peripheral organs. For candidaemia and invasive candidiasis amphotericin B or an echinocandin are recommended as first-line treatment, with extended-spectrum triazoles acceptable alternatives. Primary fluconazole resistance is uncommon but may be induced on exposure. Physicians in regions where C. tropicalis is common need to be mindful of this lesser-described pathogen.     

DNA fingerprinting elicited evolutionary trend of oral Candida tropicalis isolates from diverse geographic locales

Purpose: To study molecular profiles of oral Candida tropicalis isolates from five different geographic locales to determine the molecular diversity, clonality and evolutionary trends of this opportunistic pathogen. Methods: A total of 36 strains from five countries (China, Canada, Scotland, Japan and Tanzania) were genotyped by PCR fingerprinting with 11 separate primers. Of these, primers RSG9, RSG8, T3B and RSD12 generated complex fingerprinting patterns. Results: Three significantly dissimilar profiles were derived from the primer T3B and particularly focused on tDNA suggested the prevalence of genetic subtypes within the species. Comparison of tDNA and rDNA (RSD12) fingerprints of C. tropicalis suggested that rDNA is much more heterogeneous than the relatively distinct tDNA. Further analysis of similarity coefficient (S_AB) of gel profiles derived from computer-generated dendrograms indicated some degree of similarity in isolates from five-disparate geographic locales as well as the presence of unique isotypes in each region. Conclusions: This study demonstrates the evolutionary divergence of distinct genetic subgroups within Candida tropicalis.     

Candida tropicalis fungaemia: incidence,risk factors and mortality in a general hospital

The risk factors and clinical features of patients with Candida tropicalis fungaemia have not been fully defined. We performed a case–control study comparing 59 cases of C. tropicalis fungaemia with 177 episodes of fungaemia caused by other species of Candida in our hospital over a 24-year period (January 1985 to December 2008). Patients with C. tropicalis fungaemia were more likely to be older (median age, 67 vs. 56 years; p 0.01), to have cancer (45.5% vs. 31.6%, p 0.04), and to have the abdomen as the portal of entry (32.2% vs. 11.9%, p 0.001), and had a higher in-hospital mortality rate (61% vs. 44%, p 0.03). Multivariate analysis showed that the independent risk factors for C. tropicalis fungaemia were cancer (OR 4.5; 95% CI 1.05–3.83; p 0.03) and the abdomen as the portal of entry (OR 13.6; 95% CI 1.9–8.2; p <0.001). When survivors were compared with non-survivors, the risk factors associated with a poor outcome were neutropenia (19.4% vs. 0; p 0.03), corticosteroid treatment (36% vs. 13%; p 0.07), and septic shock (50% vs. 17.4%; p 0.01). The independent risk factors for mortality in the multivariate analysis were corticosteroid treatment (OR 8.2; 95% CI 0.9–27.7; p 0.04) and septic shock (OR 14.6; 95% CI 2.4–90.2; p 0.004), whereas urinary tract infection (OR 0.07; 95% CI 0.01–0.8; p 0.03) and catheter removal (OR 0.06; 95% CI 0.01–0.4; p 0.002) were protective factors. C. tropicalis is the fourth most common cause of fungaemia in our hospital. It is associated with underlying malignancy, the abdomen as the portal of entry, and poor outcome.     

Inactivation of the Complement Lectin Pathway by Candida tropicalis Secreted Aspartyl Protease-1

Candida tropicalis is an opportunistic fungal pathogen and is one of the most frequently isolated non-albicans species. It can cause localised as well as invasive systemic infections particularly in immunocompromised patients. Increased resistance to common anti-fungal drugs is an emerging problem. In order to establish disseminated infections, Candida has evolved several strategies to escape the host immune system. A detailed understanding of how C. tropicalis escapes the host immune attack is needed as it can help develop novel anti-fungal therapies. Secreted aspartyl proteinases (Saps) of C. albicans have been shown to be determinants of virulence and immune evasion. However, the immune evasion properties of C. tropicalis Saps have been poorly characterised. This study investigated the immune evasion properties of C. tropicalis secreted aspartic protease 1 (Sapt1). Sapt1 was recombinantly produced using a Kluyveromyces lactis yeast expression system. A range of complement proteins and immunogloublins were screened to test if Sapt1 had any proteolytic activity. Sapt1 efficiently cleaved human mannose-binding lectin (MBL) and collectin-11, which are the initiating molecules of the lectin pathway of the complement system, but not l-ficolin. In addition, Sapt1 cleaved DC-SIGN, the receptor on antigen presenting dendritic cells. Proteolysis was prominent in acidic condition (pH 5.2), a characteristic of aspartyl protease. No proteolytic activity was detected against complement proteins C1q, C3, C3b, IgG and IgA. In view of the ability of Sapt1 to cleave MBL and collectin-11, we found that Sapt1 could prevent activation of the complement lectin pathway. RT-qPCR analysis using three different C. tropicalis clinical isolates (oral, blood and peritoneal dialysis fluid) revealed relatively higher levels of mRNA expression of Sapt1 gene when compared to a reference strain; Sapt1 protein was found to be secreted by all the tested strains. Lectin pathway and its initiating components are crucial to provide front line defence against Candida infections. For the first time, we have shown that a Candida protease can proteolytically degrade the key initiating components of lectin pathway and inhibit complement activation. Findings from this study highlight the importance of exploring Sapt1 as a potential therapeutic target. We conclude that C. tropicalis secretes Sapt1 to target the complement lectin pathway, a key pattern recognition and clearance mechanism, for its survival and pathogenesis.     

Candida spp. other than Candida albicans: a major cause of fungaemia in a tertiary care centre

This study was conducted to determine the frequency of different Candida spp. isolated from different parts of the hospital, associated risk factors and mortality rate. A total of 59 cases were selected for prospective analysis over a period of one and half years. Blood samples collected were processed by BACTEC (9240) method. Candidaemia was diagnosed by positive blood culture at least from two blood culture samples or from a clinically significant single blood culture sample. Candida spp. were identified by standard techniques. Most frequent isolates were C. tropicalis (35.6%), C. parapsilosis (28.8%), C. glabrata (11.9%) and C. pelliculosa (11.9%). Candida albicans was isolated only in 3.4% cases. Neonatology department accounted for highest number of isolates (27.1%), followed by gastrointestinal surgery (15.3%) and cardiac surgery (13.6%). Mortality was noted in 16.9%. Probable risk factors determined were intensive care unit stay (74.6%), antibiotic therapy (50.8%), central line (42.4%), urinary catheter (32.2%), ventilator (23.7%), malignancy (20.3%) and abdominal surgery (15.3%).     

Interactions of Candida tropicalis pH-related antigen 1 with complement proteins C3, C3b,factor-H,C4BP and complement evasion

Candida, as a part of the human microbiota, can cause opportunistic infections that are either localised or systemic candidiasis. Emerging resistance to the standard antifungal drugs is associated with increased mortality rate due to invasive Candida infections, particularly in immunocompromised patients. While there are several species of Candida, an increasing number of Candida tropicalis isolates have been recently reported from patients with invasive candidiasis or inflammatory bowel diseases. In order to establish infections, C. tropicalis has to adopt several strategies to escape the host immune attack. Understanding the immune evasion strategies is of great importance as these can be exploited as novel therapeutic targets. C. albicans pH-related antigen 1 (CaPra1), a surface bound and secretory protein, has been found to interact strongly with the immune system and help in complement evasion. However, the role of C. tropicalis Pra1 (CtPra1) and its interaction with the complement is not studied yet. Thus, we characterised how pH-related antigen 1 of C. tropicalis (CtPra1) interacts with some of the key complement proteins of the innate immune system. CtPra1 was recombinantly produced using a Kluyveromyces lactis yeast expression system. Recombinant CtPra1, was found to bind human C3 and C3b, central molecules of the complement pathways that are important components of the innate immune system. It was also found to bind human complement regulatory proteins factor-H and C4b-binding protein (C4BP). CtPra1-factor-H and CtPra1-C4BP interactions were found to be ionic in nature as the binding intensity affected by high sodium chloride concentrations. CtPra1 inhibited functional complement activation with different effects on classical (～20 %), lectin (～25 %) and alternative (～30 %) pathways. qPCR experiments using C. tropicalis clinical isolates (oral, blood and peritoneal fluid) revealed relatively higher levels of expression of CtPra1 gene when compared to the reference strain. Native CtPra1 was found to be expressed both as membrane-bound and secretory forms in the clinical isolates. Thus, C. tropicalis appears to be a master of immune evasion by using Pra1 protein. Further investigation using in-vivo models will help ascertain if these proteins can be novel therapeutic targets.     

热带假丝酵母菌实时荧光定量PCR检测方法的建立及临床应用初步评价

目的建立、优化快速检测临床标本中热带假丝酵母菌的实时荧光定量PCR方法 ,并对其临床应用进行初步评价。方法用自行设计的高效引物、探针检测5种临床标本中的热带假丝酵母菌,对该方法的敏感性、特异性进行评价,并与真菌培养法进行比较。结果检测临床标本中热带假丝酵母菌的灵敏度可达10 copies/ml,与人类基因组、细菌及其他真菌无交叉阳性反应。与真菌培养法的检测一致性好（Kappa值为0.916,P〈0.01）。结论实时荧光定量PCR检测热带假丝酵母菌灵敏度高、特异性强,可直接用于各种临床标本中热带假丝酵母菌的检测,而且可大大缩短报告时间,为临床诊断提供可靠依据。     

Candida tropicalis biofilms: effect on urinary epithelial cells

Candida tropicalis infection is strongly associated with the presence of biofilms in urinary catheters. Thus, the aim of this work was to study the behaviour of C. tropicalis in biofilms of different ages (24-120 h) formed in artificial urine (AU) and their effect in human urinary bladder cells (TCC-SUP). Reference strain ATCC 750 and two isolates from patients with candiduria (U69 and U75) were used in this study. The adhesion to human cells was evaluated after 2 h of contact with Candida biofilms, using the Crystal violet staining method, and the human cells response was evaluated in terms of activity inhibition and cell damage. Candida tropicalis aspartyl proteinase (SAPT) gene expression was determined by real-time PCR. Candida tropicalis biofilm cells were able to adhere to TCC-SUP cells. The highest extent of yeast attachment was obtained for the 72 h old biofilm cells. Yeasts affected TCC-SUP cells, with 120 h-biofilm cells causing the highest levels of cell injury. Generally, SAPT3 was highly expressed and SAPT4 was only detected in the reference strain. Overall, it is important to highlight that C. tropicalis cells detached from biofilms are able to colonize human cells and cause some injury and reduction of metabolic activity.     

药物流出泵抑制剂联合咪康唑清除白假丝酵母菌生物被膜的研究

目的 观察咪康唑分别与两种药物流出泵抑制剂联用清除耐药株(persister)的效果.方法 白假丝酵母菌参考株YEM30,在96孔板中形成生物被膜(biofilm),CDR1抑制剂Enniatin B、CDR1/CDR2抑制剂Milbemycins α25单独或联合与咪康唑作用后,采用菌落形成单位(CFU)计数的方法统计耐药株的数量.采用SAS8.0统计软件包对数据进行q检验.结果 咪康唑分别联合两种药物流出泵抑制剂清除生物被膜耐药株的效果明显优于咪康唑单独使用(P<0.001),其中咪康唑与Enniatin B联用效果更佳.结论 咪康唑与药物流出泵抑制剂联合应用具有清除生物被膜中表现型耐药株的作用,这为提高抗真菌治疗的效果提供了一条新途径和新思路.