重症肺炎支原体肺炎肺泡灌洗液中TNF-α、IL-6、IL-8、IL-10水平观察

目的检测重症肺炎支原体肺炎患者支气管肺泡灌洗液（bronchial tube pul monary alveolus syringe fluid,BALF）中肿瘤坏死因子-α（Tumor necrosis factor-α,TNF-α）、白细胞介素-6（Interleukin-6,IL-6）、白细胞介素-8（In-terleukin-8,IL-8）白细胞介素-10（Interleukin-10,IL-10）浓度,探讨其临床意义。方法收住ICU的重症肺炎支原体肺炎患者36例,其CPIS评分〉6分为CPIS高分组共15例,CPIS评分〈6分为CPIS低分组共21例;正常对照组24例。取支气管肺泡灌洗液检测TNF-α、IL-6、IL-8、IL-10水平。结果重症肺炎支原体肺炎无论CPIS高分组和CPIS低分组支气管肺泡灌洗液中TNF-α、IL-6、IL-8、IL-10水平明显高于对照组,差异有统计学意义（P〈0.01）,CPIS高分组TNF-α、IL-6、IL-8水平高于CPIS低分组,差异有统计学意义（P〈0.01）,CPIS高分组IL-10水平稍高于CPIS低分组,差异无统计学意义（P〉0.05）。2组治疗后恢复期TNF-α、IL-6、IL-8、IL-10水平均明显下降（P〈0.01）。结论 TNF-α、IL-6、IL-8、IL-10在重症肺炎的发生发展中起重要作用,是重症肺炎支原体肺炎诊断的重要因素之一。利用支气管肺泡灌洗液检测TNF-α、IL-6、IL-8、IL-10水平变化对临床诊断治疗及预后有一定的临床价值。     

红霉素对解脲支原体肺炎的疗效及对细胞亚群和肝功能的影响

目的: 探讨红霉素对解脲支原体肺炎的疗效及对细胞亚群、肝功能的影响。方法: 收集我院2016年5月至2017年5月收治的41例解脲支原体肺炎患儿作为试验组,静脉滴注10 mg/kg红霉素与适量5%葡萄糖注射液,每天2次,疗程7～10 d,同期选取41例健康体检儿童,比较健康组与试验组治疗前后细胞亚群及肝功能,并分析试验组疗效及安全性。结果: 治疗前,试验组CD4+、CD4+/CD8+分别为（41.95±5.71）%、1.26±0.18,低于健康组的（48.13±7.21）%、1.79±0.20,试验组CD8+为（35.10±4.41）%,高于健康组的（26.56±3.13）%,试验组谷丙转氨酶、乳酸脱氢酶、总胆汁酸、胆红素、总胆固醇、甘油三酯、谷草转氨酶、谷丙转氨酶/谷草转氨酶分别为（136.71±17.20）U/L、（439.67±54.45）U/L、（28.39±3.72）U/L、（68.05±8.95）μmol/L、（7.82±0.95）mmol/L、（1.70±0.24）mmol/L、（113.37±14.25）U/L、1.24±0.14,高于健康组的（45.26±4.88）U/L、（397.98±47.20）U/L、（22.24±2.66）U/L、（36.71±4.83）μmol/L、（4.26±0.46）mmol/L、（1.24±0.17）mmol/L、（62.51±9.19）U/L、0.77±0.12;治疗后,试验组CD4+、CD4+/CD8+显著上升,CD8+、谷丙转氨酶、乳酸脱氢酶、总胆汁酸、胆红素、总胆固醇、甘油三酯、谷草转氨酶、谷丙转氨酶/谷草转氨酶显著下降,且两组比较无统计学差异（P>0.05）。结论: 红霉素对解脲支原体肺炎的疗效肯定,能够利于细胞亚群及肝功能的缓解。     

Detection of multiple Mycoplasma species in bulk tank milk samples using real-time PCR and conventional culture and comparison of test sensitivities

The objective of this study was to further validate a SYBR PCR protocol for Mycoplasma spp. by comparing it with standard microbial culture in the detection of Mycoplasma spp. in bulk tank milk samples. Additionally, we identified Mycoplasma spp. present by analysis of PCR-generated amplicons [dissociation (melt) temperature (T_m), length, and DNA sequence]. The research presented herein tests the hypothesis that the SYBR PCR protocol is as sensitive as conventional culture for the detection of Mycoplasma spp. in bulk tank milk samples. Mycoplasmas cause several important disease syndromes in cattle, including mastitis in dairy cows. The standard diagnostic method at the herd level has been microbial isolation of mycoplasmas on 1 of several specialized media and speciation through biochemical or immunological techniques; repeated sampling schemes are recommended. The development of a real-time SYBR PCR protocol offers advantages in decrease of time to detection, cost, and complexity. The T_m of the double-stranded DNA generated from the PCR reaction was used to detect the presence of and tentatively identify the species of mycoplasmas other than Mycoplasma bovis. In the SYBR PCR protocol, the presence of multiple species of mycoplasmas is indicated by an atypical dissociation curve. Gel electrophoresis and sequencing of the amplicons was used to confirm the mycoplasma species present when a non-M. bovis organism was detected (T_m not equal to M. bovis) and used to identify all the mycoplasma species present for the samples with atypical dissociation curves. Mycoplasma bovis was identified in 83% of SYBR PCR mycoplasma-positive bulk tank samples. Another mycoplasma was identified either alone or in addition to M. bovis in 25% of SYBR PCR mycoplasma-positive bulk tank milk samples. Four species of mycoplasma other than M. bovis (Mycoplasma alkalescens, Mycoplasma arginini, Mycoplasma bovigenitalium, and Mycoplasma gateae) were identified in bulk tank milk samples tested with this method. Five farms had 2 mycoplasma species occurring at different times in their bulk tanks. Two mycoplasma species were identified in the same bulk tank sample in 7 instances on 2 farms. The finding of multiple Mycoplasma spp. coexisting on a farm and even in the same bulk tank milk sample indicates that the clinical significance of multiple mycoplasma species in the pathology of intramammary infections should be investigated further. In comparison with conventional culture, the SYBR PCR protocol was slightly (but not statistically significantly) more sensitive in the detection of mycoplasmas in bulk tank milk.     

猪肺炎支原体环介导等温扩增检测方法的建立

目的建立检测猪肺炎支原体(Mycoplasma hyopneumoniae,Mh)的环介导等温扩增(Loop-mediated isothermal am-plification,LAMP)方法。方法根据GenBank中登录的猪肺炎支原体的核苷酸序列,设计4条特异性引物,用外引物进行PCR反应,对内外引物浓度、dNTP+和MgSO4浓度及Bst DNA聚合酶用量等进行优化,建立猪肺炎支原体LAMP检测方法,并对该方法进行特异性和敏感性验证。结果内外引物浓度分别为0.5和0.20 pmol/μl,dNTP+浓度为0.5 mmol/L,MgSO4浓度为3.75 mmol/L,Bst DNA聚合酶用量为8.0和9.6 U时,LAMP反应效果较好;应用该方法检测多杀性巴氏杆菌、猪胸膜肺炎放线杆菌、肺炎双球菌、支气管败血波氏杆菌、副猪嗜血杆菌和链球菌均呈阴性;当猪肺炎支原体的拷贝数低于3时,检测不到该病原体。结论已建立了猪肺炎支原体LAMP检测方法,该方法特异性较好,敏感性较高。     

Isolation of Mycoplasma spp. and serological responses in bulls prior to and following their introduction into Mycoplasma bovis-infected dairy herds

With the common use of bulls for breeding following a period of artificial insemination in seasonally bred dairy herds, it is important to consider the potential role of the bull in transmission of Mycoplasma spp. within and between herds. This study aimed to assess the prevalence of Mycoplasma spp. in a population of bulls before and after use in Mycoplasma bovis-infected herds. The frequency of subclinical infection was also measured serologically postbreeding, and the association of Mycoplasma spp. on semen quality was evaluated. Mycoplasma bovis was isolated from 4 of 118 bulls after use in 4 herds infected with M. bovis. In the bulls, M. bovis seroprevalence increased from 9% prebreeding to 46% postbreeding with a total seroconversion rate of 44% across the 4 herds, with no evidence of clinical disease. There was no association of Mycoplasma spp. in the bulls' semen and abnormal palpation characteristics (enlarged or nodular) of seminal vesicular glands or poor semen quality attributes such as semen mass activity, sperm motility, and morphology. These results demonstrate a high degree of subclinical exposure of the bulls to M. bovis in infected herds and highlight the potential for bulls to be mycoplasma carriers within and between herds. Herd biosecurity protocols and control programs should take into account the potential role of bulls in the introduction and spread of Mycoplasma spp.     

阿奇霉素针剂对小儿肺炎支原体感染治疗体会

目的探讨小儿肺炎支原体感染病例临床特点。方法将我院自2007年1月至2009年11月共治疗小儿支原体肺炎随机分成2组。治疗组80例。给予阿奇霉素粉针剂10mg/(kg·d)静脉滴注,1次/d,疗程3～5d。对照组40例。给予红霉素粉针剂20～25mg/(kg·d)静脉滴注,1次/d,疗程7～14d。结果治疗组有效率42.5%,对照组30%,2组有明显的差异性。治疗组无明显不良反应。结论阿奇霉素是治疗小儿支原体肺炎的最有效、安全的抗生素,不良反应轻。     

阿奇霉素联合盐酸氨溴索治疗小儿支原体肺炎合并急性支气管炎的临床研究

目的:观察阿奇霉素联合盐酸氨溴索治疗小儿支原体肺炎合并急性支气管炎的临床疗效和安全性。方法: 将104例小儿支原体肺炎合并急性支气管炎患儿随机分为对照组和试验组,每组52例。对照组给予阿奇霉素10 mg/kg,qd,静脉滴注;试验组给予阿奇霉素10 mg/kg+盐酸氨溴索15 mg,qd,静脉滴注,连读给药7 d。观察两组临床疗效和不良反应,比较两组患儿的血清超敏C-反应蛋白（hsCRP）、γ-干扰素（IFN-γ）、降钙素原（PCT）、免疫球蛋白A（IgA）、免疫球蛋白G（IgG）、免疫球蛋白M（IgM）水平。结果: 试验组和对照组总有效率分别为96.15%（50/52例）和82.69%（43/52例）,差异有统计学意义（P<0.05）。试验组和对照组的hsCRP分别为（13.28±4.76）和（21.11±6.42）mg/L;IFN-γ分别为（8.45±3.62）和（20.36±5.12）ng/L;PCT分别为（4.02±2.07）和（7.46±2.13）μg/L;IgA分别为（2.17±0.43）和（4.03±0.52）μg/L;IgG分别为（8.14±1.31）和（12.32±1.65）μg/L;IgM分别为（0.86±0.28）和（1.20±0.39）μg/L;差异均有统计学意义（P<0.05）。药物不良反应主要为恶心、呕吐、皮疹、腹泻、头晕、嗜睡等,试验组和对照组药物不良反应发生率分别为23.08%和19.23%（P>0.05）。 结论:阿奇霉素联合盐酸氨溴素治疗小儿支原体肺炎合并急性支气管炎临床疗效明确,可减轻患儿的炎症反应,改善免疫功能,且安全性较好。     

一株来自金雕的霉形体的形态研究

本文利用航向电镜观察了一株来自金雕的形体在培养过程中的各种形态。结果表明该霉形体的形态极其多变，主要有丝状分枝体、原质小体、较大型体和巨大型体。它们在固体培养和液体培养等条件下的形态变化各有特点和规律。同时还进一步证实，除原质小体外的细胞均有直接的分裂连续增殖，而原质小体由丝状分枝体和较大型体产生，经转主为其它形态后再增殖。     

Effect of clinical contagious agalactia on the bulk tank milk somatic cell count in Murciano-Granadina goat herds

From 19 herds of Murciano-Granadina goats, weekly bulk tank somatic cell count (BTSCC) were performed from October to April, and suspicious milk (n = 182), synovial fluid, and ocular swabs (n = 15) from diseased goats were processed for mycoplasma isolation and identification. Also BTSCC from 65 herds were determined (n = 2693). A mixed model procedure was used to establish the effect of the herd and the lactation month on the BTSCC. Monthly rolling values were calculated for each herd using data collected over the preceding 3 complete months, and 4 different BTSCC thresholds were considered: 1,750,000, 1,500,000, 1,000,000, and 750,000 cells/mL. The mean log BTSCC for the 7-mo study period was 5.89 +/- 0.28 for herds without mycoplasma detection from clinical cases, 5.91 +/- 0.31 for mycoplasma-infected herds without clinical contagious agalactia (CA), and 6.47 +/- 0.32 for the herd with clinical CA. The posthoc tests revealed that only the herd that suffered a clinical CA outbreak showed counts that were significantly higher. No significant differences were found for BTSCC between herds not showing clinical episodes of CA, regardless of whether the mycoplasma had been isolated or not. The 1,750,000-cells/mL threshold would only be surpassed by a few herds with serious mastitis problems (clinical outbreak of CA for example). Seventy percent of the goat herds studied were in compliance with the proposed European Union legal limit of 1,500,000 cells/mL for goat milk.     

纤维支气管镜联合甲泼尼龙应用于儿童RMPP中的疗效及安全性分析

目的: 探讨纤维支气管镜联合甲泼尼龙对难治性肺炎支原体肺炎（refractory mycoplasma pneumoniae pneumonia, RMPP）患儿的疗效和安全性以及可溶性B7-H3（sB7-H3）在RMPP中的作用。方法: 选取我院从2015年7月至2017年10月收治的70例难治性支原体肺炎患儿,按照随机数字表法均分为两组：支气管镜组35例,常规治疗3 d后,给予纤维支气管镜下肺泡灌洗治疗;联合治疗组35例,甲泼尼龙和常规治疗3 d后,给予纤维支气管镜下肺泡灌洗治疗。正常对照组：选取同期因异物行支气管镜的患儿35例。两组治疗4周后,观察两组的总有效率、咳嗽消失时间、发热消失时间、啰音消失时间、胸片恢复正常时间、并发症及不良反应,两组患儿和正常对照组sB7-H3和支气管肺泡灌洗液炎症因子水平,分析两组患者sB7-H3与炎症因子的相关关系。结果: 治疗4周后,联合治疗组总有效率(97.1%)明显高于支气管镜组(82.9%),差异具有统计学意义(P<0.05);联合治疗组的咳嗽消失时间、发热消失时间、啰音消失时间及胸片恢复正常时间等均明显短于支气管镜组(P<0.05);两组患者的sB7-H3及白介素1β(IL-1β)、白介素2(IL-2)、白介素6(IL-6)、干扰素γ(INF-γ)等炎症因子水平明显高于正常对照组(P<0.05);联合治疗组sB7-H3及IL-1β、IL-2明显低于支气管镜组(P<0.05);两组患者sB7-H3与IL-1β(r=0.815, P<0.001)和IL-2(r=0.629, P<0.001)呈正相关关系;sB7-H3与IL-6(r=0.295, P=0.165)和INF-γ(r=0.189, P=0.148)无明显相关关系。两组患儿均未出现严重并发症和药物不良反应。结论: 纤维支气管镜联合甲泼尼龙治疗RMPP患儿疗效显著,可明显提高临床疗效、缩短病程;同时,sB7-H3、IL-1β、IL-2、IL-6及INF-γ等因子在RMPP患儿中明显升高,联合治疗可降低sB7-H3、IL-1β及IL-2水平;此外,sB7-H3可能参与IL-1β和IL-2的释放,从而促进RMPP的发生。