冬虫夏草菌实时荧光定量PCR内参基因的筛选

以冬虫夏草单子囊孢子分离得到的菌株TZ8-1的3种菌丝形态为实验材料,提取RNA,经反转录获取cDNA,选择了11个持家基因为候选内参基因（18S rRNA、APRTase、β-TUB、RPL2、EF1-α、PGI、PGM、H⁺-ATPase、ACT1、UBQ和GAPDH）,根据该菌基因组注释结果来设计引物,采用实时荧光定量PCR（qRT-PCR）技术进行定量扩增,利用geNorm、NormFinder和BestKeeper算法程序进行表达稳定性评估,并用RefFinder对评估结果进行综合排比,最终筛选得到了最适内参基因。结果表明,所选取的11个候选内参基因均可作为冬虫夏草菌菌丝体时期的内参基因,稳定性最好的3个内参基因分别是UBQ、PGE和ACT1,稳定性较差的3个内参基因分别是GAPDH、RPL2和β-TUB。     

肺结节病合并Alternaria alternata所致皮肤链格孢病1例

患者,女,61岁,确诊肺结节病、口服糖皮质激素7月,双手背及腕部红色斑块3个月。组织病理示表皮轻度棘层增厚,真皮浅、中层中性粒细胞、淋巴、组织细胞、浆细胞、多核巨细胞混合性浸润,可见散在双轮廓厚壁酵母样结构。六铵银染色见不规则肿胀菌丝及酵母样厚壁孢子。真菌培养见棕/灰褐色菌落,镜下见棕色分隔孢子。ITS区序列分析鉴定为Alternaria alternate。口服伊曲康唑胶囊200 mg/d,联合外用硝酸舍他康唑乳膏2次/d,连续用药4个月后皮损完全消退,随访半年,未见复发。     

非阿片类镇痛复合静脉全麻在鼾症手术患者中的疗效

目的:观察三重措施预防为基础,联合非阿片镇痛药复合静脉全麻在行鼾症手术患者术后恶心呕吐的应用效果。方法:选择择期行鼾症手术男性病人80例,随机分为两组:吸入麻醉组(inhalation group, IHLA组)和静脉麻醉组(intravenous group, TIVA组),每组40例,两组均采用三重措施预防恶心呕吐,IHLA组采用以舒芬太尼为基础复合七氟烷吸入麻醉,TIVA组以氯胺酮和右美托咪定镇痛基础上丙泊酚全凭静脉麻醉。评估两组病人恶心呕吐危险系数,采用李克特量表(Likert scale),记录并分析两组患者术后6~8 h在麻醉后监测治疗室(post anesthesia care unit, PACU)及病房24 h恶心呕吐发生情况及补救用药用量。结果:两组患者一般临床资料、恶心呕吐风险评分、手术时间、术后恢复期补救用药量人数无显著差异(P>0.05);IHLA组在PACU恶心呕吐发生率为39.5%,TIVA组发生率为18.9%,两者相比有显著性差异(P<0.05);IHLA组病房24 h恶心呕吐严重程度高于TIVA组,两组术后需要补救应用抗呕吐药物用量无显著差异(P>0.05)。结论:以三重措施预防为基础,与吸入麻醉相比,非阿片类镇痛药复合静脉麻醉可以减少肥胖病人鼾症手术术后恶心呕吐发生率和严重程度,降低围术期风险,有利于患者早期恢复。     

Sustained expression of MCP-1 induced low wall shear stress loading in conjunction with turbulent flow on endothelial cells of intracranial aneurysm

Shear stress was reported to regulate the expression of AC007362, but its underlying mechanisms remain to be explored. In this study, to isolate endothelial cells of blood vessels, unruptured and ruptured intracranial aneurysm (IA) tissues were collected from IA patients. Subsequently, quantitative real-time PCR (qRT-PCR), Western blot and luciferase assay were performed to investigate the relationships between AC007362, miRNAs-493 and monocyte chemoattractant protein-1 (MCP-1) in human umbilical vein endothelial cells (HUVECs) exposed to shear stress. Reduced representation bisulphite sequencing (RRBS) was performed to assess the level of DNA methylation in AC007362 promoter. Accordingly, AC007362 and MCP-1 were significantly up-regulated while miR-493 was significantly down-regulated in HUVECs exposed to shear stress. AC007362 could suppress the miR-493 expression and elevate the MCP-1 expression, and miR-493 was shown to respectively target AC007362 and MCP-1. Moreover, shear stress in HUVECs led to the down-regulated DNA methyltransferase 1 (DNMT1), as well as the decreased DNA methylation level of AC007362 promoter. Similar results were also observed in ruptured IA tissues when compared with unruptured IA tissues. In conclusion, this study presented a deep insight into the operation of the regulatory network of AC007362, miR-493 and MCP-1 upon shear stress. Under shear stress, the expression of AC007362 was enhanced by the inhibited promoter DNA methylation, while the expression of MCP-1 was enhanced by sponging the expression of miR-493.     

SGT1 is not required for plant LRR-RLK-mediated immunity

Plant immune signalling activated by the perception of pathogen-associated molecular patterns (PAMPs) or effector proteins is mediated by pattern-recognition receptors (PRRs) and nucleotide-binding and leucine-rich repeat domain-containing receptors (NLRs), which often share cellular components and downstream responses. Many PRRs are leucine-rich repeat receptor-like kinases (LRR-RLKs), which mostly perceive proteinaceous PAMPs. The suppressor of the G2 allele of skp1 (SGT1) is a core immune regulator required for the activation of NLR-mediated immunity. In this work, we examined the requirement of SGT1 for immune responses mediated by several LRR-RLKs in both Nicotiana benthamiana and Arabidopsis. Using complementary genetic approaches, we found that SGT1 is not limiting for early PRR-dependent responses or antibacterial immunity. We therefore conclude that SGT1 does not play a significant role in bacterial PAMP-triggered immunity.     

Intestinal helminths as predictors of some malaria clinical outcomes and IL-1β levels in outpatients attending two public hospitals in Bamenda,North West Cameroon

This study aimed at determining the impact of intestinal helminths on malaria parasitaemia, anaemia and pyrexia considering the levels of IL-1β among outpatients in Bamenda. A cohort of 358 consented participants aged three (3) years and above, both males and females on malaria consultation were recruited in the study. At enrolment, patients’ axillary body temperatures were measured and recorded. Venous blood was collected for haemoglobin concentration and malaria parasitaemia determination. Blood plasma was used to measure human IL-1β levels using Human ELISA Kit. The Kato-Katz technique was used to process stool samples. Five species of intestinal helminths Ascaris lumbricoides (6.4%), Enterobius vermicularis (5.0%), Taenia species (4.2%), Trichuris trichiura (1.1%) and hookworms (0.8%) were identified. The overall prevalence of Plasmodium falciparum and intestinal helminths was 30.4% (109/358) and 17.6% (63/358) respectively. The prevalence of intestinal helminths in malaria patients was 17.4% (19/109). Higher Geometric mean parasite density (GMPD ±SD) (malaria parasitaemia) was significantly observed in patients co-infected with Enterobius vermicularis (5548 ± 2829/μL, p = 0.041) and with Taenia species (6799 ± 4584/μL, p = 0.020) than in Plasmodium falciparum infected patients alone (651 ± 6076/ μL). Higher parasitaemia of (1393 ± 3031/μL) and (3464 ± 2828/μL) were recorded in patients co-infected with Ascaris lumbricoides and with hookworms respectively but the differences were not significant (p > 0.05). Anaemia and pyrexia prevalence was 27.1% (97/358) and 33.5% (120/358) respectively. Malaria patients co-infected with Enterobius vermicularis and Ascaris lumbricoides had increased risk of anaemia (OR = 13.712, p = 0.002 and OR = 16.969, p = 0.014) respectively and pyrexia (OR = 18.07, p = 0.001 and OR = 22.560, p = 0.007) respectively than their counterparts. Increased levels of IL-1β were significantly observed in anaemic (148.884 ± 36.073 pg/mL, t = 7.411, p = 0.000) and pyretic (127.737 ± 50.322 pg/mL, t = 5.028, p = 0.000) patients than in non-anaemic (64.335 ± 38.995pg/mL) and apyretic patients (58.479 ± 36.194pg/mL). Malaria patients co-infected with each species of intestinal helminths recorded higher IL-1β levels (IL-1β > 121.68 ± 58.86 pg/mL) and the overall mean (139.63 ± 38.33pg/mL) was higher compared with levels in malaria (121.68 ± 58.86 pg/mL) and helminth (61.78 ± 31.69pg/mL) infected patients alone. Intestinal helminths exacerbated the clinical outcomes of malaria in the patients and increased levels of IL-1β were observed in co-infected patients with anaemia, pyrexia and higher parasitaemia.     

Cross-scale quality assessment of a mechanistic cation exchange chromatography model

Cation exchange chromatography (CEX) is an essential part of most monoclonal antibody (mAb) purification platforms. Process characterization and root cause investigation of chromatographic unit operations are performed using scale down models (SDM). SDM chromatography columns typically have the identical bed height as the respective manufacturing-scale, but a significantly reduced inner diameter. While SDMs enable process development demanding less material and time, their comparability to manufacturing-scale can be affected by variability in feed composition, mobile phase and resin properties, or dispersion effects depending on the chromatography system at hand. Mechanistic models can help to close gaps between scales and reduce experimental efforts compared to experimental SDM applications. In this study, a multicomponent steric mass-action (SMA) adsorption model was applied to the scale-up of a CEX polishing step. Based on chromatograms and elution pool data ranging from laboratory- to manufacturing-scale, the proposed modeling workflow enabled early identification of differences between scales, for example, system dispersion effects or ionic capacity variability. A multistage model qualification approach was introduced to measure the model quality and to understand the model's limitations across scales. The experimental SDM and the in silico model were qualified against large-scale data using the identical state of the art equivalence testing procedure. The mechanistic chromatography model avoided limitations of the SDM by capturing effects of bed height, loading density, feed composition, and mobile phase properties. The results demonstrate the applicability of mechanistic chromatography models as a possible alternative to conventional SDM approaches.     

A muscle-path-plane method for representing muscle contraction during joint movement

Traditional muscle paths (the straight-line model and the viapoint-line model) emphasise either the mechanical properties that arouse joint movement or the morphological characteristics of the muscles. To consider both the factors, a muscle-path-plane (MPP) method is introduced to model the paths of muscles during joint movement. This method is based on the premise that there is a MPP, constructed by origin, insertion and MPP control point, which represents the major direction of the muscle contraction for an arbitrary joint configuration at any time. Then, we can calculate the functions and the lengths of the muscle paths during instantaneous joint movement in MPP by mathematical approaches. Taking the triceps brachii as an example, the lengths of its paths during elbow flexion are calculated and compared with the relative studies reported in the literature. It is concluded that this method can provide an insight into the simulation of the muscle contraction.