Manipulation of host ecdysteroid hormone levels facilitates infection by the fungal insect pathogen,Metarhizium rileyi

Entomopathogenic fungi such as Metarhizium rileyi and Beauveria bassiana are widely used insect biological control agents. Little, however, is known concerning genetic or enzymatic factors that differentiate the mechanisms employed by these two fungal pathogens to infect target hosts. Infection by either of these organisms is known to increase levels of the growth and molting hormone, ecdysone, which also regulates the expression of a number of innate immune pathways. M. rileyi, but not B. bassiana, has apparently evolved an ecdysteroid-22-oxidase (MrE22O) that inactivate ecdysone. We show that deletion of MrE22O impaired virulence compared with the wild-type strain, with an increase in ecdysone titer seen in hosts that was coupled to an increase in the expression of antimicrobial genes. An M. rileyi strain engineered to overexpress MrE22O (MrE22O^OE), as well as trans-expression in B. bassiana (Bb::MrE220^OE) resulted, in strains displaying enhanced virulence and dampening of host immune responses compared with their respective wild-type parental strains. These results indicate that ecdysone plays an important role in mediating responses to fungal infection and that some insect pathogenic fungi have evolved mechanisms for targeting this hormone as a means for facilitating infection.     

高盐溶液预处理幽门螺杆菌对蒙古沙土鼠胃黏膜损伤作用

目的探讨高盐预处理的幽门螺杆菌(H.pylori)对胃黏膜的损伤作用。方法将30%高盐预处理前后的胃癌来源的H.pylori菌株(4854)灌胃蒙古沙土鼠(MGs),在灌胃后13、26和73周解剖动物,通过组织病理学检查、免疫组化染色和黏膜厚度测量,探讨高盐预处理的H.pylori对胃黏膜的损伤作用。结果与未加盐预处理的相应菌株相比,高盐预处理组小鼠的慢性炎症、黏膜变性/坏死、腺体萎缩伴肠上皮化生的发生率较低,黏膜糜烂/溃疡和黏膜上皮增生的发生率较高,差异均有统计学意义(t=8.325 6,P=0.040 8)。第73周,高盐预处理4854菌株组胃体和胃窦黏膜增生显著高于未加盐预处理组(t=12.802 4,P=0.035 1;t=16.536 0,P=0.043 8)。结论高盐预处理改变了H.pylori的体内致病性,有助于阐明H.pylori感染与高盐饮食在胃病中的相互作用模式。     

补充益生菌对功能性腹泻患者焦虑抑郁状态的影响

目的探讨益生菌对功能性腹泻患者临床症状和心理健康的影响。方法将2019年3月至2019年12月在广东省肇庆市高要区人民医院消化内科门诊收治的伴有焦虑抑郁状态的90例功能性腹泻住院病人随机分为试验组、对照组A、对照组B。三组受试者均口服匹维溴铵,试验组口服双歧杆菌四联活菌,对照组A服用氟西汀,对照组B未给其他药物治疗,疗程均为1个月。治疗前后,比较患者大便次数及性状、汉密尔顿焦虑量表(HAMA)和汉密尔顿抑郁量表(HAMD)评分。结果治疗前三组患者每周排便不同次数的人数比较,差异无统计学意义(P0.05)。治疗第3周和第4周后,与对照组A相比,对照组B和试验组的排便不同次数的人数差异具有统计学意义(P0.05)。治疗第4周后,试验组与对照组B的排便不同次数的人数比较,差异具有统计学意义(P0.05)。治疗前3组患者Bristol粪便性状评分比较差异无统计学意义(P0.05)。治疗第3和第4周后,与对照组A相比,对照组B和试验组的Bristol粪便性状评分比较,差异有统计学意义(P0.05)。治疗第4周后,试验组与对照组B的Bristol粪便性状评分比较,差异有统计学意义(P0.05)。治疗后与对照组A比较,对照组B和试验组HAMA评分和HAMD评分显著低于对照组A,且差异具有统计学意义(P0.05)。与对照组B比较,试验组HAMA评分和HAMD评分显著低于对照组B(P0.05)。结论通过补充益生菌可调节功能性腹泻患者腹泻次数,提高功能性腹泻患者生活质量,改善患者焦虑抑郁症状。     

体外诊断试剂防腐剂的选择策略

对体外诊断试剂中防腐剂的作用机理及选择原则进行了阐述,并对目前常用的防腐剂如叠氮钠、庆大霉素、硫柳汞、异噻唑啉酮类等的性能进行了比较,为体外诊断试剂的研究开发及生产中防腐剂的使用提供有益指导。     

The Transurethral Seminal Vesiculoscopy in the Diagnosis and Treatment of the Seminal Vesicle Disease

To develop a transurethral endoscopy technique of the transurethral seminal vesiculoscopy to examine and treat seminal vesicle disease. A total of 61 patients with seminal vesicle disease were diagnosed and treated with the transurethral seminal vesiculoscopy through the distal seminal tracts and vesicles. 58 cases were successfully treated using transurethral seminal vesiculoscopy via the seminal vesicles. The operation took 25 ~ 85 min, with an average of (35.6) mins. In this group, seven cases were diagnosed as ejaculatory orifice cyst, 14 cases had blood clots in the seminal vesicles, and nine patients had stones in the seminal vesicles. All patients were treated properly. Follow-up occurred at 3 months, with two cases showing post-operative discomfort in perineal region. One patient had recurrence with seminal vesiculitis, which improved with treatment. Four infertile patients had a significant increase in sperm count and ejaculation volume and two of these patients were able to naturally inseminate within seven to 18 months post-surgery. This approach enables a new endoscopic technique with the transurethral seminal vesiculoscopy to diagnose and treat seminal vesicle disease through the normal anatomic pathway which can be easily performed with few post-operative complications.     

Inhibition of tyrosinase by gastrodin: An integrated kinetic-computational simulation analysis

We studied the inhibitory effect of gastrodin on tyrosinase using inhibition kinetics and computational simulation. Gastrodin reversibly inhibited tyrosinase in a mixed-type manner with K_i = 123.8 ± 20.2 mM. Time-interval kinetics revealed the inhibition to be a first-order process with mono- and bi-phasic components. Using AutoDock Vina, we calculated a binding energy of ?6.3 kcal/mol for gastrodin and tyrosinase, and we performed a molecular dynamics simulation of the tyrosinase–gastrodin interaction. The simulation results suggested that gastrodin interacts primarily with histidine residues in the active site. A 10-ns molecular dynamics simulation showed that one copper ion in the tyrosinase active site was responsible for the interaction with gastrodin. Our study provides insight into the inhibition of tyrosinase by the hydroxyl groups of gastrodin. A combination of inhibition kinetics and computational calculations may help to confirm the inhibitory action of gastrodin on tyrosinase and define the mechanisms of inhibition.     

Efficient cloning and expression of a thermostable nitrile hydratase in Escherichia coli using an auto-induction fed-batch strategy

A nitrile hydratase (NHase) gene from Aurantimonas manganoxydans was cloned and expressed in Escherichia coli BL21 (DE3). A downstream gene adjacent to the β-subunit was necessary for the functional expression of the recombinant NHase. The structural gene order of the Co-type NHase was α-subunit beyond β-subunit, different from the order typically reported for Co-type NHase genes. The NHase exhibited adequate thermal stability, with a half-life of 1.5 h at 50 °C. The NHase efficiently hydrated 3-cyanopyridine to produce nicotinamide. In a 1-L reaction mixture, 3.6 mol of 3-cyanopyridine was completely converted to nicotinamide in four feedings, exhibiting a productivity of 187 g nicotinamide/g dry cell weight/h. An industrial auto-induction medium was applied to produce the recombinant NHase in 10-L fermenter. A glycerol-limited feeding method was performed, and a final activity of 2170 U/mL culture was achieved. These results suggested that the recombinant NHase was efficiently cloned and produced in E. coli.     

Characterization of bacteriocin bificin C6165: a novel bacteriocin

Aims

To purify and primarily characterize an anti‐Alicyclobacillus bacteriocin produced by Bifidobacterium animalis subsp. animalis CICC 6165, suggested to be named bificin C6165.

Methods and Results

During purification of the bificin C6165, optimal recovery was achieved with ammonium sulfate precipitation followed by two chromatographic steps. Mass spectrometry analyses revealed a distinctive peak corresponding to a molecular mass of 3395·1 Da. This bacteriocin was heat stable, effective after refrigerated storage and freeze–thaw cycles. The primary mode of action of bificin C6165 is most probably due to pore formation, as indicated by the efflux of K⁺ from metabolically active cells of Alicyclobacillus acidoterrestris. In the presence of 10 mmol l^?1 gadolinium, bificin C6165 did not affect cells of Alicyclobacillus acidoterrestris. This suggests that the mode of action of bificin C6165 relies on a net negatively charged cell surface.

Conclusions

Bificin C6165 is indeed a novel bacteriocin and it exhibited remarkable potency for Alicyclobacillus control.

Significance and Impact of the Study

Application of bacteriocins in preservation of fruit juices has seldom been studied. Bificin C6165 may be an alternative method to control juice spoilage by this Alicyclobacillus acidoterrestris and meet increasing consumer demand for nature and artificial chemical additive‐free food products.     

A comparative study of the atp9 gene between a cytoplasmic male sterile line and its maintainer line and further development of a molecular marker specific for male sterile cytoplasm in kenaf (Hibiscus cannabinus L.)

mtDNA was isolated from cytoplasmic male sterility (CMS) line P3A and its maintainer P3B of kenaf (Hibiscus cannabinus L.). The atp9 gene and its two flanking sequences were obtained using homology cloning and high-efficiency thermal asymmetric interlaced PCR methods. The coding sequences showed only two base pairs difference between the CMS and its maintainer, and shared a homology of over 87 % with atp9 genes from other species in GenBank. However, when comparing the flanking sequences, a 47-bp deletion was characterized at the 3′ flanking sequence of atp9 in the CMS line. Quantitative PCR analysis indicated that the expression level of atp9 in the CMS line was 0.937-fold that of its maintainer. Furthermore, the respiratory rate of anthers in the CMS line was markedly lower than that of its maintainer. The results indicated that the 47-bp deletion at the 3′ flanking sequence of atp9 and/or down-regulated expression of the atp9 gene in the CMS line might be closely related to CMS in kenaf. To confirm whether the 47-bp deletion was specific to cytoplasm of male sterile lines, another 21 varieties were used for further analysis. The results showed that the 47-bp deletion was specific to male sterile cytoplasm (MSC) of kenaf. Based on these, a specific molecular marker was developed to distinguish the MSC from male fertile cytoplasm of kenaf.