Toxoplasma gondii infection induces cell apoptosis via multiple pathways revealed by transcriptome analysis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Toxoplasma gondii is a worldwide parasite that can infect almost all kinds of mammals and cause fatal toxoplasmosis in immunocompromised patients. Apoptosis is one of the principal strategies of host cells to clear pathogens and maintain organismal homeostasis, but the mechanism of cell apoptosis induced by T. gondii remains obscure. To explore the apoptosis influenced by T. gondii, Vero cells infected or uninfected with the parasite were subjected to apoptosis detection and subsequent dual RNA sequencing (RNA-seq). Using high-throughput Illumina sequencing and bioinformatics analysis, we found that pro-apoptosis genes such as DNA damage-inducible transcript 3 (DDIT3), growth arrest and DNA damage-inducible α (GADD45A), caspase-3 (CASP3), and high-temperature requirement protease A2 (HtrA2) were upregulated, and anti-apoptosis genes such as poly(adenosine diphosphate (ADP)-ribose) polymerase family member 3 (PARP3), B-cell lymphoma 2 (Bcl-2), and baculoviral inhibitor of apoptosis protein (IAP) repeat containing 5 (BIRC5) were downregulated. Besides, tumor necrosis factor (TNF) receptor-associated factor 1 (TRAF1), TRAF2, TNF receptor superfamily member 10b (TNFRSF10b), disabled homolog 2 (DAB2)‍-interacting protein (DAB2IP), and inositol 1,4,5-trisphosphate receptor type 3 (ITPR3) were enriched in the upstream of TNF, TNF-related apoptosis-inducing ligand (TRAIL), and endoplasmic reticulum (ER) stress pathways, and TRAIL-receptor 2 (TRAIL-R2) was regarded as an important membrane receptor influenced by T. gondii that had not been previously considered. In conclusion, the T. gondii RH strain could promote and mediate apoptosis through multiple pathways mentioned above in Vero cells. Our findings improve the understanding of the T. gondii infection process through providing new insights into the related cellular apoptosis mechanisms.

DNA detection of Paragonimus westermani: Diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick.

Paragonimus westermani (P. westermani) is widely spread in Asian countries and is one of the most important causative agents for lung fluke diseases. The prevention and control of Paragonimiaisis mainly depends on the accurate diagnosis and effective treatment. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay targeted to a portion of the Ty3/gypsy-like LTR retrotransposon (Rn1) sequence coupled with a lateral flow dipstick (LFD) for the rapid detection of P. westermani-specific amplicons. The positive LAMP products were biotin-labeled and hybridized with a fluorescein isothiocyanate-labeled probe which could be visually detected by LFD. No cross-reaction were observed with other parasitic pathogens including Trichinella spiralis, Anisakis simplex, Schistosoma japonicum and Gnathostoma spinigerum, but this LAMP assay could not distinguish P. westermani with Paragonimus skrjabini and Paragonimus heterotremus. The detection limit of the LAMP assay for P. westermani was 2.7 fg/µL, while that of PCR method was 27 fg/µL. LAMP method was applied to detect P. westermani genomic DNA in blood samples form experimental infected dogs, and results showed the parasite was detectable as early as week 2. LAMP-LFD assay applicability was successfully tested in dog blood samples collected from five cities (Wenzhou, Hangzhou, Huzhou, Jiaxing and Shaoxing) in Zhejiang province. In summary, the established LAMP-LFD assay targeted to the Rn1 sequence is a rapid and convenient method for specific detection of P. westermani.

Progress in nanobody and its application in diagnosis / 生物工程学报

Nanobodies are derived from the variable domain of the heavy-chain antibodies (HCAbs) that occur naturally in the serum of Camelidae. They are the smallest antibody fragments capable to bind antigens. With the characteristics of their increased solubility, increased domain stabilities, nanomolar affinities, easy crossing the blood-brain barrier, easy generation, engineering, optimization and tailoring, easy humanization, nanobodies have extensive application prospects in diagnosis and detection. Although nanobody has demonstrated tremendous success, a number of practical challenges limit its broader applications in disease diagnosis and detection, including construction of a phage library and selection of nanobody fragments with high affinity and immunogold labeling technique. Here, we review several recent findings on the use of nanobodies in molecular diagnostics and suggest some practical strategies in resolving the current challenges in this attractive research area, particularly to optimize the affinity, solubility, humanization of nanobodies.

Characteristics of glucose metabolism in non-obese and obese women with polycystic ovarian syndrome / 中国综合临床

Objective To investigate characteristics of glucose metabolism of non-obese and obese women with polycystie ovary syndrome(PCOS).Methods From March 2008 to July 2011,118 PCOS patients were enrolled in this study and were divided into 53 cases (body mass index(BMI) ≥25 kg/m2) in obese group and 65 cases (BMI ＜ 25 kg/m2) in non-obese group.Participants' clinical data,glucose tolerance and insulin release test results were retrospectively reviewed.The prevalence of type 2 diabetes mellitus (T2DM),results of oral glucose tolerance test,impaired fasting glucose (IFG),impaired glucose tolerance (IGT) were compared between the two groups.Results (1) Blood glucose levels:at the time points of 30,60,120 and 180 minutes,the levels of fasting glucose were (5.2 ± 1.1),(8.5 ± 2.8),(8.1 ± 2.4),(6.3 ± 2.0) and (4.8 ± 1.5)mmol/L in non-obese group and(5.4 ±0.9),(9.1 ± 1.8),(9.3 ±0.6),(7.6 ± 1.0) and (5.4 ±0.8) mmol/L in obese group.Statistical difference was observed between obese and non-obese groups at each time point (t =-6.125,-4.005,-6.074,-6.751 and-4.512 respectively,P ＜0.01).(2) The level of insulin:at the time points of 30,60 and 120 min,the level of fasting insulin were (8 ± 4),(55 ± 21),(65 ± 14) and (45 ±18)mU/L in non-obese group and (13 ± 8),(85± 30),(105± 54) and (76 ± 46)mU/L in obese group.There were significant statistical difference between the two groups at each time point (t =-17.024,-12.540,-15.791 and-16.149 respectively,P ＜ 0.01).However,at the time point of 180 minutes,the level of insulin did not exhibit significant difference between obese and non-obese groups (P ＞ 0.05).(3) The prevalence of abnormal glucose metabolism:The rates of IGT were 13.85 ％ (9/65) in non-obese group and 24.53 ％ (13/53)in obese group,which also showed remarkable difference (x2 =18.446,P ＜ 0.01).The rates of T2DM were 1.54％ (1/65) in non-obese group and 7.55％ (4/53) in obese group,which reached significant difference (x2=16.005,P ＜ 0.01).Conclusion Abnormal glucose metabolism was observed more frequently in overweight or obese PCOS women.

Practice on delicacy management of hospital logistics services / 中华医院管理杂志

Delicacy management is an important issue to the hospital logistics mode innovation,which is the only way to realize a scientific hospital logistics management.This article proposes the practical ways to push on the hospital logistics delicacy management from many respects,such as working regulation,consumption measurement,contract supervision,cost accounting,structure control and convergence supporting.

Evaluation of the anthelmintic effects of artesunate against experimental Schistosoma mansoni infection in mice using different treatment protocols.

The therapeutic effects of artesunate against experimental Schistosoma mansoni infection in mice were analyzed. Previous studies showed that artesunate is highly effective against S. japonicum infection, but the action of this drug against S. mansoni remained uncovered. The present study examines the optical conditions for artesunate against S. mansoni and evaluates the effects of inhibiting the sexual maturation of adult worms. Mice infected with S. mansoni were orally administered with artesunate according to different schedules. Four consecutive administrations of 300 mg/kg of artesunate at 2-week intervals conferred almost total protection without the development of pathological lesions in the liver. The significant reduction in the number of eggs produced by surviving worms and the status of egg maturation suggested that artesunate inhibits sexual maturation. Electron microscopy revealed that artesunate caused morphological damage, especially on the worm tegument. Artesunate was also very effective in iron-deficient mice. Furthermore, the efficacy of artesunate was equal to or better than that of artemether against S. japonicum infection. Considering that artemether is more toxic, artesunate is currently one of the most efficient drugs against immature S. mansoni.

Protective effect of vaccination with Toxoplasma lysate antigen and CpG as an adjuvant against Toxoplasma gondii in susceptible C57BL/6 mice.

Infection with the intracellular protozoan parasite Toxoplasma gondii causes serious public health problems to both humans and livestock and of great economic impact worldwide. Oligodeoxynucleotides (ODN) which contain immunostimulatory CG motifs (CpG ODN) can promote Th1 responses, an adjuvant activity that is desirable for vaccination against intracellular pathogens. We investigated the feasibility of using CpG as an adjuvant combined with Toxoplasma lysate antigen (TLA) as a vaccine against toxoplasmosis. Genetically susceptible C57BL/6 mice were vaccinated with TLA with or without CpG ODN as an adjuvant and then challenged with 85 cysts of the moderately virulent RRA (Beverley) strain of T. gondii. Prior to challenge infection, immunization with TLA plus CpG ODN directed cellular and humoral immunity toward a Th1 pattern, characterized by enhanced INF gamma production by splenic cells in response to TLA, and enhanced production of toxoplasma-specific IgG and IgG (2a) antibodies. Consequently, CpG/TLA-treated mice showed prolonged survival and 64% reduction in brain parasite burden compared to non-CpG/TLA treated group. Our results suggest that CpG ODN would provide a stable and effective adjuvant for use in vaccination against toxoplasmosis.

Observation on the Change of Anti-S.japonicum Antibody Level in Population Migrated from Outside Embankment to New Town / 中国寄生虫学与寄生虫病杂志

Objective To detect the change of the anti-S. japonicum antibody level after people migrated from outside embankment to newly established town. Methods Three pilot spots were established for the investigation: one spot thut both inhabitancy and cultivation disused (A), one spot that only inhabitancy disused but farming continued (B) and the third one served as control (C). DIGFA and ELISA were used to detect the antibody level in the populations from 2002 to 2005. Results The positive rate of anti-S.japonkum antibody declined significantly from 6.63% to 3.52% by DIGFA and from 7.26% to 3.71% by ELISA at spot A (X2=5.2625, P