The most common medications for the treatment of zoonotic toxoplasmosis are pyrimethamine and sulfadiazine, which may cause serious undesirable side effects. Thus, there is an urgent need to develop novel therapeutics. Baicalein (BAI, C₁₅H₁₀O₅) has been shown to perform well against protozoan parasites including Leishmania and Cryptosporidium. In this study, the inhibition efficacy of BAI on Toxoplasma gondii was evaluated using plaque, invasion, and intracellular proliferation assays. BAI effectively inhibited T. gondii (half-maximum inhibitory concentration (IC₅₀)=6.457×10^-5 mol/L), with a reduced invasion rate (33.56%) and intracellular proliferation, and exhibited low cytotoxicity (half-maximum toxicity concentration (TC₅₀)=5.929×10^-4 mol/L). Further investigation using a mouse model shed light on the inhibitory efficacy of BAI against T. gondii, as well as the potential mechanisms underlying its anti-parasitic effects. The survival time of T. gondii-infected ICR mice treated with BAI was remarkably extended, and their parasite burdens in the liver and spleen were greatly reduced compared with those of the negative control group. Histopathological examination of live sections revealed effective therapeutic outcomes in the treatment groups, with no notable pathological alterations observed. Furthermore, alterations in cytokine levels indicated that BAI not only effectively suppressed the growth of T. gondii but also prevented excessive inflammation in mice. Collectively, these findings underscore the significant inhibitory efficacy of BAI against T. gondii, positioning it as a promising alternative therapeutic agent for toxoplasmosis.
Animals ; Toxoplasma/drug effects* ; Flavanones/therapeutic use* ; Mice ; Antiparasitic Agents/therapeutic use* ; Mice, Inbred ICR ; Toxoplasmosis/drug therapy* ; Female

Objective:To explore the curative efficacy and safety of microwave ablation(MWA)combined with percutaneous ethanol injection(PEI)in treating thyroid cystic solid nodule.Methods:A total of 62 patients with thyroid cystic solid nodule,who admitted to The Affiliated Huaian No.1 People's Hospital of Nanjing Medical University from January 2021 to December 2022,were selected,and they were divided into microwave group(n=28,only single MWA)and combination group(n=34,MWA combined with PEI)according to different treatment plans.The reduction rate of the volume of thyroid nodules,indicators of thyroid function,clinical symptom and the score of physical signs,as well as the incidence of perioperative complications,were compared between two groups after treatment.Results:After treatment,the reduction rate of the volume of thyroid nodule was(88.97±6.36)%in the combination group,which was higher than that(88.15±5.69)%in the microwave group,and the differences in those indicators between the two groups were statistically significant(t=2.465,P＜0.05).The differences of the indicators of thyroid function before and after treatment between two groups were not significant(P＞0.05).The clinical symptoms and the scores of physical signs of both groups after treatment were lower than that before treatment(P＞0.05).The incidence of perioperative complications in the combination group was 17.65%,which was lower than 42.68%in the microwave group(P＜0.05),and the difference of that between the two groups was significant(x2=4.736,P＜0.05).Conclusion:The curative efficacy of the combination of MWA and PEI is better than MWA alone,which has better safety.

Objective:To explore the curative efficacy and safety of microwave ablation(MWA)combined with percutaneous ethanol injection(PEI)in treating thyroid cystic solid nodule.Methods:A total of 62 patients with thyroid cystic solid nodule,who admitted to The Affiliated Huaian No.1 People's Hospital of Nanjing Medical University from January 2021 to December 2022,were selected,and they were divided into microwave group(n=28,only single MWA)and combination group(n=34,MWA combined with PEI)according to different treatment plans.The reduction rate of the volume of thyroid nodules,indicators of thyroid function,clinical symptom and the score of physical signs,as well as the incidence of perioperative complications,were compared between two groups after treatment.Results:After treatment,the reduction rate of the volume of thyroid nodule was(88.97±6.36)%in the combination group,which was higher than that(88.15±5.69)%in the microwave group,and the differences in those indicators between the two groups were statistically significant(t=2.465,P＜0.05).The differences of the indicators of thyroid function before and after treatment between two groups were not significant(P＞0.05).The clinical symptoms and the scores of physical signs of both groups after treatment were lower than that before treatment(P＞0.05).The incidence of perioperative complications in the combination group was 17.65%,which was lower than 42.68%in the microwave group(P＜0.05),and the difference of that between the two groups was significant(x2=4.736,P＜0.05).Conclusion:The curative efficacy of the combination of MWA and PEI is better than MWA alone,which has better safety.

OBJECTIVE To study the immunotherapeutic effect of chimeric virus-like particles(VLPs) based on hepatitis E virus(HEV) against human papillomavirus type 16(HPV 16) tumor. METHODS HPV16 E7 was inserted into the p239 protein of HEV to form the recombinant chimeric protein p239-HPV16 E7. The constructed recombinant protein was expressed by Escherichia coli, purified, and then refolded, and the protein was detected by electron microscopy and dynamic light scattering to confirm size and shape. Then, the C57B/L mice were immunized with the protein grain, and the lymphocyte differentiation of mouse spleen was detected by flow cytometry and enzyme-linked immune spot immunoassay; in addition, TC-1 tumor cells were used to construct tumor models in C57B/L mice to evaluate the anti-tumor immune effect of protein particles in mice. RESULTS After refold in vitro, the structure of chimeric protein was observed under electron microscopy, and the size of particle was 22.80 nm. The obtained protein particles induced favorable specific cellular immune response in C57B/L mice. Compared with the control group, the proportions of CD3+/CD4+ and CD3+/CD8+ in spleen lymphocytes of experimental groups were significantly different(P＜0.05), and effector T cells secreting IFN-γ interferon were also increased remarkably. At the same time, the obtained protein particles could effectively inhibit the growth of tumor cells in TC-1 tumor-bearing mice, and the mice did not die during the experimental period, while the tumors in the control mice grew rapidly and all died after 6 weeks. CONCLUSION Chimeric protein p239-HPV16E7 which was expressed in prokaryotes can form virus-like particles and effectively induce anti-tumor immunity against HPV16.

Objective:To evaluate the effect of eye-covering pretreatment on acute delirium in ophthalmology preschool-age children who underwent binocular and monocular surgery by general anesthesia.Methods:The 300 preschool-age children who underwent general anesthesia of elective ophthalmic surgery in the Second Affiliated Hospital, Zhejiang University School of Medicine, from August 2019 to February 2021 were selected as the research object. They were divided into control group and blindfold group with 150 cases each by random number-table. Children in the control group received regular education on cartoon animation videos before surgery; children in the blindfold group received eye-covering pretreatment on the basis of cartoon animation videos(monocular surgery with monocular cover, binocular surgery with binocular cover). The Modified Yale Preoperative Anxiety Scale (m-YPAS) , the Nursing Delirium Screening Scale(NU-DESC), the incidence rate of delirium and the score of postoperative nursing difficulty were compared between two groups.Results:The 271 cases were completed in this study, including 129 cases(monocular surgery 66 cases, binocular surgery 63 cases) in the blindfold group and 142 cases (monocular surgery 73 cases, binocular surgery 69 cases) in the control group. The preoperative m-YPAS score, the postoperative NU-DESC score, the incidence rate of acute delirium and postoperative nursing care difficulty score of monocular surgery in the blindfold group , monocular surgery was (40.28 ± 15.02) points, 1.00 (0.00, 2.00) points, 27.3%(18/66), 1.00 (1.00, 2.00) points,and binocular surgery was (41.69 ± 16.35) points, 1.00 (0.00, 2.00), 39.7%(25/63), 1.00(1.00, 2.00); in the control group, monocular surgery was (46.28 ± 15.76) points, 2.00 (1.00, 3.00) points, 67.1% (49/73), 2.00 (1.00, 3.00) points, and binocular surgery was (47.77 ± 14.82) points, 3.00 (2.00, 4.00) points, 82.6% (57/69) and 2.00 (1.50, 3.00) points respectively. The difference between the two groups was statistically significant ( t= -2.29, -2.24, Z values were -5.74 - -2.95, χ2= 32.94, 25.78, all P<0.05). The preoperative m-YPAS score, the postoperative NU-DESC score, the incidence rate of acute delirium and postoperative nursing care difficulty score of monocular surgery patients in the blindfold group had no significantly statistical difference with that of binocular surgery patient (all P>0.05) . Conclusions:Monocular/ binocular eye-covering pretreatment can effectively decrease the preoperative m-YPAS score, the postoperative NU-DESC score, incidence rate of acute delirium and the postoperative nursing care difficulty in preschool-age children who underwent general anesthesia both monocular or binocular surgery. There was no difference in the application effect of monocular or binocular surgery.

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.

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.
Animals ; Apoptosis ; Chlorocebus aethiops ; Gene Expression Profiling ; Humans ; Mammals/genetics* ; Toxoplasma/genetics* ; Toxoplasmosis/pathology* ; Vero Cells ; ras GTPase-Activating Proteins/genetics*

We discussed the influence of liquid nitrogen cryopreservation to survive capability of Babesia microti standard strain.The whole blood of mice infected with Babesia microti was put in liquid nitrogen to cryopreservation for 1 month,3 months,6 months,9 months,the whole blood was get out respectively and recovery at room temperature,and infected 3 mice respectively,100 μL/ mouse (the first generation after redissolution,the experiment group).In the same time,3 mice were also infected with Babesia microti as the animal conservation control group.When the infection rate was at a high level,the whole blood of the experiment group mice were injected into 3 normal BALB/c mice (the second generation after redissolution),to observe the changes of the Babesia microti form and proliferation situation,and also to observe the infection rate of the first and the second generation after redissolution in different conserving time.Compared with Babesia microti of animal subcultivation,the form of Babesia microti of liquid nitrogen cryopreservation changed a little.Small trophozoites,annular trophozoites,schizont and immature and mature merozoite and other form can also be seen.Compared with Babesia microti of animal subcultivation,the first time to see the worms and the time attaining to the high infection level were 1 to 2 days later,but for the second generation after redissolution,it is the same.There was no significant difference in different conserving time of 1,3,6,9 months.The influence of liquid nitrogen cryopreservation to survive capability and worm form of Babesia microti is a little,so liquid nitrogen cryopreservation can be a better way to conserving Babesia microti.

Objective To analyze the fractional proteins and immunoreactivity of the soluble antigens from Babesia microti (B.microti),and find the candidate antigens for diagnosis with high sensitivity and specificity.Methods BALB/c mice were inoculated with B.microti-infected red blood cells by intraperitoneal injection.The B.microti were collected from the infected red blood cells when the infection rate reached its peak (infection rate ＞70％),then the soluble antigens were extracted by repeated freezing-thawing and ultrasonic method.The mice sera before and after the infection with B.microti for 7,14,21,28,35,42,49 and 56 days were also collected.The polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze protein components of the soluble antigens of B.microti and the Western blot was used to analyze the immunoreactivity of the soluble antigens with the pooled mice sera before and after the infection.The specific positive protein bands were identified by Liquid chromatography electrospray ionisation tandem mass spectrometry (LC-ESI-MS/MS),and the amino acid sequences of the proteins were analyzed by bioinformatics tools.Results The results from SDS-PAGE analysis indicated that the soluble antigens of B.microti showed distinct protein bands with the range between 12 and 185 × 103 (kDa,relative molecular mass,Mr),among which 9 main bands and 12 minor bands were obtained.In the Western blot analysis,the protein bands with Mr at 40 and 45 kDa could be recognized by pooled mice sera 7 days after infection;the protein bands with Mr at 40,45,54 and 95 kDa could be recognized by pooled mice sera 14 days after infection;the protein bands with Mr at 27,40,45,54,95 and 110 kDa could be recognized by pooled mice sera 21 days after infection.While,the protein bands with Mr at 27,40,45,54,95,1 10 kDa and other weak-reactive bands were recognized by pooled mice sera 28-56 days after infection,and the reaction became stronger with the infection continued.There were 336 proteins,including surface antigen,heat shock protein 70,seroreactive antigen,Eta subunit of chaperonin containing t-complex polypeptide 1 and unnamed protein products,were identified as the components of soluble antigens after mass spectrometry and sequence analysis.Conclusion The 40,45 and 54 kDa protein components from the soluble antigens of B.microti may be ideal candidate antigens for diagnosis,andtheir potential applications in diagnosing of human babesiosis deserve further study.