Targeted degradation of NDUFS1 by agrimol B promotes mitochondrial ROS accumulation and cytotoxic autophagy arrest in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a global public health problem with increased morbidity and mortality. Agrimol B, a natural polyphenol, has been proved to be a potential anticancer drug. Our recent report showed a favorable anticancer effect of agrimol B in HCC, however, the mechanism of action remains unclear. Here, we found agrimol B inhibits the growth and proliferation of HCC cells in vitro as well as in an HCC patient-derived xenograft (PDX) model. Notably, agrimol B drives autophagy initiation and blocks autophagosome-lysosome fusion, resulting in autophagosome accumulation and autophagy arrest in HCC cells. Mechanistically, agrimol B downregulates the protein level of NADH:ubiquinone oxidoreductase core subunit S1 (NDUFS1) through caspase 3-mediated degradation, leading to mitochondrial reactive oxygen species (mROS) accumulation and autophagy arrest. NDUFS1 overexpression partially restores mROS overproduction, autophagosome accumulation, and growth inhibition induced by agrimol B, suggesting a cytotoxic role of agrimol B-induced autophagy arrest in HCC cells. Notably, agrimol B significantly enhances the sensitivity of HCC cells to sorafenib in vitro and in vivo. In conclusion, our study uncovers the anticancer mechanism of agrimol B in HCC involving the regulation of oxidative stress and autophagy, and suggests agrimol B as a potential therapeutic drug for HCC treatment.

Serological Comparison of Native Antigen ELISAs with Rapid ICT Test Kits for the Diagnosis of Human Alveolar and Cystic Echinococcosis in China.

BACKGROUND: The aim of this study was to compare the diagnostic performance of native antigen ELISAs and ADAMU-AE/CE commercial ICT test kits in subjects either exposed to Echinococcus infection or with clinically diagnosed alveolar (AE) or cystic (CE) echinococcosis. METHODS: A total of 370 subjects with a previous clinical confirmation of CE or AE from northwestern China were recruited. Serum samples were also obtained from 3923 children/teenagers during a community survey. All sera were tested using native antigen ELISAs. The ADAMU-AE/CE test kits were subsequently used for the serology of the 370 clinically confirmed individuals and of 251 children/teenagers that were ELISA antibody-positive for both Echinococcus species but ultrasound-negative during baseline survey. An analysis of the association between the serological tests and ultrasound classification was carried out amongst 89 AE and 164 CE cases. A Kappa consistency analysis was undertaken to compare the diagnostic performance of the native antigen ELISAs and the ADAMU kits and the ultrasound imaging results. The χ² test was also used for a comparison of the different seropositivity rates between the groups. FINDINGS: There was poor consistency (Kappa = 0.26 and 0.28 for AE and CE respectively) between the native antigen ELISAs and the ADAMU kits for the diagnosis of AE and CE among the cases and the surveyed children/teenagers, but a relatively good consistency (Kappa = 0.63) between the ADAMU-AE kit and ultrasound observations for the AE cases. Additionally, of the 251 teenagers co-positive for both AE and CE antibodies by the native antigen ELISAs, only one was found positive by the ADAMU-AE kit, verified as a new AE case on subsequent ultrasound follow-up. The remainder (N = 250) were negative by serology using the ADAMU-AE/CE kits and by ultrasound examination. The two native antigen ELISAs did not discriminate well between cases of clinically diagnosed AE and CE. In contrast, ADAMU-AE and ADAMU-CE commercial ICT test kits readily differentiated cases of AE from CE with specificities of 99% for AE and 100% for CE. CONCLUSIONS: The ADAMU-AE/CE kits proved reliable, accurate, and amenable diagnostic tools in the clinical setting for confirmation of suspected AE/CE cases. The native antigen ELISAs tests can provide useful information on the level of human exposure to Echinococcus infection.

Direct evidence of cheetah (Acinonyx jubatus) as intermediate host of Toxoplasma gondii through isolation of viable strains.

Toxoplasma gondii causes lifelong infection in most definitive and intermediate hosts. Clinical cases of toxoplasmosis in captive cheetahs have been reported. However, there are few reports of viable T. gondii strains isolated from cheetahs. Here, T. gondii infection was investigated using molecular and serological assays in cheetahs from China. Modified agglutination test (MAT) (cut-off: 1:25) indicated that all six examined cheetahs (n = 6) showed T. gondii antibodies. Toxoplasma gondii DNA was detected in three out of five cheetahs. Two viable T. gondii strains were isolated from the striated muscles of two cheetahs using mice bioassay. They were designated as TgCheetahCHn1 and TgCheetahCHn2. Genetic characterization of DNA derived from tachyzoites was performed using RFLP-PCR of 10 markers. Toxoplasma gondii TgCheetahCHn1 is ToxoDB PCR-RFLP genotype #319, and the alleles of ROP18/ROP5 types were 3/7. TgCheetahCHn2 is ToxoDB genotype #9, and the alleles of ROP18/ROP5 were 3/6. The average survival time of TgCheetahCHn1-infected Swiss mice was 22 ± 1 days (n = 23), and the mice did not have detectable T. gondii-specific antibodies until 117 ± 30 days post-inoculation (n = 8), therefore, TgCheetahCHn1 had intermediate virulence. TgCheetahCHn2 was avirulent for Swiss mice. Few brain tissue cysts (0-50) were observed in the mice inoculated with TgCheetahCHn1 or TgCheetahCHn2. The results provide direct evidence of cheetah as intermediate host of T. gondii.

Isolation and Characterization of a Viable Toxoplasma gondii from Captive Caracal (Caracal caracal).

Toxoplasma gondii is a widespread protozoan parasite that infects most warm-blooded animals, and felids can serve as intermediate and definitive hosts. Pathological diagnosis and serological and etiological investigations were conducted on a captive caracal (Caracal caracal) carcass collected from China in 2022. Pathological diagnosis revealed that cardiac insufficiency, pulmonary edema, hepatic failure, and renal insufficiency were the causes of the caracal's death. A modified agglutination test (cut-off: 1:25) revealed that IgG antibodies against T. gondii were present in the myocardium juice (1:1600), ascitic fluid (1:3200), and hydropericardium (1:800). A viable T. gondii (TgCaracalCHn2) strain was isolated from the tissue samples (heart, brain, spleen, and skeletal muscle) of this caracal using a mouse bioassay. The genotype of TgCaracalCHn2 was ToxoDB#5 (Type II variant), as determined via RFLP-PCR. The strain was avirulent in Swiss mice and matched the prediction of ROP18 and ROP5 gene alleles of TgCaracalCHn2 (2/2). Mild tissue cysts (203 ± 265) were observed in mice brains after inoculation with TgCaracalCHn2 tachyzoites. ToxoDB#5 is the dominant genotype in North American wildlife, and this is the first documented isolation of T. gondii ToxoDB#5 from China. This indicates that caracal plays an important role in the transmission of this T. gondii genotype.

Low Prevalence of Toxoplasma gondii in Sheep and Isolation of a Viable Strain from Edible Mutton from Central China.

Sheep are highly susceptible to Toxoplasma gondii, and miscarriage is the main clinical feature. This study investigated 227 sheep samples (210 myocardial tissues from slaughterhouses, 6 ewe serum samples, 3 aborted fetuses, and 8 dead lambs from veterinary clinics) from central China for T. gondii infection. Antibodies against T. gondii were detected using the modified agglutination test (MAT). PCR was performed to detect T. gondii DNA in the tissue samples. The results showed that four samples were seropositive (MAT titer ≥ 1:100), with a seroprevalence of 1.8% (4/227). The seropositive samples included two myocardial samples from a slaughterhouse, one ewe and its aborted fetus from a veterinary clinic. The results revealed that 7 out of 207 (3.4%) sheep tissue samples were PCR-positive, including two myocardial tissue samples from slaughterhouses, three aborted fetuses, and two lambs from veterinary clinics. Toxoplasma gondii vertical transmission had occurred in two of three pairs of ewes and her pups. One viable T. gondii strain (TgSheepCHn14) was isolated from the myocardial tissues of sheep from a slaughterhouse. Tachyzoites were obtained from cell cultures at 70 days following seeding in the brains and lungs of mice. This strain was non-lethal to Swiss mice. The number of parasite brain cysts in mice decreased with time post-infection (p < 0.05). Overall, the prevalence of T. gondii in the sheep samples was low. Although the samples were scattered, and not from planned collections, the current study detected T. gondii antibodies and DNA in aborted fetuses, indicating that vertical transmission could occur and maintain the parasites in sheep herds without exogenous infection.

Additional evidence of tigers (Panthera tigris altaica) as intermediate hosts for Toxoplasma gondii through the isolation of viable strains.

Toxoplasmosis is one of the most common zoonotic diseases in the world. Felines excrete Toxoplasma gondii oocysts, which play a key role in the transmission of this protozoon. Pathological diagnoses were performed on four carcasses of captive tigers collected from 2019 to 2021 in China, and T. gondii was surveyed using serology, molecular analysis, and aetiology. Striated muscle samples of the tigers (n = 4) were bioassayed in mice. DNA derived from T. gondii tachyzoites was isolated and characterized using PCR-RFLP. The pathological diagnoses revealed that ageing, declined immune function, liver, and kidney failures caused the deaths in the tigers examined. A modified agglutination test (cut-off: 1:25) revealed that IgG antibodies to T. gondii were 100% (4/4) in the captive tigers. Two viable T. gondii strains (TgTigerCHn3 and TgTigerCHn4) were isolated from tiger striated muscles and seeded on the Vero cell culture for further propagation. The genotypes of TgTigerCHn3 and TgTigerCHn4 were ToxoDB#20 and ToxoDB#2, respectively. The two strains were avirulent for Swiss mice, which matched the ROP18 and ROP5 gene alleles of TgtigerCHn3 (3/4) and TgtigerCHn4 (3/3). Few brain tissue cysts (0-213) were observed in the mice after inoculation with TgTigerCHn3 and TgTigerCHn4. This is the first documented isolation of T. gondii ToxoDB#20 and ToxoDB#2 from tigers. The results provide additional direct evidence of tiger as intermediate hosts for T. gondii. Tigers in the zoos may potentially transmit T. gondii to other animals and humans.

Impact of core strength training on young darts throwers / Impacto do treino de força no core em jovens praticantes de arremesso de dardos / Impacto del entrenamiento de fuerza en jóvenes practicantes de lanzamiento de dardos

ABSTRACT Introduction Recent research on core strength training of young athletes is vague and inconclusive. The lack of complete understanding about the relevance of core training programs still keeps some coaches wary in their training practice. Objective Explore the impact of core strength training on young athletes' physical fitness and sports ability. Methods Young athletes on the javelin throwing team underwent a bi-weekly core strengthening protocol for 16 weeks. Biomechanical changes were acquired by biomechanical kinematic analysis, and index data were worked out statistically. Results Core strength training developed the latissimus dorsi, external oblique muscles, erector spine, and hip extensor muscles of athletes and improved stability in spine motion (P<0.05). Conclusion Core strength training significantly affects developing specific skills in darts-throwing athletes. The training has been shown to increase athletes' interest in the sport. Evidence level II; Therapeutic Studies - Investigating the results.

RESUMO Introdução Pesquisas recentes sobre o treino de força no core de jovens atletas são vagas e inconclusivas. A ausência da total compreensão sobre a relevância dos programas de treinamento no core ainda mantém alguns treinadores receosos na prática de seu treino. Objetivo Explorar o impacto do treino de força no core sobre a aptidão física e a capacidade esportiva de jovens atletas. Métodos Jovens atletas da equipe de arremesso de dardos passaram por um protocolo de fortalecimento do core, bissemanal por 16 semanas. As alterações biomecânicas foram adquiridas por análise cinemática biomecânica e os dados indexadores foram trabalhados estatisticamente. Resultados O treinamento da força central desenvolveu o latíssimo do dorso, os músculos oblíquos externos, os espinhais eretos e os músculos extensores do quadril dos atletas, além de aprimorar a estabilidade no movimento da coluna vertebral (P<0,05). Conclusão O treino de força do core tem um efeito significativo sobre o desenvolvimento de habilidades específicas nos atletas de arremesso de dardos. O treino demonstrou aumentar o interesse dos atletas pela prática esportiva. Nível de evidência II; Estudos terapêuticos - Investigação de resultados.

RESUMEN Introducción Las investigaciones recientes sobre el entrenamiento de la fuerza en el core de los jóvenes atletas son vagas y no concluyentes. La falta de comprensión total sobre la relevancia de los programas de entrenamiento básico todavía mantiene a algunos entrenadores recelosos en su práctica de entrenamiento. Objetivo Explorar el impacto del entrenamiento de la fuerza del core en la aptitud física y la capacidad deportiva de jóvenes atletas. Métodos Los jóvenes atletas del equipo de lanzamiento de dardos se sometieron a un protocolo quincenal de fortalecimiento del core durante 16 semanas. Los cambios biomecánicos se adquirieron mediante un análisis cinemático biomecánico y los datos del índice se elaboraron estadísticamente. Resultados El entrenamiento de la fuerza del core desarrolló el dorsal ancho, los músculos oblicuos externos, los erectores de la columna y los músculos extensores de la cadera de los atletas, y también mejoró la estabilidad en el movimiento de la columna (P<0,05). Conclusión El entrenamiento de la fuerza del core tiene un efecto significativo en el desarrollo de habilidades específicas en los atletas que lanzan dardos. Se ha demostrado que el entrenamiento aumenta el interés de los atletas por la práctica deportiva. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

Quality improvement of sweet rice wine fermented with Rhizopus delemar on key aroma compounds content, phenolic composition, and antioxidant capacity compared to Rhizopus oryzae.

The pure culture fermentation has led to less flavorful rice wine and relatively lower bioactive substance level compared to traditional mixed culture fermentation; however, a pure strain is easily controlled by industrialized producers. The purpose of the present research was to screen a species of Rhizopus for improving the flavor deficiency and antioxidant function of sweet rice wine. Seven rice wine samples fermented with isolated strains were analyzed for their total phenolics, total flavonoids, peptide and antioxidant activity using spectrophotometry, as well as ethanol, ethyl acetate, ß-phenethyl alcohol, and volatile alcohol contents measured by headspace gas chromatography (HS-GC), the further principal component analysis determined Rhizopus delemar rice wine better on aroma and antioxidant capacity. A comparison of phenolics profile between R. delemar and R. oryzae rice wines was made based on the measurement data of ultrahigh-performance liquid chromatography coupled with Q-exactive orbitrap mass spectrometry (UHPLC-QE-MS) data. Thirty-two phenolics were identified in sweet rice wine. Overall, the results presented in this study showed that a strain of R. delemar is available for the improvement of flavor and antioxidant activity in sweet rice wine, which has the great potential to be applied to industrialized products. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05250-x.

Toxoplasma gondii Rhoptry Protein 7 (ROP7) Interacts with NLRP3 and Promotes Inflammasome Hyperactivation in THP-1-Derived Macrophages.

Toxoplasma gondii is a common opportunistic protozoan pathogen that can parasitize the karyocytes of humans and virtually all other warm-blooded animals. In the host's innate immune response to T. gondii infection, inflammasomes can mediate the maturation of pro-IL-1ß and pro-IL-18, which further enhances the immune response. However, how intercellular parasites specifically provoke inflammasome activation remains unclear. In this study, we found that the T. gondii secretory protein, rhoptry protein 7 (ROP7), could interact with the NACHT domain of NLRP3 through liquid chromatography-mass spectrometry analysis and co-immunoprecipitation assays. When expressing ROP7 in differentiated THP-1 cells, there was significant up-regulation in NF-κB and continuous release of IL-1ß. This process is pyroptosis-independent and leads to inflammasome hyperactivation through the IL-1ß/NF-κB/NLRP3 feedback loop. The loss of ROP7 in tachyzoites did not affect parasite proliferation in host cells but did attenuate parasite-induced inflammatory activity. In conclusion, these findings unveil that a T. gondii-derived protein is able to promote inflammasome activation, and further study of ROP7 will deepen our understanding of host innate immunity to parasites.

High-intensity physical ability training of teenager basketball players / Treino de alta intensidade do preparo físico de jogadores de basquete adolescentes / Entrenamiento de alta intensidad de la preparación física de jugadores de baloncesto adolescentes

ABSTRACT Introduction: The antagonism of basketball is very prominent during the game. Athletes with good physical fitness will have great advantages in basketball games. Objective: To study the influence of physical exercise on the physical fitness of young athletes. Methods: This article uses mathematical statistics to investigate and analyze the physical training status of young basketball players. Results: After studying various training methods, we found that impedance training can improve the physical fitness of young athletes. Basketball physical training includes strength, speed, endurance, sensitivity, etc. Conclusion: Young basketball players need to adopt a variety of training methods. Group sports can effectively improve the physical fitness of young basketball players and increase the winning probability of the team in the game. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: O antagonismo é muito presente em partidas de basquete. Atletas em boas condições físicas terão grande vantagem nessas partidas. Objetivos: Estudar a influência da atividade física no preparo físico de jovens atletas. Métodos: Esse artigo utiliza estatísticas matemáticas para investigar e analisar o estado do treinamento físico de jovens jogadores de basquete. Resultados: Após o estudo de vários métodos de treinamento, descobrimos que o treino de impedância pode aprimorar o preparo físico de jovens atletas. O treino físico do basquete inclui força, velocidade, resistência, sensibilidade, etc. Conclusão: Jogadores de basquete jovens precisam adotar diversos métodos de treinamento. Esportes de grupo podem, efetivamente, aprimorar o preparo físico de jovens jogadores de basquete, aumentando as chances de vitória de seus times nas partidas. Nível de evidência II; Estudos terapêuticos - investigação do resultado de tratamentos.

RESUMEN Introducción: El antagonismo está muy presente en partidos de baloncesto. Atletas en buenas condiciones físicas tendrán una gran ventaja en estos partidos. Objetivos: Estudiar la influencia de la actividad física en la preparación física de jóvenes atletas. Métodos: Este artículo utiliza estadísticas matemáticas para investigar y analizar el estado del entrenamiento físico de jóvenes jugadores de baloncesto. Resultados: Después del estudio de varios métodos de entrenamiento, descubrimos que el entrenamiento de impedancia puede mejorar la preparación física de jóvenes atletas. El entrenamiento físico del baloncesto incluye fuerza, velocidad, resistencia, sensibilidad, etc. Conclusión: Jugadores de baloncesto jóvenes necesitan adoptar diversos métodos de entrenamiento. Deportes de grupo pueden, efectivamente, mejorar la preparación física de jóvenes jugadores de baloncesto, aumentando las oportunidades de victoria de sus equipos en los partidos. Nivel de evidencia II; Estudios terapéuticos - investigación del resultado de tratamientos.

Isolation, genotyping and virulence determination of a Toxoplasma gondii strain from non-human primate from China.

Toxoplasma gondii infects almost all warm-blooded animals, including humans and non-human primates. Many cases of T. gondii infection in non-human primates have been reported worldwide. In this study, 15 monkeys were collected from zoos in Henan Province between 2016 and 2019. A modified agglutination test (MAT) (cut-off: 1:8) showed that 46.7% (7/15) of the heart juices had T. gondii IgG antibody transformation. One viable T. gondii strain was successfully isolated from the myocardium of a rhesus monkey by bioassay in mice. This strain was designated as TgMonkeyCHn1. The DNA of T. gondii tachyzoites was obtained using cell cultures, and the genotype of this strain was determined by PCR-RFLP with 10 markers and the virulence genes ROP5 and ROP18. The genotype and ROP18/ROP5 (3/6) of TgMonkeyCHn1 did not match any known genotypes. In addition, the TgMonkeyCHn1 formed low number of tissue cysts and was non-lethal to mice. To our knowledge, this is the first T. gondii strain isolated from Old World monkeys. Rhesus monkey is a new host record for T. gondii.

Asymmetric Total Synthesis of (+)-Quinocarcinamide.

The first asymmetric total synthesis of (+)-quinocarcinamide (3), an enantiomer of the natural oxidation product from antitumor antibiotic (-)-quinocarcin (1), is described. Key steps include an iridium-catalyzed asymmetric allylic amidation of racemic alcohol 9, olefin cross-metathesis followed by a SN2' to forge tetrahydroisoquinoline, and stereocontrolled 1,3-dipolar cycloaddition between a facilely generated azomethine ylide and tert-butyl acrylate to construct the diazabicyclo[3.2.1]octane ring.

Comparative Immunological Response and Pathobiology of Mice Inoculated with Toxoplasma gondii Isolated from Different Hosts.

Toxoplasma gondii is an obligate intracellular parasite that has a worldwide distribution and can infect almost all warm-blood animals. Serological tests are the main detection methods for T. gondii infection in animals and humans. Little is known of biological behavior, antibody responses, and virulence of T. gondii strains in mice from China. Here we document antibody responses, tissue cyst burden, and mouse virulence of T. gondii strains isolated from different hosts in China. All T. gondii strains formed tissue cysts in the brains of mice and positively correlated with the T. gondii antibody titer (R2 = 0.3345). These results should aid in the diagnosis and characterization of T. gondii isolates.

Morinda officinalis polysaccharides improve meat quality by reducing oxidative damage in chickens suffering from tibial dyschondroplasia.

Tibial dyschondroplasia (TD) is the common leg disease in commercial broilers. However, the effects of TD on meat quality and the protective of Morinda officinalis polysaccharide (MOP) are largely unknown. Three hundred broiler chicks (one-day-old) were equally allocated into control (CON), TD and MOP-treated groups for 15 days. The results indicated that TD influenced morphology and meat quality-related parameters of the breast muscle, and changed the activity and mRNA expression of antioxidant enzymes in plasma and breast muscles. Moreover, metabolomics profiling of breast muscle revealed that the main altered metabolites 4-guanidinobutyric acid and chenodeoxycholic acid, which are related to meat quality and oxidative stress. Additionally, 500 mg/L MOP effectively restored the content of meat metabolites and oxidative damage. These findings suggest that oxidative damage caused by TD may affect meat quality in broilers by changing the content of breast muscle metabolites and that MOP supplementation has a restorative effect.

Isolation and characterization of Toxoplasma gondii from captive caracals (Caracal caracal).

Toxoplasma gondii infects most warm-blooded animals, including humans. Felids can serve as both intermediate and definitive hosts for T. gondii. However, there is no direct evidence to prove the caracal (Caracal caracal) is an intermediate host for T. gondii. Here, we report T. gondii infection in two caracals in a zoo from China. Antibodies against T. gondii were found in both caracals by modified agglutination test (MAT) (cut-off titer: 1:25). Tissue cysts were observed in the leg and tongue muscles of caracal case# 1. These cysts were confirmed as T. gondii by immunohistochemical staining and T. gondii was detected by polymerase chain reaction (PCR). Viable T. gondii strain was isolated from the striated muscles of caracal case# 2 and designated as TgCaracalCHn1. DNA from tachyzoites obtained from cell cultures was characterized by RFLP-PCR using ten markers (SAG1, SAG3, SAG2, BTUB, c22-8, GRA6, c29-2, PK1, L358, and Apico) and the virulence genes (ROP5 and ROP18). The results indicate that this isolate belongs to ToxoDB genotye #2 (Type III). The virulence of this isolate was evaluated in BALB/c mice. A dose of 104 TgCaracalCHn1 tachyzoites was non-lethal to mice. Tissue cysts were found in brain tissues of infected mice. This result confirmed that the TgCaracalCHn1 is non-virulent to mice. Current study documents first isolation of viable T. gondii strain from caracal and also indicates that caracal can act as new intermediate host for T. gondii.

Construction and immunogenicity of a gE/gI/TK-deleted PRV based on porcine pseudorabies virus variant.

Pseudorabies (PR) caused by re-emerging pseudorabies virus (PRV) variant has outbroken among PRV vaccine-immunized swine herds on many Chinese pig farms, with severe socioeconomic consequences since late 2011. Here, a gE/gI/TK-deleted recombinant virus (rPRV NY-gE-/gI-/TK-) was constructed based on PRV NY strain from 2012 through homologous DNA recombination and gene-editing technology termed clustered regularly interspaced palindromic repeats (CRISPR)/associated (Cas9) system. The rPRV NY-gE-/gI-/TK- strain showed similar growth kinetics to the parental PRV NY strain in vitro, and was safe for mice. Sixty mice were injected subcutaneously (s.c.) twice with 106.0 TCID50 of rPRV NY-gE-/gI-/TK- and DMEM, respectively, with two-week interval. The levels of PRV gB antibodies and neutralizing antibodies against PRV NY in mice immunized with rPRV NY-gE-/gI-/TK- were higher than those in the DMEM control group. The number of T lymphocyte subclasses CD3+, CD4+ and CD8+ in rPRV NY-gE-/gI-/TK--immunized mice was higher than that in DMEM-injected mice. After challenge with 106.0 TCID50 PRV NY at 42 dpi, all rPRV NY-gE-/gI-/TK--immunized mice survived without exhibiting any pathological lesions in different tissues and intranuclear eosinophilic inclusions of the brain, and the viral genomic copy numbers in various organs of mice were obviously lower than DMEM group. These results showed the rPRV NY-gE-/gI-/TK- could be a promising next-generation vaccine to control now epidemic PR in China.

Isolation and pathogenicity testing of avian reticuloendotheliosis virus from layer chickens in China.

Reticuloendotheliosis virus (REV) can cause runting, immunosuppression, acute reticulum cell neoplasia, and chronic lymphoid tumors in a variety of domestic and wild birds. We diagnosed a case of reticuloendotheliosis with obvious tumors in liver and kidney. We isolated and sequenced the virus and performed pathogenicity testing of the REV strain. Immunohistochemistry and PCR confirmed that the diseased layer chickens were infected with REV. The strain, named BJ1503, was successfully isolated from the case by inoculation of tissue homogenates onto chicken embryo fibroblasts. The length of the proviral REV genome is 8,293 nucleotides. The isolate had 99.7% identity with REV-HA9901 (AY842951.1), which was isolated from Jiangsu, China, in 1999. The chickens infected with REV-BJ1503 had depressed weight gain and lymphoid atrophy. Our findings suggest that REV isolate BJ1503 was phylogenetically close to the earlier strain found in China, with minor variations, and the virus was associated with severe production problems.

Toxoplasma gondii in lambs of China: Heart juice serology, isolation and genotyping.

Toxoplasmosis is one of the most common foodborne diseases in the world. The objective of this study was to determine Toxoplasma gondii infection in lambs from Henan province, China. A total of 166 lamb hearts were collected from 2017 to 2019. T. gondii infection was determined by the Modified Agglutination Test (MAT) using heart juice of lambs. 11 isolates (TgSheepCHn3 - TgSheepCHn13) were obtained from samples with MAT titers ≥1:100. The rate of T. gondii isolation increased with antibody titer against T. gondii (P < 0.05). No isolate was obtained from samples with titer 1:25 and 1:50, suggesting the cut-off titer for MAT is better set at 1:100. With cut-off value of 1:100, IgG antibodies to T. gondii were found in 25.3% (42/166) of the lambs by MAT. T. gondii parasite was not found in IHC and HE-stained tissue sections of lamb hearts (0/166). Sixty-seven heart tissues with ≥1:25 MAT titers were subjected to acid pepsin digestion and detected T. gondii by PCR. Only 7.5% (5/67) of DNA amplified products were found in heart tissues by the primer TOX5/TOX8. Brain tissue cysts were observed in all mice infected with the 11 isolates at day 60 post infection, suggesting these isolates are non-lethal to mice. PCR-RFLP analysis revealed that 7 isolates belonged to ToxoDB#2, 4 isolates belonged to ToxoDB#4. This is the first isolation of ToxoDB#2 and ToxoDB#4 from lambs in China. Interestingly, none of these isolates belongs to the ToxoDB#9 that is common in China. Our results suggest that the genetic diversity and population structure of T. gondii from China maybe more abundant and magical than previous speculation.

Can Prohibitin 1 be a Safeguard against liver disease?

Prohibitin (PHB) 1 is involved in multiple regulatory pathways in liver disease to protect hepatocytes, and its function is associated with subcellular localization. PHB1 located in the nucleus, cytoplasm and the mitochondrial inner membrane has anti-oxidative stress and anti-inflammatory effects in hepatitis and cirrhosis, which can protect liver cells from damage caused by inflammatory factors and reactive oxygen species (ROS) stimulation. The low expression of PHB1 located in the nucleus of liver cancer cells inhibits the proliferation and metastasis of liver cancer; thus, PHB1 exhibits the function of a tumor suppressor gene. Understanding the mechanisms of PHB1 in liver diseases may be useful for further research on the disease and may provide new ideas for the development of targeted therapeutic drugs in the future. Therefore, this review puts forward an overview of the role of PHB1 and its protective mechanism in liver diseases.

Evidence of red panda as an intermediate host of Toxoplasma gondii and Sarcocystis species.

Toxoplasma gondii has been found to infect almost all warm-blooded animals; however, some hosts lack direct evidence of T. gondii infection. The red panda (Ailurus fulgens) is an endangered species that mainly lives in temperate forests of South Asia. Here, T. gondii infection in red pandas from zoos in China were reported. Antibodies to T. gondii were found in 14.3% (2/14) of red pandas via the modified agglutination test (MAT) with a cut-off titer of 1:25. One viable T. gondii strain was isolated from tissues of red panda and designated as TgRedpandaCHn1. DNA from tachyzoites obtained from cell culture was characterized by PCR-RFLP with 10 markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) and virulence genes of ROP5 and ROP18. The results indicate that this isolate belonged to ToxoDB genotype #20. The ROP18/ROP5 genotype combination predicated that this strain is non-lethal to mice, which is supported by the infection in mice. T. gondii tissue cysts were readily formed and mice survived. Tissue cysts observed in the histopathological sections of the tongue and diaphragm of one red panda were speculated as sarcocysts, but not T. gondii base on morphological characteristics. To our knowledge, this study is the first to report on the isolation of T. gondii from red panda. Additionally, this report provides direct evidence of red panda as an intermediate host of T. gondii and Sarcocystis species.