To perform the phylogenetic characterization of an isolated porcine rotavirus(PoRV)and investigate its pathogenicity in suckling mice and piglets.A G4P[23]genotype PoRV strain JSJR2023 was successfully isolated from the diarrheic piglet feces through propagation in MA104 cells.The viral proliferation kinetics were analyzed using TCID50 assays,followed by complete genome sequencing through Sanger sequencing platforms.Comprehensive genotyping and phylogenetic reconstruction were conducted using MEGA7.0 with maximum likelihood algorithms.Pathogenicity was assessed in the following animal models:5-day-old C57BL/6 mice and 3-day-old piglets.Multidimensional evaluation included clinical monitoring(diarrhea scoring,growth parameters),virological detection,and histopathological analysis of intestinal tissues.The virus strain JSJR2023 could replicate efficiently in MA104 cells,achieving peak titers of 107.5 TCID50/mL.Whole genome genotype analysis showed that the strain belonged to G4-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1.Phylogenetic analysis indicated that the VP3 and NSP4 genes of JSJR2023 strain were most closedrelated to human species rotaviruses,suggesting genetic reassortment between human and porcine RV strains.The animal experiments in suckling mice showed that the JSJR2023 strain infection caused diarrhea symptoms,intestinal edema and congestion,and shedding of intestinal villus epithelial cells.The pathogenicity experiments in piglets showed that compared with the control group,the challenged group of pig-lets had severe diarrhea symptoms,accompanied by reduced appetite and listlessness.Post-mortem examination revealed that the intes-tines were significantly thinner,congested,and filled with yellow watery contents.The challenged piglets showed typical pathological changes such as thinning of the intestinal wall and shortening and shedding of intestinal villi.In conclusion,this study successfully iso-lated a human-porcine recombinant G4P[23]PoRV strain and established the infection models in suckling mice and piglets,providing important tools for investigating the pathogenic mechanism of PoRV,evaluating vaccines and developing antiviral drug.

To perform the phylogenetic characterization of an isolated porcine rotavirus(PoRV)and investigate its pathogenicity in suckling mice and piglets.A G4P[23]genotype PoRV strain JSJR2023 was successfully isolated from the diarrheic piglet feces through propagation in MA104 cells.The viral proliferation kinetics were analyzed using TCID50 assays,followed by complete genome sequencing through Sanger sequencing platforms.Comprehensive genotyping and phylogenetic reconstruction were conducted using MEGA7.0 with maximum likelihood algorithms.Pathogenicity was assessed in the following animal models:5-day-old C57BL/6 mice and 3-day-old piglets.Multidimensional evaluation included clinical monitoring(diarrhea scoring,growth parameters),virological detection,and histopathological analysis of intestinal tissues.The virus strain JSJR2023 could replicate efficiently in MA104 cells,achieving peak titers of 107.5 TCID50/mL.Whole genome genotype analysis showed that the strain belonged to G4-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1.Phylogenetic analysis indicated that the VP3 and NSP4 genes of JSJR2023 strain were most closedrelated to human species rotaviruses,suggesting genetic reassortment between human and porcine RV strains.The animal experiments in suckling mice showed that the JSJR2023 strain infection caused diarrhea symptoms,intestinal edema and congestion,and shedding of intestinal villus epithelial cells.The pathogenicity experiments in piglets showed that compared with the control group,the challenged group of pig-lets had severe diarrhea symptoms,accompanied by reduced appetite and listlessness.Post-mortem examination revealed that the intes-tines were significantly thinner,congested,and filled with yellow watery contents.The challenged piglets showed typical pathological changes such as thinning of the intestinal wall and shortening and shedding of intestinal villi.In conclusion,this study successfully iso-lated a human-porcine recombinant G4P[23]PoRV strain and established the infection models in suckling mice and piglets,providing important tools for investigating the pathogenic mechanism of PoRV,evaluating vaccines and developing antiviral drug.

ObjectiveTo explore the mechanism of Bushen Huoxue enema in treating the rat model of kidney deficiency and blood stasis-thin endometrium （KDBS-TE） by transcriptome sequencing. MethodThe rat model of KDBS-TE was established by administration of tripterygium polyglycosides tablets combined with subcutaneous injection of adrenaline. The pathological changes of rat endometrium in each group were then observed. Three uterine tissue specimens from each of the blank group， model group， and Bushen Huoxue enema group were randomly selected for transcriptome sequencing. The differentially expressed circRNAs， lncRNAs， and miRNAs were screened， and the disease-related specific competitive endogenous RNA （ceRNA） regulatory network was constructed. Furthermore， the gene ontology （GO） functional annotation and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment were performed for the mRNAs in the network. ResultCompared with the blank group， the model group showed endometrial dysplasia， decreased endometrial thickness and endometrial/total uterine wall thickness ratio （P<0.01）， and differential expression of 18 circRNAs， 410 lncRNAs， and 7 miRNAs. Compared with the model group， the enema and estradiol valerate groups showed improved endometrial morphology and increased endometrial thickness and ratio of endometrial to total uterine wall thickness （P<0.05）. In addition， 21 circRNAs， 518 lncRNAs， and 17 miRNAs were differentially expressed in the enema group. The disease-related specific circRNA-miRNA-mRNA regulatory network composed of 629 nodes and 664 edges contained 2 circRNAs， 34 miRNAs， and 593 mRNAs. The lncRNA-miRNA-mRNA regulatory network composed of 180 nodes and 212 edges contained 5 lncRNAs， 10 miRNAs， and 164 mRNAs. The mNRAs were mainly enriched in Hippo signaling pathway， autophagy-animal， axon guidance， etc. ConclusionBushen Huoxue enema can treat KDBS-TE in rats by regulating specific circRNAs， lncRNAs， and miRNAs in the uterus and the ceRNA network.

Background::The correlation between metals and hypertension, such as sodium, zinc, potassium, and magnesium, has been confirmed, while the relationship between aluminum and hypertension is not very clear. This study aimed to evaluate the correlation between plasma aluminum and hypertension in electrolytic aluminum workers by the Bayesian networks (BN).Methods::In 2019, 476 male workers in an aluminum factory were investigated. The plasma aluminum concentration of workers was measured by inductively coupled plasma mass spectrometry. The influencing factors on the prevalence of hypertension were analyzed by the BN.Results::The prevalence of hypertension was 23.9% in 476 male workers. The risk of hypertension from plasma aluminum in the Q2, Q3, and Q4 groups was 5.20 (1.90-14.25), 6.92 (2.51-19.08), and 7.33 (2.69-20.01), respectively, compared with that in the Q1 group. The risk of hypertension from the duration of exposure to aluminum of >10 years was 2.23 (1.09-4.57), compared without aluminum exposure. Area under the curve was 0.80 of plasma aluminum and the duration of exposure to aluminum was based on covariates, indicating that aluminum exposure had important predictive value in the prevalence of hypertension in the occupational population. The results of the study using the BN model showed that if the plasma aluminum of all participants was higher than Q4 (≥47.86 μg/L) and the participants were drinking, smoking, diabetes, central obesity, dyslipidemia, and aged >50 years, the proportion of hypertension was 71.2%.Conclusions::The prevalence of hypertension increased significantly with the increase of plasma aluminum level.

Background::The correlation between metals and hypertension, such as sodium, zinc, potassium, and magnesium, has been confirmed, while the relationship between aluminum and hypertension is not very clear. This study aimed to evaluate the correlation between plasma aluminum and hypertension in electrolytic aluminum workers by the Bayesian networks (BN).Methods::In 2019, 476 male workers in an aluminum factory were investigated. The plasma aluminum concentration of workers was measured by inductively coupled plasma mass spectrometry. The influencing factors on the prevalence of hypertension were analyzed by the BN.Results::The prevalence of hypertension was 23.9% in 476 male workers. The risk of hypertension from plasma aluminum in the Q2, Q3, and Q4 groups was 5.20 (1.90-14.25), 6.92 (2.51-19.08), and 7.33 (2.69-20.01), respectively, compared with that in the Q1 group. The risk of hypertension from the duration of exposure to aluminum of >10 years was 2.23 (1.09-4.57), compared without aluminum exposure. Area under the curve was 0.80 of plasma aluminum and the duration of exposure to aluminum was based on covariates, indicating that aluminum exposure had important predictive value in the prevalence of hypertension in the occupational population. The results of the study using the BN model showed that if the plasma aluminum of all participants was higher than Q4 (≥47.86 μg/L) and the participants were drinking, smoking, diabetes, central obesity, dyslipidemia, and aged >50 years, the proportion of hypertension was 71.2%.Conclusions::The prevalence of hypertension increased significantly with the increase of plasma aluminum level.

Objective:To compare dosimetric and radiobiological parameters between automatic and manual uARC plans in the treatment of esophageal cancer patients, aiming to provide reference for clinical application.Methods:High-quality uARC plans of 100 patients with esophageal cancer were selected, and the mean values of the dosimetric parameters in the target area and organs at risk (OAR) were counted, and the goal table of uRT-TPOIS intelligent plan was established. Automatic and manual uARC plans were generated with UIH (United Imaging) treatment planning system (TPS) for 21 esophageal cancer patients. The differences in mean dose (D mean), approximate minimum (D 98%) and maximum (D 2%) dose of planning target volume (PTV), homogeneity index (HI) and conformity index (CI), dose of OAR, mean planning time, monitor unit (MU), tumor control probability (TCP) and normal tissue complication probability (NTCP) were compared between automatic and manual uARC plans. Normally distributed data between two groups were compared by paired t-test, and non-normally distributed data were assessed by nonparametric Wilcoxon test. Results:The D 98% (PTV 60 Gy: P<0.001, PTV 54 Gy: P=0.001) , CI (PTV 60 Gy: P<0.001, PTV 54 Gy: P=0.002) and target volume of area covered by prescription dose (V 54 Gy: P<0.001) of the automatic uARC plans were better than those of manual uARC plans (all P<0.05). There was no significant difference in D mean or HI between the two plans [PTV 54 Gy (59.32±1.87) Gy vs. (59.13±1.64) Gy, (0.19±0.02) vs. (0.18±0.02), all P>0.05]. The D mean and D max of spinal cord of the automatic plan were better than those of the manual plan [(13.22±4.27) Gy vs. (13.75±4.44) Gy, P=0.020 and (36.99±1.67) Gy vs. (38.14±1.31) Gy, P=0.011]. There was no significant difference in the mean dose of V 20 Gy of the lung between two plans ( P>0.05), whereas the mean doses of V 5 Gy and V 10 Gy of the lung of the manual plan were less than those of the automatic plan ( both P<0. 001). Automatic uARC plan had a significantly shorter mean planning time than manual uARC plan [(11.79±1.71) min vs. (53.36±8.23) min, P<0.001]. MU did not significantly differ between two plans [(762.84±74.83) MU vs. (767.41±80.63) MU, P>0.05]. The TCP of the automatic plan was higher than that of the manual plan (PTV 60 Gy 89.15%±0.49% vs. 86.75%±6.46%, P=0.004 and PTV 54 Gy 79.79%±3.48% vs. 77.51%±5.04%, P=0.006). However, manual plan had a lower NTCP of the lung than automatic uARC plan (0.46%±0.40% vs. 0.35%±0.32%, P<0.001). There was no significant difference in NTCP of heart and spinal cord between two plans (all P>0.05). Conclusion:It is feasible to generate automatic uARC plan with uRT-TPOIS TPS for esophageal cancer patients, which can increase the target CI and shorten the plan design time.

Translationally controlled tumor proteins (TCTP) and SNF1- related protein kinase (SnRK1) are conserved and widely present in eukaryotic cells. TCTP regulates cell division, plant growth and development, and mediates plant resistance against pathogen infection. SnRK1 participates in a range of physiological processes including sugar metabolism and resistance to abiotic and biotic stresses. Previous work in our laboratory demonstrated that wheat TCTP can respond to Puccinia triticina infection and induce host defense responses. In order to further investigate the mechanism of TaTCTP in wheat resistance to Puccinia triticina infection, we used TAP (tandem affinity purification) and mass spectrometry to screen the potential interactants of TaTCTP. A SNF1- related protein kinase (SnRK1) was identified as a potential interacting protein of TaTCTP. The results of yeast two-hybrid assay showed that TCTP could interact with SnRK1 in yeast, and the yeast carrying TCTP and SnRK1 could grow on SD/-Leu/-Trp/-His/-Ade (SD/-LWHA) medium. The fluorescence signal of the interaction between TCTP and SnRK1 was found to be distributed in the cytoplasm in the Bi-fluorescense complementation experiment. Co-IP experiments further showed that TCTP and SnRK1 could interact in plant cells. This study lays an important foundation for further studying the mechanism of TaTCTP in the interaction between wheat and Puccinia triticina, and it play a great influence on further improving the molecular mechanism of wheat resistant to Puccinia triticina.
Basidiomycota ; Humans ; Neoplasms ; Protein Biosynthesis ; Protein-Serine-Threonine Kinases ; Triticum

Objective To discuss the correlation between shock index (SI) and severity and the values to forecast the prognosis in patients with septic shock. Methods 127 patients with septic shock admitted to intensive care unit (ICU) of Zhejiang Provincial People's Hospital from January 1st, 2016 to October 31st, 2017 were enrolled, and they were divided into survival group and death group according to the outcomes after 28-day hospitalized. The vital signs, laboratory indexes, acute physiology and chronic health evaluation Ⅱ(APACHEⅡ), sequential organ failure assessment (SOFA), lactate clearance rate (LCR) of 3 hours after fluid resuscitation, and shock index on admission (SI1) and shock index of 3 hours after fluid resuscitation (SI2) were compared between the two groups. The correlation among SI and APACHE Ⅱ, SOFA, LCR was analyzed. The receiver operating characteristic curve (ROC) was drawn to evaluate the prognostic value of SI in patients with septic shock. Results A total of 127 patients were included, 52 in survival group (40.9%) and 75 in death group (59.1%). The SI1, SI2, APACHE Ⅱ and SOFA in the death group were significantly higher than those in the survival group (SI1: 1.62±0.46 vs. 1.35±0.32, SI2: 1.36±0.24 vs. 0.93±0.15, APACHEⅡ:17.5±4.0 vs. 13.6±3.5,SOFA: 9.5±2.3 vs. 6.3±1.5), and LCR was significantly lower than that in the survival group [(14.4±5.2)% vs. (28.6±8.6)%], with statistically significant differences (all P < 0.01). The correlation analysis showed that SI1was significantly positively correlated with APACHEⅡ (r = 0.458, P = 0.000) and SOFA (r = 0.535, P = 0.000), SI2was also significantly positively correlated with APACHEⅡ (r = 0.624, P = 0.000) and SOFA (r = 0.656, P = 0.000), while SI1and SI2were significantly negatively correlated with LCR (r values were -0.348, -0.435, both P = 0.000), and the SI2were more remarkable. ROC curve analysis showed that the area under ROC curve (AUC) of SI1for predicting the prognosis of septic shock was 0.720 [95% confidence interval (95%CI) = 0.620-0.831, P < 0.05]; when SI1= 1.68, the sensitivity, specificity, Yoden index, positive predictive value and negative predictive value were 79.5%, 65.6%, 0.451, 0.759 and 0.636, respectively. The AUC of the SI2to predict prognosis of septic shock was 0.826 (95%CI =0.739-0.826, P < 0.05); when SI2= 1.37, the sensitivity, specificity, Yoden index, positive predictive value and negative predictive value were 85.7%, 87.6%, 0.733, 0.893 and 0.902, respectively. Conclusion Compared with SI1, SI2was more correlated with the severity in patients with septic shock and it had more values to predict prognosis.

It is not fully clear why there is a higher contribution of pluripotent stem cells (PSCs) to the chimera produced by injection of PSCs into 4-cell or 8-cell stage embryos compared with blastocyst injection. Here, we show that not only embryonic stem cells (ESCs) but also induced pluripotent stem cells (iPSCs) can generate F0 nearly 100% donor cell-derived mice by 4-cell stage embryo injection, and the approach has a "dose effect". Through an analysis of the PSC-secreted proteins, Activin A was found to impede epiblast (EPI) lineage development while promoting trophectoderm (TE) differentiation, resulting in replacement of the EPI lineage of host embryos with PSCs. Interestingly, the injection of ESCs into blastocysts cultured with Activin A (cultured from 4-cell stage to early blastocyst at E3.5) could increase the contribution of ESCs to the chimera. The results indicated that PSCs secrete protein Activin A to improve their EPI competency after injection into recipient embryos through influencing the development of mouse early embryos. This result is useful for optimizing the chimera production system and for a deep understanding of PSCs effects on early embryo development.
Activins ; metabolism ; Animals ; Cells, Cultured ; Embryonic Development ; Germ Layers ; metabolism ; Mice ; Pluripotent Stem Cells ; cytology ; metabolism