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Background Bacterial blight of cotton (BBC), which is caused by the bacterium Xanthomonas citri pv. malvacearum ( Xcm ), is a destructive disease in cotton. Transcription activator-like effectors (TALEs), encoded by tal -genes, play critical roles in the pathogenesis of xanthomonads. Characterized strains of cotton pathogenic Xcm harbor 8–12 different tal genes and only one of them is functionally decoded. Further identification of novel tal genes in Xcm strains with virulence contributions are prerequisite to decipher the Xcm -cotton interactions. Results In this study, we identified six tal genes in Xss-V 2 –18, a highly-virulent strain of Xcm from China, and assessed their role in BBC. RFLP-based Southern hybridization assays indicated that Xss-V 2 –18 harbors the six tal genes on a plasmid. The plasmid-encoded tal genes were isolated by cloning Bam HI fragments and screening clones by colony hybridization. The tal genes were sequenced by inserting a Tn 5 transposon in the DNA encoding the central repeat region (CRR) of each tal gene. Xcm TALome evolutionary relationship based on TALEs CRR revealed relatedness of Xss-V 2 –18 to MSCT1 and MS14003 from the United States. However, Tal2 of Xss-V 2 –18 differs at two repeat variable diresidues (RVDs) from Tal6 and Tal26 in MSCT1 and MS14003, respectively, inferred functional dissimilarity. The suicide vector pKMS1 was then used to construct tal deletion mutants in Xcm Xss-V 2 –18. The mutants were evaluated for pathogenicity in cotton based on symptomology and growth in planta . Four mutants showed attenuated virulence and all contained mutations in tal2 . One tal2 mutant designated M2 was further investigated in complementation assays. When tal2 was introduced into Xcm M2 and expressed in trans , the mutant was complemented for both symptoms and growth in planta , thus indicating that tal2 functions as a virulence factor in Xcm Xss-V 2 –18. Conclusions Overall, the results demonstrated that Tal2 is a major pathogenicity factor in Xcm strain Xss-V 2 –18 that contributes significantly in BBC. This study provides a foundation for future efforts aimed at identifying susceptibility genes in cotton that are targeted by Tal2.

Bacterial blight of cotton (BBC) caused by Xanthomonas citri pv. malvacearum (Xcm) is an important and destructive disease affecting cotton plants. Transcription activator‐like effectors (TALEs) released by the pathogen regulate cotton resistance to the susceptibility. In this study, we sequenced the whole genome of Xcm Xss‐V2‐18 and identified eight tal genes: seven on the plasmids and one on the chromosome. Deletion and complementation experiments of Xss‐V2‐18 tal genes demonstrated that Tal1b is required for full virulence on cotton. Transcriptome profiling coupled with TALE‐binding element prediction revealed that Tal1b targets GhSWEET14A04/D04 and GhSWEET14D02 simultaneously. Expression analysis confirmed the independent inducibility of GhSWEET14A04/D04 and GhSWEET14D02 by Tal1b, whereas GhSWEET14A04/D04 is additionally targeted by Tal1. Moreover, β‐glucuronidase and Xa10‐mediated hypersensitive response assays indicated that the effector‐binding element (EBEs) are required for the direct and specific activation of the candidate targets by Tal1 and Ta1b. These insights enhance our understanding of the underlying mechanisms of bacterial blight in cotton and might lead to improved resistance through EBEs disruption or a TALE‐trap strategy. Tal1b and Tal1 effectors of Xanthomonas citri pv. malvacearum activate GhSWEET14 genes to promote water soaking in cotton.

Copper (Cu) is listed as one of the main heavy metal pollutants, which poses potential health risks to humans. Excessive intake of Cu has shown toxic effects on the organs of many animals, and the liver is one of the most important organs to metabolize it. In this study, pigs, the mammal with similar metabolic characteristics to humans, were selected to assess the effects of long-term exposure to Cu on mitochondria-mediated apoptosis, which are of great significance for studying the toxicity of Cu to humans. Pigs were fed a diet with different contents of Cu (10, 125, and 250 mg/kg) for 80 days. Samples of blood and liver tissue were collected on days 40 and 80. Experimental results demonstrated that the accumulation of Cu in the liver was increased in a dose-dependent and time-dependent manner. Meanwhile, the curve of pig’s body weight showed that a 125 mg/kg Cu diet promoted the growth of pigs during the first 40 days and then inhibited it from 40 to 80 days, while the 250 mg/kg Cu diet inhibited the growth of pigs during 80 days of feeding. Additionally, the genes and protein expression levels of Caspase-3, p53, Bax, Bak1, Bid, Bad, CytC, and Drp1 in the treatment group were higher than that in the control group, while Bcl-2, Bcl-xL, Opa1, Mfn1, and Mfn2 were decreased. In conclusion, these results indicated that long-term excessive intake of Cu could inhibit the growth of pigs and induced mitochondria-mediated apoptosis by breaking the mitochondrial dynamic balance. Graphical abstract Synopsis: Long-term exposure to high doses of Cu could lead to mitochondrial dysfunction by breaking the mitochondrial dynamic balance, which ultimately induced mitochondria-mediated apoptosis in the liver of pigs. This might be closely related to the growth inhibition and liver damage in pigs.

Bacterial blight of cotton (BBC), which is caused by the bacterium Xanthomonas citri pv. malvacearum (Xcm), is a destructive disease in cotton. Transcription activator-like effectors (TALEs), encoded by tal-genes, play critical roles in the pathogenesis of xanthomonads. Characterized strains of cotton pathogenic Xcm harbor 8-12 different tal genes and only one of them is functionally decoded. Further identification of novel tal genes in Xcm strains with virulence contributions are prerequisite to decipher the Xcm-cotton interactions. In this study, we identified six tal genes in Xss-V -18, a highly-virulent strain of Xcm from China, and assessed their role in BBC. RFLP-based Southern hybridization assays indicated that Xss-V -18 harbors the six tal genes on a plasmid. The plasmid-encoded tal genes were isolated by cloning BamHI fragments and screening clones by colony hybridization. The tal genes were sequenced by inserting a Tn5 transposon in the DNA encoding the central repeat region (CRR) of each tal gene. Xcm TALome evolutionary relationship based on TALEs CRR revealed relatedness of Xss-V -18 to MSCT1 and MS14003 from the United States. However, Tal2 of Xss-V -18 differs at two repeat variable diresidues (RVDs) from Tal6 and Tal26 in MSCT1 and MS14003, respectively, inferred functional dissimilarity. The suicide vector pKMS1 was then used to construct tal deletion mutants in Xcm Xss-V -18. The mutants were evaluated for pathogenicity in cotton based on symptomology and growth in planta. Four mutants showed attenuated virulence and all contained mutations in tal2. One tal2 mutant designated M2 was further investigated in complementation assays. When tal2 was introduced into Xcm M2 and expressed in trans, the mutant was complemented for both symptoms and growth in planta, thus indicating that tal2 functions as a virulence factor in Xcm Xss-V -18. Overall, the results demonstrated that Tal2 is a major pathogenicity factor in Xcm strain Xss-V -18 that contributes significantly in BBC. This study provides a foundation for future efforts aimed at identifying susceptibility genes in cotton that are targeted by Tal2.

ABSTRACT The 25-hydroxyvitamin D [25(OH)D] level in serum is currently considered the best indicator of vitamin D supply to the body from cutaneous synthesis and nutritional intake. Few reports are available regarding the effects of vitamin D and its metabolites on heart development in swine. The effect of 25(OH)D3 and Ca2+ supplementation in diets on heart development in neonatal piglets during pregnancy were examined in this study. Total 40 sows of 7 gestational age with good health and nutritional condition were divided into four groups (n=10): the control group, low calcium, 25(OH)D3 group and low calcium with 25(OH)D3 group. Each groups consist of 5 piglets, randomly selected and euthanized within 2 h after birth. After euthanasia, heart was collected for histopathological examination and IGF, IGF1, IGF2, IGFBP3, PCNA and ki67 expression were analyzed. The results showed that low calcium supplementation decreased the cardiac index and development of myocardial muscle fiber, while vitamins D3 can improve the decrease of cardiac index and development of myocardial fiber due to low calcium supplementation. The expression profile of IGF1, IGF2, IGFBP3, PCNA and ki67 genes were down regulated in LCa group as compared to control group; while the vitamin D3 supplementation significantly up regulated the above genes expression. In conclusion, our findings suggested that 25(OH)D3 and Ca2+ supplementation in sows' diets has an important role in the development of heart and differentiation of myocardial fibers in neonatal piglets by changing the expression of the IGF pathway and myocardial cells proliferation.

Bacterial Blight of Cotton (BBC) caused by Xanthomonas citri pv. malvacearum (Xcm) is an important and destructive disease affecting cotton plants. Transcription activator-like effectors (TALEs) released by the pathogen regulate cotton resistance to the susceptibility. In this study, we sequenced the whole genome of Xcm Xss-V2-18 and identified eight tal genes; seven on the plasmids and one on the chromosome. Deletion and complementation experiments of Xss-V2-18 tal genes demonstrated that Tal1b is required for full virulence on cotton. Transcriptome profiling coupled with TALE-binding element prediction revealed that Tal1b targets GhSWEET15A04/D04 and GhSWEET15D02 simultaneously. Expression analysis confirmed the independent inducibility of GhSWEET15A04/D04 and GhSWEET15D02 by Tal1b, whereas GhSWEET15A04/D04 is additionally targeted by Tal1. Moreover, GUS (β-glucuronidase) and Xa10-mediated HR (hypersensitive response) assays indicated that the EBEs are required for the direct and specific activation of the candidate targets by Tal1 and Ta1b. These findings may advance our understanding of the dynamics between TALEs and EBEs, and decipher a simple and effective DNA-binding mechanism that could lead to the development of more efficient methods for gene editing and transgenic research.

Background Drug closed-loop management reflects the level of hospital management and pharmacist service. It is a challenge for hospital pharmacists to realize the whole-process closed-loop management of drugs in hospital pharmacies. Therefore, this study aimed to evaluate the operational effect of using mobile technology to build a closed-loop drug management system. Methods Using mobile technology, replacing the traditional paper dispensing model and constructing a multinode information collection system according to the Healthcare Information and Management Systems Society Standard, we reformed the hospital information system and inpatient pharmacy workflow and then evaluated the new approach using statistical methods. Results After the transformation, the entire process of drug data can be traced. Closed-loop management, as well as real-time data verification and control, thereby improves the work efficiency and reduces the drug dispensing time. By reducing the work error rate, the number of dispensing errors decreased from 5 to 1 case/month. The comprehensive dispensing process can achieve the whole workflow of paperless operation and reduce the use of paper A4 by 180,000 pieces per year. Conclusions Mobile technology can improve the service level of pharmacies, enhance the level of drug management and hospital quality management, ensure the safety of medication for inpatients, and significantly reduce the amount of paper used.

Background Antimicrobial Stewardship Pharmacy Consultation (ASPC) in China has been shown to reduce patients' length of stay (LOS). However, prolonged LOS remains a challenge, resulting in unnecessary psychological and financial burden for patients. Objective This study aimed to develop a prediction model using ASPC parameters to identify high-risk pediatric patients with infectious diseases. These patients received ASPC interventions but still experienced prolonged LOS, which defined their high-risk status. Methods Predictors for the ASPC model were selected using lasso regression, a nomogram was developed using multivariate logistic regression, and internal validation was performed using tenfold cross-validation. The data set consisted of 474 electronic medical records of pediatric patients with infectious diseases from two hospitals. LOS was dichotomized at the median, and patients with LOS greater than the median were considered to have achieved the outcome. Results The proportion of outcome events was set at 50% by design. Five independent predictors were identified in the ASPC model: (1) the suggestions from the crucial consultation (OR: 1.74; 95% CI: 1.10 to 2.74), (2) weight (OR: 0.98; 95% CI: 0.97 to 1.00), (3) whether the patient received first aid (OR: 0.54; 95% CI: 0.3 to 1.00), (4) the aim of the crucial consultation (OR: 0.15; 95% CI: 0.03 to 0.66), and (5) whether the patient was critically ill (OR: 0.22; 95% CI: 0.12 to 0.41). The ASPC model showed good discrimination with a C-statistic of 0.772 (95% CI: 0.748 to 0.797) and good calibration performance with intercept and slope values of 0.00 (95% CI: -0.12 to 0.12) and 0.93 (95% CI: 0.82 to 1.04), respectively, under tenfold cross-validation. Conclusions The antimicrobial stewardship pharmacy consultation model has good discrimination and calibration, and effectively identifies patients at risk for prolonged length of stay. Key messages Extensive research on clinical pharmacy practice has highlighted the important role that clinical pharmacists play in patient care and their valuable contributions to personalized medical treatment. However, the quality of pharmacy consultations in China still requires improvement to gain wider recognition from clinicians. Our study examined the impact of Antimicrobial Stewardship Pharmacy Consultations (ASPCs) on the length of stay (LOS) among pediatric patients with infectious diseases and developed a risk prediction model to identify the high-risk patients who received ASPC interventions yet still experienced prolonged LOS. This model has the potential to enhance the quality of ASPC, reduce patients' psychological and financial burdens, and support the advancement of clinical pharmacy practice in China.