Bioinformatics-based design of a fusion vaccine with CTLA-4 variable region to combat Brucella

Brucellosis has become a global zoonotic disease, seriously endangering the health of people all over the world. Vaccination is an effective strategy for protection against Brucella infection in livestock in developed countries. However, current vaccines are pathogenic to humans and pregnant animals, which limits their use. Therefore, it is very important to improve the safety and immune protection of Brucella vaccine. In this study, different bioinformatics approaches were carried out to predict the physicochemical properties, T/B epitope, and tertiary structure of Omp2b and Omp31. Then, these two proteins were sequentially linked, and the Cytotoxic T lymphocyte associated antigen-4 (CTLA-4) variable region was fused to the N-terminal of the epitope sequence. In addition, molecular docking was performed to show that the structure of the fusion protein vaccine had strong affinity with B7 (B7-1, B7-2). This study showed that the designed vaccine containing CTLA-4 had high potency against Brucella, which could provide a reference for the future development of efficient brucellosis vaccines.

Impact of inorganic salts on vase life and postharvest qualities of the cut flower of Perpetual Carnation / Investigar os efeitos dos sais inorgânicos na vida dos vasos e nas qualidades pós-colheita da flor de corte do Cravo Perpétuo

Abstract This study was carried out in the laboratory of Shangqiu Institute of Technology, Henan to investigate the effect of a different combination of inorganic salt on the quality and physiological characteristics of cut flowers (CFs) of Perpetual Carnation. Furthermore, to find out the best preservation solution of inorganic salt that can enhance the ornamental value of CFs of Carnation and prolong its vase life. Sucrose, 8-hydroxyquinoline, paclobutrazol, salicylic acid and different kinds of inorganic salts were added as a preservation solution. And the same amount of distilled water was used as control. The effects of these various inorganic salts on the morphological characteristics including vase life, changes in flower stems, fresh weight (FW) and water balance and the physiological characteristics including contents of malondialdehyde (MDA), cell membrane permeability and the contents of proline of carnation were investigated. The CFs placed in vase solution with inorganic salts showed significant changes in its morphology and physiological characteristics as compared to control. The changes in flower diameter (FD), FW, malondialdehyde and cell membrane permeability showed an increasing trend first and then decreasing. The value of water balance was observed with a downward trend. However, the vase life, FD, the contents of malondialdehyde, contents of proline and FW of CFs held in the preservative solution containing inorganic salts were increased than that of control. The fresh preservative solution contained sucrose 3% + 8-hydroxyquinoline (8-HQ) (200 mg·L‾1) + paclobutrazol (100 mg·L‾1) + salicylic acid (SA) (25 mg·L‾1) + CaCl2 (100 mg·L‾1) has the best effect on longevity (34 days), FW and FD of carnation CFs. This solution has improved the ornamental and physiological characteristics of fresh carnation CFs.

Resumo Este estudo foi realizado no laboratório do Instituto de Tecnologia de Shangqiu, Henan, para investigar o efeito de diferentes combinações de sal inorgânico na qualidade e características fisiológicas de flores cortadas do Cravo Perpétuo. Além disso, para descobrir a melhor solução de preservação de sal inorgânico que pode aumentar o valor ornamental das flores cortadas de Cravo e prolongar a vida do vaso. Sucrose, 8-hidroxiquinolina, paclobutrazol, ácido salicílico e diferentes tipos de sais inorgânicos foram adicionados como uma solução de preservação. E a mesma quantidade de água destilada foi usada como controle. Os efeitos destes vários sais inorgânicos sobre as características morfológicas incluindo a vida dos vasos, alterações nos caules, peso fresco e balanço hídrico e as características fisiológicas incluindo conteúdo de malondialdeído (MDA), permeabilidade da membrana celular e conteúdo de prolina de cravo foram investigados. As flores de corte colocadas em solução de vaso com sais inorgânicos apresentaram mudanças significativas em sua morfologia e características fisiológicas em relação ao controle. As alterações no diâmetro das flores, no peso fresco, no malondialdeído e na permeabilidade da membrana celular mostraram uma tendência crescente primeiro e depois decrescendo. O valor do balanço hídrico foi observado com tendência de queda. No entanto, a vida útil do vaso, o diâmetro da flor, o conteúdo de malondialdeído, o conteúdo de prolina e o peso fresco de flores cortadas mantidos na solução preservativa contendo sais inorgânicos foram aumentados em relação ao controle. A solução conservante fresca continha sacarose 3% + 8-hidroxiquinolina (8-HQ) (200 mg·L‾1) + paclobutrazol (100 mg·L‾1) + ácido salicílico (SA) (25 mg·L‾1) + CaCl2 (100 mg·L‾1) tem o melhor efeito na longevidade (34 dias), peso fresco e diâmetro de flor de flores cortadas de cravo. Esta solução melhorou as características ornamentais e fisiológicas das flores frescas de cravo.

Knocking down LSD1 inhibits the stemness features of colorectal cancer stem cells

As a top leading cause of cancer death in many countries, colorectal cancer (CRC) has drawn increasing attention to the study of the pathological mechanism. According to the "cancer stem cell hypothesis", malignancies originate from a small fraction of cancer cells that show self-renewal properties to initiate and sustain tumor growth and tumor metastasis. Therefore, these cancer stem cells (CSC) probably play important roles in tumor recurrence, metastasis, and drug resistance. Previous research reported that lysine-specific histone demethylase 1 (LSD1) maintains cancer stemness through up-regulating stemness markers SOX2 and OCT4. CD133 is believed to be the most robust surface marker for CRC stem cells, however the regulatory effect of LSD1 on stemness of CD133+ CRC has never been reported. In this study, our objectives included: 1) to isolate pure CD133+ and CD133− cells from SW620 cell line; 2) to investigate the effect of LSD1 on the characteristics of CD133+ stem cancer cells by knocking down the target gene. Results suggested that the SW620 cell line had both CD133+ and CD133− subsets. The CD133+ subset exhibited more CSC-like characteristics compared with the CD133− subset with higher viability, colony formation rate, migration and invasion rate, resistance to anti-cancer drugs, and apoptosis in vitro. The CD133+ also induced faster tumor formation and larger tumors in vivo. In the LSD1-knockdown CD133+ cells, the CSC-like characteristics had been all weakened. We conclude that LSD1 was important for CSCs to maintain their "stemness" features, which could be a potential therapeutic target of CRC.

Impact of inorganic salts on vase life and postharvest qualities of the cut flower of Perpetual Carnation

Abstract This study was carried out in the laboratory of Shangqiu Institute of Technology, Henan to investigate the effect of a different combination of inorganic salt on the quality and physiological characteristics of cut flowers (CFs) of Perpetual Carnation. Furthermore, to find out the best preservation solution of inorganic salt that can enhance the ornamental value of CFs of Carnation and prolong its vase life. Sucrose, 8-hydroxyquinoline, paclobutrazol, salicylic acid and different kinds of inorganic salts were added as a preservation solution. And the same amount of distilled water was used as control. The effects of these various inorganic salts on the morphological characteristics including vase life, changes in flower stems, fresh weight (FW) and water balance and the physiological characteristics including contents of malondialdehyde (MDA), cell membrane permeability and the contents of proline of carnation were investigated. The CFs placed in vase solution with inorganic salts showed significant changes in its morphology and physiological characteristics as compared to control. The changes in flower diameter (FD), FW, malondialdehyde and cell membrane permeability showed an increasing trend first and then decreasing. The value of water balance was observed with a downward trend. However, the vase life, FD, the contents of malondialdehyde, contents of proline and FW of CFs held in the preservative solution containing inorganic salts were increased than that of control. The fresh preservative solution contained sucrose 3% + 8-hydroxyquinoline (8-HQ) (200 mg·L1) + paclobutrazol (100 mg·L1) + salicylic acid (SA) (25 mg·L1) + CaCl2 (100 mg·L1) has the best effect on longevity (34 days), FW and FD of carnation CFs. This solution has improved the ornamental and physiological characteristics of fresh carnation CFs.

Resumo Este estudo foi realizado no laboratório do Instituto de Tecnologia de Shangqiu, Henan, para investigar o efeito de diferentes combinações de sal inorgânico na qualidade e características fisiológicas de flores cortadas do Cravo Perpétuo. Além disso, para descobrir a melhor solução de preservação de sal inorgânico que pode aumentar o valor ornamental das flores cortadas de Cravo e prolongar a vida do vaso. Sucrose, 8-hidroxiquinolina, paclobutrazol, ácido salicílico e diferentes tipos de sais inorgânicos foram adicionados como uma solução de preservação. E a mesma quantidade de água destilada foi usada como controle. Os efeitos destes vários sais inorgânicos sobre as características morfológicas incluindo a vida dos vasos, alterações nos caules, peso fresco e balanço hídrico e as características fisiológicas incluindo conteúdo de malondialdeído (MDA), permeabilidade da membrana celular e conteúdo de prolina de cravo foram investigados. As flores de corte colocadas em solução de vaso com sais inorgânicos apresentaram mudanças significativas em sua morfologia e características fisiológicas em relação ao controle. As alterações no diâmetro das flores, no peso fresco, no malondialdeído e na permeabilidade da membrana celular mostraram uma tendência crescente primeiro e depois decrescendo. O valor do balanço hídrico foi observado com tendência de queda. No entanto, a vida útil do vaso, o diâmetro da flor, o conteúdo de malondialdeído, o conteúdo de prolina e o peso fresco de flores cortadas mantidos na solução preservativa contendo sais inorgânicos foram aumentados em relação ao controle. A solução conservante fresca continha sacarose 3% + 8-hidroxiquinolina (8-HQ) (200 mg·L1) + paclobutrazol (100 mg·L1) + ácido salicílico (SA) (25 mg·L1) + CaCl2 (100 mg·L1) tem o melhor efeito na longevidade (34 dias), peso fresco e diâmetro de flor de flores cortadas de cravo. Esta solução melhorou as características ornamentais e fisiológicas das flores frescas de cravo.

High mobility group box 1 as a mediator of endotoxin administration after hemorrhagic shock-primed lung injury

High mobility group box 1 (HMGB1) was discovered as a novel late-acting cytokine that contributes to acute lung injury (ALI). However, the contribution of HMGB1 to two-hit-induced ALI has not been investigated. To examine the participation of HMGB1 in the pathogenesis of ALI caused by the two-hit hypothesis, endotoxin was injected intratracheally in a hemorrhagic shock-primed ALI mouse model. Concentrations of HMGB1 in the lung of the shock group were markedly increased at 16 h (1.63 ± 0.05, compared to the control group: 1.02 ± 0.03; P < 0.05), with the highest concentration being observed at 24 h. In the sham/lipopolysaccharide group, lung HMGB1 concentrations were found to be markedly increased at 24 h (1.98 ± 0.08, compared to the control group: 1.07 ± 0.03; P < 0.05). Administration of lipopolysaccharide to the hemorrhagic shock group resulted in a notable HMGB1 increase by 4 h, with a further increase by 16 h. Intratracheal lipopolysaccharide injection after hemorrhagic shock resulted in the highest lung leak at 16 h (2.68 ± 0.08, compared to the control group: 1.05 ± 0.04; P < 0.05). Compared to the hemorrhagic shock/lipopolysaccharide mice, blockade of HMGB1 at the same time as lipopolysaccharide injection prevented significantly pulmonary tumor necrosis factor-alpha, interleukin-1beta and myeloperoxidase. Lung leak was also markedly reduced at 16 h; blockade of HMGB1 24 h after lipopolysaccharide injection failed to alter lung leak or myeloperoxidase at 48 h. Our observations suggest that HMGB1 plays a key role as a late mediator when lipopolysaccharide is injected after hemorrhagic shock-primed ALI and the kinetics of its release differs from that of one-hit ALI. The therapeutic window to suppress HMGB1 activity should not be delayed to 24 h after the disease onset.

Studies on the use of cobalt-60 irradiation to control infectivity of Toxoplasma gondii cysts.

Mouse brains harboring the Chinese NT strain of Toxoplasma gondii cysts were homogenized with normal saline and irradiated with cobalt-60 gamma rays at various doses. The homogenate was introduced intraperitoneally into NIH mice or per os into kittens. Loss of infectivity was confirmed according to the following criteria: no cyst found in mouse brain impression smears on the 50th day after inoculation; no oocyst found in feces of kittens 3-15 days after inoculation; subinoculation in mice and a negative IHA test. All bioassays, parasitological examinations and serological tests in the control group gave positive results. Activity of radioactive source: 10 KCi; uniform dosage: 1238 rad/min; dose range of irradiation: 0.1-1.0 KGy. Minimal effective dose of gamma rays to control infectivity of T. gondii cysts was 0.55 KGy. Infectivity of bradyzoites irradiated with gamma rays at a dose of 0.45 KGy decreased by 10,000 times. Minimal effective dose of gamma rays to control infectivity of American ME-49 and Ts-2 strain, is slightly higher (0.6KGy) than that of NT strain. These studies present useful data for practical use of cobalt-60 to control infectivity of T. gondii in meat products.