RESUMO
Streptococcus agalactiae (Group B Streptococcus, GBS) is associated with diverse diseases in aquatic animals. The capsule polysaccharide (CPS) encoded by the cps gene cluster is the major virulence factor of S. agalactiae; however, limited information is available regarding the pathogenic role of the CPS of serotype Ia piscine GBS strains in fish. Here, a non-encapsulated mutant (Δcps) was constructed by insertional mutagenesis of the cps gene cluster. Mutant pathogenicity was evaluated in vitro based on the killing of whole blood from tilapia, in vivo infections, measuring mutant survival in tilapia spleen tissues and pathological analysis. Compared to wild-type (WT) GBS strain, the Δcps mutant had lower resistance to fresh tilapia whole blood in vitro (p < 0.01), and more easily cleared in tilapia spleen tissue, and was highly attenuated in tilapia and zebrafish. Additionally, compared to the Δcps mutant, numerous GBS strains and severe tissue necrosis were observed in the tilapia spleen tissue infected with WT strains. These results indicated that the CPS is essential for GBS pathogenicity and may serve as a target for attenuation in vaccine development. Gaining a better understanding of the role, the GBS pathogenicity in fish will provide insight into related pathogenesis and host-pathogen interactions.
Assuntos
Cápsulas Bacterianas/metabolismo , Ciclídeos , Doenças dos Peixes/microbiologia , Infecções Estreptocócicas/veterinária , Streptococcus agalactiae/patogenicidade , Animais , Cápsulas Bacterianas/genética , Doenças dos Peixes/sangue , Mutagênese Insercional , Polissacarídeos/genética , Polissacarídeos/metabolismo , Baço/microbiologia , Baço/patologia , Infecções Estreptocócicas/patologia , Streptococcus agalactiae/química , Streptococcus agalactiae/genética , Fatores de Virulência/genética , Peixe-ZebraRESUMO
BACKGROUND: The electroacupuncture (EA) with different number of points significantly affected its efficacy on knee osteoarthritis (KOA), and the severity of KOA also influenced its response to treatments. Hence, we prospectively compared the clinical efficacy of EA on KOA with different severities. METHODS: A total of 132 KOA patients recruited from 181st Central Hospital of The Chinese People's Liberation Army between March 2014 and March 2015 were classified into 4 KOA stages according to Kellgren Lawrence grading scale. They were allocated into three treatment groups, including two-point group, four-point group and six-point group. Patients in the six-point group received treatment at six-points including ST34, SP10, SP9, ST36, ST35 and EX-LE4. Patients in the four-point group received treatment at ST34, SP10, ST35 and EX-LE5, while patients in the two-point group received treatment at ST35 and EX-LE4. Visual Analog Scale (VAS), McMaster Universities Osteoarthritis Index (WOMAC) and self-assessment questionnaire of patients were assessed after treatment. RESULTS: Three kinds of EA treatments all have significant clinical effects on KOA patients with down-regulated scores of VAS and WOMAC. Regarding post-treatment efficacy, the six point group exhibited lower VAS score and higher WOMAC score compared with the other two groups. For patients with different KOA grades, patients with higher KOA grades were associated with lower grade of treatment efficacy. CONCLUSIONS: Patients with KOA, especially those with lower KOA stages, could gain beneficial efficacies from EA treatments with two, four and six points, respectively.
Assuntos
Eletroacupuntura , Osteoartrite do Joelho/patologia , Osteoartrite do Joelho/terapia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Medição da Dor , Estudos Prospectivos , Resultado do TratamentoRESUMO
BACKGROUND: Lumbar disc herniation (LDH) is a common clinical disease of the skeletal system, and its prevalence has been on a rise. OBJECTIVE: To evaluate the efficacy of Huoxue Tongluo decoction plus acupuncture in the treatment of lumbar disc herniation and its effectiveness in improving the functional recovery of the patients' affected joints and mitigating their pain. METHODS: In this prospective study, 110 patients with lumbar disc herniation enrolled in our Hospital from June 2019 to June 2021 were collected and randomized to receive either conventional treatment (control group) or Huoxue Tongluo Decoction plus acupuncture (study group). RESULTS: Huoxue Tongluo Decoction plus acupuncture resulted in more rapid mitigation of lower extremity symptoms and lumbar symptoms versus conventional treatment (P< 0.05). Patients receiving traditional Chinese medicine (TCM) showed milder inflammatory responses than those with conventional medication, as evidenced by the lower serum concentrations of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and high-sensitivity C-reactive protein (hs-CRP) (P< 0.05). After treatment, the study group had higher Japanese Orthopedic Association (JOA) scores and lower visual analogue scale (VAS) scores than the control group (P< 0.05), suggesting that the combination of the herbal decoction and acupuncture provided better functional recovery of the affected joints and pain mitigation for the patients. Furthermore, the lower Pittsburgh sleep quality index (PSQI) scores in patients in the study group indicated better sleep quality of patients after TCM intervention than after conventional treatment (P< 0.05). Huoxue Tongluo Decoction plus acupuncture was associated with a significantly higher efficacy (94.55%) versus conventional treatment (80%) (P< 0.05). CONCLUSIONS: Huoxue Tongluo Decoction combined with acupuncture significantly offers a viable treatment alternative for lumbar disc herniation with promising treatment outcomes, mitigates patients' limb pain, and improves their lumbar function and sleep quality. Further trials are, however, required prior to general application in clinical practice.
Assuntos
Terapia por Acupuntura , Medicamentos de Ervas Chinesas , Deslocamento do Disco Intervertebral , Humanos , Medicamentos de Ervas Chinesas/uso terapêutico , Deslocamento do Disco Intervertebral/terapia , Vértebras Lombares , Dor , Estudos Prospectivos , Resultado do TratamentoRESUMO
High-amylose cereal starch has a great benefit on human health. Acid modification is very helpful for application of high-amylose starch in food and non-food industries. In this study, the ordered structure of acid-modified high-amylose rice starch was investigated by GPC, HPAEC, (13)C CP/MAS NMR and XRD. Acid preferentially degraded the amylose, then A chain and short B chain of amylopectin. Relative double helix content and crystallinity both initially increased sharply and then progressively with acid hydrolysis. The relative crystallinity of starches obtained from (13)C CP/MAS NMR was higher than that from XRD. The onset gelatinisation temperature decreased, while the peak and conclusion temperatures increased with increasing hydrolysis time. The endothermic value initially increased and then decreased with acid hydrolysis. The swelling power decreased while solubility increased after acid hydrolysis. These results add to our understanding of the effect of acid hydrolysis on the high-amylose rice starch.
Assuntos
Amilose/química , Oryza/química , Ácidos/química , Cristalização , Temperatura Alta , Hidrólise , Estrutura Molecular , SolubilidadeRESUMO
High-amylose cereal starch has a great benefit on human health through its resistant starch (RS) content. Enzyme hydrolysis of native starch is very helpful in understanding the structure of starch granules and utilizing them. In this paper, native starch granules were isolated from a transgenic rice line (TRS) enriched with amylose and RS and hydrolyzed by α-amylase. Structural properties of hydrolyzed TRS starches were studied by X-ray powder diffraction, Fourier transform infrared, and differential scanning calorimetry. The A-type polymorph of TRS C-type starch was hydrolyzed faster than the B-type polymorph, but the crystallinity did not significantly change during enzyme hydrolysis. The degree of order in the external region of starch granule increased with increasing enzyme hydrolysis time. The amylose content decreased at first and then went back up during enzyme hydrolysis. The hydrolyzed starches exhibited increased onset and peak gelatinization temperatures and decreased gelatinization enthalpy on hydrolysis. These results suggested that the B-type polymorph and high amylose that formed the double helices and amylose-lipid complex increased the resistance to BAA hydrolysis. Furthermore, the spectrum results of RS from TRS native starch digested by pancreatic α-amylase and amyloglucosidase also supported the above conclusion.
Assuntos
Amilose/análise , Bacillus/enzimologia , Oryza/química , Amido/química , Amido/metabolismo , alfa-Amilases/metabolismo , Glucana 1,4-alfa-Glucosidase/metabolismo , Hidrólise , alfa-Amilases Pancreáticas/metabolismo , Plantas Geneticamente Modificadas/química , Sementes/químicaRESUMO
A high-amylose transgenic rice line (TRS) modified by antisense RNA inhibition of starch branching enzymes revealed a resistant starch-rich quality. Compound starch granules in whole grains of the regular rice cultivar Teqing (TQ) were readily split during fracturing, whereas the starch granules in TRS were structurally intact and showed large voluminous, non-angular rounded bodies and elongated, filamentous structures tolerant of fracturing. In isolated preparation, TQ starch granules broke up into separate polygonal granules, whereas TRS starch granules kept their intactness. TRS starch granules consisted of packed smaller subgranules, some of which located at the periphery of starch granules were fused to each other with adjacent ones forming a thick band or wall encircling the entire circumference of the granules. TQ starch granules had a high concentration of amylose in the concentric hilum, whereas TRS starch granules showed a relatively even distribution of amylose with intense amylose in both hilum and band.
Assuntos
Enzima Ramificadora de 1,4-alfa-Glucana/genética , Amilose/análise , Oryza/química , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/química , RNA Antissenso/genética , Amido/química , Enzima Ramificadora de 1,4-alfa-Glucana/metabolismo , Oryza/enzimologia , Oryza/genética , Oryza/ultraestrutura , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/ultraestrutura , Amido/metabolismo , Amido/ultraestruturaRESUMO
High-amylose starch is a source of resistant starch (RS) which has a great benefit on human health. A transgenic rice line (TRS) enriched amylose and RS had been developed by antisense RNA inhibition of starch branching enzymes. In this study, the native starch granules were isolated from TRS grains as well as the wild type, and their crystalline type was carefully investigated before and after acid hydrolysis. In high-amylose TRS rice, the C-type starch, which might result from the combination of both A-type and B-type starch, was observed and subsequently confirmed by multiple physical techniques, including X-ray powder diffraction, solid-state nuclear magnetic resonance, and Fourier transform infrared. Moreover, the change of starch crystalline structure from C- to B-type during acid hydrolysis was also observed in this RS-rich rice. These data could add to our understanding of not only the polymorph structure of cereal starch but also why high-amylose starch is more resistant to digestion.
Assuntos
Enzima Ramificadora de 1,4-alfa-Glucana/genética , Oryza/química , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/química , Interferência de RNA , RNA Antissenso/genética , Amido/química , Enzima Ramificadora de 1,4-alfa-Glucana/metabolismo , Amilose/química , Oryza/enzimologia , Oryza/genética , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , RNA Antissenso/metabolismo , Amido/isolamento & purificação , Amido/metabolismo , Difração de Raios XRESUMO
Cereal starch granules with high-amylose and resistant starch (RS) always show irregular morphology and special crystalline structure, but their formation during grain development is not yet clear. In our previous studies, we had generated a transgenic rice line (TRS) enriched with amylose and RS, which contained semi-compound starch showing a C-type crystalline structure. In this study, the formation of semi-compound C-type starch granule during TRS endosperm development was carefully investigated with light, scanning electron, and transmission electron microscopes and X-ray powder diffraction. The results showed that the TRS starch subgranules, each with a central hilum, were individually initiated in amyloplast and showed an A-type crystal at the early stage of starch granule development, which was similar to that in its wild type. However, with the endosperm development, the amylose content in TRS endosperm starch increased and the B-type starch crystal was deposited in the periphery of subgranules; then, the adjacent subgranules fused together and finally formed a continuous outer layer band surrounding the entire circumference of the starch granule. Accordingly, a mechanistic model for the formation of semi-compound C-type starch granules is proposed.
Assuntos
Amilose/análise , Oryza/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/metabolismo , RNA Antissenso/genética , Amilose/metabolismo , Oryza/química , Oryza/enzimologia , Oryza/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , RNA Antissenso/metabolismo , Amido/biossíntese , Amido/químicaRESUMO
C-type starch, which is a combination of both A-type and B-type crystal starch, is usually found in legumes and rhizomes. We have developed a high-amylose transgenic line of rice (TRS) by antisense RNA inhibition of starch branching enzymes. The starch in the endosperm of this TRS was identified as typical C-type crystalline starch, but its fine granular structure and allomorph distribution remained unclear. In this study, we conducted morphological and spectroscopic studies on this TRS starch during acid hydrolysis to determine the distribution of A- and B-type allomorphs. The morphology of starch granules after various durations of acid hydrolysis was compared by optical microscopy, scanning electron microscopy, and transmission electron microscopy. The results showed that amorphous regions were located at the center part of TRS starch subgranules. During acid hydrolysis, starch was degraded from the interior of the subgranule to the outer surface, while the peripheral part of the subgranules and the surrounding band of the starch granule were highly resistant to acid hydrolysis. The spectroscopic changes detected by X-ray powder diffraction, 13C cross-polarization magic-angle spinning NMR, and attenuated total reflectance Fourier transform infrared showed that the A-type allomorph was hydrolyzed more rapidly than the B-type, and that the X-ray diffraction profile gradually changed from a native C-type to a CB-type with increasing hydrolysis time. Our results showed that, in TRS starch, the A-type allomorph was located around the amorphous region, and was surrounded by the B-type allomorph located in the peripheral region of the subgranules and the surrounding band of the starch granule. Thus, the positions of A- and B-type allomorphs in the TRS C-type starch granule differ markedly from those in C-type legume and rhizome starch.