RESUMO
A novel acidic polysaccharide named AeP-P-1 was prepared from Abelmoschus esculentus L. Moench (okra). AeP-P-1 is a heteropolysaccharide with a molecular weight of 3.02 × 103 kDa and is composed of L-rhamnose, D-galactose, and D-galacturonic acid in the ratio 1.87 : 3.58 : 1.00. Structural characterization based on methylation and 1D/2D NMR analyses indicated that AeP-P-1 is composed of T-linked-Rhap, T-linked-Galp, 1,2,4-linked-Rhap, 1,4-linked-Galp, 1,6-linked-Galp, and 1,3,4-linked-Galp in a molar ratio of 2.42 : 3.36 : 6.46 : 13.31 : 3.12 : 1, respectively. The hypoglycemic effect and mechanism of AeP-P-1 on type 2 diabetes mellitus were also explored. Firstly, AeP-P-1 can reduce blood lipids and liver and kidney damage caused by T2DM. Finally, AeP-P-1 induces the phosphorylation of GSK3ß, maintains the activity of glycogen synthase (GCS), and promotes glycogen synthesis by regulating the expression of insulin/PI3K/Akt pathway proteins. These results indicated that AeP-P-1 could be developed as a potential ingredient in immunostimulatory agents.
Assuntos
Abelmoschus , Diabetes Mellitus Tipo 2 , Abelmoschus/química , Hipoglicemiantes/farmacologia , Diabetes Mellitus Tipo 2/tratamento farmacológico , Fosfatidilinositol 3-Quinases , Polissacarídeos/farmacologia , Polissacarídeos/química , Carboidratos da DietaRESUMO
BACKGROUND: Glycolysis caused by hypoxia-induced abnormal activation of hypoxia inducible factor-1α (HIF-1α) in the immune microenvironment promotes the progression of hepatocellular carcinoma (HCC), leading to enhanced drug resistance in cancer cells. Therefore, altering the immunosuppressive microenvironment by imp-roving the hypoxic state is a new goal in improving cancer treatment. AIM: To analyse the role of HIF-1α, which is closely related to tumour proliferation, invasion, metastasis, and angiogenesis, in the proliferation and invasion of liver cancer, and to explore the HIF-1α pathway-mediated anti-cancer mechanism of sirolimus (SRL) combined with Huai Er. METHODS: Previous studies on HCC tissues identified the importance of HIF-1α, glucose transporter 1 (GLUT1), and lactate dehydrogenase A (LDHA) expression. In this study, HepG2 and Huh7 cell lines were treated, under hypoxic and normoxic conditions, with a combination of SRL and Huai Er. The effects on proliferation, invasion, cell cycle, and apoptosis were analysed. Proteomics and genomics techniques were used to analyze the HIF-1α-related signalling pathway during SRL combined with Huai Er treatment and its inhibition of the proliferation of HCC cells. RESULTS: High levels of HIF-1α, LDHA, and GLUT-1 were found in poorly differentiated HCC, with lower patient survival rates. Hypoxia promoted the proliferation of HepG2 and Huh7 cells and weakened the apoptosis and cell cycle blocking effects of the SRL/Huai Er treatment. This was achieved by activation of HIF-1α and glycolysis in HCC, leading to the upregulation of LDHA, GLUT-1, Akt/mammalian target of rapamycin (mTOR), vascular endothelial growth factor (VEGF), and Forkhead box P3 and downregulation of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and p27. The hypoxia-induced activation of HIF-1α showed the greatest attenuation in the SRL/Huai Er (S50 + H8) group compared to the drug treatments alone (P < 0.001). The S50 + H8 treatment significantly downregulated the expression of mTOR and HIF-1α, and significantly reduced the expression of VEGF mRNA. Meanwhile, the combined blocking of mTOR and HIF-1α enhanced the downregulation of Akt/mTOR, HIF-1α, LDHA, and GLUT-1 mRNA and resulted in the downregulation of PTEN, p27, and VEGF mRNA (P < 0.001). CONCLUSION: SRL increases the anti-cancer effect of Huai Er, which reduces the promotion of hypoxia-induced HIF-1α on the Warburg effect by inhibition of the PI3K/Akt/mTOR-HIF-1α and HIF-1α-PTEN signalling pathways in HCC.
Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Transportador de Glucose Tipo 1/genética , Transportador de Glucose Tipo 1/metabolismo , Glicólise , Hipóxia , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Lactato Desidrogenase 5 , Neoplasias Hepáticas/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro/metabolismo , Sirolimo , Tensinas/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
AeP-P-2, a pectic polysaccharide, was extracted from the fruit pod of okra. It composed of rhamnose (Rha), arabinose (Ara), glucose (Glc), galactose (Gal) and galacturonic acid (GalA) with the ratio of 4.75:2.01:1.00:4.91:7.24. The main structural feature of AeP-P-2 are 1,4-linked galacturonan units (homogalacturonan backbone) and (1 â 2) and (1 â 2,4) linked Rha (rhamnogalacturonan I region). And the other side chains contained â1)-linked Ara, (1 â 5)-linked Ara, (1 â 4)-linked Glc, (1 â 6)-linked Gal, (1 â 4)-linked Rha, (1 â 2,4)-linked Rha, â1)-linked Ara and â1)-linked Gal. When the concentration of AeP-P-2 was 3.2 mg/mL, the scavenging rates on DPPH·, ABTS, O2-· and ·OH reached to 61.88%, 87.10%, 52.17% and 60.32%, respectively. AeP-P-2 also could protect PC12 cells from the damage of H2O2 and reduce apoptosis caused by oxidative damage by decreasing the level of ROS. The findings indicated that okra was a functional vegetable and AeP-P-2 was worth studying and developing into antioxidant component.
Assuntos
Abelmoschus , Abelmoschus/química , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Peróxido de Hidrogênio , Células PC12 , Polissacarídeos/química , Polissacarídeos/farmacologia , RatosRESUMO
The purpose was aimed to establish a simple computational model to predict tumor prognosis by combining neutrophil to lymphocyte Ratio (NLR) and biomarkers of oncological characteristics in patients undergoing vascular reconstructive radical resection of PDAC. The enrolled patients was divided into high or low NLR group with the cutoff value determined by the receiver operator characteristic (ROC) curve. Different vascular anastomoses were selected according to the Chaoyang classification of PDAC. Survival rates were calculated using the Kaplan-Meier and evaluated with the log-rank test. Cox risk regression model was used to analyze the independent risk factors for prognostic survival. The optimal cut-off value of NRL was correlated with the differentiation, tumor size, TNM stage and distant metastasis of advanced PDAC. A curative resection with vascular reconstructive of advanced PDAC according to Chaoyang classification can obviously improve the survival benefits. Cox proportional hazards demonstrated higher evaluated NLR, incisal margin R1 and lymphatic metastasis were the independent risk predictor for prognosis with the HR > 2, meanwhile, age beyond 55, TNM stage of III-IV or Tumor size > 4cm were also the obvious independent risk predictor for prognosis with the HR ≤ 2. The advanced PADC patients marked of RS group (3 < RS ≤ 6) showed no more than 24 months of survival time according to RS model based on the six independent risk predictors. Vascular reconstruction in radical resection of advanced PDAC improved survival, higher elevated NLR (>2.90) was a negative predictor of DFS and OS in those patients accompanying portal system invasion.
RESUMO
Background: This presented study was aimed to evaluate the diagnostic and prognostic value of PD-L1+Neutrophils (PD-L1+NEUT) and neutrophil to lymphocyte ratio (NLR) based on our previous experience of Foxp3+Treg in transplantation. Methods: the NLR cutoff value of 1.79 was used to include 136 cases from the 204 patients with hepatocellular carcinoma (HCC) confirmed by clinical pathology, which were divided into highly-moderately and poorly differentiated HCC groups. The expressions of PD-L1+NEUT and Foxp3+Treg in peripheral blood and cancer tissue were detected with flow cytometry, meanwhile, PD-L1 and Foxp3 expressed in carcinoma and para-carcinoma tissues were marked by immunohistochemistry. Survival rates, including overall survival and disease-free survival, were calculated by the Kaplan-Meier curve and evaluated with the log-rank test. Finally, Cox risk regression model was used to analyze the independent risk factors for prognostic survival. Results: The level of PD-L1+NEUT, Foxp3+Treg, and NLR in peripheral blood of patients with poorly differentiated HCC were significantly increased (all P < .001). Both PD-L1+NEUT and NLR were positively correlated with Foxp3+Treg (r = 0.479, P = .0017; r = 0.58, P < .0001). The level of PD-L1+NEUT and Foxp3+Treg as well as PD-L1 and Foxp3 in cancer tissue and patients with poorly differentiated HCC were obviously increased (all P < .01), respectively. Cox regression analysis indicated that PD-L1+NEUT, NLR, and Foxp3+Treg were independent risk factors for the prognosis (P = .000, .000, .006) with a RR and 95%CI of 2.704-(2.155-3.393), 3.139-(2.361-4.173), 1.409-(1.105-1.798), respectively. Conclusion: PD-L1+NEUT, NLR, and Foxp3+Treg are independent risk factors for prognosis which maybe new marker of lower survival benefits.
Assuntos
Antígeno B7-H1/metabolismo , Carcinoma Hepatocelular/mortalidade , Fatores de Transcrição Forkhead/metabolismo , Regulação Neoplásica da Expressão Gênica , Linfócitos/patologia , Neutrófilos/patologia , Linfócitos T Reguladores/imunologia , Adolescente , Adulto , Idoso , Apoptose , Antígeno B7-H1/genética , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Proliferação de Células , Feminino , Fatores de Transcrição Forkhead/genética , Humanos , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Taxa de Sobrevida , Células Tumorais Cultivadas , Adulto JovemRESUMO
Steam explosion (SE) was a friendly environmentally pretreatment method. In this study, the effect of steam explosion (SE) pretreatment on structure and α-glucosidase inhibitory activity of Ampelopsis grossedentata polysaccharides was evaluated. Two novel polysaccharides (AGP and AGP-SE) were extracted, isolated, purified and analyzed by NMR, FT-IR and methylation. The results indicated that AGP mainly consisted of Rha, Xyl, Glc, and Ara with a molecular weight of 2.74 × 103 kDa and AGP-SE mainly consisted of Man, Ara, and Gal with a molecular weight of 2.14 × 103 kDa. Furthermore, the backbone of AGP and AGP-SE were mainly composed of 5)-Araf-(1â, -Glcp-(1â, 6)-Glcp-(1â, 6)-Galp-(1â, 3,6)-Manp-(1â, and 2,3,6)-Glcp-(1â. Finally, we demonstrated that all polysaccharides exhibited obviously α-glucosidase inhibition activity and mixed type inhibition. AGP-SE had better α-glucosidase inhibition activity and the binding affinity KD on α-glucosidase by using Surface Plasmon Resonance (SPR) than AGP. Overall, SE pretreatment is an effective method for extracting polysaccharide and provides a new idea into the improvement of biological activity.
Assuntos
Ampelopsis/química , Inibidores de Glicosídeo Hidrolases/farmacologia , Polissacarídeos/farmacologia , alfa-Glucosidases/metabolismo , Sequência de Carboidratos , Inibidores de Glicosídeo Hidrolases/química , Inibidores de Glicosídeo Hidrolases/isolamento & purificação , Humanos , Metilação , Peso Molecular , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Espectroscopia de Infravermelho com Transformada de Fourier , Vapor , Ressonância de Plasmônio de SuperfícieRESUMO
OBJECTIVE: To investigate the influence of tolerance dendritic cells (tolDCs), generated from Bone marrow mesenchymal stem cells (BM-MSCs) treated with rapamycin (Rapa) on liver allograft survival in a rat acute liver transplantation model. METHODS: Different GM-CSF induction project was used to obtain immature DCs (imDCs), mature DCs (matDCs) or tolDCs from BM-MSCs. First, MLR was performed to analyze the activity of tolDCs on polyclonaly stimulated total T cells. Then, co-cultured imDCs, matDCs and tolDCs with CD8+T cells isolated by magnetic activated cell sorting to analyze the influence on its regulatory characteristic. Last, the established rat acute liver transplantation model were adoptive transfused with imDCs, matDCs or tolDCs isolated by anti-CD11c immunomagnetic beads. The phenotype of DC cells and level of CD8+Treg in the culture system and in vivo, the expression of CD8 and CD45RC in the tissues were analyzed by flow cytometry and immunohistochemistry, respectively. RESULTS: The loGM-CSF plus IL-4 decreased the costimulatory molecules of CD80/86 and MHC class II of DCs comparison with hiGM-CSF from BM-MSCs no matter whether stimulation by LPS (P<0.05). Rapa treated not only reduced the expression of CD80/86 and MHC class II but also down-regulated the expression of CD11c after LPS stimulation which was more obviously in tolDCs by loGM-CSF project (P<0.05). Moreover, tolDCs displayed a rather higher level of IL-10 and low level of IL-12p70 than others (P<0.01), which shown a rather lower stimulative effect on the proliferation of T cells comparison with matDCs and imDCs. Co-cultured with CD8+Treg showed an improvement on induction of CD8+TCR+CD45RC-T cells (CD8+Treg) in ex vivo. The rats transfused with tolDCs has a delayed survival benefits with high level of CD8+Tregs (P<0.01) and high expression of CD45RC in liver tissue (P<0.01) and spleen when comparison with other groups. The infused tolDCs improved a mean survival time (MST) of 32 days comparison with a MTS of 9.5 days and 15.75 days displayed by rat that per-infused with matDCs and imDCs, respectively. CONCLUSION: Rapa modified tolDCs derived from BM-MSCs reversed graft rejection by improve tolerance characteristics of CD8+CD45RC-Treg in acute liver rat transplantation.
Assuntos
Medula Óssea/efeitos dos fármacos , Linfócitos T CD8-Positivos/efeitos dos fármacos , Células Dendríticas/efeitos dos fármacos , Antígenos Comuns de Leucócito/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Sirolimo/farmacologia , Linfócitos T Reguladores/efeitos dos fármacos , Animais , Medula Óssea/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Células Dendríticas/metabolismo , Rejeição de Enxerto/tratamento farmacológico , Rejeição de Enxerto/metabolismo , Tolerância Imunológica/efeitos dos fármacos , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Transplante de Fígado/métodos , Células-Tronco Mesenquimais/metabolismo , Ratos , Linfócitos T Reguladores/metabolismo , Transplante Homólogo/métodosRESUMO
BACKGROUND: To demonstrate the surgical procedures and techniques of the robotic anatomical isolated complete caudate lobectomy. METHODS: A retrospective analysis was performed on the demographic, operative, postoperative outcomes of seven patients who underwent robotic anatomical isolated complete caudate lobectomy at our department from January 2018 to November 2019. Mobilization of the left lateral and Spiegel lobe, dissection of the short hepatic veins and liver parenchyma transection from the dorsal plane of middle and right hepatic vein were crucial procedures for the robotic left-side approach. Anatomic complete caudate lobectomy was defined as total removal of the caudate lobe, in which the dorsal middle and right hepatic vein, the inferior vena cava and its right side were fully exposed on the raw surface. RESULTS: All patients successfully underwent the robotic anatomical isolated caudate lobectomy with a left-side approach without conversion to laparotomy, and without Clavien-Dindo Grade III or higher complications. The average tumor diameter was 65.00 ± 10.61 mm, the average operation time was 212.00 ± 74.53 min, the median bleeding loss was 100 mL, and the average postoperative hospital stay was 8.71 ± 4.89 d, respectively. There were four patients with primary hepatocellular carcinoma, one with tumor recurrence five months after surgery and three patients were free of recurrence. All patients survived at the last follow-up. CONCLUSION: Robotic anatomical isolated complete caudate lobectomy with a left-sided approach is safe and feasible for selected patients.
Assuntos
Carcinoma Hepatocelular/cirurgia , Hepatectomia/métodos , Neoplasias Hepáticas/cirurgia , Fígado/anatomia & histologia , Fígado/cirurgia , Procedimentos Cirúrgicos Robóticos/métodos , Adulto , Idoso , Perda Sanguínea Cirúrgica/estatística & dados numéricos , Carcinoma Hepatocelular/patologia , Feminino , Veias Hepáticas/cirurgia , Humanos , Fígado/irrigação sanguínea , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Duração da Cirurgia , Estudos Retrospectivos , Resultado do Tratamento , Veia Cava Inferior/cirurgiaRESUMO
A novel polysaccharide from Siraitia grosvenorii residues (SGP, molecular weight 1.93 × 103 KDa) was isolated and purified. SGP was composed of α-L-Arabinose, α-D-Mannose, α-d-Glucose, α-D-Galactose, Glucuronic acid, and Galacturonic acid with the ratio of 1: 1.92: 3.98: 7.63: 1.85: 7.34. The backbone of SGP was consist of galactoses and linked by α-(1,4)-glycosidic bond. The branch chains including α-1,6 linked glucose branch, α-1,6 linked mannose branch, α-1,3 linked galactose branch and arabinose branched (α-L-Ara(1â). The results of bioactivity experiments suggested that SGP had antioxidant in vitro, especially on scavenging DPPH radicals. Besides, SGP resulted in the decrease of ROS and the percentage of apoptotic and necrotic cells in a dose-dependent manner in H2O2 oxide injury PC12 cells. This research could help to develop the potential value and utilization of Siraitia grosvenorii.
Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Cucurbitaceae/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Fluorescência , Peróxido de Hidrogênio/toxicidade , Radical Hidroxila/química , Espectroscopia de Ressonância Magnética , Metilação , Monossacarídeos/análise , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Células PC12 , Ratos , Rotação , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Superóxidos/químicaRESUMO
BACKGROUND: Radical resection is an important treatment method for hepatic echinococcosis. The posterosuperior segments of the liver remain the most challenging region for laparoscopic or robotic hepatectomy. AIM: To demonstrate the safety and preliminary experience of robotic radical resection of cystic and alveolar echinococcosis in posterosuperior liver segments. METHODS: A retrospective analysis was conducted on the clinical data of 5 patients with a median age of 37 years (21-56 years) with cystic and alveolar echinococcosis in difficult liver lesions admitted to two centers from September to December 2019. The surgical methods included total pericystectomy, segmental hepatectomy, or hemihepatectomy. RESULTS: Among the 5 patients, 4 presented with cystic echinococcosis and 1 presented with alveolar echinococcosis, all of whom underwent robotic radical operation successfully without conversion to laparotomy. Total caudate lobectomy was performed in 2 cases, hepatectomy of segment VII in 1 case, total pericystectomy of segment VIII in 1 case, and right hemihepatectomy in 1 case. Operation time was 225 min (175-300 min); blood loss was 100 mL (50-600 mL); and postoperative hospital stay duration was 10 d (5-19 d). The Clavien-Dindo complication grade was I in 4 cases and II in 1 case. No recurrence of echinococcosis was found in any patient at the 3 mo of follow-up. CONCLUSION: Robotic radical surgery for cystic and selected alveolar echinococcosis in posterosuperior liver segments is safe and feasible.
Assuntos
Equinococose Hepática/cirurgia , Hepatectomia/métodos , Complicações Pós-Operatórias/epidemiologia , Procedimentos Cirúrgicos Robóticos/efeitos adversos , Adulto , Perda Sanguínea Cirúrgica/estatística & dados numéricos , Equinococose Hepática/parasitologia , Estudos de Viabilidade , Feminino , Hepatectomia/efeitos adversos , Humanos , Tempo de Internação/estatística & dados numéricos , Fígado/parasitologia , Fígado/cirurgia , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/etiologia , Recidiva , Estudos Retrospectivos , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Current reports on robotic hepatic caudate lobectomy are limited to Spiegel lobectomy. This study aimed to compare the safety and feasibility of robotic isolated partial and complete hepatic caudate lobectomy. METHODS: Clinical data of 32 patients who underwent robotic resection of the hepatic caudate lobe in our department from May 2016 to January 2020 were retrospectively analyzed. The patients were divided into three groups according to the lobectomy location: left dorsal segment lobectomy (Spiegel lobectomy), right dorsal segment lobectomy (caudate process or paracaval portion lobectomy), and complete caudate lobectomy. General information and perioperative results of the three groups were compared and analyzed. RESULTS: Among the 32 patients, none had conversion to laparotomy, three received intraoperative blood transfusion (9.38%), and none had complications of Clavien-Dindo grade III or higher or died in the perioperative period. Among them, 17 patients (53.13%) underwent Spiegel lobectomy, 7 (21.88%) underwent caudate process or paracaval portion lobectomy, and 8 (25.00%) underwent complete caudate lobectomy. The operative time and blood loss in the left dorsal segment lobectomy group were significantly better than those in the right dorsal segment lobectomy and complete caudate lobectomy groups (operative time: P = 0.010 and P = 0.005; blood loss: P = 0.005 and P = 0.017, respectively). The postoperative hospital stay in the left dorsal segment lobectomy group was significantly shorter than that in the complete caudate lobectomy group (P = 0.003); however, there was no difference in the postoperative hospital stay between the left dorsal segment lobectomy group and right dorsal segment lobectomy group (P = 0.240). CONCLUSIONS: Robotic isolated partial and complete caudate lobectomy is safe and feasible. Spiegel lobectomy is relatively straightforward and suitable for beginners.
Assuntos
Hepatectomia , Procedimentos Cirúrgicos Robóticos , Adulto , Idoso , Idoso de 80 Anos ou mais , Pequim , Perda Sanguínea Cirúrgica , Feminino , Hepatectomia/efeitos adversos , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Estudos Retrospectivos , Procedimentos Cirúrgicos Robóticos/efeitos adversos , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Investigate immunoregulation and anti-tumor immunity of FoxP3+Tregs after treatment with rapamycin (RAPA/SRL) plus thymalfasin (Zadaxin) and Huaier extract (PS-T) in a hepatocellular carcinoma (HCC) rat model simulating HCC relapse after liver transplant (LT). METHODS: We successfully established a rat model simulating HCC relapse after LT using an optimized chemical induction method with TACROLIMUS, methylprednisolone, and diethylnitrosamine as identified by visible liver nodules and hematoxylin-eosin staining. The model rats were then treated with RAPA, Zadaxin, and PS-T. Immune status changes were analyzed by flow cytometry, and protein expression of Akt and mTOR was determined by western blotting. Cytokines were measured by ELISAs. RESULTS: Combined therapy by RAPA plus Zadaxin and PS-T obviously alleviated hepatic pathological changes and significantly decreased the levels of FoxP3+Tregs in peripheral blood, the spleen, and the liver (P<0.05) and expression of mTOR protein (P<0.01) in the liver, obviously improved survival time (P=0.02). Moreover, the levels of CD8+T cells were increased significantly to almost normal levels (P<0.05) in comparison with no SRL monotherapy protocols. Inhibitory cytokines were also decreased in accordance with FoxP3+Tregs. Significant decreases of IL-10 and TGF-ß were observed after SRL-based therapy (P<0.01) in comparison with the other groups. Serum alpha fetoprotein (AFP) and vascular endothelial growth factor (VEGF) levels were also decreased significantly (P<0.05). FoxP3+Tregs showed a negative correlation with CD8+ and CD4+/CD8+T cells and a positive correlation with AFP, and VEGF (P<0.05). CONCLUSIONS: SRL-based therapy reduces FoxP3+Tregs to decrease secreted inhibitory cytokines which may enhancement the viability and number of CD8+T cells to exert anti-tumor effects that are mainly mediated through the AKT-mTOR signaling pathway.
RESUMO
GIBP, a homogeneous polysaccharide extracted from Glycyrrhiza inflata Batalin with a molecular weight 1.96 × 103 kDa, had a triple helix structure, smooth and sheet-like structure. Comprehensive analysis showed that the main chain of GIBP was composed of α-D-1,4 linked glucose, branch points were composed of α-D-1,3,6 and α-D-1,2,3,6 linked glucoses, and side chains were composed of α-D-1,3 and ß-D-1,6 linked galactose, ß-L-1,2 linked arabinose, α-D-1,3 and ß-D-1,3 mannose. The scavenging abilities of GIBP (3 mg/mL) against DPPH radical, OH radical, O2- radical and ABTS were 50.75 ± 0.13% and 52.32 ± 0.13, 25.84 ± 0.35% and 44.57 ± 0.15% and it also demonstrated an obvious dose-effect relationship. The inhibitory activity of α-glucosidase showed that the inhibitory effect of GIBP was enhanced with the increase of concentration. When the concentration reached 6 mg/mL, the inhibition rate of α-glucosidase activity reached 64.77%. And the ka, kd and KD were 6.472 × 104 1/Ms., 2.934 × 10-3 1/s and 4.534 × 10-8 M.
Assuntos
Antioxidantes/farmacologia , Sequestradores de Radicais Livres/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Glycyrrhiza/metabolismo , Polissacarídeos/química , Polissacarídeos/farmacologia , alfa-Glucosidases/química , Antioxidantes/química , Sequestradores de Radicais Livres/química , Inibidores de Glicosídeo Hidrolases/química , Peso Molecular , alfa-Glucosidases/metabolismoRESUMO
The crude polysaccharide was extracted from Cordyceps militaris. Material ratio of powder and water was 1:10. The polysaccharide was successively purified by Sevag and chromatography on Sephadex G-100 column to produce a polysaccharide fraction termed CBPS-II. The average molecular weight of CBPS-II was 1.273 × 103 kDa. The study was conducted to investigate the hypoglycemic effect of Cordyceps militaris polysaccharide on diabetic mice. Analysis of the clinical chemistry of the serum samples included serum creatinine (CRE), urea nitrogen (BUN), triglyceride (TG) and total cholesterol (TC). Results revealed that a certain dose of polysaccharide can alleviate the symptoms of metabolic disorders of diabetes, contributing to the body to restore the normal levels. The metabolic profiling method was adopted to find the related biomarkers and the metabolic pathway of diabetes. Moreover, results showed that 100 mg·kg-1 of Cordyceps polysaccharides can effectively reduce the blood glucose level of diabetic mice, thus regulating the metabolism of their energy, amino acids and intestinal microbes. The biomarkers noted in their metabolism were glucose, lactic acid, 3-hydroxy butyric acid, creatine, glutamate, valine, leucine, isoleucine and very low density lipoprotein (VLDL).
Assuntos
Cordyceps/química , Diabetes Mellitus Experimental/tratamento farmacológico , Hipoglicemiantes/isolamento & purificação , Metabolômica/métodos , Polissacarídeos/isolamento & purificação , Animais , Biomarcadores/sangue , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Nitrogênio da Ureia Sanguínea , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/metabolismo , Hipoglicemiantes/química , Hipoglicemiantes/metabolismo , Hipoglicemiantes/farmacologia , Camundongos , Polissacarídeos/química , Polissacarídeos/farmacologia , Polissacarídeos/uso terapêutico , Espectroscopia de Prótons por Ressonância Magnética , EstreptozocinaRESUMO
A polysaccharide was obtained from Lepidium meyenii Walp by hot water extraction and purification by Millipore (100 kD) and Sephadex G-200. The content of polysaccharide was examined to be 89.9% with phenol-sulfuric acid method. Its average molecular weight was estimated to be 2.213 × 106 Da by High Performance Gel Permeation Chromatography (HPGPC). Monosaccharide analysis showed that the polysaccharide was composed of arabinose, mannose, glucose and galactose with the molar ratio of 2.134: 1: 2.78: 2.82. After Smith degradation, methylation, infrared spectroscopy and NMR, the primary structure of the polysaccharide was identified. The backbone of the polysaccharide was composed of â4)-ß-D-Galp-(1â and â4)-α-D-Galp-(1â, while the branches were comprised of â6)-ß-D-Glup-(1â, â5)- ß-D-Araf-(1â, â3,6)-α-D-Manp-(1â, â3)-α-D-Galp-(1â, and α-D-Glup-(1â. The anti-fatigue effect of the polysaccharide was evaluated using exhaustive swimming test and biochemical indexes. The results indicated the polysaccharide has anti-fatigue effect.
Assuntos
Fadiga/tratamento farmacológico , Lepidium/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Animais , Sequência de Carboidratos , Cromatografia em Gel , Avaliação Pré-Clínica de Medicamentos/métodos , Galactose/análise , Glucose/análise , Espectroscopia de Ressonância Magnética , Masculino , Camundongos , Peso Molecular , Monossacarídeos/análise , Extratos Vegetais/química , Polissacarídeos/isolamento & purificação , Espectroscopia de Infravermelho com Transformada de Fourier , NataçãoRESUMO
A polysaccharide named PNP was extracted and purified from Pholiota nameko. The total sugar content of PNP was 95.29% and the molecular weight was 1.89 × 103 kDa. The structural features of PNP were investigated by the combination of chemical and instrumental analysis such as UV spectrophotometer, specific rotation determination, FT-IR, methylisation analysis and Congo red. The results showed that the optical rotation of PNP was +120° and that it had a triple-helical structure. Besides, PNP was mainly composed of glucose and mannose at the molar ratio of 4.24:1.00. The backbone of PNP was composed of (1â3)-linked-Glc and (1â3)-linked-Man whereas the branches of (1â3,6)-linked- Glc, (1â3,6)-linked-Man and T- Glc. Consistenting with the results of UV-Vis spectra, FT-IR spectroscopy and 1H NMR, indicated that PNP was a complex of polysaccharides and polyphenols. In vitro antioxidant results suggested that PNP was processed with certain scavenging capacity.
Assuntos
Antioxidantes/farmacologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Pholiota/química , Antioxidantes/química , Vermelho Congo/química , Carpóforos/química , Polissacarídeos Fúngicos/isolamento & purificação , Glucose/análise , Espectroscopia de Ressonância Magnética , Manose/análise , Metilação , Peso Molecular , Ácido Periódico/química , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
This study investigates the effect of fermentation conditions on the structure and anti-tumor activity of intracellular polysaccharides (IPS) of Cordyceps gunnii (C. gunnii) in submerged fermentation. The environmental and nutritional conditions are determined in a shaker flask by a single factor test. The inhibition of IPS on S180 cells was as an optimization index. The results show that the optimal fermentation conditions of C. gunnii are an initial pH value of 6, a temperature of 25 °C, a rotation speed of 150 rpm, 4% glucose, and 1.0% peptone. Under these conditions, the macro molecular weight (M w) polysaccharide content and anti-tumor activity of IPS are significantly higher than that in the basal culture medium. GC, HPGPC, periodate oxidation-Smith degradation, NMR, and FT-IR determine the structural characteristics of CPS-JC and CPS-YH (pure IPS cultured in basal culture medium and optimal culture medium, respectively). The results indicate that CPS-JC is mainly composed of α-d-glucans, whereas CPS-YH primarily contain α-d-glucans with a trace amount of ß-d-glucans.
RESUMO
Although liver transplantation (LT) lengthens the survival time of patients with hepatocellular carcinoma (HCC), LT patients exhibit a high recurrence rate; particularly those that had advanced HCC associated with the tumor biological characteristics and long-term application of immunosuppressants. A consensus on optimal prophylaxis and treatment for recurrent HCC following LT does not currently exist. The present study retrospectively analyzed data from 36 non-University of California at San Francisco criteria-eligible patients with advanced HCC who underwent LT, and then treated them with sirolimus (SRL)-based therapy with thymalfasin and huaier granules (SRL+, n=18), or with tacrolimus-based therapy (controls; n=18). The SRL+ group had significantly longer recurrence times (P=0.008) and survival times (P<0.0001) (OS, 1-year: 100%, 3-year: 94.4%, 5-year: 77.8%; DFS, 1-year: 88.9%, 3-year: 55.6%, 5-year: 50.0%). Furthermore, compared with pre-LT values and the control group, the SRL+ group had significantly lower serum α-fetoprotein (AFP) levels (both P<0.0001) and percentage of Forkhead box P3 (FoxP3)+ Treg lymphocytes (P<0.001) during the first year. In the SRL+ group, FoxP3+/cluster of differentiation (CD)8+ Treg lymphocyte percentages decreased significantly following LT (P<0.001); however, CD8+/CD3+ T-cells significantly increased (P<0.001). Levels of serum AFP and FoxP3+ Treg cells increased when tumors relapsed, and decreased to near-normal when relapse foci were cured or stabilized. SRL+ therapy may decrease AFP and Treg levels, while increasing CD8+ T cells, indicating an associated mechanism among them. In conclusion, SRL+ therapy appears to be safe and effective in preventing HCC recurrence following LT with no significant adverse events, and warrants further investigation.
RESUMO
A new glycoside, named as CG-1, was separated from Imperial Chrysanthemum with silica gel column chromatography. The purity was detected by TLC and HPLC. The crystal shape of CG-1 was consisting of a quadrangular and two rectangular pyramids. The analysis of DSC and TGA showed that the melting point of CG-1 crystal was 150.22⯰C and had good thermal stability. The monosaccharide conformation analysis showed that it was d-glucose. The structure characteristics were compared by FT-IR, NMR spectroscopy and LC-MS and a molecular structure has been deduced which consistent with spectroscopic results. In vitro antioxidant results suggested that the glycoside extracted from Imperial Chrysanthemum could be effectively employed as natural antioxidant in health or functional food. This work is of significance to keep the antioxidant activity in the processing and application of Imperial Chrysanthemum.
Assuntos
Antioxidantes/química , Chrysanthemum/química , Glicosídeos/química , Antioxidantes/isolamento & purificação , Relação Dose-Resposta a Droga , Glicosídeos/isolamento & purificação , Estrutura Molecular , Tamanho da Partícula , Relação Estrutura-Atividade , Propriedades de SuperfícieRESUMO
There is currently no consensus on the most suitable therapeutic approach for psoriasis (PS) co-existing with posthepatic cirrhosis (PCs) and hepatocellular carcinoma (HCC) following liver transplantation (LT). The present study provides an analysis of the therapeutic experience of such patients. Five LT recipients (two with PC and three with HCC) with accompanying PS were included. The induction program consisted of methylprednisolone plus basiliximab treatment. The initial postoperative treatment scheme consisted of tacrolimus (FK506) plus mycophenolate mofetil (MMF) and hormone; the latter was withdrawn 1 week after LT. The patients with PC had been using FK506 with or without a postoperative MMF program; the patients with HCC and recurrence of PS had been switched to a sirolimus (SRL)-based replacement therapy. Furthermore, all patients received anti-hepatitis B virus (HBV) therapy. The patients were followed up after 8.3±1.5 years. There was a positive correlation between HBV-DNA copy numbers, and psoriatic area and severity index (PASI) scores (r=0.97; P=0.006). The PASI scores were decreased significantly at 6 months following surgery compared with pre-transplantation (P<0.05). The patients who had received the FK506-based treatment experienced PS recurrence two years post-transplantation. The PASI scores increased significantly (P<0.05) and then declined gradually, maintaining a stable level (P<0.05) by 1 year after switching to the SRL-based treatment. The patients who had received the SRL-based treatment exhibited no recurrence of PS. The results of the present study suggest that SRL therapy provides a promising novel treatment method for patients with PS following LT that may be superior to tacrolimus treatment. When co-existing HBV is present pre-transplantation, regular injection of human hepatitis B immunoglobulin should be used to prevent the HBV from relapsing or aggravating the PS.