Morusin-Cu(II)-indocyanine green nanoassembly ignites mitochondrial dysfunction for chemo-photothermal tumor therapy.

Nanoscale drug delivery systems derived from natural bioactive materials accelerate the innovation and evolution of cancer treatment modalities. Morusin (Mor) is a prenylated flavonoid compound with high cancer chemoprevention activity, however, the poor water solubility, low active pharmaceutical ingredient (API) loading content, and instability compromise its bioavailability and therapeutic effectiveness. Herein, a full-API carrier-free nanoparticle is developed based on the self-assembly of indocyanine green (ICG), copper ions (Cu2+) and Mor, termed as IMCNs, via coordination-driven and π-π stacking for synergistic tumor therapy. The IMCNs exhibits a desirable loading content of Mor (58.7 %) and pH/glutathione (GSH)-responsive motif. Moreover, the photothermal stability and photo-heat conversion efficiency (42.8 %) of IMCNs are improved after coordination with Cu2+ and help to achieve photothermal therapy. Afterward, the released Cu2+ depletes intracellular overexpressed GSH and mediates Fenton-like reactions, and further synergizes with ICG at high temperatures to expand oxidative damage. Furthermore, the released Mor elicits cytoplasmic vacuolation, expedites mitochondrial dysfunction, and exerts chemo-photothermal therapy after being combined with ICG to suppress the migration of residual live tumor cells. In vivo experiments demonstrate that IMCNs under laser irradiation could excellently inhibit tumor growth (89.6 %) through the multi-modal therapeutic performance of self-enhanced chemotherapy/coordinated-drugs/ photothermal therapy (PTT), presenting a great potential for cancer therapy.

Pharmacokinetic study of deufruquintinibs in rats by HPLC-MS/MS after oral administration.

A sensitive and specific high-performance liquid chromatography-tandem mass spectrometry method was established to quantitatively determine the pharmacokinetics of fruquintinib (HMPL-013) and its derivatives [deufruquintinib-3D (HMPL-013-3D), deufruquintinib-6D (HMPL-013-6D) and deufruquintinib-9D (HMPL-013-9D)] in rats. Detection was performed on a triple quadrupole mass spectrometer in multiple reaction monitoring mode. The method established in this assay was successfully applied to a pharmacokinetic study of HMPL-013 and HMPL-013-Ds after oral administration. These results showed that HMPL-013-Ds had longer half-life and larger area under the plasma concentration-time curve than HMPL-013, especially HMPL-013-6D, which provides a significant basis for innovative ideas for drug structure transformation to reduce drug administration frequency and dosage.

Effect of ultra­wide­band electromagnetic pulses on blood­brain barrier permeability in rats.

The restrictive nature of the blood brain barrier (BBB) brings a particular challenge to the treatment of central nervous system (CNS) disorders. The effect of ultra­wide band electromagnetic pulses (UWB­EMPs) on BBB permeability was examined in the present study in order to develop a safe and effective technology that opens the BBB to improve treatment options for CNS diseases. Rats were exposed to a single UWB­EMP at various field strengths (50, 200 or 400 kV/m) and the BBB was examined using albumin immunohistochemistry and Evans blue staining at different time periods (0.5, 3, 6 and 24 h) after exposure. The expression and distribution of zonula occludens 1 (ZO­1) were evaluated using western blotting to identify a potential mechanism underlying BBB permeability. The results showed that the BBB permeability of rats exposed to UWB­EMP increased immediately following UWM­EMP treatment and peaked between 3 and 6 h after UWB­EMP exposure, returning to pre­exposure levels 24 h later. The data suggested that UWB­EMP at 200 and 400 kV/m could induce BBB opening, while 50 kV/m UWB­EMP could not. The levels of ZO­1 in the cerebral cortex were significantly decreased at 3 and 6 h after exposure; however, no change was observed in the distribution of ZO­1. The present study indicated that UWB­EMP­induced BBB opening was field strength­dependent and reversible. Decreased expression of ZO­1 may be involved in the effect of UWB­EMP on BBB permeability.

A Method for Preparation of an Internal Layer of Artificial Vascular Graft Co-Modified with Salvianolic Acid B and Heparin.

Studies have shown that salvianolic acid B (SAB), which is derived from Chinese salvia ( Salvia miltiorrhiza), a plant used in traditional Chinese medicine, can promote the proliferation and migration of endothelial cells. The inner layer of an artificial vascular graft was fabricated using the coaxial electrospinning method and was loaded with the anticoagulant heparin and SAB. The release of heparin and SAB was sustained for almost 30 days and without an initial burst release of SAB. Furthermore, the combined effect of SAB and heparin contributed to promoting human umbilical vein endothelial cell (HUVEC) growth and improved the blood compatibility of the graft. In addition, upregulation of GRP78 by SAB protected human endothelial cells from oxidative stress-induced cellular damage. In vivo evaluation through Masson's trichrome and H&E staining was performed after the graft was subcutaneously embedded in SD rats for 2 weeks and indicated that the graft possessed satisfactory biocompatibility and did not cause a significant immune response. Hence, the functional inner layer is promising for preventing acute thrombosis and promotes rapid endothelialization of artificial vascular grafts.

[Efficacies of neoadjuvant chemotherapy plus nutritional support in advanced gastric cancer complicated with pylori obstruction].

OBJECTIVE: To explore the efficacies of neoadjuvant chemotherapy plus nutritional supports for gastric cancer complicated with pyloric obstruction. METHODS: Retrospective analyses were performed for a total of 116 patients of gastric cancer complicated with pyloric obstruction undergoing exploratory laparotomy from January 2004 to June 2013. RESULTS: Sixty-two patients (group A) received neoadjuvant chemotherapy (regimen of FOLFOX) plus preoperative nutritional support. And parenteral (PN, n = 30) and enteral (EN, n = 32) nutritional supports were provided. Another 54 patients (group B) underwent exploratory laparotomy alone. The serum level of albumin and score of quality of life in group A at the last preoperative day improved significantly. And EN was better than PN. The rate of excision/radical excision of group A (85.5%, 45.2%) was much higher than group B (64.8%, 18.5%) (both P < 0.05). CONCLUSION: Nutritional support, especially EN, can improve the nutritional status and quality of life in patients with gastric cancer complicated with pyloric obstruction. And nutritional support plus neoadjuvant chemotherapy increase the rate of tumor excision.

An open-labeled, randomized, multicenter phase IIa study of gambogic acid injection for advanced malignant tumors.

BACKGROUND: Gambogic acid is a pure active compound isolated from the traditional Chinese medicinal plant gamboge (Garcinia morella Desv.). Based on the preliminary results of a phase I study, this phase IIa study compared the efficacy and safety of different dosage schedules of gambogic acid in patients with advanced malignant tumors. METHODS: Patients with advanced or metastases cancer who had not received any effective routine conventional treatment or who had failed to respond to the existing conventional treatment were randomly assigned to receive either 45 mg/m(2) gambogic acid intravenously from Days 1 to 5 of a 2-week cycle (Group A), or 45 mg/m(2) every other day for a total of five times during a 2-week cycle (Group B). The primary endpoint was objective response rate (ORR). RESULTS: Twenty-one patients assigned to Group A and 26 to Group B were included in the final analysis. The ORRs were 14.3% in Group A and 0% in Group B. It was not possible to analyze the significant difference because one of the values was zero. The disease control rates (DCRs) were 76.2% in Group A and 61.5% in Group B (P = 0.0456). The observed adverse reactions were mostly Grades I and II, and occurred in most patients after administration of the trial drug. There was no significant difference in the incidence of adverse reactions between the two arms. CONCLUSIONS: The preliminary results of this phase IIa exploratory study suggest that gambogic acid has a favorable safety profile when administered at 45 mg/m(2). The DCR was greater in patients receiving gambogic acid on Days 1 - 5 of a 2-week cycle, but the incidence of adverse reactions was similar irrespective of the administration schedule.

[Effect of early enteral nutrition supplemented with glutamine on postoperative intestinal mucosal barrier function in patients with gastric carcinoma].

OBJECTIVE: To investigate the effect of early enteral nutrition (EEN) supplemented with glutamine on postoperative intestinal mucosal barrier function of patients with gastric carcinoma. METHODS: Eighty patients with gastric carcinoma who underwent intraoperative peritoneal hyperthermic chemotherapy(IPHC) were randomized into two groups: EEN+glutamine (EEN+Gln) group(n=40) and EEN group(n=40). Intestinal mucosal barrier function was evaluated by serum diamine oxidase (DAO), ratio of lactulose to mannitol(L/M), endotoxin lipopolysaccharides(LPS), and tumor necrosis factor-α(TNF-α) at 1 day before operation, 1 day, 7 days, 12 days after operation. Time to first flatus and tolerance to EEN were recorded as well. RESULTS: There were no significant differences in the two groups in demographics(all P>0.05). Two cases(5%) in the EEN+Gln group and 1 case (2.5%) in the EEN group could not tolerate well(P>0.05). On postoperative day 1, there were no differences in serum DAO, L/M ratio, LPS, TNF-α between the two groups (P>0.05). On postoperative day 7, all the parameters for mucosal barrier function were significantly lower in the EEN+Gln group. On postoperative day 12, the urinary L/M and DAO, LPS, and TNF-α were still significantly lower in the EEN+Gln group, however, urinary L/M was comparable between the two groups. There were no differences between the two groups in the time to first flatus (P>0.05). CONCLUSION: The immunologic tolerance of enteral nutrition supplemented with glutamine is favorable, which provides protective effect on intestinal mucosal barrier in patients with gastric carcinoma undergoing IPHC.

[Nutritional therapy of duodenocutaneous fistula].

OBJECTIVE: To summarize the experience in nutritional support for the management of duodenocutaneous fistula. METHODS: Data of 32 patients with duodenocutaneous fistula in Zhejiang provincial people's hospital from January 1999 to December 2009 were analyzed retrospectively. RESULTS: The mean duration of nutritional support was 35.6 days (range, 8-82 days). Eight received total parenteral nutrition, 2 total enteral nutrition, and 22 parenteral nutrition combined with enteral nutrition respectively. Succus entericus reinfusion with enteral nutrition was used in 11 cases, glutamine-enriched nutritional support in 28 cases, somatostatin in 12 cases. In these patients, the healing rate was 75.0% after conservative treatment. In the 8 patients who underwent surgery, 6 were cured and 2 died (due to severe abdominal infection and multiple organ failure). A total of 30 patients had the fistulas cured and discharged. CONCLUSIONS: Parenteral nutrition combined with enteral nutrition, succus entericus reinfusion combined with enteral nutrition, glutamine-enriched nutritional support and somatostatin are important factors for the healing of duodenocutaneous fistulas.

[Studies on effect enhancement of the Schistosoma japonicum DNA vaccine pVIV02-IL12-Sj23 by vegetal polysaccharides].

OBJECTIVE: To enhance the immunogenicity of the recombinant pVIVO2-IL12-Sj23 vaccine of Schistosoma japonicum by using mixed vegetal polysaccharides as adjuvant. METHODS: The plasmid pVIVO2-IL12-Sj23 was constructed. 3 groups of BALB/C mice were injected intramuscularly with normal saline (Group A), pVIVO2-IL12-Sj23 plasmid DNA (B), and pVIVO2-IL12-Sj23 plus mixed vegetal polysaccharides (C) respectively, and challenged with S. japonicum cercariae on the 4th week after immunization. Mice were killed to calculate the worm reduction rate and egg reduction rate in liver tissue on the 6th week after infection. Before and 4 weeks after immunization blood samples were collected. RESULTS: The worm reduction rate and egg reduction rate were 64.3% and 79.9%, respectively in group C, 45.5% and 58.4%, respectively in group B, showing a remarkable difference hetween them (P < 0.05). ELISA analysis showed a significantly higher level of IgG specific for Sj23 4 weeks after vaccination in groups B and C (P < 0.05). However, there was no significant difference in IgG level between groups C and B (P > 0.05). CONCLUSION: When mixed vegetal polysaccharides are used as adjuvant, the effect of the vaccine pVIVO2-IL12-Sj23 can he considerably enhanced.

[Protective effect of Gastrodia elata and E-gelatin on lead-induced damage to the structure and function of rat hippocampus].

OBJECTIVE: To observe the counteraction of Gastrodia elata and E-gelatin on the effect of subchronic lead poisoning on the ability of learning and memory and the ultrastructure in hippocampus. METHODS: Subchronic lead acetate exposure was given to rats (0.2 g x kg(-1) x d(-1)). Single and combined administration of Gastrodia elata (4 g x kg(-1) x d(-1)) and E-gelatin (1 g x kg(-1) x d(-1)) were conducted at the same time. Pb concentration in blood, and the ability of learning and memory (Y-maze test) of rats were measured. Ultrastructure of CA3 pyramidal cells in hippocampus under transmission electron microscope was observed. RESULTS: Blood Pb concentrations in each group (Pb group: 690.6 micro g/L, Pb + Gastodiae eleta group: 688.8 micro g/L, Pb + E-gelatin group: 663.8 micro g/L, Pb + combined group: 667.2 micro g/L) were higher than that in the control (28.24 micro g/L, P < 0.01). But there was no significant difference (P > 0.05) among these groups. In Y-maze test, the number of electric stroke in Pb group is higher than that in control (P < 0.01). Gastrodia elata or E-gelatin used singly could significantly reduce the number of electric stroke of lead-exposed groups (P < 0.05 in the first month and P < 0.01 in the second and third month). And the effect of combined use of them was more efficient than single use (P < 0.01). Under electron microscope, no anomaly was seen in the pyramidal cells of CA3 area in hippocampus of control group. But there was significant anomaly such as neucleus separation in the cells of Pb exposed group. In Pb-Gastrodia elata group, there were some stress response phenomena such as the occurrence of huge mitochondria while in Pb-E-gelatin group, the anomaly was mild. The cells in the CA3 area in hippocampus of the pb-Gastrodia-E-gelatin group were almost normal, and also showed some stress response phenomena. CONCLUSION: Gastrodia elata and E-gelatin may protect the neurons in CA3 area of hippocampus against damage induced by lead to improve the ability of learning and memory, and they have synergistic effect.

[The antagonistic action of Gastrodia elata combined with E-gelatin on lead-induced down regulation of c-fos expression in rat brain].

OBJECTIVE: To observe the effect of lead on the expression of c-fos mRNA in hippocampus and cerebellum and the ability of learning and memory in rat, and to observe the antagonistic action of Gastrodia elata and E-gelatin on the effect mentioned above. METHODS: Rats were exposed to lead acetate (0.2 g x kg(-1) x d(-1) and 0.1 g x kg(-1) x d(-1)). The single and combined administration of Gastrodia elata (4 g x kg(-1) x d(-1)) and E-gelatin (1 g x kg(-1) x d(-1)) were conducted at the same time. Blood lead concentration in rats were measured. The ability of learning and memory by Y-maze test were examined. The expression of c-fos mRNA in hippocampus and cerebellum during Y-maze test were observed by in situ hybridization (ISH). RESULTS: (1) Blood lead concentrations were significantly increased in both high and low doses of lead-exposed rats (P < 0.01). But there were no differences among high dose groups or low dose groups (P > 0.05). (2) In Y-maze test, the number of electric stroke during learning in lead-exposed group was significantly increased (P < 0.01) while that in Gastrodia elata and E-gelatin groups was significantly decreased (P < 0.05), and that in combined use group was more significantly decreased (P < 0.01). (3) There were much more deep-colored c-fos positive cells in CA3 area of hippocampus in low dose of Pb-exposed with Gastrodia elata + E-gelatin group. These cells were also found in high Pb with combined use or single use group, but which were not so densely distributed as the former. And they were hardly found in high and low doses of Pb alone groups. The changes of expression of c-fos cells in cerebellam were similar to those in hippocampus. CONCLUSION: The down regulation of c-fos expression may be one of the molecular mechanism of lead-induced impairment of learning and memory. Gastrodia elata and E-gelatin may antagonize the effect of lead on c-fos expression, and combined use of both drugs may potentiate the antagonism.

[The antagonism of rhizoma gastrodiae to lead-induced damage of hippocampus in rats].

OBJECTIVE: To observe and explore the antagonism of rhizoma gastrodiae (RGT) to the impairment of learning and memory ability induced by lead in rats. METHODS: 36 Wistar rats were randomly divided into 3 groups, 12 rats in every group, (1) control group: treated with distilled water; (2) lead group: treated with lead acetate (0.1 g.kg-1.d-1); (3) lead-RGT group: lead acetate (0.1 g.kg-1.d-1) + RGT(4.0 g.kg-1.d-1). The ability of learning and memory of the rats was measured monthly by swimming test; 3 months later, the rats were decapitated and nitric oxide(NO) and total antioxidative capacity(TAOC) in hippocampus were measured immediately and the examination for pathology was also made. RESULTS: (1) In swimming test, the number of seeking for anchorage in lead group(1, 2, 3 month: 10.10 +/- 1.10, 7.80 +/- 1.32, 5.40 +/- 0.97 respectively) were significantly decreased, compared with the control(P < 0.01); the number of seeking for anchorage in RGT-lead group(1, 2, 3 month: 11.90 +/- 0.95, 10.90 +/- 0.95, 9.70 +/- 0.96 respectively) were significantly increased, compared with lead group(P < 0.01). (2) NO (0.733 +/- 0.015) mumol/g pro and TAOC (0.945 +/- 0.017) U/mg pro in hippocampus of lead group were significantly decreased compared with the control(P < 0.01) whereas NO(0.769 +/- 0.021) mumol/g pro and TAOC(0.986 +/- 0.010) U/mg pro in hippocampus of RGT-lead group were significantly increased compared with lead group(P < 0.01). (3) Pathological examination showed that in lead group, marked atrophy in hippocampus, cellular denaturation and necrosis, dissolution and disappearance in axon were found. In lead-RGT group, the atrophy of hippocampus was not obvious, the cell morphology was nearly normal, no obvious abnormal changes were seen. CONCLUSION: RGT is antagonistic to the impairment of hippocampus and of learning and memory ability induced by lead.