Saponin-enriched sea cucumber extracts exhibit an antiobesity effect through inhibition of pancreatic lipase activity and upregulation of LXR-β signaling

Context: Sea cucumbers have been consumed as tonic, food, and nutrition supplements for many years.

Objective: The objective of this study is to investigate the antiobesity and lipid-lowering effects of sea cucumber extracts in in vitro and in vivo models and elucidate the mechanism of action of the extracts on obesity and dyslipidemia.

Materials and methods: The 60% ethanol extracts from the body walls of 10 different sea cucumbers were investigated for the inhibition of pancreatic lipase (PL) activity in vitro. The optimal active extract (SC-3) was further chemically analyzed by LC-MS and UV. And 0.1% and 0.2% of SC-3 was mixed with a high-fat diet to treat C57/BL6 mice for 6 weeks or 2 weeks as preventive and therapeutic study. The body weight, serum, and liver lipid profile in the mice were investigated.

Results: The crude extract of Pearsonothuria graeffei Semper (Holothuriidae) inhibited the PL activity by 36.44% of control at 0.5?μg/mL. SC-3 and echinoside A inhibited PL with an IC₅₀ value at 2.86?μg/mL and 0.76?μM. 0.1% of SC-3 reduced the body weight (23.0?±?0.62 versus 26.3?±?0.76 g), the serum TC (2.46?±?0.04 versus 2.83?±?0.12?mmol/L), TG (0.19?±?0.08 versus 0.40?±?0.03?mmo/L), and LDL-c (0.48?±?0.02 versus 0.51?±?0.02?mmol/L), and liver TC (1.19?±?0.17 versus 1.85?±?0.13?mmol/mg) and TG (6.18?±?0.92 versus 10.87?±?0.97?mmol/mg) contents of the obese C57BL/six mice on a high-fat diet.

Discussion and conclusion: Sea cucumber may be used for developing antiobesity and antihyperlipidemia drugs.     

Melatonin mitigates neomycin-induced hair cell injury in zebrafish

Context: Ototoxicity due to medications, such as aminoglycosides, is irreversible, and free radicals in the inner ear are assumed to play a major role. Because melatonin has an antioxidant property, we hypothesize that it might mitigate hair cell injury by aminoglycosides.

Objective: The objective of this study was to evaluate whether melatonin has an alleviative effect on neomycin-induced hair cell injury in zebrafish (Danio rerio).

Methods: Various concentrations of melatonin were administered to 5-day post-fertilization zebrafish treated with 125?μM neomycin for 1?h. Surviving hair cells within four neuromasts were compared with that of a control group. Apoptosis was assessed via terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. The changes of ultrastructure were confirmed using a scanning electron microscope.

Results: Melatonin alleviated neomycin-induced hair cell injury in neuromasts (neomycin?+?melatonin 100?μM: 13.88?±?0.91 cells, neomycin only: 7.85?±?0.90 cells; n?=?10, p?Conclusion: Melatonin is effective in alleviating aminoglycoside-induced hair cell injury in zebrafish. The results of this study demonstrated that melatonin has the potential to reduce apoptosis induced by aminoglycosides in zebrafish.     

Mineral pitch stimulates humoral,cellular and innate immune responses in mice

Abstract

Context: Mineral pitch (MP), a traditional medicine, is proposed to boost immunity in conditions that suppress Th1 cytokines such as AIDS/HIV, tuberculosis, leishmaniasis and cancer.

Objective: This study investigates the immunoregulatory mechanisms of MP in innate, humoral and cell-mediated immunity.

Materials and methods: Mice given MP (100, 200, 300 or 400?mg/kg, orally) for 10 consecutive days were immunized intravenously with goat RBC or ovalbumin, and investigated for plaque-forming cells (PFC), hemagglutination titer, hypersensitivity response, lymphocyte proliferation and macrophage function.

Results: MP increased PFC (330.2 versus 182.2/10⁶ splenocytes) in mice immunized with goat RBC and elicited ovalbumin-specific IgG titer at 400?mg/kg. Increase in Th1 immunity was correlated with the increased level of IFN-γ (724 versus 470?pg/ml) and decreased IL-4 (96 versus 178?pg/ml). CD4⁺/CD3⁺ ratio and delayed-type hypersensitivity response also increased to, respectively, 20.62?±?0.59 (versus 16.47?±?0.72) and 1.59?±?0.12 (versus 0.87?±?0.10?mm) in MP-treated mice. MP increased lymphocyte proliferation (11.14?±?0.60 versus 5.81?±?0.40 SI) and macrophage phagocyte response (0.24?±?0.02 versus 0.15?±?0.009), expressed as absorbance at 570?nm, but decreased nitrite production (17.4?±?1.10 versus 24.3?±?1.30?µM/10⁶ cells). We also observed an increased bone marrow cellularity (24.5?±?1.10 versus 17.10?±?0.70 cells/femur) and WBC count (12?667?±?377 versus 9178?±?213 cells/mm³) following MP treatment. There was no sign of toxicity at 400?mg/kg, 1/12th of reported LD₅₀.

Conclusion: MP elicits a dose-dependent Th1 immune response.     

Effects of garlic polysaccharide on alcoholic liver fibrosis and intestinal microflora in mice

Context: Alcoholic liver fibrosis (ALF) is treatable and reversible consequence of liver disease. Intestinal microflora plays an important role in the progression of liver disease. Garlic (Allium sativum L. [Amaryllidaceae]) has been consumed as a traditional medicine to treat liver injury.

Objective: To investigate the effects of garlic polysaccharide (GP) on ALF and intestinal microflora in mice.

Materials and methods: KM mice were orally administered with alcohol (56%, 6?mL/kg) for 30?d to establish ALF model, and divided into four groups together with control group (water only). Hugan tablet (60?mg/kg) or GP (250 and 150?mg/kg) were given 5?h after each dose of alcohol. Biochemical markers in serum and liver homogenate were determined with kits. Alteration of intestinal microflora, and protein expressions of TGF-β1, TNF-α and decorin were detected.

Results: In GP-H group, ALT and AST decreased to 18.85?±?4.71?U/L and 40.84?±?7.89?U/L. MDA, TC, TG and LDL-C decreased to 2.32?±?0.86?mmol/mg, 0.21?±?0.12?mmol/L, 0.96?±?0.31?mmol/L and 0.084?±?0.027?mmol/L. SOD, GSH-Px and GSH increased to 118.32?±?16.32?U/mg, 523.72?±?64.20?U/mg and 0.56?±?0.05?mg/g. Ratios of TGF-β1 and TNF-α decreased to 0.608?±?0.170 and 1.057?±?0.058, decorin increased to 2.182?±?0.129. Lachnospiraceae and Lactobacillus increased, Facklamia and Firmicutes decreased with GP pretreatment.

Discussion and conclusions: Intestinal microflora provides novel insight into the mechanisms of GP that may be used to treat ALF and intestinal microflora dysbiosis.     

Potential of Vitex negundo roots in the treatment of ulcerative colitis in mice

Context: Vitex negundo Linn. (Verbenaceae) is an indigenous tree species in India. This tree species has been of interest to researchers because traditionally its roots are reported in the treatment of ulcer and colic pain.

Objective: The present work was undertaken to validate its folk use in the treatment of ulcerative colitis (UC) by using the method of acetic acid-induced colitis in mice.

Materials and methods: Ethanol and aqueous extracts of roots of V. negundo (100?mg/kg) were screened for use in the treatment of UC by the method of acetic acid-induced UC in mice. Macroscopical study of the colon, level of myeloperoxidase (MPO), malondialdehyde (MDA) in colon tissue and blood and histopathology of the colon tissue were studied for the assessment.

Results: Ethanol extract (100?mg/kg) reduced the level of MPO in blood from 355?±?0.39 to 240?±?0.36?U/mL and from 385?±?0.35 to 257?±?0.36?U/mg in tissue. Similarly, it reduced the level of MDA in blood from 9.40?±?0.42 to 6.10?±?0.36 nmol/mL and from 9.38?±?0.56 to 5.89?±?0.56?U/mg in tissue. Both the results are comparable with the standard drug, prednisolone (5?mg/kg). This preventive effect was observed by morphological and histopathological study.

Discussion and conclusion: Results showed that ethanol extract of V. negundo root is effective in the treatment of UC and results are comparable with the standard drug, prednisolone, and thus possessing a great potential in the treatment of UC.     

Acute and chronic in vivo effects of exposure to nicotine and propylene glycol from an E-cigarette on mucociliary clearance in a murine model

Objective: To determine the effect of an acute (1 week) and chronic (3 weeks) exposure to E-cigarette (E-cig) emissions on mucociliary clearance (MCC) in murine lungs.

Methods: C57BL/6 male mice (age 10.5?±?2.4 weeks) were exposed for 20?min/day to E-cigarette aerosol generated by a Joyetech 510-T^® E-cig containing either 0% nicotine (N)/propylene glycol (PG) for 1 week (n?=?6), or 3 weeks (n?=?9), or 2.4% N/PG for one week (n?=?6), or 3 weeks (n?=?9), followed by measurement of MCC. Control mice (n?=?15) were not exposed to PG alone, or N/PG. MCC was assessed by gamma camera following aspiration of ^99mtechnetium aerosol and was expressed as the amount of radioactivity removed from both lungs over 6?hours (MCC6hrs). Venous blood was assayed for cotinine levels in control mice and in mice exposed for 3-weeks to PG alone and N/PG.

Results: MCC6hrs in control mice and in mice acutely exposed to PG alone and N/PG was similar, averaging (±1 standard deviation) 8.6?±?5.2%, 7.5?±?2.8% and 11.2?±?5.9%, respectively. In contrast, chronic exposure to PG alone stimulated MCC6hrs (17.2?±?8.0)% and this stimulation was significantly blunted following chronic exposure to N/PG (8.7?±?4.6)% (p?Conclusions: In this murine model, a chronic, daily, 20?min-exposure to N/PG, but not an acute exposure, slowed MCC, compared to exposure to PG alone and led to systemic absorption of nicotine.     

A molecular connection of Pterocarpus marsupium,Eugenia jambolana and Gymnema sylvestre with dipeptidyl peptidase-4 in the treatment of diabetes

Context: Pterocarpus marsupium (PM) (Leguminosae), Eugenia jambolana (EJ) (Myrtaceae) and Gymnema sylvestre (GS) (Asclepiadaceae) are the most important medicinal plants in the Indian system of traditional medicine for the treatment of hyperglycemia.

Objectives: Dipeptidyl peptidase-4 (DPP-4) inhibitors are the emerging class of anti-diabetic agents. However, only few compounds are commercially available. Therefore, in the present study we tried to explore the naturally occurring PM, EJ and GS semi-standardized extracts for their potential DPP-4 inhibition in vitro and in vivo.

Materials and methods: DPP-4 inhibition was evaluated by in vitro inhibitory assay, and enzyme kinetics were calculated using one-phase exponential decay equation. Glucose load (2?g/kg) was administered to control and diabetic rats 30?min following extract administration (100, 200 and 400?mg/kg) orally once, and blood samples were withdrawn at 0, 0.5, 1, 1.5, 2 and 3?h to measure plasma active glucagon-like peptide-1 (GLP-1) levels.

Results: PM and EJ inhibit DPP-4 potently with IC₅₀ values of 273.73?±?2.96 and 278.94?±?6.73?µg/mL, respectively, compared to GS (773.22?±?9.21?µg/mL). PM, EJ and GS exhibit long duration of action with enzyme inhibitory half-lives of 462.3, 317.2 and 153.8?min, respectively. Extracts significantly increase GLP-1 levels compared to negative control groups and peak GLP-1 level was observed at 2?h for PM and EJ, whereas for GS it was at 1.5?h

Discussion and conclusion: Taken together, results suggest the extracts may have potent DPP-4 inhibitory action, and their hypoglycemic action attributed through an increase in plasma active GLP-1 levels.     

Phytochemical screening and analgesic profile of the lyophilized aqueous extract obtained from Chrysobalanus icaco leaves in experimental protocols

Context: Chrysobalanus icaco L. (Chrysobalanaceae) has been used for the treatment of abdominal pain and cramps.

Objective: Assess the chemical and pharmacological profile of the lyophilized aqueous extract from C. icaco leaves (AEC).

Materials and methods: Chromatographic methods were used to assess compounds from AEC. Mice were treated with vehicle (control group) or AEC (100, 200 or 400?mg/kg, p.o.) (group with 7–8 mice) and the analgesic profile was assessed employing the acetic acid-induced writhing, formalin, hot plate tests and hyperalgesia induced by carrageenan (CG) or tumour necrosis factor-alpha. The animal motor performance was assessed using rota-rod and grip strength tests.

Results: The chromatographic profile of AEC demonstrated the presence of terpenoid compounds. The acute pretreatment with AEC, at all doses, produced a significant (p?<?0.01) inhibition of painful bahaviour (11.4?±?3.6; 10.3?±?2.8; 11.3?±?2.2) when compared to the control group (24.7?±?4.7) in acetic acid-induced writhing test. In the formalin test, AEC were effective in the second phase (p?<?0.01) (57.2?±?10.3; 56.3?±?9.2; 54.7?±?8.9) when compared to control group (121.9?±?18.5). No response was observed in the hot plate test. The higher dose of AEC produced a significant (p?<?0.01 or p?<?0.05) inhibitory effect on the mechanical hyperalgesia test. AEC did not affect the motor performance of the mice.

Discussion: The terpenoids from AEC are known for its analgesic and anti-inflammatory properties. So, these results corroborate the experiments using the AEC in inflammatory pain protocols.

Conclusion: Our results suggest that AEC act against inflammatory pain.     

Alteration of pentylenetetrazole-induced seizure threshold by chronic administration of ginger (Zingiber officinale) extract in male mice

Abstract

Context: Zingiber officinale Roscoe (Zingiberaceae), or ginger, used in traditional Chinese medicine, has antioxidant activity and neuroprotective effects. The effects of this plant on clonic seizure have not yet been studied.

Objective: The present study evaluated the anticonvulsant effect of ginger in a model of clonic seizures induced with pentylenetetrazole (PTZ) in male mice.

Materials and methods: The anticonvulsant effect of Z. officinale was investigated using i.v. PTZ-induced seizure models in mice. Different doses of the hydroethanolic extract of Z. officinale (25, 50, and 100?mg/kg) were administered intraperitonal (i.p.), daily for 1 week before induction of PTZ. Phenobarbital sodium (30?mg/kg), a reference standard, was also tested for comparison. The effect of ginger on to the appearance of three separate seizure endpoints, e.g., myoclonic, generalized clonic, and tonic extension phase, was recorded.

Results: Hydroethanolic extract of Z. officinale significantly increased the onset time of myoclonic seizure at doses of 25–100?mg/kg (55.33?±?1.91 versus 24.47?±?1.33?mg/kg, p?<?0.001) and significantly prevented generalized clonic (74.64?±?3.52 versus 47.72?±?2.31?mg/kg, p?<?0.001) and increased the threshold for the forelimb tonic extension (102.6?±?5.39 versus 71.82?±?7.82?mg/kg, p?<?0.01) seizure induced by PTZ compared with the control group.

Discussion and conclusion: Based on the results, the hydroethanolic extract of ginger has anticonvulsant effects, possibly through an interaction with inhibitory and excitatory systems, antioxidant mechanisms, and oxidative stress inhibition.     

Levels of urinary human chorionic gonadotrophin (hCG) following conception and variability of menstrual cycle length in a cohort of women attempting to conceive

ABSTRACT

Objectives: To define the variability of menstrual cycle length and contribution of follicular and luteal phases to overall cycle variability, and to examine the rise in urinary hCG in early pregnancy.

Methods: Menstrual cycle study. Urine samples from 101 women (recruited from two south-east counties in the UK) were assayed to determine day of luteinising hormone (LH) surge, lengths of follicular and luteal phases and correlations with total menstrual cycle length. HCG study. Daily urine samples collected from 86 women prior to conception until 43 days post-conception were assayed for hCG and examined versus time since LH surge, determined using fertility test kits.

Results: Mean menstrual cycle length was 27.7?±?3.4 days, mean follicular phase length was 14.5?±?3.4 days and mean luteal phase length was 13.2?±?1.9 days. Total cycle lengths varied between and within women. There was a significant correlation (r²?=?0.70) between follicular phase length and total cycle length; luteal phase length was less variable and showed no association with total cycle length. Concentrations of hCG were significantly similar between women when referenced against the day since LH surge. Three thresholds were determined to indicate time since conception as 1–2 weeks, 2–3 weeks and 3+ weeks.

Conclusions: Total cycle length variation is mainly determined by follicular phase variation and predicting menses onset to estimate time of pregnancy testing is unreliable. Evaluating concentrations of hCG relative to LH surge results in consistent increases between women up to 21 days after conception. Therefore, urinary hCG concentration can be used to accurately estimate time since conception.     

Combination therapy of Chinese herbal medicine Fructus Ligustri Lucidi with high calcium diet on calcium imbalance induced by ovariectomy in mice

Abstract

Context: Our previous biological study demonstrated that Fructus Ligustri Lucidi (FLL), the fruit of Ligustrum lucidum Ait. (Oleaceae), could be used to maintain calcium balance and prevent age-related osteoporosis since it effectively decreased calcium loss and increased calcium retention in rats.

Objective: This study investigates the combination effect of the Chinese herbal medicine FLL and a high calcium diet on calcium imbalance induced by ovariectomy in mice.

Materials and methods: The ovariectomized (OVX) mice were orally treated with vehicle, FLL extract (700?mg/kg), milk powder (5?g/mice) fortified with calcium (1.0% Ca) and the combination of FLL with milk powder. After 6 weeks of treatment, urine, serum, and tibia were preserved for biochemical analysis and kidneys were taken for gene expression analysis.

Results: The combination treatment of FLL and a high calcium diet significantly increased bone calcium content (6.80?±?0.34?mg) by 22% (p?<?0.05) and decreased urine calcium excretion (0.099?±?0.009?mg/mg) by 62% (p?<?0.01) as compared with those of the OVX group (bone Ca, 5.57?±?0.31?mg; urine Ca/Cr, 0.261?±?0.017?mg/mg). The mRNA expression of renal calcium-binding protein-9k (CaBP-9k) and calcium-sensing receptor (CaSR) in combination treatment group was significantly up-regulated and down-regulated, respectively, as compared with those of the OVX group.

Conclusion: The beneficial effects of this combination therapy on calcium balance of OVX mice were, at least partially, attributed to its regulation on mRNA expression of CaBP-9k and CaSR in kidney.     

Essential oil of Psidium cattleianum leaves: Antioxidant and antifungal activity

Abstract

Context: Psidium cattleianum Sabine (Myrtacea) is rich in vitamin C and phenolic compounds, including epicatechin and gallic acid as the main components.

Objective: To evaluate the antifungal and antioxidant capacity in vitro of the essential oil of araçá (EOA). The acute toxicity of the EOA also was evaluated in mice.

Materials and methods: The leaves of the P. cattleianum were extracted by steam distillation. The antioxidant capacity was evaluated by in vitro tests [1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS), ferric ion reducing antioxidant power (FRAP), linoleic acid oxidation, thiobarbituric acid reactive species (TBARS)], and ex vivo analysis [TBARS, δ-aminulevunilate dehydratase (δ-Ala-D) and catalase activity, non-protein thiols (NPSH), and ascorbic acid levels]. The toxicity was studied in mice by a single oral administration of EOA; and the antifungal activity was performed with five strains of fungi.

Results: The EOA exhibited antioxidant activity in the FRAP assay and reduced lipid peroxidation in the cortex (I_max?=?32.90?±?2.62%), hippocampus (IC₅₀?=?48.00?±?3.00?µg/ml and I_max?=?32.90?±?2.62%), and cerebellum (I_max?=?45.40?±?14.04%) of mice. Acute administration of the EOA by the oral route did not cause toxicological effects in mice (LD₅₀?>?500?µg/ml). The EOA also showed antifungal activity through of the determination minimum inhibitory concentration (MIC) values ranging from 41.67?±?18.04 to 166.70?±?72.17?µg/ml for tested strains.

Conclusion: The results of present study indicate that EOA possess antioxidant properties, antifungal and not cause toxicity at tested doses.     

The anti-angiogenic herbal composition Ob-X from Morus alba,Melissa officinalis,and Artemisia capillaris regulates obesity in genetically obese ob/ob mice

Context: The growth and development of adipose tissue leading to obesity is suggested to depend on angiogenesis. Our previous study showed that Melissa officinalis L. (Labiatae), Morus alba L. (Moraceae), and Artemisia capillaris Thunb. (Compositae) are involved in the regulation of angiogenesis. We hypothesized that Ob-X, a mixture of three herbs, M. alba, M. officinalis, and A. capillaris, can regulate obesity.

Objective: To investigate the inhibitory effect of Ob-X on obesity in genetically obese ob/ob mice.

Materials and methods: The effect of Ob-X on angiogenesis was measured using a mouse Matrigel plug assay. The effects of Ob-X on obesity were investigated in ob/ob mice.

Results: Ob-X inhibited angiogenesis in a dose-dependent manner, as evidenced by decreased blood vessel density in a mouse matrigel plug assay. Administration of Ob-X to ob/ob mice for 5 weeks produced a significant reduction in body weight gain by 27% compared with control (12.1?±?3.01 vs. 16.6?±?2.24?g, respectively). Ob-X also significantly decreased visceral adipose tissue mass by 15% (0.87?±?0.12 vs. 1.02?±?0.15?g, respectively). The size of adipocytes in visceral adipose tissue was reduced by 46% in Ob-X-treated mice. Ob-X treatment inhibited hepatic lipid accumulation and significantly decreased circulating glucose levels compared with controls (197?±?56.5 vs. 365?±?115?mg/dL, respectively).

Discussion and conclusion: These results suggest that Ob-X, which has an anti-angiogenic activity, reduces body weight gain and visceral adipose tissue mass in genetically obese mice, providing evidence that obesity can be prevented by angiogenesis inhibitors.     

Effectiveness of a new tobramycin (0.3 % ) and dexamethasone (0.05 % ) formulation in the treatment of experimental Pseudomonas keratitis

ABSTRACT

Objective: To quantitatively determine, in a Pseudomonas keratitis model, the anti-inflammatory and bactericidal properties of a new formulation of tobramycin (0.3?%?) and dexamethasone (0.05?%?) that utilizes a xanthan gum vehicle.

Research methods: In a randomized and masked fashion, rabbit corneas (n?≥?16 eyes per group) were intrastromally injected with 10³ colony-forming units (CFU) of P. aeruginosa. Eyes were untreated or were administered a single drop every 15?min between 16 and 17?h postinfection (PI) and then a single drop every 30?min between 17 and 22?h PI, a total of 15 drops of either 0.1?%?dexamethasone and 0.3?%?tobramycin (TobraDex; Tdex) or a new formulation 0.3?%?tobramycin and 0.05?%?dexamethasone with xanthan gum (TobraDex ST; ST). Slit lamp examination scores (SLE?±?SEM) were derived from grading seven parameters at 22?h PI. Rabbits were sacrificed at 23?h PI and the log CFU?±?SEM per cornea was determined.

Results: Untreated eyes had SLE scores of 11.11?±?0.43 and had log CFU of 7.27?±?0.06. Eyes treated with Tdex, as compared to the untreated eyes, had significantly lower SLE scores (7.39?±?0.21, p?<?0.0001) and significantly fewer bacteria (6.32?±?0.29 log CFU, p?=?0.0213). Eyes treated with ST had a SLE score (6.56?±?0.19) that was significantly lower than both the untreated eyes (p?<?0.0001) and the eyes treated with Tdex (p?=?0.0124). Furthermore, eyes treated with ST had significantly fewer log CFU (5.78?±?0.30) than untreated eyes (p?=?0.0001) or eyes treated with Tdex (p?=?0.0434).

Conclusions: The ST formulation with xanthan gum demonstrated statistically superior anti-inflammatory and bactericidal properties as compared to Tdex.

Limitations: Variations in inoculation procedures produced limited eye-to-eye differences in the infection.     

Effects of polysaccharide from Physalis alkekengi var. francheti on liver injury and intestinal microflora in type-2 diabetic mice

Context: Diabetic liver injury is a serious diabetic complication. The alterations of intestinal microbiota play an important role in induction and promotion of liver injury progression. Physalis alkekengi L. var. francheti (Mast.) Makino (Solanaceae) has been used as a water decoction for treating diabetes.

Objective: To study the effects of a polysaccharide (PPSB) from Physalis alkekengi var. francheti on liver injury and intestinal microflora in type-2 diabetic mice.

Materials and methods: Streptozotocin (160?mg/kg) was injected i.p. for 3?days to build model. The diabetic mice were randomly divided into four groups together with control group (10 mice in each group). The doses of PPSB were 50 and 100?mg/kg, respectively. After 5 weeks administration, level of blood glucose, ALT and AST were measured. Alterations of intestinal microflora, and protein expression of TGF-β1, TNF-α and DCN were detected.

Results: Level of blood glucose decreased from (25.38?±?2.21) mmol/L to (18.01?±?2.53) mmol/L, ALT and AST decreased to (24.67?±?4.86) U/L and (30.84?±?7.50) U/L in PPSB-H group. Lactobacillus, Clostridium butyricum, and Bacteroides increased remarkably with increasing concentration of PPSB, but Enterobacter was inhibited. The relative expression of TGF-β1 and TNF-α decreased to (0.70?±?0.17) and (0.39?±?0.06), and the expression of DCN increased to (0.65?±?0.13).

Discussion and conclusions: Probiotics have been promoted by PPSB, and protein expressions have been modulated in the progression of liver injury. PPSB could be used as a natural agent for treating diabetic liver injury and intestinal microflora imbalance.     

5-Fluorouracil shell-enriched solid lipid nanoparticles (SLN) for effective skin carcinoma treatment

Context: The effective treatment of skin carcinoma is warranted for targeting the chemotherapeutic agents into tumor cells and avoiding unwanted systemic absorption.

Objective: This work was dedicated to the purpose of engineering highly penetrating shell-enriched nanoparticles that were loaded with a hydrophilic chemotherapeutic agent, 5-fluorouracil (5-FU).

Methods: Varying ratios of lecithin and poloxamer188 were used to produce shell-enriched nanoparticles by enabling the formation of reversed micelles within this region of the SLN. The localization of 5-FU within the shell region of the SLN, was confirmed using 5-FU nanogold particles as a tracer. SLN were introduced within sodium carboxy methylcellulose hydrogel, and then applied onto the skin of mice-bearing Ehrlich’s ascites carcinoma. The mice were treated with the gel twice daily for 6 weeks.

Results: The transmission electron microscope (TEM) revealed the formation of uniform nanoparticles, which captured reversed micelles within their shell region. The SLNs’ had particle size that ranged from 137?±?5.5?nm to 800?±?53.6, zeta potential of ?19.70?±?0.40?mV and entrapment efficiency of 47.92?±?2.34%. The diffusion of the drug-loaded SLN (269.37?±?10.92?μg/cm²) was doubled when compared with the free drug (122?±?3.09?μg/cm²) when both diffused through a hydrophobic membrane. SLN-treated mice exhibited reduced inflammatory reactions, with reduced degrees of keratosis, in addition to reduced symptoms of angiogenesis compared to 5-FU-treated mice.

Conclusion: SLN possesses the capacity to be manipulated to entrap and release hydrophilic antitumor drugs with ease.     

Comparison of lenograstim and filgrastim on haematological effects after autologous peripheral blood stem cell transplantation with high-dose chemotherapy

SUMMARY

Objective: To compare the efficacy of lenograstim and filgrastim on haematological recovery following an autologous peripheral blood stem cell transplantation (PBSCT) with high-dose chemotherapy.

Methods: A retrospective case-controlled study.

Results: Absolute neutrophil count (ANC) recovery above 0.5?×?10⁹?l^?1 and white blood cell (WBC) recovery above 4?×?10⁹?l^?1 for 3 consecutive days was achieved earlier with filgrastim than with lenograstim ((13.2?±?8.0 vs 19.0?±?10.0 days, p?=?0.004), (16.9?±?9.7 vs 29.9?±?16.6 days, p?=?0.001), respectively). The platelet recovery above 20 x 10⁹/l was also achieved earlier with filgrastim than with lenograstim (19.5?±?11.6 vs

27.2?±?13.8 days, p?=?0.006). Furthermore, filgrastim-treated patients received fewer days of granulocyte colony simulating factor (G-CSF) administration (12.5?±?7.0 vs 18.6?±?8.5 days, p?=?0.001) and spent less time in hospital (23.7?±?10.9 vs 32.0?±?17.6 days, p?=?0.009). Duration of antibiotic administration was also significantly shorter in the filgrastim group (13.6?±?7.6 vs 29.1?±?19.8 days, p?=?0.001). Conclusion: In patients undergoing PBSCT following high-dose chemotherapy, filgrastim significantly reduced the duration of neutropenia, thrombocytopenia and days of G-CSF administration, and led to earlier hospital discharge compared with lenograstim.     

A novel nanogel formulation of methotrexate for topical treatment of psoriasis: optimization,in vitro and in vivo evaluation

Context: Although several formulation strategies have been developed for the treatment of psoriasis, there is an unmet need for optimization of its therapy.

Objective: The objective was to develop a nanogel composed of methotrexate (MTX)-loaded nanostructured lipid carrier (MTX-NLC) and to evaluate its potential in imiquimod-induced psoriasis model to ameliorate symptoms of psoriasis.

Materials and methods: MTX-NLC nanogel was prepared by hot-homogenization method and optimized by Design of Experiments. Particle size, polydispersity index (PDI) and entrapment efficiency were selected as the critical quality attributes. Antipsoriatic potential of MTX-NLC nanogel was evaluated by Psoriatic Area and Severity Index (PASI) score and histopathological examination in the imiquimod-induced psoriasis model.

Results and discussion: Optimized MTX-NLC exhibited particle size of 278?±?10?nm, PDI of 0.231?±?0.05 and EE of 22.29?±?1.23%. At the end of 48?h, MTX-NLC gel exhibited slow and prolonged release of MTX (47.32?±?0.94% versus 94.23?±?0.79%) compared to MTX gel. Furthermore, it significantly reduced the PASI score with recovery of normalcy of the mice’s skin, while the MTX gel exhibited signs of hyper and parakeratosis at the end of the study.

Conclusion: The developed MTX-NLC gel formulation can be a promising alternative to existing MTX formulation in treating psoriasis.     

Crataegus Aronia protects and reverses vascular inflammation in a high fat diet rat model by an antioxidant mechanism and modulating serum levels of oxidized low-density lipoprotein

Context: Crataegus aronia (Willd.) Bosc (Rosaceae) (syn. Azarolus L) is traditionally used to treat cardiovascular disorders.

Objectives: To investigate C. aronia protection against a high-fat diet (HFD)-induced vascular inflammation in rats.

Materials and methods: Wistar Male rats (180–220?g) were divided (n?=?10/group) as control fed a standard diet (STD), STD + C. aronia (200?mg/kg, orally), HFD, HFD + C. aronia and HFD post-treated with C. aronia. Simvastatin (20?mg/kg) was co- or post-administered as a positive control drug. HFD was given for 8?weeks, and all other treatments were administered for 4?weeks.

Results: Most significantly, co-administration of C. aronia to HFD-fed rats reduced the thickness of aorta tunica media (90?±?5 vs. 160?±?11.3?µm) and adventitia (54.3?±?3.8 vs. 93.6?±?9.4?µm). It also lowered protein levels of TNF-α (0.51?±?0.15 and 0.15?±?0.16 vs. 0.1?±?0.09%) and IL-6 (0.52?±?0.19 vs. 1.0?±?0.2%) in their aorta or serum (5.9?±?0.91 vs. 12.98?±?1.3?ng/mL and 78.1?±?6.7 vs. 439?±?78?pg/mL, respectively). It also lowered all serum lipids and increased aorta levels of GSH levels (70.4?±?4.0 vs. 40.7?µM) and activity of SOD (5.7?±?0.7 vs. 2.9?±?0.6?U/mg) and decreased serum levels of ox-LDL-c (566.7?±?46 vs. 1817?±?147?ng/mL). Such effects were more profound than all other treatments.

Conclusions: C. aronia inhibits the HFD-induced vascular inflammation and its use in clinical trials is recommended.     

Nanovesicular carrier-based formulation for skin cancer targeting: evaluation of cytotoxicity,intracellular uptake,and preclinical anticancer activity

Context: Skin cancer has turned into global epidemic leading to higher incidences among cancer stricken population.

Objective: The aim of the present investigation is to evaluate the anticancer potential and intracellular uptake of a novel nanovesicular formulation of 5-FU.

Materials and methods: Detailed intracellular uptake study in conjunction with estimation of intracellular reactive oxygen species was done using skin melanoma cell lines (A375) along with cytotoxicity studies. To further obtain the mechanistic insights into inhibition of tumor cell proliferation, cell-cycle arrest studies were conducted. The preclinical anticancer activity was carried out employing in vivo DMBA–croton oil-induced skin cancer model in mice.

Results and discussion: Significant reduction in the number of papillomas was observed in skin cancer-bearing mice on treatment with nanovesicular formulation (51.4?±?3.2%) in comparison with marketed formulation (21.3?±?2.1%) of 5-FU. Tumor volume was found to be reduced to 46.3?±?3.5% with prepared formulation, whereas the marketed formulation-treated group showed the reduction of 18.6?±?1.8% in comparison with the control (untreated) group.

Conclusion: The results of present study demonstrated that nanovesicular formulation of 5-FU possessed the enhanced anticancer activity which could be attributed to better intracellular uptake, cellular retention, and sustained release of drug.