Objective: To evaluate a novel polyherbal formulation (BSVT) containing the standardized extracts from the leaves of Boerhavia diffusa, Solidago virgaurea, Vitex negundo, and thymoquinone in CCl₄ induced hepatorenal toxicity in rats. Methods: A total of 36 rats were divided into six groups including normal control, CCl₄ (2 mL/kg, i.p.), CCl₄ (2 mL/kg, i.p.) + Cystone^® (750 mg/kgp.o.), CCl₄ (2 mL/kg, i.p.) + BSVT (25 mg/ kg, p.o.), CCl₄ (2 mL/kg, i.p.) + BSVT (50 mg/kg, p.o.), and CCl₄ (2 mL/kg, i.p.) + BSVT (100 mg/kg,p.o.). All treatments were given for four weeks. Serum levels of aspartate transaminase, alanine transaminase, alkaline phosphatase, cholesterol, total protein, serum urea, blood urea nitrogen and creatinine were assessed. Superoxide dismutase, malondialdehyde, and glutathione peroxidase were evaluated in tissue homogenate. The histopathological study of liver and kidney tissues was also done. Results: Aspartate transaminase, alanine transaminase, alkaline phosphatase, cholesterol, serum urea, blood urea nitrogen and creatinine were significantly elevated (P<0.001) while total protein was considerably reduced in the CCl₄ group as compared to the normal control (P<0.001), which indicated hepatorenal toxicity. In addition, superoxide dismutase and glutathione peroxidase activities were significantly decreased (P<0.001) while malondialdehyde levels were increased markedly (P<0.001). Treatment with BSVT formulation recovered these parameters towards a normal level in a dose-dependent manner. Conclusions: BSVT formulation ameliorates the hepatorenal toxicity in a dose-dependent manner. Furthermore, clinical studies are required to confirm its efficacy.

Objective: To evaluate the effect of free and liposome form of gallic acid on bone regeneration in critical defects in Wistar rats. Methods: Thirty-two female Wistar rats were divided into four study groups: group 1, negative control; group 2, positive control; group 3, gallic acid powder; group 4, gallic acid liposome. A critical-sized defect was created in all rats. Groups 2 to 4 had xenograft, autograft and membrane placement while negative control rats did not receive any treatment. The defect area was sutured and rats were kept alive for 30 d. At the end of the study, a bone specimen including the defect area was removed from calvaría. All specimens were evaluated under the stereomicroscope, then underwent histological analysis. Inflammatory cell counts, osteoblast, osteoclast counts, receptor activator of nuclear factor κ-B (RANKL), osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2), bone morphogenetic protein-2 (BMP-2), and alkaline phosphatase were determined. Results: The biggest unhealed defect area was observed in the negative control group and the smallest was observed in the gallic acid liposome group. There were no differences between the positive control group vs. the gallic acid powder group and the gallic acid powder group vs. the gallic acid liposome group. The severity of inflammation was the highest in the negative control group and the lowest in the gallic acid liposome group with significant differences between the groups. All groups had similar osteoblast counts while osteoclast counts were the highest in the positive control group. Gallic acid groups had a lower number of osteoclasts compared with the positive control group. Runx2 and alkaline phosphatase levels were similar in the groups while OPG and BMP-2 levels exhibited a significant increase compared with the negative control group and the positive control group. RANKL was similar in the negative control group, the positive control group, and the gallic acid powder groups but decreased in the gallic acid liposome group. Conclusions: Gallic acid powder and liposome significantly improve bone regeneration in Wistar rats with calvarial defects. The improvement in healing is evident with decreased inflammation and RANKL expressions and increased OPG and BMP-2 expressions.

Objective: To enhance the dissolution rate and oral bioavailability of Terminalia arjuna bark extract by formulating its nanosuspension. Methods: Nanoprecipitation approach was used for the formulation of nanosuspension using polysorbate-80 as a stabilizer. The formulated nanosuspension was assessed for particle size, polydispersity index, zeta potential value and for in vitro dissolution study. Oral bioavailability studies were carried out in Wistar male albino rats by administering a single dose (50 mg/kg. b. wt) of the formulated nanosuspension and coarse suspension. The storage stability of the formulated nanosuspension was determined after three months of storage at room temperature and under the refrigerated condition. Mutagenicity assay was carried out to evaluate the toxicity of the formulated nanosuspension using two mutant strains (Salmonella typhimurium TA100 and Salmonella typhimurium TA98). Results: The mean particle size of the formulated nanosuspension was 90.53 nm with polydispersity index and zeta potential values of 0.175 and -15.7 mV, respectively. Terminalia arjuna nanosuspension showed improved dissolution rate and 1.33 fold higher oral bioavailability than its coarse suspension. The formulated nanosuspension also showed better stability under the refrigerated condition and was non-mutagenic against both strains. Conclusions: Our study demonstrates that nanosuspension technology can effectively enhance the dissolution rate and oral bioavailability of Terminalia arjuna bark extract.

Objective: To examine the effect of Rumex crispus (R. crispus) and Rumex sanguineus (R. sanguineus) plant extracts against isolates of Acinetobacter baumannii (A. baumannii) from wounds, including multidrug-resistant strains. Methods: Six prepared Rumex extracts were subjected to liquid chromatography-tandem mass spectrometry. Antimicrobial activity of extracts and pure compounds (catechin, quercetin, isoquercitrin, emodin, and gallic acid) was examined by a microtiter plate method, while for determination of compound binary combinations activity a checkerboard method was applied. Active fractions of extracts were detected by agar-overlay high-performance thin- layer chromatography-bioautography assay followed by liquid chromatography - diode array detection - mass spectrometry analysis. Results: A total of 28 compounds were detected in two extracts of R. crispus and 26 compounds in four different R. sanguineus extracts, with catechin as a dominant component. Anti-A. baumannii activity was confirmed for all six R. sanguineus and R. crispus extracts at the concentration range from 1 to 4 mg/mL. Neither examined single compounds nor their binary combinations exhibited an anti-A. baumannii activity (MIC>256 μg/mL). The bioautography showed that fractions with the most prominent anti-A. baumannii activity tended to contain more polar compounds, predominantly flavonol (quercetin and kaempherol) glycosides; but also fractions containing flavanone (eriodictyol) glycosides and anthraquinone (emodin) glycosides; and less polar eriodictyol aglycone. Conclusions: The results justify and elucidate the traditional application of R. sanguineus and R. crispus extracts for wound healing, indicating the necessity for their further examination in combat against multidrug-resistant A. baumannii isolates from wounds.

Objective: To investigate the inhibitory effect on Burkholderia pseudomallei (B. pseudomallei) strain HNBP001 of a bacillomycin D-like cyclic lipopeptide compound named bacillomycin DC isolated from Bacillus amyloliquefaciens HAB-2. Methods: The antibacterial effect of bacillomycin DC on B. pseudomallei was determined using the disk diffusion method. The minimum inhibitory concentrations were evaluated by microdilution assay. In addition, transmission electron microscopy was performed and quantitative real-time polymerase chain reaction assay was carried out to determine the expression of MexB, OprD2, and qnrS genes. Results: Bacillomycin DC produced an inhibition zone against B. pseudomallei with minimum inhibitory concentration values of 12.5 μg/mL 24 h after treatment and 50 μg/mL at 48 and 72 h. Transmission electron microscopy showed that bacillomycin DC resulted in roughening cell surface and cell membrane damage. Quantitative real-time polymerase chain reaction analysis showed low expression of MexB, OprD2 and qnrS genes. Conclusions: Bacillomycin DC inhibits the growth of B. pseudomallei and can be a new candidate for antimicrobial agents of B. pseudomallei.

Objective: To investigate the prevalence of Calodium hepaticum (C. hepaticum) in rodents and insectivores from Wuhan section of the Yangtze River in China, and to provide evidence for the prevention and treatment of hepatic Calodium infection. Methods: Rodents and insectivores were captured from three selected Yangtze River beaches using mousetraps. The three survey sites were divided into six areas according to natural conditions, with 60 mousetraps placed in each area. The liver lesions in the captured rodents were observed by the naked eye and the eggs in the liver tissue were observed by microscopic examination. Results: A total of 1 080 mousetraps were placed, and 1 075 mousetraps were retrieved, with the retrieve rate as 99.5%. A total of 101 Apodemus agrarius, 12 Rattus norvegicus, and 9 Crocidura attenuata were caught. The average density of rodents and insectivores was 10.5% and 0.8%, respectively. DNA of egg nodules from infected rodents showed 98% similarity with that of C. hepaticum 18S rRNA (LC425008.1). One Rattus norvegicus was infected with C. hepaticum, with an infection rate of 3.23% in the Erqi river beach; the other two beaches did not show the incidence of C. hepaticum. Conclusions: The monitoring of C. hepaticum in the Yangtze River beaches should be strengthened to reduce the risk of human C. hepaticum infection.