In Vivo Therapeutic Effect of Some Medicinal Plants' Methanolic Extracts on the Growth and Development of Secondary Hydatid Cyst Infection.

PURPOSE: The current study aimed to explore the in vivo therapeutic effects of the methanolic extracts of Citrullus colocynthis, Ruta graveolens, and Peganum harmala against hydatid cyst secondary infection. METHODS: Aerial parts of P. harmala and R. graveolens, including leaves and stems, and seeds of C. colocynthis were collected and extracted using absolute methanol. Rats that are infected with secondary infection of hydatid cysts were treated orally and intraperitoneally according to the determined lethal doses for 30 days. Histological, hematological, and biochemical investigations were done 8 months after the infection. RESULTS: Compared to Albendazole drug, C. colocynthis, and P. harmala, the methanol extract of R. graveolens showed higher and significant (P < 0.05) therapeutic effects on the secondary hydatid cysts growth. Those effects were represented by the reduction in the cysts' number, size, and weight; as well as the significant changes (P < 0.05) in values of hematological and biochemical parameters, the elevation of IFN-γ levels, and the decline of IL-10 and IL-4 cytokines, compared to the negative control group in both routes of treatment (oral and IP). Moreover, the histological sections showed that R. graveolens has a clear damaging effect on the hydatid cysts GL in the infected rats represented by the detachment of GL from LL and AL. CONCLUSION: This study can open an avenue to find new therapeutics for secondary hydatid cyst infections using the studied plant extracts, especially the extract of R. graveolens.

[Effects of persistent Echinococcus multilocularis infections on hepatic fibrosis in mice].

OBJECTIVE: To investigate the effects of persistent Echinococcus multilocularis infections on hepatic fibrosis in mice, so as to provide insights into the understanding of liver fibrogenesis induced by E. multilocularis infections and the treatment of alveolar echinococcosis. METHODS: Hepatic stellate HSC-T6 and LX-2 cells were exposed to the sera (25, 50 and 100 µL) from Meriones unguiculatus infected with E. multilocularis, and E. multilocularis, germinal layer cells (GCs) and protoscoleces (PSCs) for 48 hours, respectively. The cell proliferation was measured using a CCK-8 assay, and the levels of collagen 1 (Col1) and α-smooth muscle actin (α-SMA) were measured in the culture supernatant of HSC-T6 cells using ELISA. In addition, the serum and liver samples were collected 1, 2, 4, 6, 8 months post-infection with E. multilocularis, respectively. The serum Col1 and α-SMA concentrations were measured using enzyme-linked immunosorbent assay (ELISA), and the deposition of collagen fibers was examined in mice livers using Sirius red staining. RESULTS: The sera of E. multilocularis-infected gerbils promoted the proliferation of HSC-T6 and LX-2 cells in vitro, and there were significant differences seen in the proliferative rate of HSC-T6 (FHSC-T6 = 126.50, P < 0.05) and LX-2 cells (FLX-2 = 201.50, P < 0.05) among different serum groups, with the highest proliferative rate of HSC-T6 (573.36% ± 206.34%) and LX-2 cells (940.38% ± 61.65%) found following exposure to 100 µL mouse sera. Exposure to serum from E. multilocularis-infected gerbils resulted in an increase in the Col1 and α-SMA levels in the culture supernatant of HSC-T6 cells, with the greatest Col1 (20.99 ng/mL ± 2.01 ng/mL) and α-SMA levels (305.52 pg/mL ± 16.67 pg/mL) measured following exposure to 100 µL sera. The metacestodes (142.65% ± 9.17% and 189.99% ± 7.75%), GCs (118.55% ± 8.96% and 122.54% ± 0.21%) and PSCs of E. multilocularis (156.34% ± 17.45% and 160.59% ± 31.41%) all promoted the proliferation of HSC-T6 and LX-2 cells in vitro, and there were significant differences in the proliferative rates of HSC-T6 (FHSC-T6 = 11.24, P < 0.05) and LX-2 cells among groups (FLX-2 = 47.72, P < 0.05). Exposure to E. multilocularis resulted in an increase in Col1 and α-SMA levels in the culture supernatant of HSC-T6 cells, and the highest Col1 (4.43 ng/mL ± 2.23 ng/mL) and α-SMA levels (285.20 pg/mL ± 90.67 pg/mL) were detected following treatment with E. multilocularis metacestodes. In addition, a persistent increase was seen in the deposition of collagen fibers in mice livers 1 to 8 months post-infection with E. multilocularis, with the greatest Col1 level (280.26 ng/mL ± 23.04 ng/mL) seen 6 months post-infection and the highest α-SMA level (33.68 ng/mL ± 4.45 ng/mL) detected 8 months post-infection, respectively. CONCLUSIONS: Persistent E. multilocularis infections promote hepatic stellate cell proliferation, induce an increase in mouse serum Col1 and α-SMA levels, and cause elevated deposition of collagen fibers in mice livers. The infective stage of E. multilocularis is a critical period for inducing hepatic fibrosis of alveolar echinococcosis.

Commentary on "The Significance of the Granular Layer of the Cerebellum: a Communication by Heinrich Obersteiner (1847-1922) Before the 81st Meeting of the Society of German Natural Scientists and Physicians in Salzburg, September 1909".

This commentary highlights a "cerebellar classic" by Heinrich Obersteiner (1847-1922), the founder of Vienna's Neurological Institute. Obersteiner had a long-standing interest in the cerebellar cortex, its development, and pathology, having provided one of the early accurate descriptions of the external germinal layer (sometimes called the "marginal zone of Obersteiner" or "Obersteiner layer"). In his communication before the 81st meeting of the Society of German Natural Scientists and Physicians in Salzburg in September 1909, Obersteiner placed special emphasis on the histophysiology of the granule cell layer of the cerebellum and covered most of the fundamental elements of the cerebellar circuitry, on the basis of Ramón y Cajal's neuronism. Those elements are discussed in a historic and a modern perspective, including some recent ideas about the role of granule cells, beyond the mere relay of sensorimotor information from mossy fibers to the Purkinje cells, in learning and cognition.

Effects of persistent Echinococcus multilocularis infections on hepatic fibrosis in mice / 中国血吸虫病防治杂志

Objective To investigate the effects of persistent Echinococcus multilocularis infections on hepatic fibrosis in mice, so as to provide insights into the understanding of liver fibrogenesis induced by E. multilocularis infections and the treatment of alveolar echinococcosis. Methods Hepatic stellate HSC-T6 and LX-2 cells were exposed to the sera (25, 50 and 100 μL) from Meriones unguiculatus infected with E. multilocularis, and E. multilocularis, germinal layer cells (GCs) and protoscoleces (PSCs) for 48 hours, respectively. The cell proliferation was measured using a CCK-8 assay, and the levels of collagen 1 (Col1) and α-smooth muscle actin (α-SMA) were measured in the culture supernatant of HSC-T6 cells using ELISA. In addition, the serum and liver samples were collected 1, 2, 4, 6, 8 months post-infection with E. multilocularis, respectively. The serum Col1 and α-SMA concentrations were measured using enzyme-linked immunosorbent assay (ELISA), and the deposition of collagen fibers was examined in mice livers using Sirius red staining. Results The sera of E. multilocularis-infected gerbils promoted the proliferation of HSC-T6 and LX-2 cells in vitro, and there were significant differences seen in the proliferative rate of HSC-T6 (FHSC-T6 = 126.50, P < 0.05) and LX-2 cells (FLX-2 = 201.50, P < 0.05) among different serum groups, with the highest proliferative rate of HSC-T6 (573.36% ± 206.34%) and LX-2 cells (940.38% ± 61.65%) found following exposure to 100 μL mouse sera. Exposure to serum from E. multilocularis-infected gerbils resulted in an increase in the Col1 and α-SMA levels in the culture supernatant of HSC-T6 cells, with the greatest Col1 (20.99 ng/mL ± 2.01 ng/mL) and α-SMA levels (305.52 pg/mL ± 16.67 pg/mL) measured following exposure to 100 μL sera. The metacestodes (142.65% ± 9.17% and 189.99% ± 7.75%), GCs (118.55% ± 8.96% and 122.54% ± 0.21%) and PSCs of E. multilocularis (156.34% ± 17.45% and 160.59% ± 31.41%) all promoted the proliferation of HSC-T6 and LX-2 cells in vitro, and there were significant differences in the proliferative rates of HSC-T6 (FHSC-T6 = 11.24, P < 0.05) and LX-2 cells among groups (FLX-2 = 47.72, P < 0.05). Exposure to E. multilocularis resulted in an increase in Col1 and α-SMA levels in the culture supernatant of HSC-T6 cells, and the highest Col1 (4.43 ng/mL ± 2.23 ng/mL) and α-SMA levels (285.20 pg/mL ± 90.67 pg/mL) were detected following treatment with E. multilocularis metacestodes. In addition, a persistent increase was seen in the deposition of collagen fibers in mice livers 1 to 8 months post-infection with E. multilocularis, with the greatest Col1 level (280.26 ng/mL ± 23.04 ng/mL) seen 6 months post-infection and the highest α-SMA level (33.68 ng/mL ± 4.45 ng/mL) detected 8 months post-infection, respectively. Conclusions Persistent E. multilocularis infections promote hepatic stellate cell proliferation, induce an increase in mouse serum Col1 and α-SMA levels, and cause elevated deposition of collagen fibers in mice livers. The infective stage of E. multilocularis is a critical period for inducing hepatic fibrosis of alveolar echinococcosis.

Cystic echinococcosis: Development of an intermediate host rabbit model for using in vaccination studies.

The aims of this study were an establishment of the domestic rabbit as an intermediate host for cystic echinococcosis (CE) and to evaluate the potency of the crude germinal layer and the protoscoleces antigens to protect against the CE. Firstly; Two groups of white Newzeland rabbits were infected orally either by 5000 active oncospheres or viable protoscoleces separately. After 20 weeks, the slaughtered rabbits showed the presence of hydatid cysts at different internal organs. Molecular detection of the resulted cysts was conducted. Secondly; 27 rabbits were divided into nine groups (n = 3). Groups 1 and 2 were immunized with the crude germinal layer antigen while the groups 3 and 4 were immunized with the crude protoscoleces antigen. Groups 5 and 6 received the adjuvant mineral oil. Groups 7 and 8 were used as positive control. The last 9 group was kept as a negative control. The obtained results showed a significant high protection percentage of 83.4% and high antibody titer was recorded in groups that received the crude germinal layer antigen comparing with the groups that immunized with the crude protoscoleces antigen as their protection percentage was 66.7% with lower IgG response. In conclusion, the domestic rabbits could be used as a laboratory model for CE. Developing of the germinal layer antigen is more immunogenic than the protoscoleces one and could be used as a promising vaccine. Attention should be directed towards the existing rabbit in the environment adjacent to infected dogs as it could be a part of Echinococcus life cycle.

Ultrastructural changes in hydatid cyst walls obtained from human cases, exposed to different therapeutic approaches.

Recurrence of cystic echinococcosis as a result of treatment failure is frequently reported to cause a major problem in management of such serious parasitic infection. The deeply seated innermost germinal layer of hydatid cysts is a relatively delicate layer, yet responsible for viability maintenance of this parasitic stage. In this study, a trial was done to explore the ultrastructural changes in germinal and laminated layer of the hydatid cyst for the first time in human cases exposed to different therapeutic approaches which were done earlier to the final open surgical intervention. Four groups were included: group 1 did not receive any earlier form of treatment; group 2 was previously treated with only medical therapy; group 3 was treated with a single course of medical treatment, plus a single PAIR technique; group 4 was treated with multiple courses of medical treatment plus multiple PAIR techniques. Complete alteration of ultrastructural features of germinal and laminated layers were observed only with samples from group 4, indicating a kind of failure of the therapeutic approaches used in group, 1, 2, and 3, unless repeated in group 4 to achieve a real change regarding the fitness of the parasitic cystic lesions. Searching for more effective, safe, therapeutic method is highly recommended which may end the suffering of the affected patients.

A Experimental Pathological Study of Different Frequency Extracorporeal Shock Wave Induced Tibia Osteogenesis in Rabbits / 中国运动医学杂志

Objective To investigate the osteogenesis capability of different frequency extracorporeal shock wave.Methods 39 rabbits received different frequency extracorporeal shock wave at the middle potion of tibia for 3 or 7 days,these rabbits were then sac rificed and the tibia bones were collected to process for HE and toluidine blue staining,the pathological changes were observed under the light microscope.Results After different frequency extracorporeal shock wave treatment,the typical periosteal reaction were observed,external periosteum bleeded and thickened but there was no reaction at internal periosteum,marrow cavity opened and fibrosed.the osteoblast-1iking cell proliferated,however,no cartilage cells were observed;The rabbits received 7 days shock wave treatment showed more severe reaction than those for 3 days.The shock wave at lower frequency showed more severe reaction than higher frequency.Conclusion shock wave induced osteogenesis through the periosteal reaction of external periosteum;the osteogenesis capability of different frequency extracorporeal shock wave were affected by the frequency.Higher frequency of shock wave was not the ideal way to promote osteogenesis.