Identification of hub genes in calcific aortic valve disease.

Calcific aortic valve disease (CAVD) is a heart valve disorder characterized primarily by calcification of the aortic valve, resulting in stiffness and dysfunction of the valve. CAVD is prevalent among aging populations and is linked to factors such as hypertension, dyslipidemia, tobacco use, and genetic predisposition, and can result in becoming a growing economic and health burden. Once aortic valve calcification occurs, it will inevitably progress to aortic stenosis. At present, there are no medications available that have demonstrated effectiveness in managing or delaying the progression of the disease. In this study, we mined four publicly available microarray datasets (GSE12644 GSE51472, GSE77287, GSE233819) associated with CAVD from the GEO database with the aim of identifying hub genes associated with the occurrence of CAVD and searching for possible biological targets for the early prevention and diagnosis of CAVD. This study provides preliminary evidence for therapeutic and preventive targets for CAVD and may provide a solid foundation for subsequent biological studies.

Exploration of animal models to study the life cycle of Fasciola hepatica.

The parasite Fasciola hepatica is an important zoonotic parasite. The development of an animal model of F. hepatica's life cycle is critical for studying the biological characteristics of the parasite in snails and mammals. Eggs of F. hepatica of bovine origin were cultured, and metacercariae were obtained after infection of Galba pervia snails. The life cycle system of F. hepatica was initiated in 2 different animals by orally infecting rabbits, SD rats and Kunming mice with the metacercariae. The animals' survival after infection, parasite migration in the animals and pathological damage to the liver were observed. We discovered that rabbits died due to acute suppurative hepatitis 60­69 days after infection, and eggs were found in the feces on day 63 of infection. The liver of SD rats showed punctate lesions on day 3 of infection, and further changes occurred as the infection progressed. However, liver repair was observed at week 9. SD rats survived for more than a year after infection and continued the F. hepatica life cycle. The liver lesions in Kunming mice after infection were similar but more severe than those in SD rats. Death was observed on the 31st post-infection day. We discovered that while rabbits, SD rats and Kunming mice can all be used as animal models of F. hepatica, SD rats are more suitable experimental animals in terms of tolerance and pathological response.

MicroRNA-140 inhibits skeletal muscle glycolysis and atrophy in endotoxin-induced sepsis in mice via the WNT signaling pathway.

Sepsis is a systemic inflammatory response syndrome resulting from infection. This study aimed at exploring the role of microRNA-140 (miR-140) in septic mice. Wnt family member 11 (WNT11) was verified to be a target gene of miR-140 after bioinformatic prediction and dual luciferase reporter gene assay. Importantly, miR-140 negatively regulated WNT11. We initially induced the model of sepsis by endotoxin, and then ectopic expression and knockdown experiments were performed to explore the functional role of miR-140 in sepsis. Additionally, cross-sectional areas of muscle fiber, lactic acid production, 3-methylhistidine (3-MH) and tyrosine (Tyr) production in extensor digitorium longus (EDL) muscles, and serum levels of inflammatory factors were examined. The effect of miR-140 on the expression of WNT signaling pathway-related and apoptosis-related factors in skeletal muscle tissue was determined. The experimental results indicated that upregulated miR-140 or silenced WNT11 increased cross-sectional areas of muscle fiber while decreasing lactic acid production, skeletal muscle cell apoptosis [corresponding to downregulated B cell lymphoma 2 (Bcl-2)-associated X protein (Bax) and caspase-3 and upregulated Bcl-2], and the proteolytic rate of Tyr and 3-MH. Also, overexpressed miR-140 or silenced WNT11 reduced inflammation as reflected by decreased serum levels of IL-6, IL-10, and TNF-α. Furthermore, overexpression of miR-140 was shown to suppress the activation of the WNT signaling pathway, accompanied by decreased expression of WNT11, ß-catenin, and GSK-3ß. Taken together, upregulation of miR-140 could potentially inhibit skeletal muscle lactate release, an indirect measure of glycolysis, and atrophy in septic mice through suppressing the WNT signaling pathway via inhibiting WNT11 expression.

[Formation and Characteristics of Fasciola hepatica Metacercariae].

The Fasciola hepatica miracidia were used to infect 10 Galba pervia in a random manner. Beginning from day 44 after infection on which 10 metacercariae were found and a total of 495 metacercariae found in 24 days. No signs of cercaria escape or metacercaria formation in the early morning observed during 8 : 00-24 : 00, regardless of the environmental change. Metacercaria formation occurred mainly in the early morning. The metacercariae could attach to any object in water, but were easy to detach themselves. The findings suggest that F. hepatica cercaria escape and metacercaria formation mainly occur in the early morning in Dali.

[Experimental infection of Galba pervia, Radix swinhoei and Physa acuta with Fasciola hepatica in Dali, Yunnan].

OBJECTIVE: To determine the intermediate host of Fasciola hepatica in Dali of Yunnan Province, and investigate its development and characteristics. METHODS: F. hepatica eggs from cattle were collected from July 2012 to July 2013, and placed in 28 degrees C water bath for incubation. Galba pervia, Radix swinhoei, and Physa acuta were collected from Dali, and used to be infected with F. hepatica in the laboratory. Trematode infections were excluded from the snails before experiment. All the snails were infected with F. hepatica miracidia, reared in mud pots. Dead snails were dissected for observing the development of F. hepatica. The metacercariae were collected and identified by PCR amplification of partial sequence of mitochondrial cytochrome oxidase subunit I (COX1) gene. RESULTS: A total of 1 146 R. swinhoei, 996 P. acuta, and 3 307 G. pervia snails were infected with F. hepatica, respectively. Mother rediae were found in two R. swinhoei snails, but no child rediae were observed in the snails. No larval forms were found in P. acuta. G. pervia was infected by F. hepatica with an infection rate of 27.2% (900/3307). The miracidium escaped from the egg and penetrated into G. pervia at temperature 22 degrees C, developed into a sporocyst after 7-15 days, which transformed into mother redia at the 11 st-20th day post-infection. The mother redia developed into daughter redia at the 30th-37th day, and produced cercaria with longtail, and became metacercaria at the 42nd-55th day. PCR confirmed that the metacercariae were that of F. hepatica, with an obvious band (approximately 500 bp). CONCLUSION: Among the three potential intermediate hosts in Dali, G. pervia is experimentally infected with F. hepatica.

[Molluscicidal effect of WPN and MNSC on Lymnaea].

OBJECTIVE: To evaluate the molluscicidal effect of wettable powder of 50% niclosamide ethanolamide salt (WPN) and suspension concentrate of 26% metaldehyde and niclosamide (MNSC) on Lymnaea. METHODS: WPN and MNSC were prepared as a series of solutions containing the active ingredient concentrations of 0.06, 0.13, 0.25, 0.50, 1.00, 2.00 mg/L and 4.00 mg/L, and the adult Lymnaea snails were soaked in the above mentioned series of solutions in the laboratory, and the LC50 values were calculated. The doses of active ingredient concentrations of 0.03, 0.06, 0.13, 0.25, 0.50, 1.00 g/m2 and 2.00 g/m2 of WPN and MNSC were adopted to spray on Lymnaea snails in the laboratory, and the LC50 values were calculated. A series of solutions containing the active ingredient concentrations of 1.00 mg/L, 0.50 mg/L, 0.25 mg/L, and 0.13 mg/L of WPN and MNSC were prepared, and the adult Lymnaea snails were put into the bowls with each concentration solution above mentioned and the climbing situation of the snails was observed at different time. RESULTS: By the immersion method, LC50 values of WPN at 48 h and 72 h were 0.93 mg/L and 0.64 mg/L respectively; LC50 values of MNSC at 48 h and 72 h were 0.74 mg/L and 0.51 mg/L respectively; by the spray method, when active ingredient concentrations of WPN and MNSC were 1.00 g/m2 or more, the death rates were both 100% after 3 days. In the climbing test, the Lymnaea snails did not climb in the solutions containing the active ingredient concentration of 1.00 mg/L of WPN and MNSC, however, a few snails climbed in the low concentrations. CONCLUSIONS: WPN and MNSC both have the effect of killing Lymnaea snails and inhibiting their climbing. By using the immersion method in the field, the active ingredient concentration of 2.00 mg/L of WPN and MNSC for 48 h is appropriate; by using the spray method, the active ingredient content of 1.00 g/m2 of WPN and MNSC for 3 days is appropriate.

[Survey of the number of eggs in Taenia asiatica gravid proglottids in Dali Prefecture of Yunnan Province].

OBJECTIVE: To observe the number of eggs in the gravid proglottids of Taenia asiatica. METHODS: Twenty gravid proglottids at each end of two adults of T. asiatica were digested by 1% pepsin. Then, the collected eggs were observed and counted by a microscope. RESULTS: The number of eggs in each gravid proglottid were not the same, with the maximum of 132 500, minimum of 44 180, and the average number of 90 051, and there was a significant difference among the different gravid proglottids (t = -3.487, P = 0.003). CONCLUSION: The number of eggs in different gravid proglottids of T. asiatica is evidently different.