Parasitological and molecular detection of human fascioliasis in a young man from Guizhou, China

@#A 24-year-old man born in Guizhou province was diagnosed with obstructive jaundice and bile duct stones in 2013. Four living trematodes were found during laparotomy and cholecystectomy. Based on the morphology and molecular genetics analysis of internal transcribed spacer and pcox1 genes of the flatworm specimens, the trematodes from the patient were confirmed to be Fasciola hepatica. This report provided the clinical and molecular diagnosis information on human fascioliasis, which is an emerging sanitary problem still ignored in China. Human fascioliasis constantly occurs due to climatic changes and frequency of human travel. Therefore, it deserves more attention from physicians working in both developing and developed countries.

Etanercept protects rat cardiomyocytes against hypertrophy by regulating inflammatory cytokines secretion and cell apoptosis

We aimed to investigate the effect of etanercept, a tumor necrosis factor-α (TNF-α) inhibitor, on rat cardiomyocyte hypertrophy and its underlying mechanism. Primary neonatal rat cardiomyocytes were isolated from Sprague-Dawley rats. The model of rat cardiomyocyte hypertrophy was induced by endothelin, and then treated with different concentrations of etanercept (1, 10, and 50 μM). After treatment, cell counts, viability and cell apoptosis were evaluated. The mRNA levels of myocardial hypertrophy marker genes, including atrial natriuretic factor (ANF), matrix metalloproteinase (MMP)-9 and MMP-13, were detected by qRT-PCR, and the expressions of apoptosis-related proteins (Bcl-2 and Bax) were measured by western blotting. The protein levels of transforming growth factor-β1 (TGF-β1), interleukin (IL)-1β, IL-6, leukemia inhibitory factor (LIF) and cardiotrophin-1 (CT-1) were determined using enzyme linked immunosorbent assay (ELISA) kits. In the present study, TNF-α level in cardiomyocytes with hypertrophy was significantly enhanced (P<0.05). Compared to the model group, cell number and viability were significantly increased and ratio of apoptotic cells was reduced by etanercept (P<0.05, P<0.01, or P<0.001). In addition, etanercept remarkably reduced the mRNA levels of ANF, MMP-9 and MMP-13, inhibited the expression of Bax, and increased the expression of Bcl-2 compared to the model group (P<0.05). ELISA results further showed that etanercept lowered the levels of IL-1β, IL-6, LIF and CT-1 but not TGF-β1 compared to the model group (P<0.05). Etanercept may protect rat cardiomyocytes from hypertrophy by inhibiting inflammatory cytokines secretion and cell apoptosis.

Edaravone protects endotoxin-induced liver injury by inhibiting apoptosis and reducing proinflammatory cytokines

Studies have shown that edaravone may prevent liver injury. This study aimed to investigate the effects of edaravone on the liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in female BALB/c mice. Edaravone was injected into mice 30 min before and 4 h after GalN/LPS injection. The survival rate was determined within the first 24 h. Animals were killed 8 h after GalN/LPS injection, and liver injury was biochemically and histologically assessed. Hepatocyte apoptosis was measured by TUNEL staining; proinflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] in the liver were assayed by ELISA; expression of caspase-8 and caspase-3 proteins was detected by Western blot assay; and caspase-3 activity was also determined. Results showed that GalN/LPS induced marked elevations in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Edaravone significantly inhibited elevation of serum AST and ALT, accompanied by an improvement in histological findings. Edaravone lowered the levels of TNF-α and IL-6 and reduced the number of TUNEL-positive cells. In addition, 24 h after edaravone treatment, caspase-3 activity and mortality were reduced. Edaravone may effectively ameliorate GalN/LPS-induced liver injury in mice by reducing proinflammatory cytokines and inhibiting apoptosis.

Airway provocation test with ragweed pollen extract in Chinese asthmatics.

As ragweed becomes widespread over China, ragweed pollenosis tends to be more frequent. Incidence of ragweed pollenosis is about 1.04% in Qingdao district. To elucidate ragweed as an allergen in the development of bronchial asthma, ragweed pollen extract inhalation airway provocation tests (APT) were studied in 55 patients with ragweed pollenosis. A single dosage of 9,000 PNU ragweed pollen extract was applied and positive response was defined as more than 35% fall in SGrs at the dose of PT35-Grs. Among 55 patients, positive APTs were found in 27 cases (49%), dual late-phase responses in 11 patients, single late-phase responses in 2 patients in the complicated asthma group. Late-phase responses were related to the increased specific IgE and strong positive skin test response (wheal > 11 mm). The results suggested ragweed pollen was one of the important allergens causing bronchial asthma. There was airway hyperresponsiveness to ragweed pollen in some patients (20%) with ragweed pollenosis without asthmatic symptoms.

Strain complex of Schistosoma japonicum in the mainland of China.

The present paper deals with studies on the characteristics of Schistosoma japonicum isolated from five localities in the mainland of China. The following items were observed and compared including morphometric data, susceptibility of six mammalian hosts, prepatent period, compatibility between larvae and snail hosts, size of hepatic granuloma produced by eggs, immunoreactions in experimental animals, sensitivity to praziquantel, SDS-PAGE protein pattern and its antigenicity analysis, DNA hybridization and genetic variation and differentiation by analysis with multilocus enzyme electrophoresis. By means of these multidisciplinary methods, from morphological to molecular level, the following conclusions may be drawn from our results. The evidence indicates firstly that S. japonicum in the mainland of China comprises a strain complex with several components of geographically distributed strains. At least four distinct strains exist, ie Yunnan, Guangxi, Sichuan and Anhui-Hubei. Characteristics of each strain are distinct and the results of these studies lead to discussion on the problem of the intraspecific and interstrain differentiation of S. japonicum in the mainland of China.

Sensitivity of different isolates of Schistosoma japonicum from China to praziquantel.

Groups of C57BL inbred mice infected with each of the 4 different isolates, (Anhui, Hubei, Sichuan and Yunnan) of Schistosoma japonicum from the mainland of China were treated with praziquantel (PZQ) and the parasiticidal effects were compared. Worm reduction rate was recorded to assess systematically the sensitivity of 4 different isolates to PZQ in the mouse. Three dosage-levels of PZQ, ie 150, 230 and 310 mg/kg body weight in single doses were used. The worm development rates of control groups infected with schistosomes from Anhui, Hubei, Sichuan and Yunnan were 75.5, 81.8, 81.5, and 83.0%, respectively. At the dosage-level of 150 mg/kg, the worm reduction rates for the 4 different isolates were 36.0, 33.9, 25.5 and 35.6%, respectively. At the dosage-level of 230 mg/kg, the rates were 47.1, 46.0, 38.1 and 47.7%, while at the dosage-level of 310 mg/kg, they were 59.3, 58.6, 50.8 and 61.7%, respectively. The results indicated that the worm reduction rate of the Sichuan isolate was lower than that of the other three isolates, however, the differences were not statistically significant, suggesting that schistosomes of Anhui, Hubei, Sichuan and Yunnan isolates bear resemblance in drug response.

Parasitological and histopathological studies on rhesus monkeys infected with Chinese mainland strain of Schistosoma japonicum.

Fifteen rhesus monkeys were infected by cutaneous exposure each with 200 or 300 cercariae of Schistosoma japonicum. The dynamic distribution of schistosomula in the skin showed that 77-90% of them were found in the connective tissue, while 10-23% migrated in the hair follicles and sebaceous glands at different time intervals after cercarial penetration. Dead schistosomula recovered from the skin varied from 8.7% to 28.7%. The average rate of adult worm recovery was 74.4% and 61.3% in the 6th and 15th weeks of infection, thereafter the rate decreased to 32.3% and 9.0% in the 19th and 42nd weeks, respectively. The mean length of mature pair-worms was 13.2 +/- 2.3 mm in male and 18.0 +/- 1.9 mm in female 6 weeks of worm age. Afterwards the body length of females and their sexual gland diminished markedly. The mean prepatent period was 35.0 +/- 0.6 days. The average size of mature eggs in the feces was 86.6 +/- 5.4 x 64.3 +/- 3.6 microns, and the peak of eggs passage in the feces occurred between 7th and 15th weeks after infection, later on the number of eggs markedly decreased. Skin reaction to the primary infection was slight. The pathological changes observed in liver were chiefly cellular infiltration of portal spaces and the lesions produced by egg granulomas. The mean volume of single-egg granulomas of the productive stage in liver was 22.7 +/- 10.5 mm3 x 10(-3). The most intensive damages in the gastro-intestinal tract were observed in the large intestine.(ABSTRACT TRUNCATED AT 250 WORDS)

Characteristics of different isolates of Schistosoma japonicum from China in the final hosts.

The present paper deals with the susceptibility of common laboratory animals, such as mouse, rat, hamster, jird, rabbit and rhesus monkey, to infection with different isolates of Schistosoma japonicum in the mainland of China under laboratory conditions. With the exception of the rat, all the animals under study were permissive hosts for different isolates though their worm recovery rates varied. The mean body length of pair-worms of the Yunnan isolate was considerably smaller than that of the Anhui, Hubei, Guangxi and Sichuan isolates, and the percentage of male specimens with 7 testes in the Yunnan isolate was also significantly less than that in the other 4 isolates. Judging from the egg index (width/length x 100), the eggs of the Sichuan isolate were broad and short in shape, giving a high index; those of Guangxi and Hubei isolates were oblong, giving the lowest index; the other two isolates from Yunnan and Anhui, lay between these two extremes. The mean prepatent periods were longer in mice, hamsters and rhesus monkeys infected with Yunnan and Guangxi isolates, than those with Sichuan isolate. A dendrogram of the 5 isolates of S. japonicum was constructed on the basis of similarity coefficients by means of fuzzy cluster analysis on the biological characters mentioned above. Our results provide evidence of the existence of different strains of S. japonicum in the mainland of China as shown by comparative studies of their characteristics in the final hosts.

Compatibility between Oncomelania hupensis and different isolates of Schistosoma japonicum in China.

Oncomelania hupensis from six localities were used for infection with different isolates of Schistosoma japonicum in the mainland of China, ie Anhui in the east, Hubei in the center, Guangxi in the south, Sichuan in the West, Yunnan in the southwest and Fujian in the southeast. Snails from Anhui and Hubei were readily infected with the local isolate of S. japonicum and cross infection also took place readily between the snails and the schistosomes from these two places. Snails from Sichuan and Yunnan were refractory to infection with schistosome isolates from Hubei and Anhui, but the isolates from Sichuan and Yunnan were able to develop in snails from Hubei and Anhui. Though the Guangxi isolate developed readily in both Anhui and Guangxi snails, the average precercarial period in the former was significantly longer than in the latter. None of the other snails from Sichuan, Yunnan and Fujian became infected. On the other hand, snails from Guangxi infected with Anhui parasites also had a longer precercarial period than that in Anhui snails. Snails from Fujian were readily infected with the isolates from Anhui and Yunnan. The present results suggest that there might be different geographic strains of S. japonicum and their Oncomelania snail hosts in the mainland of China.