The Analysis of Gene Expression on Fertility Decline in Caenorhabditis elegans after the Treatment with 5-Fluorouracil.

BACKGROUND: 5-Fluorouracil could lead to a decline in fertility in Caenorhabditis elegans. The aim of this study was to describe the mechanisms underlying such an altered fertility phenotype and to illustrate the specific genes and pathways that are involved in the related phenotypic changes in C. elegans. METHODS: We isolated total RNA from the samples and used a new method called Digital Gene Expression (DGE), which can rapidly identify genes with altered transcript levels. The random genes were confirmed by real-time RT-PCR. RESULTS: We analyzed the results of two methods to draw conclusions based on a comparison between C. elegans and other harmful parasites. Compared with controls, 1147 genes were up-regulated, and 1067 were down-regulated. Overall, 101 up-regulated genes had a log2 ratio higher than 8, whereas the log2 ratio of 141 down-regulated genes was higher than 8. After mapping to the reference database, 4 pathways were confirmed to be involved in this phenomenon, with statistically significant participation from 19 genes. CONCLUSION: For the first time, the transcript sequence of 5-Fu-treated worms and controls was detected. We found that 4 possible pathways, i.e., ECM-receptor interaction pathway, TGF-beta signaling pathway, Focal adhesion and Hypertrophic cardiomyopathy, may be involved in the number decline in the embryos of C. elegans. Specifically, the ECM-receptor interaction pathway and Focal adhesion may be very important pathways that alter the reproduction of C. elegans.

A Newly Discovered Epidemic Area of Echinococcus multilocularis in West Gansu Province in China.

Alveolar echinococcosis (AE) is a lethal parasitic disease. In Gansu Province of China, all AE cases reported in literature were from Zhang and Min Counties, the southern part of the province. Here, we report the discovery of nine AE cases and one cystic echinococcosis (CE) case from Nanfeng Town of Minle County, in the middle of Hexi Corridor in west Gansu Province. The diagnosis of these cases were confirmed by serology, histopathology, computed tomography, B-ultrasound, immunohistochemistry method, DNA polymerase chain reaction and sequencing analysis. Because eight of nine AE cases came from First Zhanglianzhuang (FZLZ) village, we conducted preliminary epidemiological analyses of 730 persons on domestic water, community and ecology such as 356 dogs' faeces of FZLZ, in comparison with those of other five villages surrounding FZLZ. Our studies indicate that Nanfeng Town of Minle County is a newly discovered focus of AE in China as a CE and AE co-epidemic area. Further research of Echinococcus multilocularis transmission pattern in the area should be carried for prevention of this parasitic disease.

Incidence of Japanese encephalitis, visceral leishmaniasis and malaria before and after the Wenchuan earthquake, in China.

The objective of this study was to investigate the incidence of insect-borne diseases from 2005 to 2011, before and after the 2008 Wenchuan earthquake in Longnan City, Gansu Province, China. The data include Japanese encephalitis, Kala-azar and malaria cases from 2005 to 2011 that occurred in Longnan City. We calculated the incidence rates and analyzed the epidemiological characteristics of the diseases before and after the Wenchuan earthquake. During 2005-2011, 212 Japanese encephalitis cases were reported in Longnan City, and the average incidence was 1.11/100,000. Compared with any year from 2005 to 2010 the incidence of Japanese encephalitis in Longnan City in 2011 was not significantly different (P≥0.05). From 2005 to 2011, there were 719 Kala-azar cases in Longnan City, the annual incidence was 3.77/100,000, and the incidence in males was higher than females (P<0.001). Compared with 2011, there was no significant difference in incidence of Kala-azar in 2009 or 2010 (P≥0.05). There were seven total cases of malaria from 2005 to 2011, and the annual incidence was 0.07/100,000. Wudu District and Wen County were the main endemic areas of insect-borne diseases in Longnan City. The results showed that Japanese encephalitis and Kala-azar were common insect-borne infectious diseases in Longnan City, and that the incidence of insect-borne disease did not increase after the Wenchuan earthquake. It is possible that vector control measures implemented after the earthquake prevented an increase in such diseases.

[Experimental treatment on alveolar hydatid disease in mice with multi-frequency focused ultrasound].

OBJECTIVE: To investigate the efficacy of multi-frequency focused ultrasound (MfFU) in the treatment of alveolar hydatid disease in mice. METHODS: Thirty Kunming mice infected subcutaneously with alveolar protoscoleces were divided into 3 groups randomly of 10 mice each and irradiated with different intensity of MfFU. Mice in the experiment groups B and C were irradiated only once for 5 min and group A served as control. The irradiation power of the 3 transducers in the low-power group (group B) was 4 W + 4 W + 5 W; that in the high-power group (group C) was 10 W + 11 W + 10 W. After the irradiation, the morphological change of alveolar tissues was observed with transmission electron microscope. The survival rate of protoscoleces was evaluated with methylene blue staining. The mitochondrial content in the alveolar tissues was detected with laser confocal microscope. The Coomassie brilliant blue staining and succinate dehydrogenase (SDH) kit were applied for measuring the amount of general proteins and the activity of succinate dehydrogenase of the alveolar hydatid cyst respectively. RESULTS: After irradiated by MfFU, transmission electron microscopy showed that in groups B and C the cells on the germinal layer decreased. The mitochondria swelled or broke. The endo cytoplasmic reticulum became swollen markedly. The karyotheca looked unclear. The microvilli shortened or disappeared. All the damages in group C displayed more seriously than in group B. The survival rate of protoscoleces in groups B (70.50%) and C (59.83%) was statistically lower than that of group A (82.33%) (P < 0.05). And there was also a statistical difference between groups B and C (P < 0.05). The amount of general proteins and the activity of SDH in groups B (3.07 mg/ml and 2.15 U/mg respectively) and C (2.87 mg/ml and 1.87 U/mg) were lower significantly than those in group A (3.83 mg/ml and 3.50 U/mg) (P < 0.05), but no statistical difference between B and C (P > 0.05). The mitochondria content in groups B (105.46 a.u/a) and C (70.90 a.u/a) was lower than that in group A (133.45 a.u/a). Group C showed statistical difference than A and B (P < 0.05), but there was no statistical difference between A and B (P > 0.05). CONCLUSION: It is evident that the cysts of Echinococcus multilocularis in mice can be damaged by MfFU which shows certain curative effect.

[In vitro influence of multi-frequency focused ultrasound on the ultrastructure of Echinococcus granulosus hydatid cysts].

OBJECTIVE: To investigate the change of Echinococcus granulosus hydatid cysts in vitro treated with single-frequency and multi-frequency focused ultrasound. METHODS: Mice were inoculated with echinococcus protoscolices from infected sheep, and sacrificed after one year. Forty cysts (about 2 cm in diameter) were taken from infected mice, and randomly divided into four groups. Cysts in the control group were immediately opened, fixed with 3% glutaraldehyde solution, and frozen. Cysts in groups A, B and C were treated with transducer No. 2 (4W), transducer No. 2 (4W) +No. 3 (5W), and transducer No. 1 (4W) +No. 2 (4W) +No. 3 (5W), respectively. After irradiated for 1 minute, the cysts were all fixed and refrigerated, then observed by naked eyes, transmission electron microscopy and scanning electron microscopy. RESULTS: The wall of hydatid cyst in the experimental groups became thicker and less transparent. Images from the transmission electron microscopy showed more serious damage of cyst wall with the increase in ultrasonic power and irradiation time. The fiber of the laminated layer became thicker and disordered, the nucleus in germinal layer became swollen and broken, the mitochondria were destroyed remarkably and the microvillus got shorter, fractured or even disappeared. In some occasions, only cell debris and broken nucleolus left. Scanning electron microscopy also demonstrated that with the increase in ultrasound power and irradiation time, the damage of the cyst wall became more serious even the normal structure was completely destroyed. CONCLUSION: Multi-frequency focused ultrasound causes significant damage of the Echinococcus granulosus hydatid in vitro, and the damage level is related to the ultrasound power and irradiation time.

[In vitro observation on albendazole sulfoxide and its enantiomers against Echinococcus granulosus protoscolex].

OBJECTIVE: To investigate in vitro anti-hydatid efficacy on Echinococcus granulosus protoscolex (EgPSC) by using albendazole sulfoxide (ASOX) and its two enantiomeric antipodes, L-ASOX and D-ASOX. METHODS: Eg protoscoleces were divided into eight groups and cultured in the DMEM culture media under two concentrations(50 microg/ml and 100 microg/ml) of ASOX, L-ASOX and D-ASOX respectively. The appropriate controls included (i) a culture containing an equal amount of DMSO and (ii) a culture medium alone. The mortality of EgPSC in each group was daily counted until 100% EgPSC death in some groups. RESULTS: Significant difference of EgPSC mortality was found among the three drugs with various concentrations compared to control group (P < 0.01), and a significant difference between L-ASOX group and D-ASOX group (P < 0.05). There were no statistical difference between ASOX group and D-ASOX group (P > 0.05), but between ASOX group and L-ASOX group (P < 0.05). On the 9th day of culture, the mortality of protoscoleces with the concentration of 50 microg/ml was 93.6%, 56.2% and 99.0% in ASOX, L-ASOX and D-ASOX groups respectively, and those under the concentration of 100 microg/ml were 100%, 74.5% and 100% respectively. The mortality was 19.1% and 22.5% respectively in the control and solvent groups. CONCLUSION: ASOX, L-ASOX and D-ASOX demonstrate significant effect of anti- Eg protoscolex in vitro. D-ASOX shows stronger effect than L-ASOX.

[Effect of combined pentoxifylline and albendazole against echinococcus multilocularis infection in mice].

OBJECTIVE: To observe the effect of pentoxifylline (PTX), albendazole (ABZ), and a combination of PTX and ABZ in mice infected with Echinococcus multilocularis (E.M). METHODS: The first part of the experiment was to observe the in vitro effect of PTX on the cultured E.M protoscolex. In the second part, mice were infected by abdominal inoculation of E.M and divided into groups given by ABZ 50 mg/kg-d, PTX 360 mg/kg-d, PTX 180mg/kgxd, and a combined regimen ABZ 50 mg/(kgxd)+PTX 180 mg/(kgxd). Another infected group and a uninfected group served as controls which received normal saline only. 100 days post-treatment, the mice were sacrificed for further observation. Indicators included wet weight of the cyst, cyst inhibition rate, level of serum cytokines TGF-beta determined by ELISA, IL-2 and IL-10 determined by radio-immunoassay (RIA). RESULTS: The inhibition rate on cysts of the combined ABZ and PTX was 88%, considerably higher than 58 % of the group ABZ. The serum TGF-beta and IL-10 decreased and IL-2 increased after treatment in comparison to the controls. CONCLUSION: The PTX and ABZ combination shows better effect on E.multilocularis infection than that of single ABZ. PTX might help increase immunity of the mice.