Molecular characterization and analysis of a novel calcium-dependent protein kinase from Eimeria tenella.

The calcium-dependent protein kinases (CDPKs) are unique enzymes found only in plants, green algae, ciliates and apicomplexan parasites. In this study, a novel CDPK gene of Eimeria tenella, designed EtCDPK3, was cloned using rapid amplification of cDNA ends (RACE) based on the expressed sequence tag (EST). The entire cDNA of EtCDPK3 contained 1637 nucleotides encoding 433 amino acids and the deduced EtCDPK3 protein had canonical characteristic domains identified in other CDPKs, including a well-conserved amino-terminal kinase domain and a carboxy-terminal calmodulin-like structure with 4 EF-hand motifs for calcium binding. The expression profiles of the EtCDPK3 gene in different development stages were investigated by real-time quantitative PCR. Messenger RNA levels from the EtCDPK3 gene were higher in sporozoites than in other stages (unsporulated oocysts, sporulated oocysts and merozoites). Western blot analysis showed that rabbit antiserum against recombinant EtCDPK3 could recognize a native 49 kDa protein band of parasite. Indirect immunofluorescent antibody labelling revealed dispersed localization of EtCDPK3 during the first schizogony and intense specific staining. EtCDPK3 was located at the apical end of the sporozoites after early infection of DF-1 cells and the protein was highly expressed. Inhibition of EtCDPK3 function using specific antibodies reduced the ability of E. tenella to invade host cells. These results suggested that EtCDPK3 may be involved in invasion and survival of the parasite intracellular stages of E. tenella. Because this kinase family is absent from hosts, it represents a valid target that could be exploited for chemotherapy against Eimeria spp.

Neuroprotective effects of LBP on brain ischemic reperfusion neurodegeneration.

AIM: The present study was conducted to investigate whether LBP had a protective effect on cerebral ischemic reperfusion injury and to determine the possible mechanisms. MATERIALS AND METHODS: Male Kunming (KM) mice were used to make the model cerebral artery occlusion/reperfusion (MCAO/R). The behavioral test was used to measure neurological deficit scores for evaluation of ischemic reperfusion damage of brain. The change of electroencephalograph (EEG) was monitored by Model SMUP-E Bio-electric Signals Processing System. The infarction area of brain was assessed in brain slices with 2% solution of 2,3,5-triphenyl tetrazolium chloride (TTC). Spectrophotometric assay was used to determine the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and lactate dehydrogenase (LDH), contents of malondialdehyde (MDA) and adenosine triphosphate (ATP) of the brain. RESULTS: The results showed that LBP at doses of 20 and 40 mg/kg markedly decreased the neurological deficit scores and the infarction area in MCAO/R mice. At the same time, LBP significantly decreased MDA content, and increased SOD, GSH-Px, CAT, LDH activities in ischemic reperfusion brain. CONCLUSIONS: These suggest that LBP might act as a potential neuroprotective agent against the cerebral reperfusion-induced injury in the brain through reducing lipid peroxides, scavenging free radicals, and improving the energy metabolism.

[Oncomelania hupensis control strategy during the stage moving towards elimination of schistosomiasis in China].

Oncomelania hupensis is the only intermediate host of Schistosoma japonicum, and O. hupensis control is an important measure for schistosomiasis control. With the progress of national schistosomiasis control program, the prevalence of schistosomiasis is low in China; however, there are still multiple challenges for O. hupensis breeding and schistosomiasis transmission risk. Considering the target of the national schistosomiasis elimination program and environmental protection in the new era, the introduction of precision identification, precision interventions and precision assessment into O. hupensis control may facilitate the progress towards elimination of schistosomiasis in China.

Accuracy of CT-guided biopsies in 158 patients with thoracic spinal lesions.

BACKGROUND: Inconsistent accuracies of CT-guided thoracic spinal biopsies have been reported in previous studies. PURPOSE: To determine the accuracy of CT-guided thoracic spinal biopsy, to compare the results with those previously reported, and to determine if there are any factors that influence the accuracy of CT-guided thoracic spinal biopsy. MATERIAL AND METHODS: In total, 158 consecutive CT-guided percutaneous thoracic spine procedures (performed at the Department of Spinal Surgery, Xi'an Red Cross Hospital between April 2000 and July 2010) were reviewed. The 158 lesions were categorized by location and radiographic features. Pathological and clinical follow-up were used to determine accuracy. RESULTS: The diagnostic accuracy of CT-guided thoracic spinal biopsy was 90.5% overall. Biopsy of metastatic bone disease (98.2%) was significantly more accurate than biopsies of primary tumors (80.9%) and of hematological malignancies (47.0%) (P < 0.05 and P < 0.005, respectively). The diagnostic accuracy of CT-guided thoracic spinal biopsy was significantly higher for the lower thoracic spine (97.6%) than for the middle (90.0%) or upper thoracic spine (80.4%) (P < 0.05 and P < 0.025, respectively). The diagnostic accuracy was significantly higher for lytic lesions (96.4%) than for sclerotic lesions (81.3%) (P < 0.010). The accuracy of biopsies performed using the transpedicular approach (91.0%) was not significantly different from that of biopsies performed using posterolateral approaches (91.5%) (0.25 < P < 0.5). CONCLUSION: Percutaneous CT-guided thoracic spinal biopsy is a viable alternative to open surgical biopsy. The diagnostic accuracy was not affected by any of the variables except for lesion level, histology, and radiographic features.

[Complete mitochondrial genomes of Paragonimus westermani in China and phylogenetic analysis of various geographical isolates].

OBJECTIVE: To compare the complete mitochondrial genome sequences of two phenotypes of Paragonimus westermani isolated from Fujian Province with different sizes of metacercariae, and perform a phylogenetic analysis of various geographical isolates of P. westermani from Asia, so as to identify the possible genetic characteristics associated with the P. westermani phenotypes. METHODS: P. westermani metacercariae with different sizes (large metacercariae, 380-420 µm in diameter; small metacercariae, 320-340 µm) isolated from freshwater crabs were used to infect dogs, and the eggs and adult worms of P. westermani were collected from the dog stool samples and lung tissues. Then, the egg size and morphology were compared. In addition, genomic DNA was extracted from the adult worms of the two phenotypes of P. westermani and used for the PCR amplification to yield the complete mitochondrial genome sequence. Sequence structure and phylogenetic analyses were performed based on the complete mitochondrial genome of P. westermani. RESULTS: Following infection with large and small P. westermani metacercariae, the adult worms recovered from the dog lung had a thick body, and had oral and ventral suckers. The ventral sucker was located slightly in front of the midline of the body, and testes, ovary and vitelline gland were seen in the adult worms. Following fixation, the adults appeared oval, with an approximately 1.7∶1 of the length-width ratio. The length and width of the eggs isolated from the fecal samples of dogs infected with large and small P. westermani metacercariae varied significantly, and the large metacercariae produced bigger eggs than the smaller metacercariae. Based on the morphological features of adults and eggs and the ITS2 sequences, both phenotypes were identified as P. westermani. The complete mitochondrial genome sequence analysis of adults showed almost consistent sequences in the protein-coding region of the mitochondrial genome of adult worms derived from large and small metacercariae, with a major variation seen in the former non-coding region. Sliding window analysis revealed the most polymorphic region within the ND4 gene across the mitochondrial genome from various geographical isolates of P. westermani, and phylogenetic analysis showed that both phenotypes were clustered into the Chinese branch of P. westermani, which was close to the Japanese branch and distinct from the South/Southeast Asian branch. CONCLUSIONS: The genetic distance between the phenotypes of P. westermani isolated from Fujian Province is near at a mitochondrial genome level, with no remarkable genetic differentiation seen; however, the mutation and structural changes in the non-coding region may result in the phenotypic variations. In addition, there is a distinct variation of the evolutionary rate in the mitochondrial coding genes, suggesting the selection of appropriate molecular markers during the phylogenic researches.

Combined neuroform intracranial stent and bioactive matrix detachable coil for embolization of a broad-necked persistent primitive trigeminal artery aneurysm. A case report.

SUMMARY: We report a patient with a wide-necked aneurysm arising at the bifurcation of the right internal carotid artery and the persistent primitive trigeminal artery (PPTA) treated successfully by Matrix detachable coil occlusion and assisted by a Neuroform intracranial stent. First, a Neuroform self-expanding intracranial stent was delivered via a 5-F Guider Softtip XP and placed as desired, then the aneurysm dome was embolized with two Matrix detachable coils through the interstices of the stent. The aneurysm was 80% occluded angiographically and the parent artery was patent. DSA imaging six months after the procedure showed the aneurysm to be obliterated at angiography and the neck tissue thickness of the aneurysm to be increased, but the parent artery diameter was not impacted. We describe the case in detail and discuss our preliminary experience of using the Neuroform stent and Matrix detachable coils for the treatment of a PPTA wide-necked aneurysm.

Complication analysis of 469 brain arteriovenous malformations treated with N-butyl cyanoacrylate.

SUMMARY: We independently assessed the frequency, severity and determinants of neurological deficits after endovascular embolization with NBCA of brain arteriovenous malformations (BAVMs) to have a better basis for making treatment decisions. All the charts of 469 BAVMs patients who underwent embolization with NBCA were reviewed. We analyzed the complications and their relation to angiographic features. The 469 patients were treated with 1108 endovascular procedures. Each met one to eight times, average 2.3 times. Eleven patients showed treatment-related complications, including four haemorrhagic and seven ischemic complications. Of these 11 cases, two died, two had persistent disabling deficits, and another seven suffered transient neurological deficits. Our finding suggests a low rate of disabling treatment complications for embolization of brain AVMs with NBCA in this center. The management of AVM patients who have high risk of embolization therapy should be treated by special strategy.

[Construction of cDNA library of Eimeria tenella sporulated oocysts].

A lambda ZAP express cDNA library was constructed using mRNA from Eimeria tenella sporulated oocysts. Total RNA was isolated by the TRIzol from Eimeria tenella sporulated oocysts, mRNA was further purified through oligo(dT)-cellulose columns. The first-strand cDNA was synthesized by using MMLV reverse transcriptase with oligo(dT)18 primers containing Xho I restriction site. After the second strand cDNA replacement synthesized, the uneven termini of the double-stranded cDNA were filled in with cloned Pfu DNA polymerase and EcoR I adapters were ligated to the blunt ends. Then the double-strand cDNA was digested with Xho I restriction enzyme. The fragments of 0.5 kb-4 kb were collected by agarose gel fraction method. After ligation of the cDNA with the lambda ZAP Express vector, the cDNA library was packaged using Gigapack III Gold Packaging extract. According to the phage plaques bright selection, the cDNA library contained 6 x 10(6) clones and the titer of the amplified library was 1 x 10(11) pfu/mL. By using PCR identification, the cDNA library contained approximately 96% recombinant phages.

Diarrhoea surveillance in children aged under 5 years in a rural area of Hebei Province, China.

A surveillance of diarrhoeal diseases in children aged under five years was carried out from March 1986 to February 1987 in a rural area of Hebei Province, China. Using a cluster sampling method, 270 study children were selected from 30 villages to represent a population of 105,405. Parents noted episodes of diarrhoea in children on a calender using a given definition of diarrhoea. The findings were verified by trained village doctors and were recorded monthly in the county Health and Anti-epidemic Stations. Stools of diarrhoea-affected children were examined to detect causal agents of diarrhoea, once in each of four seasons of the year. Of the 270 children, 218 (80.74%) suffered from one or more than one attacks of diarrhoea with a 95% confidence interval between 75.94% and 85.54%. There was no death due to diarrhoea. The diarrhoeal incidence in children of 3-4-year age group was significantly less than that in the children of less than 3 years (p less than 0.05). In the children with diarrhoea, the number of mean episodes was 3.15 per child/year, and the total mean course of suffering was 14 days per child/year. The ratio of diarrhoeal episodes in spring, summer, autumn, and winter was about 2.5:5:2:1. July was the month of highest occurrence of diarrhoea (146; 21.28% of all episodes). The epidemiological parameters of the disease declined with the growing age of the children, and there was no gender-specific preponderance in the number of episodes. During the whole year, 21.7% of the diarrhoeal episodes were due to bacterial pathogens, 4% of the episodes were due to rotavirus, and 3.65% episodes were associated with the detection of Trichomonas hominis.(ABSTRACT TRUNCATED AT 250 WORDS)