Novel study on the prevalence of Trichinella spiralis in farmed American minks (Neovison vison) associated with exposure to wild rats (Rattus norvegicus) in China.

Minks and brown rats are reservoir hosts for many endoparasites including those of the genus Trichinella, a group of parasite nematodes with a worldwide distribution. However, little is known about the prevalence of Trichinella sp. infection in the American mink (Neovison vison) and rats (Rattus norvegicus) in China. Therefore, we aimed to examine the prevalence of Trichinella sp. infection in farmed minks in Weihai city, Shandong province, China and infer the possible route for Trichinella transmission to farmed American minks. In total, 289 muscle samples from minks and 102 carcasses of rats were collected from Weihai City. The appearance of Trichinella sp. was examined using the pooled artificial HCl-pepsin digestion method. The results showed that muscle larvae were detected in 20 of 289 minks (6.92%) and 2 of 102 synanthropic rats (1.96%). The larval density of Trichinella sp. in mink samples ranged from 0.025 to 0.815 larvae per gram (lpg), while the average larval burden in rats was 0.17 lpg. The isolates derived from minks and rats were identified at the species level using multiplex polymerase chain reaction (PCR), which revealed that the size of the two PCR products matched that of T. spiralis at 173 bp. Furthermore, sequence analysis showed 100% identity of the 5S rDNA inter-gene spacer regions of the two isolates to that of T. spiralis. This study presents a novel report of T. spiralis-mediated infection in minks and synanthropic rats in China. We highlight the vulnerability of farmed minks to Trichinella infection through exposure to synanthropic rats, which may raise a public health concern of potential zoonotic risks for domestic animals.

Diabetic kidney disease-predisposing proinflammatory and profibrotic genes identified by weighted gene co-expression network analysis (WGCNA).

Diabetic kidney disease (DKD) is one of the most serious microvascular complications of diabetes. Despite enormous efforts, the underlying underpinnings of DKD remain incompletely appreciated. We sought to perform novel and informative bioinformatic analysis to explore the molecular mechanism of DKD. The gene expression profiles of GSE142025, GSE30528, and GSE30529 datasets were downloaded from the Gene Expression Omnibus database. After the GSE142025 data set was preprocessed, a gene co-expression network was constructed by weighted gene co-expression network analysis (WGCNA), and hub genes were selected in the key modules. Meanwhile, differentially expressed genes (DEGs) upregulated commonly were identified between the GSE30528 and GSE30529 datasets. Then, pathway and process enrichment analysis were performed for hub genes and commonly upregulated DEGs. Next, candidate targets were identified by comparing hub genes to commonly upregulated DEGs. Finally, reverse-transcription quantitative polymerase chain reaction (RT-qPCR) was carried out to validate the expression of candidate targets, and protein-protein interaction (PPI) network was constructed. A total of 17 modules were clustered by WGCNA, and the most significant turquoise module was selected. Based upon MM > 0.7 and GM > 0.7, 313 hub genes were screened out in turquoise module. Functional analysis of these 313 genes demonstrated their enrichment in pathways involved in leukocyte differentiation, cell morphogenesis, lymphocyte activation, vascular development, collagen synthesis, chemotaxis, and chemokine signaling. A total of 115 commonly upregulated DEGs were identified between the GSE30528 and GSE30529 datasets. Intriguingly, a total of six proinflammatory and profibrotic candidate targets were selected and validated in DKD mice in vivo, including CCR2, MOXD1, COL6A3, COL1A2, PYCARD, and C7. Based on WGCNA and DEG analysis of DKD datasets, six DKD-predisposing candidate targets were uncovered. The data suggest that inflammation and fibrosis are key mechanisms of DKD, and future studies may determine the causal link between the six proinflammatory and profibrotic genes and DKD.

Evaluation of potential anti-toxoplasmosis efficiency of combined traditional herbs in a mouse model.

Toxoplasma gondii is a worldwide spread protozoan and is able to infect almost all warm-blood animals. No effective drugs are available clinically on toxoplasmosis. Chinese traditional herbal medicines have provided remedies for many health problems. There exists a possibility that Chinese herbs may provide protection against T. gondii. This work aims to assess the protective efficacy of combined Chinese herbs against T. gondii. We screened five herbal medicines that have different pharmacological effects and combined them into a prescription according to the traditional Chinese medicine compatibility principle. The drug potential and protective efficacy were evaluated through a mouse model by determining the survival time, the parasite load in blood and tissues, the change of cell proportions in blood and histological detection. The results showed that the survival time of mice in the 500 mg Chinese herbs group and sulfadiazine group was significantly longer than that of the PBS control group. Also the parasite load in blood and tissues of 500 mg Chinese herbs and sulfadiazine groups was significantly lower than that of PBS group at 7 days post infection (dpi), which was in accordance with the result of histological detection. Monocyte and neutrophil of infected mice were remarkably increased while lymphocyte was dramatically decreased compared to that of blank group at 7 dpi. The results demonstrated that the 500 mg dosage of our Chinese herbs could slow down the replication of T. gondii and prolong the survival time of mice and could be considered as possible candidate drug against toxoplasmosis.

Efficacy and safety of ticagrelor versus clopidogrel with different dosage in high-risk patients with acute coronary syndrome.

BACKGROUND: Dual antiplatelet therapy is recommended as a standard antiplatelet strategy in acute coronary syndrome. For those with reduced pharmacologic response to clopidogrel, strengthening antiplatelet therapy (clopidogrel 150mg daily) may reduce adverse clinical events. Ticagrelor is a direct-acting inhibitor of the adenosine diphosphate receptor P2Y12 that has a more rapid onset and offset than clopidogrel. METHODS: In this retrospective study, we compared ticagrelor (180mg loading dose 90mg twice daily thereafter), clopidogrel (300mg loading dose, 75mg or 150mg daily thereafter) for the prevention of cardiovascular events in 273 high-risk patients admitted to coronary care unit with acute coronary syndrome. RESULTS: The rate of IST in hospital was significantly reduced in patients of ticagrelor group comparing with those receiving clopidogrel 75mg (0.69% vs 8.2%, p=0.009). Moreover, the TVR rate was less in the ticagrelor group than clopidogrel 75mg group (2.7% vs 13.1%, p=0.007) 6months follow-up. The incidence of MACCE has no difference between the two clopidogrel groups. Kaplan-Meier analysis of MACCE-free indicated that there was no difference between the three groups. Ticagrelor significantly increased the rate of minor bleeding compared with clopidogrel 75mg daily during hospital (45.5% vs 26.2%,p=0.012) and 6-month follow-up (66.9% vs 45.9%,p=0.004).Bleeding-free prognosis was significantly better in the clopidogrel 75mg daily group. CONCLUSIONS: In patients with acute coronary syndrome undergoing PCI, the rate of in-stent thrombosis and TVR were significantly reduced treated with ticagrelor compared with clopidogrel 75mg daily, without an increase of overall major bleeding, but with an increase of minor bleeding.

[Value of coronary CT angiography in assessment of bifurcation lesions].

OBJECTIVE: To evaluate the value of coronary CT angiography in assessment of bifurcation lesions. METHODS: The original image of 79 established and suspected coronary artery disease patients who underwent both coronary CT angiography and conventional artery angiography (CAG) sequentially were included in this analysis. Bifurcation lesions were assessed on primary and secondary vessels with diameter ≥ 2.0 mm, bifurcation lesions were graded according to Chen's classification. CAG was used as golden standard. The sensitivity, specificity, positive predictive value and negative predictive value were calculated. Spearman's test and Kappa test were used to evaluate the correlation and classification identity of the two methods. RESULTS: CAG evidenced 177 bifurcation lesions out of 445 bifurcation vessels and coronary CT detected 168 bifurcation lesions out of 404 bifurcation vessels with satisfactory imaging quality and 390 bifurcation vessels could be analyzed by both CAG and coronary CT. Sensitivity, specificity, positive predictive value and negative predictive value of coronary CT angiography were 94.2%, 94.6%, 90.7%, 96.1%, respectively. The results for the lesions at LM-LAD/LCX + LAD/Mid, LAD/Diag, RCA/PDA were more satisfactory and the sensitivity and specificity were as high as: 97.1% and 94.2%, 95.7% and 89.5%, 92.3% and 98.7%, respectively. There were significant correlations for evaluating the narrow degree of the opening of the bifurcation branch with these two methods (r = 0.799 58, P < 0.01) and for identifying I, II, III type bifurcation lesions (Kappa coefficient = 0.7959, P < 0.01) as well as for identifying the subtype bifurcation lesions (Kappa coefficient = 0.6328, P < 0.01) using the two methods. CONCLUSION: Coronary CT angiography is efficient in identifying the bifurcation lesions and offers a reasonable indication for bifurcation lesion classification.

[Effects of smoke on clinical prognosis of patients with acute ST-segment elevation myocardial infarction].

OBJECTIVE: To explore the effects of smoke on the clinical prognosis of patients with acute ST-segment elevation myocardial infarction (ASTEMI). METHODS: A total of 1213 consecutive ASTEMI patients were admitted into 20 hospitals in Liaoning province between May 2009 and May 2010. They were stratified into smoke (n = 588) and non-smoke (n = 625) groups. Basic demographic profiles, treatment data and clinical outcomes were compared between two groups. The primary endpoint was cardiac death and the secondary endpoints included non-fatal myocardial infarction, stroke and revascularization. Cox proportional hazard analyses were performed. RESULTS: The proportion of percutaneous coronary intervention (PCI) in the smoke group was significantly higher than that in the non-smoke group (40.8% vs 22.1%, P < 0.001). During the follow-up period, the medication rate was significantly higher in the smoke group than that in the non-smoke group (aspirin: 75.3% vs 62.2%, P < 0.001; clopidogrel: 40.5% vs 32.2%, P = 0.003; ß receptor blockade: 45.4% vs 36.0%, P = 0.001; angiotensin-converting-enzyme inhibitor/angiotensin II receptor blocker (ACEI/ARB): 38.3% vs 32.2%, P = 0.026; statins: 57.3% vs 44.2%, P < 0.001). During the follow-up period, the rate of cardiac death was lower in the smoke group than that in the non-smoke group (10.2% vs 24.2%, P < 0.001). No significant differences existed between two groups. During the follow-up period, the rate of cardiac death was significantly correlated with smoke (HR 2.777, 95%CI 1.113 - 6.928, P = 0.029), PCI (HR 0.208, 95%CI 0.062 - 0.700, P = 0.011), age (HR 1.049, 95%CI 1.005 - 1.095, P = 0.028), aspirin (HR 0.165, 95%CI 0.061 - 0.446, P < 0.001) and statins (HR 0.382, 95%CI 0.317 - 0.462, P < 0.001). CONCLUSION: Among the ASTEMI patients, the rate of cardiac death is significantly lower in the smoke group than that in the non-smoke group. And it is significantly correlated with such independent risk factors as smoke, PCI, age, aspirin and statins.

[Impact of high-density lipoprotein levels at hospital admission on the outcome of patients with acute ST-segment elevation myocardial infarction].

OBJECTIVE: To analyze the impact of high-density lipoprotein cholesterol (HDL-C) levels at hospital admission on the incidence of major adverse cardiovascular events (MACCE) in patients with acute ST segment elevation myocardial infarction (ASTEMI). METHODS: 1067 patients with ASTEMI who were admitted to the 20 hospitals in Liaoning region and with lipid profile tested within the 24 hours of admission from May 2009 until May 2010, were enrolled. Data on basic demographic, clinical, status on admission and method of treatment were collected. Rate on various medical use and MACCE (cardiovascular death, non-fatal myocardial infarction, revascularization and stoke) were compared between the two groups through follow-up observation. Cox proportional hazard analysis was estimation. RESULTS: The median HDL-C level was 1.27 mmol/L, with 587 patients having HDL-C below and 489 patients HDL-C above the median level. The incidence rates of non-fatal myocardial infarction and MACCE at one-year follow-up period, was higher in low HDL-C group (4.8% vs. 0.9%, P<0.001; 23.7% vs. 18.1%, P=0.03, respectively). At one month follow-up, the incidence rate of non-fatal myocardial infarction was higher in low HDL-C group (1.4% vs. 0.0%, P=0.01). At six month follow-up, the incidence rates of non-fatal myocardial infarction and MACCE on one-year follow-up was higher in low HDL-C group (2.8% vs. 0.4%, P=0.003; 18.3% vs. 13.7%, P=0.04, respectively). RESULTS: from Cox proportional hazards analysis indicated that age (HR=1.02, 95%CI: 1.006-1.035, P=0.005), diabetes (HR=1.05, 95%CI: 1.053-2.171, P=0.03), HDL-C level (HR=0.56, 95%CI: 0.340-0.921, P=0.02) were significantly related to the incidence of MACCE. CONCLUSION: The incidence rates of one year and six month MACCE (mainly non-fatal myocardial infarction) and one month non-fatal myocardial infarction were significant higher in patients with low than high HDL-C levels at admission while kept on the ascending along with time. Age, diabetes, HDL-C level were independent risk factors related to the incidence of MACCE.

Anticoccidial effect of halofuginone hydrobromide against Eimeria tenella with associated histology.

Halofuginone (stenorol) has been used as an effective anticoccidial reagent for decades but very little is known about its mode of action. In this study, chickens were inoculated with Eimeria tenella oocysts on 14-day-old and medicated with halofuginone at days 0, 1, 2, 3, 4, 5 and 6 post inoculations (groups 0, 1, 2, 3, 4, 5 and 6, respectively). Chickens in group 7 were taken as challenge-unmedicated control and in group 8 unchallenged-unmedicated control. The survival rate, body weight gains (BWG), oocysts production, cecal scores, bloody diarrhea and histological examinations were analyzed to evaluate the anticoccidial efficacy of halofuginone and to initially elucidate its mechanisms. Results showed that halofuginone which acted as a coccidiostatic can significantly enhance the BWG, and decrease both the oocyst shedding and cecal destruction caused by E. tenella infection. The histological slide examination noted that halofuginone was effective when provided 0-2 days post inoculation but only partially effective when applied 3-7 days post infection. The second-generation schizonts treated with halofuginone appeared vacuolated and degenerated. It is concluded that halofuginone can inhibit the parasite's invasion of host cecal hypothetical cell at the early stages of life cycle and later disturb the parasite's development by vacuolation of the schizonts. The resulting abnormal schizonts could not divide into schizoites and were eventually eliminated by the host's immune response.

[The impact of body mass index on the presentation, treatment and clinical outcomes of patients with ST-segment elevated myocardial infarction].

OBJECTIVE: To analyze the impact of body mass index (BMI) on the presentation, treatment, and clinical outcomes of patients with ST-segment elevated myocardial infarction (STEMI). METHODS: 1414 patients with STEMI who were admitted to the 20 hospitals in Liaoning region from May 2009 until May 2010 were enrolled. Patients were stratified according to the BMI levels as normal weight group (18.5 kg/m(2) ≤ BMI < 24.0 kg/m(2)) (n = 485), overweight (24.0 kg/m(2) ≤ BMI < 28.0 kg/m(2)) (n = 736), or obesity (BMI ≥ 28.0 kg/m(2)) (n = 193). Presentation, treatment and mortality during hospitalization, MACCE (cardiovascular death, non-fatal myocardial infarction, revascularization and stroke) were compared between the three groups at 3-month and 1-year follow-up. RESULTS: Obesity in patients with STEMI was associated with younger age (P < 0.001), being male (P < 0.001), with diabetes (P = 0.013) or hypertension (P < 0.001) and hyperlipidmia (P < 0.001). A higher prevalence of reperfusion treatment (P = 0.018), mainly percutaneous coronary intervention (PCI) (P < 0.001) was seen during the period of hospitalization. Rates of using other kinds of medicines as well as the mortalities during hospitalization, were similar among the groups with different BMI categories. At 3-month and 1-year follow-up, more use of asprin (3-months: P = 0.018; 1-year: P = 0.002) and ß-receptor blockers were seen in the obesity group (3-months: P = 0.025; 1-year: P = 0.030) while the use of other drugs were not significantly different among the three groups. The incidence rates of MACCE were not significantly different among the BMI categories while the cumulative survival rate was similar between obese group and normal weight group. Results from the Cox proportional hazards analysis indicated that factors as age (HR = 1.045, 95%CI: 1.028-1.062, P < 0.001), diabetes (HR = 1.530, 95%CI: 1.107 - 2.301, P = 0.041), hyperlipidmia (HR = 2.127, 95%CI: 1.317 - 3.435, P = 0.002), urgent PCI (HR = 0.473, 95%CI: 0.307 - 0.728, P = 0.001) and the use of ß-receptor blockers at 3-months follow-up period (HR = 0.373, 95%CI: 0.195 - 0.713, P = 0.003) were significantly related to the incidence of MACCE at 1-year follow-up period. CONCLUSION: Despite the fact that patients with obesity presented with STEMI at younger age and having received active treatment of reperfusion and medicine, both the 3-month and 1-year outcomes did not show significant difference among the BMI categories.

Seroprevalence of Toxoplasma gondii infection in pigs, in Zhejiang Province, China.

Seroprevalence of Toxoplasma gondii infection in pigs in 10 regions of Zhejiang Province, China, was obtained by enzyme-linked immunosorbent assay (ELISA). Anti- T. gondii antibodies were found in 53.4% (434/813) of pigs. Results were analyzed by a chi-square (χ(2)) test. Differences were observed according to farm size, animal age, and sampling regions. Seroprevalences in pigs raised on small farms (71.4%) were significantly higher than that (42.7%) on large farms (P < 0.05), and seroprevalence increased progressively with age. The seroprevalence ranged from 28.1% to 66.0% in different regions, with Jiaxing having the lowest level (28.1%), followed by Hangzhou (36.0%) and Taizhou (42.0%). This is the first study on seroprevalence of T. gondii infection in pigs in Zhejiang Province.

Ginsenoside Rg1 enhances immune response induced by recombinant Toxoplasma gondii SAG1 antigen.

Ginsenoside, the most important component isolated from Panax ginseng, exhibits a variety of biological activities. Particularly, ginsenoside Rg1 is known to have immune-modulating activities such as increase of immune activity of T helper (Th) cells. In the present study, we evaluated the immunomodulatory potentials of the Rg1 at three dose levels on the cellular and humoral immune responses of ICR mice against T. gondii recombinant surface antigen 1 (rSAG1). ICR mice were immunized subcutaneously with 50 µg Rg1 alone, 100 µg rSAG1 alone or with 100 µg rSAG1 dissolved in saline containing ginsenoside Rg1 (10 µg, 50 µg or 100 µg). After immunization, we evaluated the immune response using lymphoproliferative assay, cytokine and antibody measurements, and the survival times of mice challenged lethally. The results showed that the groups immunized with rSAG1 and Rg1 (50 µg, 100 µg) developed a high level of specific antibody responses against T. gondii rSAG1, a strong lymphoproliferative response, and significant levels of cytokine production, compared with the other groups. After lethal challenge, the mice immunized with the rSAG1 and Rg1 (50 µg, 100 µg) showed a significantly increased survival time compared with control mice which died within 6 days of challenge. Our data demonstrate that by addition of ginsenoside Rg1, the rSAG1 triggered a stronger humoral and cellular response against T. gondii, and that Rg1 is a promising vaccine adjuvant against toxoplasmosis, worth further development.

Cellular mechanisms underlying the laxative effect of flavonol naringenin on rat constipation model.

BACKGROUND & AIMS: Symptoms of constipation are extremely common, especially in the elderly. The present study aim to identify an efficacious treatment strategy for constipation by evaluating the secretion-promoting and laxative effect of a herbal compound, naringenin, on intestinal epithelial anion secretion and a rat constipation model, respectively. METHODS/PRINCIPAL FINDINGS: In isolated rat colonic crypts, mucosal addition of naringenin (100 microM) elicited a concentration-dependent and sustained increase in the short-circuit current (I(SC)), which could be inhibited in Cl- free solution or by bumetanide and DPC (diphenylamine-2-carboxylic acid), but not by DIDS (4, 4'- diisothiocyanatostilbene-2, 2'-disulfonic acid). Naringenin could increase intracellular cAMP content and PKA activity, consisted with that MDL-12330A (N-(Cis-2-phenyl-cyclopentyl) azacyclotridecan-2-imine-hydrochloride) pretreatment reduced the naringenin-induced I(SC). In addition, significant inhibition of the naringenin-induced I(SC) by quinidine indicated that basolateral K+ channels were involved in maintaining this cAMP-dependent Cl- secretion. Naringenin-evoked whole cell current which exhibited a linear I-V relationship and time-and voltage- independent characteristics was inhibited by DPC, indicating that the cAMP activated Cl- conductance most likely CFTR (cystic fibrosis transmembrane conductance regulator) was involved. In rat constipation model, administration of naringenin restored the level of fecal output, water content and mucus secretion compared to loperamide-administrated group. CONCLUSIONS: Taken together, our data suggest that naringenin could stimulate Cl- secretion in colonic epithelium via a signaling pathway involving cAMP and PKA, hence provide an osmotic force for subsequent colonic fluid secretion by which the laxative effect observed in the rat constipation model. Naringenin appears to be a novel alternative treatment strategy for constipation.

Cellular mechanism of adrenalin stimulated chloride secretion via beta-adrenoceptor in T84 cells.

In the present study, the intracellular regulatory pathways involved in the adrenalin-stimulated chloride secretion across T84 cells were investigated. Biphasic characteristics were observed in the Isc response to the basolateral addition of adrenalin (0.25 nM-100 microM). The biphasic response was almost abolished by removing ambient Cl(-). Chloride secretion was found to depend on the activities of basolaterally located Na+-K+-2Cl(-) cotransporters and K+ channels. The alpha-adrenoceptor antagonist phentolamine did not have any effect on either phase of adrenalin-induced Isc, while after pretreatment of the beta-adrenoceptor antagonist propranolol, the adrenalin-induced Isc was substantially abolished, suggesting the biphasic response may be mediated by the beta-adrenoceptor. Under whole cell patch-clamp conditions, T84 cells responded to adrenalin with a rise in inward current. The current, which exhibited a linear I-V relationship and time- and voltage-independent characteristics, was inhibited by the chloride channel blocker DPC and the reverse potential was close to the equilibrium potential for Cl(-) (0 mV), implying that the current was Cl(-) selective. When preloaded with a Ca2+-chelating agent, BAPTA/AM did not affect the Isc response to adrenalin, whereas the Isc was destroyed by pretreating the cells with an adenyl cyclase inhibitor, MDL12330A. These observations were further supported by the intracellular [cAMP] measurement experiment, indicating that adrenalin induced chloride secretion could be mediated by a beta-adrenoceptor only involving cAMP as an intracellular second messenger.