EEG microstate analysis reveals large-scale brain network alterations in depressed adolescents with suicidal ideation.

BACKGROUND: Accumulating evidence showed abnormalities in brain network connectivity in depressive individuals with suicidal ideation (SI). We aimed to investigate the large-scale brain network dynamics in adolescents with SI and major depressive disorder (MDD). METHODS: We recruited 47 first-episode drug-naïve adolescents with MDD and SI, 26 depressed adolescents without SI (noSI), and 26 age-matched healthy controls (HC). The Columbia Suicidal Ideation Severity Scale (C-SSRS) was utilized to assess suicide ideation. We acquired 64-channel resting-state EEG recordings from all subjects and used microstate analysis to investigate the large-scale brain network dynamics. RESULTS: We observed a significant reduction in the occurrence and coverage of microstate B within the SI group when contrasted with the noSI group. Conversely, there was a significant increase in the occurrence and coverage of microstate A in the SI group as compared to the HC group. Additionally, we observed heightened transition probabilities from microstates D and C to microstate A in the SI group; meanwhile, transitions from microstate D to B were more prevalent in the noSI group. Furthermore, the noSI group exhibited a significant decline in the transition probabilities from microstate D to microstate C. LIMITATIONS: The cross-sectional nature limits the capacity to determine whether microstate dynamics have prognostic significance for SI. CONCLUSION: We provided evidence that depressed adolescents with SI have a distinct pattern in microstate dynamics compared to those without SI. These findings suggest that microstate dynamics might serve as a potential neurobiomarker for identifying SI in depressed adolescents.

A resting-state electroencephalographic microstates study in depressed adolescents with non-suicidal self-injury.

Neuroimaging studies have revealed abnormal brain activities in depressed teenagers who engage in non-suicidal self-injury (NSSI). We used resting-state electroencephalography (EEG) microstate analysis, which indicates the brief overlap of brain network activation for exploring the characteristics of large-scale cortical activities in depressed adolescents engaged with NSSI to clarify the underlying temporal mechanism. A modified k-means cluster algorithm was used to segment 64-channel resting-state EEG data into microstates. Data from 27 healthy adolescents, 37 adolescents with major depressive disorder (MDD), and 53 adolescents with both MDD and NSSI were examined in this study. The resting-state microstate parameters were compared among groups using the one-way ANOVA and Spearman correlation. Then the associations between significantly different microstate parameters and the depressive severity and self-harming data in the patient groups were further analyzed. The MDD group had higher contribution (p < 0.01), occurrence (p < 0.01) of microstate A, and higher microstate E→A transition (p < 0.05) than the HC and the NSSI group. The MDD group showed a distinctly longer duration (p < 0.05) of microstate A and microstate A→C transition than the HC. The transition probability from B to C was increased in the NSSI group compared to the HC. In the MDD group, the HAMD correlated with the duration of microstate A (Spearman's rho = 0.34, p = 0.044), as the PHQ-9 correlated with its occurrence (Spearman's rho = 0.37, p = 0.028). This research revealed that whereas depressive adolescents with NSSI and MDD displayed similar patterns with healthy controls in EEG microstate, the MDD group did not. Additionally, the non-random transition from microstate E→A may protect against recent self-harm in adolescents with MDD.

Neurophysiological biomarkers for depression classification: Utilizing microstate k-mers and a bag-of-words model.

Microstates are analogous to characters in a language, and short fragments consisting of several microstates (k-mers) are analogous to words. We aimed to investigate whether microstate k-mers could be used as neurophysiological biomarkers to differentiate between depressed patients and normal controls. We utilized a bag-of-words model to process microstate sequences, using k-mers with a k range of 1-10 as terms, and the term frequency (TF) with or without inverse-document-frequency (IDF) as features. We performed nested cross-validation on Dataset 1 (27 patients and 26 controls) and Dataset 2 (34 patients and 30 controls) separately and then trained on one dataset and tested on the other. The best area under the curve (AUC) of 81.5% was achieved for the model with L1 regularization using the TF of 4-mers as features in Dataset 1, and the best AUC of 88.9% was achieved for the model with L1 regularization using the TF of 9-mers as features in Dataset 2. When Dataset 1 was used as the training set, the best AUC of predicting Dataset 2 was 74.1% for the model with L2 regularization using the TF-IDF of 9-mers as features, while the best AUC of predicting Dataset 1 was 70.2% for the model with L1 regularization using the TF of 8-mers as features. Our study provided novel insights into the potential of microstate k-mers as neurophysiological biomarkers for individual-level classification of depression. These may facilitate further exploration of microstate sequences using natural language processing techniques.

Altered Neural Reactivity in Adolescents With Nonsuicidal Self-Injury During Exposure to Self-Injury Related Cues: Electrophysiological Evidence From a Two-Choice Oddball Paradigm.

Background: Nonsuicidal self-injury (NSSI) may be a type of addiction, that is characterized by cue reactivity. We aimed to explore the behavioral performance and neural reactivity during exposure to self-injury cues in adolescents with NSSI and major depressive disorder (MDD). Methods: Eighteen MDD patients, 18 MDD patients with NSSI, and 19 healthy controls (HC) were recruited to perform a two-choice oddball paradigm. All subjects were 12-18 years old. Neutral cues and self-injury related cues separately served as deviant stimuli. Difference waves in N2 and P3 (N2d and P3d) were derived from deviant waves minus standard waves. Accuracy cost and reaction time (RT) cost were used as behavioral indexes, while the N2d and P3d were used as electrophysiological indexes; the N2d reflects early conflict detection, and the P3d reflects the process of response inhibition. Results: No significant main effects of group or cue or an effect of their interaction were observed on accuracy cost and P3d latency. For RT cost, N2d amplitude, and N2d latency, there was a significant main effect of cue. For P3d amplitude, there was a significant main effect of cue and a significant group × cue interaction. In the NSSI group, the P3d amplitude with self-injury cues was significantly larger than that with neutral cues. However, there was no such effect in the MDD and HC groups. Conclusions: Adolescents with NSSI showed altered neural reactivity during exposure to self-injury cue. Further studies with larger sample sizes are needed to confirm our results.

Prediction of Pathological Subtypes of Lung Adenocarcinoma with Pure Ground Glass Nodules by Deep Learning Model / 中国医学科学院学报

To make a preliminary pathological classification of lung adenocarcinoma with pure ground glass nodules(pGGN)on CT by using a deep learning model. CT images and pathological data of 219 patients(240 lesions in total)with pGGN on CT and pathologically confirmed adenocarcinoma were collected.According to pathological subtypes,the lesions were divided into non-invasive lung adenocarcinoma group(which included atypical adenomatous hyperplasia and adenocarcinoma in situ and micro-invasive adenocarcinoma)and invasive lung adenocarcinoma group.First,the lesions were outlined and labeled by two young radiologists,and then the labeled data were randomly divided into two datasets:the training set(80%)and the test set(20%).The prediction Results of deep learning were compared with those of two experienced radiologists by using the test dataset. The deep learning model achieved high performance in predicting the pathological types(non-invasive and invasive)of pGGN lung adenocarcinoma.The accuracy rate in pGGN diagnosis was 0.8330(95% =0.7016-0.9157)for of deep learning model,0.5000(95% =0.3639-0.6361)for expert 1,0.5625(95% =0.4227-0.6931)for expert 2,and 0.5417(95% =0.4029-0.6743)for both two experts.Thus,the accuracy of the deep learning model was significantly higher than those of the experienced radiologists(=0.002).The intra-observer agreements were good(Kappa values:0.939 and 0.799,respectively).The inter-observer agreement was general(Kappa value:0.667)(=0.000). The deep learning model showed better performance in predicting the pathological types of pGGN lung adenocarcinoma compared with experienced radiologists.

The Effect of Clostridium butyricum on Gut Microbiota, Immune Response and Intestinal Barrier Function During the Development of Necrotic Enteritis in Chickens.

Necrotic enteritis (NE) causes huge economic losses to the poultry industry. Probiotics are used as potential alternatives to antibiotics to prevent NE. It is known that Clostridium butyricum can act as a probiotic that can prevent infection. However, whether or not it exerts a beneficial effect on NE in chickens remains elusive. Therefore, we investigated the impact of C. butyricum on immune response and intestinal microbiota during the development of NE in chickens, including experimental stages with basal diets, high-fishmeal-supplementation diets, and Clostridium perfringens challenge. Chickens were divided into two groups from day 1 to day 20: one group had its diet supplemented with C. butyricum supplementation and one did not. At day 20, the chickens were divided into four groups: C. perfringens challenged and unchallenged chickens with and without C. butyricum supplementation. All groups were fed a basal diet for 13 days and thereafter a basal diet with 50% fishmeal from day 14 to 24. Chickens were infected with C. perfringens from day 21 to 23. At days 13, 20 and 24, samples were collected for analysis of the relative expression of immune response and intestinal mucosa barrier-related genes and intestinal microbes. The results show that C. butyricum can inhibit the increase in IL-17A gene expression and the reduction in Claudin-1 gene induced-expression caused by C. perfringens challenge. Moreover, C. butyricum was found to increase the expression of anti-inflammatory IL-10 in infected chickens. Although C. butyricum was found to have a significant beneficial effect on the structure of intestinal bacteria in the basal diet groups and decrease the abundance of C. perfringens in the gut, it did not significantly affect the occurrence of intestinal lesions and did not significantly correct the shift in gut bacterial composition post C. perfringens infection. In conclusion, although C. butyricum promotes the expression of anti-inflammatory and tight junction protein genes and inhibits pro-inflammatory genes in C. perfringens-challenged chickens, it is not adequate to improve the structure of intestinal microbiota in NE chickens. Therefore, more effective schemes of C. butyricum supplementation to prevent and treat NE in chickens need to be identified.

Overview of Experimental Methods for Antimicrobial Activity of Natural Drugs / 中国实验方剂学杂志

The antibiotics have obvious antibacterial and anti-inflammatory effects, but their toxic side effect, secondary infection and bacterial resistance have become a global problem. Traditional Chinese medicine(TCM) has always been the treasure of traditional culture and national characteristics in China since ancient times. It also has remarkable effect on inhibiting the growth of bacteria and killing pathogenic bacteria. The research on bacteriostatic experiment of TCM has gradually become a hot topic. Sensitivity experiments for such natural medicines have gradually become a research hotspot, but the complexity and particularity of natural medicines will vary with different methods. Therefore, different methods of drug sensitivity experiments should be matched with different natural drugs. By collecting and sorting out the relevant literature at home and abroad, this paper systematically summarizes the commonly used in vitro, in vivo and their combination bacteriostasis experimental methods of natural medicine activity, analyses the advantages and disadvantages of each method in the process of application, finds that different kinds of natural drugs have different applicable methods, and puts forward suggestions for the operation of each experimental method, in order to provide ideas for the selection of antibacterial susceptibility research experiments of natural medicines. It also provides a reliable reference method for solving the problem of antibiotic abuse and the development and utilization of natural medicines.

Temporal Effects of High Fishmeal Diet on Gut Microbiota and Immune Response in Clostridium perfringens-Challenged Chickens.

Necrotic enteritis (NE) caused by Clostridium perfringens is responsible for huge financial losses in the poultry industry annually. A diet highly supplemented with fishmeal is one factor predisposing chickens to the development of clinical NE. However, the effects of fishmeal-rich diets on the gut microbiota and immune response in chickens with C. perfringens challenge over the long-term are not well-understood. Here, a chicken NE model was established in which chickens were fed high fishmeal diet and subsequently infected with C. perfringens (FM/CP). Two control groups of chickens, one that was not infected and had a high fishmeal feeding (FM) and another group only infected with C. perfringens with basic diets (CP), were used as comparators. We analyzed the gut microbiota and immune response of the three groups at the age of 20, 24 [1 day post-infection (dpi)] and 30 days (7 dpi) using 16S rDNA sequencing and real-time PCR, respectively. We found that the composition of the gut microbiota had significant shifted in both the CP and FM/CP groups, although the CP group did not have intestinal lesions. The structure of the gut microbiota in C. perfringens-challenged chickens, independent of a high fishmeal diet, had the tendency to return to their non-infection state if the chickens no longer received C. perfringens challenge. Gut microbiota variation with time in challenged chickens with high fishmeal diet feeding was superimposed upon that of non-infected chickens with high fishmeal feeding. For the immune response, the relative expression of IL-8 in the ileum was significantly higher in infected chickens independent of high fishmeal feeding than in non-infected chickens. However, the expression of alpha 1-acid glycoprotein (AGP) and serum amyloid A (SAA) genes in chicken liver were significantly increased in FM/CP compared to the other groups. In conclusion, high fishmeal feeding induced significant changes to the structure of chicken gut microbiota over time and such changes provided an opening for C. perfringens infection to progress to NE. The relative expression of AGP and SAA in liver tissue may be used as diagnostic biomarkers for poultry NE but such an indication requires further investigation.

Effect of Huaier granule on recurrence after curative resection of HCC: a multicentre, randomised clinical trial.

OBJECTIVE: There is little evidence that adjuvant therapy after radical surgical resection of hepatocellular carcinoma (HCC) improves recurrence-free survival (RFS) or overall survival (OS). We conducted a multicentre, randomised, controlled, phase IV trial evaluating the benefit of an aqueous extract of Trametes robinophila Murr (Huaier granule) to address this unmet need. DESIGN AND RESULTS: A total of 1044 patients were randomised in 2:1 ratio to receive either Huaier or no further treatment (controls) for a maximum of 96 weeks. The primary endpoint was RFS. Secondary endpoints included OS and tumour extrahepatic recurrence rate (ERR). The Huaier (n=686) and control groups (n=316) had a mean RFS of 75.5 weeks and 68.5 weeks, respectively (HR 0.67; 95% CI 0.55 to 0.81). The difference in the RFS rate between Huaier and control groups was 62.39% and 49.05% (95% CI 6.74 to 19.94; p=0.0001); this led to an OS rate in the Huaier and control groups of 95.19% and 91.46%, respectively (95% CI 0.26 to 7.21; p=0.0207). The tumour ERR between Huaier and control groups was 8.60% and 13.61% (95% CI -12.59 to -2.50; p=0.0018), respectively. CONCLUSIONS: This is the first nationwide multicentre study, involving 39 centres and 1044 patients, to prove the effectiveness of Huaier granule as adjuvant therapy for HCC after curative liver resection. It demonstrated a significant prolongation of RFS and reduced extrahepatic recurrence in Huaier group. TRIAL REGISTRATION: NCT01770431; Post-results.

A Retrospective Study to Compare Five Induction Chemotherapy Regimens Prior to Radiotherapy in the Reduction of Regional Lymph Node Size in Patients with Nasopharyngeal Carcinoma.

BACKGROUND Combined chemotherapy and radiation therapy are used to treat nasopharyngeal carcinoma (NPC). Previous studies have shown that induction chemotherapy, given before radiotherapy, is beneficial in patients with local lymph node metastases. The aim of this study was to evaluate regional lymph node size in patients with NPC and the efficacy of five induction chemotherapy regimens given before radiotherapy. MATERIAL AND METHODS Between December 2007 and June 2011, 190 patients were included in this study, who had regionally advanced NPC (Stages II-IV). Five induction chemotherapy regimens were given prior to radiation: 98 patients (51.6%) received the TPF regimen (docetaxel, cisplatin, and fluorouracil); 56 patients (29.5%) received PF regimen (cisplatin and fluorouracil); 26 patients (13.7%) received the TP regimen (cisplatin and docetaxel); seven patients (3.7%) received combined nimotuzumab with TPF; three patients (1.6%) received a combination of the novel modified recombinant human endostatin (Endostar) with PF. The length and width of the regional lymph nodes were measured using neck B-mode (high-resolution grey scale) ultrasonography before chemotherapy and on the second day following completion of chemotherapy. Gastrointestinal tract and bone marrow suppression were also monitored during and after chemotherapy. RESULTS The TPF chemotherapy induction regimen resulted in an improved early response of lymph node size reduction, compared with the PF and TP chemotherapy induction regimens. The combined use of nimotuzumab with the TPF regimen improved efficacy by 15%. The combined use of Endostar improved the efficacy of the PF regimen by 56% (P<0.05). CONCLUSIONS In a retrospective study in patients with NPC, different induction chemotherapy regimens had different effects on lymph node size before radiation therapy.

Acoustic Radiation Force Impulse (ARFI) Elastography and Serological Markers in Assessment of Liver Fibrosis and Free Portal Pressure in Patients with Hepatitis B.

BACKGROUND The aim of this study was to investigate the feasibility of using acoustic radiation force impulse (ARFI) elastography, AST-to-platelet ratio index (APRI), and FIB-4 in assessing liver fibrosis and free portal pressure in patients with hepatitis B. MATERIAL AND METHODS We enrolled 126 patients with hepatitis B who underwent liver surgery at the General Surgery Department of the First Affiliated Hospital of Shihezi University Medical School from February 2013 to August 2015. Preoperatively, shear wave velocity (SWV) of the liver was measured with the Siemens S2000 ultrasound system to reflect liver stiffness. Serological markers were collected and fibrosis indices APRI and FIB-4 were calculated. Intraoperatively, liver tissues were harvested and free portal pressure (FPP) was measured. Postoperatively, fibrosis of liver tissues was pathologically staged. RESULTS The results of SWV, APRI, FIB-4, and FPP were all correlated with the degree of liver fibrosis (Spearman correlation coefficients: r=0.777, P<0.001; r=0.526, P<0.001; r=0.471, P<0.001; p<0.000; r=0.675, p<0.000). Receiver operating characteristic curve (ROC) analysis showed that the areas under the curve (AUC) of ARFI, APRI, and FIB-4 in diagnosing liver fibrosis were 0.830, 0.768, and 0.717, respectively, for stage F≥1; 0.861, 0.773, and 0.754, respectively, for stage F≥2; 0.941, 0.793, and 0.779, respectively, for stage F≥3; and 0.945, 0.783, and 0.754, respectively, for stage F=4. SWV, APRI, and FIB-4 were all correlated with FPP (Pearson correlation coefficients: 0.387, P<0.001; 0.446, P<0.001; 0.419, P<0.001). CONCLUSIONS ARFI, APRI, and FIB-4 can assess liver fibrosis in patients with hepatitis B when assessing the portal venous pressure. The difference in diagnostic efficacy between the 3 was not significant.

Usefulness of Contrast-Enhanced Ultrasonography for Predicting Esophageal Varices in Patients with Hepatitis B Virus (HBV)-Related Cirrhosis.

BACKGROUND The aim of this study was to investigate the usefulness of contrast-enhanced ultrasonography (CEUS) in predicting of esophageal varices (EV) and assessing high-risk EV in patients with hepatitis B virus (HBV)-related cirrhosis. MATERIAL AND METHODS Patients with HBV-related cirrhosis who had undergone endoscopy were prospectively recruited. Hepatic dynamic CEUS was performed. Regions of interest (ROI) were drawn on the hepatic artery, hepatic vein, portal vein, and liver parenchyma to measure the corresponding features, such as arrival times. Spearman's correlation analysis was used to determine the relations between several dynamic CEUS features and the degree of EV. Receiver operating characteristics (ROC) curves were constructed to investigate the diagnostic performance of CEUS in assessing the presence of EV and high-risk EV. RESULTS Fifty-eight patients (44 men; mean age 51.3 years) were included in this study. Of these, 18 (31.0%), 12 (20.7%), 11 (19.0%), and 17 (29.3%) of patients had grade 0, 1, 2, and 3 EV, respectively. Grade 2 and grade 3 EV were considered high-risk EV. Among the CEUS features, the area under the ROC curves of intrahepatic transit time (HV-HA, i.e., the difference between hepatic vein arrival time and hepatic artery arrival time) both for assessment of the presence of EV and high-risk EV (0.883 and 0.915, respectively) were larger than the other indices. HV-HA was negatively correlated with the grade of EV. An HV-HA of under 8.2 s indicated the presence of EV and under 7 s indicated high-risk EV. CONCLUSIONS Dynamic CEUS imaging is useful in assessing the presence of EV and high-risk EV in patients with HBV-related cirrhosis.

Multiple differential expression networks identify key genes in rectal cancer.

BACKGROUND: Rectal cancer is an important contributor to cancer mortality. OBJECTIVE: The objective of this paper is to identify key genes across three phenotypes (fungating, polypoid and polypoid & small-ulcer) of rectal cancer based on multiple differential expression networks (DENs). METHODS: Differential interactions and non-differential interactions were evaluated according to Spearman correlation coefficient (SCC) algorithm, and were selected to construct DENs. Topological analysis was performed for exploring hub genes in largest components of DENs. Key genes were denoted as intersections between nodes of DENs and rectal cancer associated genes from Genecards. Finally, we utilized hub genes to classify phenotypes of rectal cancer on the basis of support vector machines (SVM) methodology. RESULTS: We obtained 19 hub genes and total 12 common key genes of three largest components of DENs, and EGFR was the common element. The SVM results revealed that hub genes could classify phenotypes, and validated feasibility of DEN methods. CONCLUSIONS: We have successfully identified significant genes (such as EGFR and UBC) across fungating, polypoid and polypoid & small-ulcer phenotype of rectal cancer. They might be potential biomarkers for classification, detection and therapy of this cancer.

Diagnostic Accuracy of SuperSonic Shear Imaging for Staging of Liver Fibrosis: A Meta-analysis.

OBJECTIVES: The purpose of this study was to assess the performance of SuperSonic shear imaging (SuperSonic Imagine SA, Aix-en-Provence, France) for diagnosis of liver fibrosis. METHODS: Literature databases were searched to identify relevant studies from inception to February 28, 2015. Sensitivity, specificity, and other information were extracted from the studies. Pooled data were calculated by a bivariate mixed-effects binary regression model. Subgroup and sensitivity analyses were performed. Publication bias was tested by funnel plots. RESULTS: Twelve studies were included in this meta-analysis and reported on 1635 patients. The pooled sensitivity and specificity were 0.78 (95% confidence interval [CI], 0.69-0.85) and 0.95 (95% CI, 0.75-0.99), respectively, for fibrosis stages F≥1, 0.84 (95% CI, 0.81-0.86) and 0.81 (95% CI, 0.74-0.87) for F≥2, 0.89 (95% CI, 0.85-0.93) and 0.84 (95% CI, 0.77-0.89) for F≥3, and 0.88 (95% CI, 0.82-0.91) and 0.86 (95% CI, 0.81-0.90) for F=4. The areas under the summary receiver operating characteristic curves were 0.87 (95% CI, 0.84-0.90) for F≥1, 0.85 (95% CI, 0.81-0.88) for F≥2, 0.93 (95% CI, 0.91-0.95) for F≥3, and 0.93 (95% CI, 0.90-0.95) for F=4. No significant publication bias was found. CONCLUSIONS: SuperSonic shear imaging could be used for staging of liver fibrosis. Especially, it has high diagnostic accuracy for severe fibrosis and cirrhosis.

Efficacy of Albendazole-Chitosan Microsphere-based Treatment for Alveolar Echinococcosis in Mice.

This study aimed to investigate the pharmacology and anti-parasitic efficacy of albendazole-chitosan microspheres (ABZ-CS-MPs) for established intraperitoneal infections of Echinococcus multilocularis metacestodes in an experimental murine model. Male outbred Kunming mice infected with E. multilocularis Metacestodes were administered with three ABZ formulations, namely, ABZ-CS-MPs, Liposome-Albendazole (L-ABZ), and albendazole tablet (ABZ-T). Each of the ABZ formulations was given orally at three different doses of 37.5, 75, and 150 mg/kg, three times a week for 12 weeks postinfection. After administering the drugs, we monitored the pharmacological performance and anti-parasitic efficacy of ABZ-CS-MPs compared with L-ABZ, and ABZ-T treated mice. ABZ-CS-MPs reduced the weight of tissues containing E. multilocularis metacestodes most effectively compared with the ABZ-T group and untreated controls. Metacestode grown was Highly suppressed during treatment with ABZ-CS-MPs. Significantly higher plasma levels of ABZ metabolites were measured in mice treated with ABZ-CS-MPs or L-ABZ compared with ABZ-T. In particular, enhanced ABZ-sulfoxide concentration profiles were observed in the mice given 150 mg/kg of ABZ-CS-MPs, but not in the mice treated with L-ABZ. Histological examination showed that damages caused disorganization of both the germinal and laminated layers of liver hyatid cysts, demolishing their characteristic structures after treatment with ABZ-CS-MPs or L-ABZ. Over time, ABZ-CS-MPs treatment induced a shift from Th2-dominant to Th1-dominant immune response. CS-MPs As a new carrier exhibited improved absorption and increased bioavailability of ABZ in the treatment of E. multilocularis infections in mice.

[Efficacy of albendazole chitosan microspheres against Echinococcus granulosus infection in mice].

OBJECTIVE: To observe the therapeutic effect of albendazole chitosan microspheres (ABZ-CS-MPs) on cystic echinococcosis in mice. METHODS: Two hundred male kunming mice were each infected by intraperitoneal inoculation of about 5 000 viable protoscoleces of Echinococcus granulosus. Another 20 mice were kept as blank control. After 12 weeks post infection, the mice were randomly divided into four groups named as infection control group (n = 20), ABZ-CS-MPs group, albendazole liposome (L-ABZ) group, and albendazole tablet group. The latter three treatment groups were then each divided into three subgroups (n = 20) by given the dose of 37.5, 75.0, and 150.0 mg/kg for three times per week, respectively. After 12 weeks of treatment, all mice were sacrificed. The weight of hydatid cysts was measured and the inhibition rate were calculated. Mouse liver was observed. The histopathological changes of E. granulosus were observed by microscopy. The concentration of albendazole sulfoxide in plasma and liver tissues was determined by high-performance liquid chromatography. RESULTS: Compared with the other treatment groups, the turbidity of contained fluid, the consolidation level and calcification level of hydatid cysts in ABZ-CS-MPs group were higher. The average weight of hydatid cysts in each treatment group was lower than that of infection control group [(3.19 +/- 2.94) g] (P < 0.05). The cyst weight in 37.5, 75.0, and 150.0 mg/kg ABZ-CS-MPs group [(0.28 +/- 0.28), (0.24 +/- 0.22), and (0.20 +/- 0.19) g, respectively] was lower than that of albendazole tablet groups [(0.77 +/- 0.74), (0.55 +/- 0.42), (0.76 +/- 0.35) g] (P < 0.05). Among the same dosage groups, the inhibition rate in ABZ-CS-MPs group (from low to high dosage sub-group: 91.1%, 92.6%, and 93.7%, respectively) was highest. In 75.0 mg/kg ABZ-CS-MPs group, there were 15 mice with class I (degeneration) and II (necrosis) pathological changes of E. granulosus hydatid. The number of mice with class I and II pathological changes in each dosage ABZ-CS-MPs sub-group and L-ABZ sub-group was more than that of albendazole tablet group (P<0.05). Plasma concentration of albendazole sulfoxide in 75.0 and 150.0 mg/kg ABZ-CS-MPs sub-groups [(0.83 +/- 0.39), (0.80 +/- 0.5) microg/ml] were higher than that of L-ABZ sub-groups [(0.34 +/- 0.03), (0.43 +/- 0.15) microg/ml] and albendazole tablet sub-groups [(0.31 +/- 0.02), (0.40 +/- 0.10) microg/ml] (P < 0.05). Compared with 37.5, 75.0, and 150.0 mg/kg albendazole tablet sub-groups [(0.04 +/- 0.02), (0.07 +/- 0.04), (0.04 +/- 0.0) microg/g], the albendazole sulfoxide concentration in liver tissue was higher in ABZ-CS-MPs sub-groups [(0.33 +/- 0.06), (0.45 +/- 0.31), (0.50 +/- 0.30) microg/g] (P < 0.05). In 37.5 mg/kg dosage sub-group, the albendazole sulfoxide concentration in liver tissue in ABZ-CS-MPs group was higher than that of L-ABZ group [(0.14 +/- 0.19) microg/g] (P < 0.05). CONCLUSION: ABZ-CS-MPs can reduce the weight of hydatid cyst and increase the concentration of al-bendazole sulfoxide in plasma and liver tissue of mice.

[Advances in research on the MAPK signal transduction pathway of Echinococcus].

Mitogen-activated protein kinase (MAPK) is an important signaling transduction molecules, which can enter the nucleus and activate target gene when it was stimulated and become phosphorylation. MAPK signaling pathway is closely associated with various diseases. Recent studies have indicated that MAPK signaling transduction pathway is also involved in the growth and development of Echinococcus. This review summarizes the progress on the relationship between MAPK signal pathway and Echinococcus.

[Expression of tumor necrosis factor-alpha and caspase-3 protein in monocytes adjacent to the invaded Echinococcus multilocularis in liver].

OBJECTIVE: To investigate the expression and its significance of tumor necrosis factor-alpha (TNF-alpha) and caspase-3 protein in monocytes adjacent to the invaded Echinococcus multilocularis in liver. METHODS: 40 female Kunming mice were randomly divided into experimental group (n=20) and sham operation (control) group (n=20). Mice in experimental group were infected with 20% E. multilocularis suspension (0.1 ml per mouse) through abdominal opening injection in liver and the mice in control group were injected with equal physiological saline. The mice were sacrificed at 6 months post-infection for observing the growth and metastasis of E. multilocularis. Pathological changes were observed by HE staining. The expression of TNF-alpha and caspase-3 protein in hydatid cyst and metastasis tissue were detected by immunohistochemistry staining and the apoptosis of the monocytes was measured by TUNEL. RESULTS: After 6 months post-infection, E. multilocularis were spread over the liver of the mice in experimental group. Metastasis rate of lymph nodes was 45.0% (9/20). Infiltration of monocytes was observed around E. multilocularis in liver and lymph nodes with metastasis by HE staining. Immunohistochemistry showed that the positive expression rate of TNF-alpha and caspase-3 protein in monocytes was 100% and 100%, and 95% and 100% respectively around the cyst in experimental group, while the expression rate was only 5% and 0 respectively in the liver of the control mice (P<0.01). The monocytes showed significant apoptosis by TUNEL in experimental group with a positive expression rate of 100%, with a significant difference between experiment group and the control (P<0.01). CONCLUSION: In the process of alveolar Echinococcus infection, the high expression of TNF-alpha protein might be associated with the apoptosis of monocytes, which may inhibit the host immunological function.

[Effect of anti-osteopontin antibody on expression of IL-2 and IL-5 in hepatic alveolar hydatid tissue of gerbil].

OBJECTIVE: To observe the effect of anti-osteopontin antibody on the level of IL-2 and IL-5 in the liver of gerbil infected with Echinococcus multilocularis (Em). METHODS: 180 gerbils were infected with echinococcus protoscoleces (approximately 400 for each gerbil) by abdominal opening inoculation in liver. The gerbils were randomly divided into three groups with 60 each: experiment group (group A, with anti-osteopontin antibody interference), control group (group B, with normal rabbit serum injection) and model group (group C, with no interference). Ten gerbils from each group were sacrificed at 20, 60, 100, 140, 180, and 220 days post-infection respectively. The liver tissue with hydatid cysts was collected and the expression of IL-2 and IL-5 was observed after immunohistochemistry staining (SP method). RESULTS: E. multilocularis hydatid tissue spreaded over the liver and abdominal cavity. The positive expression rate of IL-2 in the tissue showed no statistical difference among the three groups (P > 0.05). On the days 140 and 180, however, the positive expression rate of IL-5 in group A was 40% and 20% respectively, considerably lower than that in group B (100% and 90%) and group C (90% and 80% respectively). CONCLUSION: The anti-osteopontin antibody can reduce Th2 type cytokine response in the Em-infected gerbils, which may strengthen the immunity of the host.

Pericyst may be a new pharmacological and therapeutic target for hydatid disease.

BACKGROUND: Most hydatid cysts with calcified walls are biologically and clinically silent and inactive. Transforming growth factor-beta 1 (TGF-ß1) plays a critical role in the calcification process of cells. The aim of this study was to assess the effect of modulating TGF-ß1 signaling on the calcification of hydatid cysts. METHODS: Pericyst cells isolated from hepatic hydatid cysts were cultured with osteogenic media. These cells were assessed for alkaline phosphatase activity and mineralization capacity using Alizarin Red staining. Cells were also treated with recombinant human TGF-ß1 and TGF-ß inhibitor, and the expression profiles of osteoblast markers (RUNX2, osterix, and osteocalcin) were analyzed using Western blotting. The effects of inhibiting TGF-ß1 signaling on calcification of pericyst walls were assessed using different doses of TGF-ß inhibitor for 7 weeks in a preclinical disease model of liver cystic echinococcosis. RESULTS: Cells within the pericyst displayed high levels of alkaline phosphatase activity and mineralized nodule formation, as induced by osteogenic media. These activities, as well as expression profiles of osteoblast markers (RUNX2, osterix, and osteocalcin) could be inhibited by addition of recombinant human TGF-ß1 (rhTGF-ß1) and enhanced by TGF-ß inhibitor. In the animal model of cystic echinococcosis, inhibition of TGF-ß1 signaling increased calcification of the pericyst wall, which was associated with decreased cyst load index and lower viability of protoscoleces. CONCLUSIONS: Cells within the pericysts adopt an osteoblast-like phenotype and have osteogenic potential. Inhibition of TGF-ß1 signaling increases hydatid cyst calcification. Pharmacological modulation of calcification in pericysts may be a new therapeutic target in the treatment of hydatid disease.