Skeletal muscle-secreted myokine interleukin-6 induces white adipose tissue conversion into beige adipose tissue in myostatin gene knockout pigs.

Myostatin (MSTN) is primarily expressed in skeletal muscle and plays an important role in the regulation of muscle growth and development as well as fat deposition; however, little is known about the molecular mechanism through which MSTN regulates body fat deposition. Therefore, in this study, we sought to identify the signaling pathways through which MSTN regulates fat accumulation in pigs. MSTN knockout (MSTN-/-) pigs showed increased muscle mass, decreased fat mass, and a leaner body composition. In this study, we found that the adipose tissue of MSTN-/- pigs exhibits the characteristics of beige adipose tissue, and the mRNA expression levels of beige adipose marker genes, including UCP3, Cidea, and CD137, were significantly increased. Remarkably, the observed beige phenotype was not adipocyte autonomous but rather caused by muscle-secreted myokine interleukin (IL)-6. This occurrence results in increased AMP-activated protein kinase (AMPK) phosphorylation in adipose tissue, which subsequently activates peroxisome proliferator-activated receptor gamma coactivator 1α and the conversion of white adipocytes to beige in pigs. Therefore, we concluded that MSTN deficiency leads to increased IL-6 secretion in skeletal muscle and activates AMPK in adipocytes, thereby increasing the beige adipose tissue in MSTN-/- pigs.

Comparison of contrast-enhanced ultrasound and contrast-enhanced computed tomography for the diagnosis of cervical lymph node metastasis in squamous cell carcinoma of the oral cavity.

Early detection of cervical lymph node metastasis (CLNM) from squamous cell carcinoma (SCC) of the oral cavity remains challenging. This prospective study was performed to evaluate the ability of contrast-enhanced ultrasound (CEUS) to detect CLNM from oral cavity SCC. Additionally, its diagnostic value was compared with that of contrast-enhanced computed tomography (CECT). Forty-eight consecutive patients with SCC of the oral cavity were enrolled. All subjects were examined preoperatively with both CEUS and CECT. Subsequently, neck dissections were performed for these patients, and cervical lymph nodes separated from the surgical specimens were assessed histologically. The diagnostic performance of these two examinations was compared based on the results of histopathology. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value, and Youden index for CEUS and CECT were 69.39% vs. 44.90%, 94.71% vs. 97.12%, 89.88%% vs. 87.16%, 75.56% vs. 78.57%, 92.92% vs. 88.21%, and 64.10% vs. 42.02%, respectively. A significant difference was observed in terms of sensitivity (P=0.024) and Youden index (rate difference 22.08%, 95% confidence interval 2.72-41.44%). Therefore, CEUS appears to be a promising diagnostic tool that is superior to CECT for detecting CLNM from SCC of the oral cavity, with a higher sensitivity.

Difference in resting-state fractional amplitude of low-frequency fluctuation between bipolar depression and unipolar depression patients.

OBJECTIVE: To investigate the difference in fractional amplitude of low-frequency fluctuation (fALFF) of localized brain activities in the resting-state between bipolar depression and unipolar depression patients and to find biological markers that differentiate the two groups of patients. PATIENTS AND METHODS: Thirteen patients with bipolar depression, 15 patients with unipolar depression, and 16 healthy control subjects that were matched in age and years of education were subjected to 3.0 T resting-state functional magnetic resonance scans. The values of whole brain fALFF were calculated and statistical analysis was performed. RESULTS: The fALFF-values of the right inferior temporal gyrus, left cerebellar posterior lobe, right middle temporal gyrus, left inferior frontal gyrus/insula, right inferior frontal gyrus/insula, left lingual gyrus and right middle temporal gyrus of the three groups showed significant differences (p < 0.05). Compared with the healthy control (HC) group, the fALFF-values of the unipolar depression (UD) patient group significantly increased in the right superior temporal gyrus, left insula, left inferior frontal gyrus, right inferior frontal gyrus, right supramarginal gyrus and right medial frontal gyrus but significantly decreased in the right medial occipital gyrus, left frontal lobe, right superior parietal lobule; the fALFF-values of the bipolar depression (BD) patient group significantly decreased in the left cerebellum posterior lobe, right lingual gyrus, left lingual gyrus, right middle temporal gyrus, left middle temporal gyrus, and left superior frontal gyrus and significantly increased in the right inferior frontal gyrus and left insula compared to those of the HC group; compared with those of the UD group, the fALFF-values of the BD group significantly decreased in the left middle occipital gyrus, right middle temporal gyrus, left middle frontal gyrus, and left medial frontal gyrus. CONCLUSIONS: The brain activities of BD and UD patients in the resting-state exhibit abnormalities, which differ between the two groups of patients.

Histone deacetylase inhibitor M344 significantly improves nuclear reprogramming, blastocyst quality, and in vitro developmental capacity of cloned pig embryos.

M344 is a novel histone deacetylase inhibitor. There is no report on the effect of M344 treatment on the development of pig embryos after somatic cell nuclear transfer (SCNT). In the present study, we investigated the effect of M344 on the blastocyst formation rate in cloned embryos, acetylation level of histone H4 lysine 12 (AcH4K12), and the expression of pluripotency-related genes , , and . Our results indicated that treatment with 5 µ M344 for 6 h improved the development of porcine embryos, in comparison with the untreated group (25.1% ± 5.0 vs. 10.9% ± 2.4; < 0.05). Moreover, M344-treated embryos had increased average fluorescence intensity of AcH4K12 at the pseudo-pronuclear stage ( < 0.05). However, no differences exist in Oct4, NANOG, and SOX2 expression in M344-treated and untreated SCNT blastocysts. In evaluating the effect of M344 on in vivo development, 845 M344-treated embryos were transferred into 3 surrogates, 1 of whom became pregnant and developed 3 fetuses. These findings suggested that M344 elevated the level of histone acetylation, facilitated the nuclear programming, and subsequently improved the developmental competence of pig SCNT embryos.

Application of digital surgical guides in mandibular resection and reconstruction with fibula flaps.

Surgical guides have been used widely in maxillofacial surgery. Details of the types of digital surgical guide used in mandibular resection and reconstruction with fibula flaps at the authors' institution are presented in this article. Three patients diagnosed with mandibular ameloblastoma underwent mandibular resection and reconstruction with fibula flaps using surgical guides for assistance. These digital surgical guides included a mandibular osteotomy guide, a fibular osteotomy guide, and a mandibular fixation guide. Surgical guides are helpful in improving the accuracy of operations and are appropriate for many types of mandibular resection and reconstruction.

TissueCypher(™): A systems biology approach to anatomic pathology.

BACKGROUND: Current histologic methods for diagnosis are limited by intra- and inter-observer variability. Immunohistochemistry (IHC) methods are frequently used to assess biomarkers to aid diagnoses, however, IHC staining is variable and nonlinear and the manual interpretation is subjective. Furthermore, the biomarkers assessed clinically are typically biomarkers of epithelial cell processes. Tumors and premalignant tissues are not composed only of epithelial cells but are interacting systems of multiple cell types, including various stromal cell types that are involved in cancer development. The complex network of the tissue system highlights the need for a systems biology approach to anatomic pathology, in which quantification of system processes is combined with informatics tools to produce actionable scores to aid clinical decision-making. AIMS: Here, we describe a quantitative, multiplexed biomarker imaging approach termed TissueCypher™ that applies systems biology to anatomic pathology. Applications of TissueCypher™ in understanding the tissue system of Barrett's esophagus (BE) and the potential use as an adjunctive tool in the diagnosis of BE are described. PATIENTS AND METHODS: The TissueCypher™ Image Analysis Platform was used to assess 14 epithelial and stromal biomarkers with known diagnostic significance in BE in a set of BE biopsies with nondysplastic BE with reactive atypia (RA, n = 22) and Barrett's with high-grade dysplasia (HGD, n = 17). Biomarker and morphology features were extracted and evaluated in the confirmed BE HGD cases versus the nondysplastic BE cases with RA. RESULTS: Multiple image analysis features derived from epithelial and stromal biomarkers, including immune biomarkers and morphology, showed significant differences between HGD and RA. CONCLUSIONS: The assessment of epithelial cell abnormalities combined with an assessment of cellular changes in the lamina propria may serve as an adjunct to conventional pathology in the assessment of BE.

The alteration of Id-1 and TSP-1 expression in mucoepidermoid carcinoma associated with its clinical features and prognosis.

Expression of Id-1 (inhibitors of DNA binding/differentiation protein 1) and TSP-1 (thrombospondin-1) in mucoepidermoid carcinoma and their relationship to pathological features and prognosis was studied. Moderately and poorly differentiated groups had significantly higher Id-1 positive expression rate (p<0.05) than well differentiated carcinoma. Stages III-IV showed significant increase of Id-1 positive expression rate (p<0.05) compared with stages I and II. Id-1 positive expression was significantly higher in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After that, patients with negative Id-1 expression had significantly higher tumor-free survival than patients with positive expression (p<0.05). Correlation between the expression of Id-1 and TSP-1 in mucoepidermoid carcinoma was negative (p<0.05). Poorly differentiated groups show significantly lower TSP-1 positive expression rate than well differentiated groups (p<0.05). No significant differences of TSP-1 positive expression were detected with clinical stage. TSP-1 positive expression was significantly lower in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After 5 years, patients with positive TSP-1 expression had significantly higher tumor-free survival than patients with negative TSP-1. Positive Id-1 expression is associated with high malignancy/poor prognosis; positive TSP-1 expression is associated with low malignancy/good prognosis. Protein expression status may help assess tumor malignancy and patient prognosis.

Intratumoral lymphangiogenesis in oral squamous cell carcinoma and its clinicopathological significance.

Lymphatic metastasis has always been regarded as a major prognostic indicator for disease progression and as a guide for therapeutic strategies to oral squamous cell carcinoma (OSCC), but to date, how tumor cells access and spread via the lymphatics have not been fully elucidated. Whether tumor cells metastasize by expansion and invasion of pre-existing peritumoral lymphatics or by the induction and invasion of newly formed lymphatics within tumors is controversial. In order to address this issue and find out the clinicopathological significance of intratumoral lymphangiogenesis, we investigated 86 archival specimens from patients with OSCC, quantitating lymph vessels by immunostaining with D2-40. We also quantified lymphatic invasion and examined the possible associations of all the above parameters with clinicopathological features and outcome. Higher intratumoral lymphatic density (ILD) and peritumoral lymphatic density (PLD) were both significantly associated with the presence of lymph node metastasis at the time of diagnosis and the outcome of the post-operation biopsy of 77 patients (P = 0.001). Higher ILD was significantly associated with a higher incidence of intratumoral lymphatic invasion, peritumoral lymphatic invasion and recurrence of tumor (P = 0.001 and P = 0.041 and P = 0.001, respectively). Patients with higher ILD exhibited shorter 5-year cumulative and disease-free survival (P = 0.001). Thus, lymphangiogenesis indeed occurs in oral squamous cell carcinoma; ILD might be used as an index to inflect the aggression of the disease, to evaluate the status of lymphatic metastasis, to separate patients at higher risk of an adverse clinical outcome.

Odontogenic tumours: a retrospective study of 1642 cases in a Chinese population.

A total of 1642 odontogenic tumour cases retrieved from the files of the College of Stomatology, Sichuan University, China were retrospectively analyzed for gender, age, tumour site and relative frequency of various types, and the data compared with that of previous reports. The final diagnosis in each case was based on the WHO 2005 histopathological classification of odontogenic tumours. Of these tumours 1592 (97.0%) were benign and 50 (3.0%) were malignant. Ameloblastoma (40.3%) was the most frequent type, followed by keratocystic odontogenic tumour (35.8%), odontoma (4.7%) and odontogenic myxoma (4.6%). The mean age of the patients was 32.1, with a wide range (3-84 years). The male-female ratio and maxilla-mandible ratio were 1.4:1 and 1:4.0, respectively. Ameloblastoma and keratocystic odontogenic tumours, important indications of extensive surgical procedures, are not considered rare in this Chinese population, whereas odontoma is uncommon.

[Pathologic changes of lymphatic capillaries after injection of U14 cells in rat tongue perineoplastic area].

OBJECTIVE: 1. To establish a model of neck lymphatic metastasis of cancer by injecting tumor cells into the rat tongue; 2. To observe the relationship between pathologic changes of perineoplastic lymphatic in different stages of implanted tumor growth and lymphatic metastasis of tumor cells. METHODS: 60 Kunming rats with injection of 5 x 10(6) U14 tumor cells into each rat tongue were chosen and randomized into 6 groups. In every 5 days, specimens of rat tongues with implanted tumor and neck lymph nodes with possible metastatic tumor cells were removed from each group of rats. Cryosections of rat tongue were processed with a modified enzyme-histochemical staining technique, using 5'-nucleotidase with HE to demonstrate lymphatic capillaries. Attention was paid to pathologic change of lymphatic capillaries in perineoplastic area of rat tongue in different growth stages of implanted tumor. Routing wax sections of lymph nodes processed with HE staining were examined under light microscopy to determine whether there were any metastasis tumor cells. RESULTS: 1. Metastasis to neck lymph nodes could be observed 15 days later after injection of U14 tumor cells into rat tongue. The situation of metastasis of lymph nodes varied greatly from tumor cells in bundle within marginal sinus to complete occupation of lymph nodes by U14 cells. 2. Tumor cells penetrated the lymphatic capillary by passing singly or in clumps between endothelial cells or through some destroyed endothelia. A great number of lymphatic with dilated cavity was found nearby the tumor tissue which facilitated tumor cells to enter the lymphatic cavity. 3. There were much more amount of lymphatic capillaries with dilated cavity around tumor mass comparing with that of normal tongue of the control group rats, and the number of lymphatic capillaries with opening cavity tend to increase with growth of implanted tumor. CONCLUSION: Direct and objective view of lymphatic capillaries can be obtained by means of the modified enzyme-histochemical staining technique, using 5'-nucleotidase with HE to demonstrate lymphatic capillaries; A great number of lymphatic capillaries with dented cavity in normal physical state are forced to be dilated during process of tumor growth, thus in turn increase the possibility for tumor cells to penetrate the lymphatic capillary by passing singly or in clumps between endothelial cells.

[Immunohistochemical findings on liver tissue in patients with hepatitis C].

OBJECTIVE: To investigate the distributions and infer relations of HCV antigens in the liver tissues. METHODS: Immunohistochemical techniques as polyclonal antibody to HCV, monoclonal antibody to HCV - NS3 and monoclonal antibody to HCV - NS5 were used to detect HCV antigens in 107 patients with hepatitis C. RESULTS: HCV antigens in liver tissues were identified on these three kinds of antibodies. The positive rates were 38.3%, 28.0% and 52.3% respectively. The positive granules were located in the cytoplasma of liver cells. The distributions of positive cells were scattered, diffusive and clustered. No obvious anatomic association was observed between the expression of HCV - NS3 and inflammatory reaction. HCV - NS5 positive cells were more commonly seen around or in the flammatory focci. CONCLUSION: The expressions of various components of HCV were demonstrated in different concentrations. The expressions of HCV - NS5 might be related to the pathogenesis of hepatitis C.

[Study of superinfection of HBV and HCV].

OBJECTIVES: To understand the situation in hepatitis B patients coinciding with HCV and to explore its influence on HCV on the replication of HBV. METHODS: Using ELISA, 712 hepatitis B patients were tested for serum anti-HCV and markers of HBV. RESULTS: Of the 712 patients, anti-HCV positive rate was 14.47% with the highest 48.98% in patients with severe hepatitis and the lowest 3.25% in patients with acute hepatitis. Markedly different anti-HCV positive rates (P < 0.001) in patients of different clinical stages were discovered. The more severe the case with longer the course, the higher the anti-HCV positive rates. In patients with superinfection of HBV and HCV, serum HBsAg, HBeAg and anti-HBcIgM positive rates were lower than those in patients with hepatitis B (P < 0.001, P < 0.001 and P < 0.05) but the anti-HBe positive rates were higher. All the differences showed an obvious statistical significance. CONCLUSION: Hepatitis B coinciding with HCV infection is responsible for the deterioration of the disease and towards its formation of its chronic phase as well as for the inhibition of HBV replication.

[Study on hepatitis G virus infection].

OBJECTIVE: To find out the situation of HGV infection in Shandong Province, and to explore the relations between HGV infection and HCV or HBV infection. METHODS: Enzyme linked immunosorbent assay (ELISA) was used to determine the serum anti-HGV in 1,082 patients with viral hepatitis, 77 patients with non A-E hepatitis and 361 blood donors. RESULTS: 53 patients whose serum anti-HGVs were positive (positive rate: 3.49%) were noticed. The anti-HGV positive rate (8.93%) in patients with Hepatitis C was remarkably higher than that (3.32%) in patients with Hepatitis B (chi 2 = 8.80, P < 0.01). The anti-HGV positive rate (4.82%) in patients with chronic hepatitis was significantly higher than that (0.79%) in patients with acute hepatitis (chi 2 = 10.79, P < 0.01). The anti-HGV positive rate (8.00%) in patients with severe hepatitis was obviously higher than that in patients with acute hepatitis (chi 2 = 10.23, P < 0.01). CONCLUSION: The manifestations of HGV infection can be expressed as virus-carriers, subclinical infection or various clinical types. Patients with Hepatitis C were more subjective to be overlapped with HGV than the patients with Hepatitis B; moreover, HCV or HBV infection superinfected with HGV is associated with exacerbation of patients' condition and the formation of chronic infection.

Odontogenic tumors. A demographic study of 759 cases in a Chinese population.

Seven hundred fifty-nine cases of odontogenic tumors retrieved from the files of College of Stomatology, West China University of Medical Sciences were classified according to the World Health Organization's Histological Classification of Odontogenic Tumors and compared with similar reports from other countries. Among these cases, 93.9% of the tumors were benign and 6.1% were malignant. Ameloblastomas predominated (58.6%) with a predilection for the mandible, while odontomas, generally regarded as the most frequent odontogenic tumor in North America, only accounted for 6.7%, the fourth most common tumor in this series. The mandible and the maxilla were divided into eight anatomic regions, and the distribution of each odontogenic tumor type amongst these regions was recorded. The relative incidence of each tumor type, patient age and gender were also compared with data from other selected large series. Geographic differences were noted in the relative incidence of ameloblastoma, odontoma and malignant odontogenic tumors among the Chinese/African, North American and Turkish/German groups. Ameloblastoma and malignant odontogenic tumors are not considered rare in a Chinese population.

[Study of HDV infection in Shandong province].

To understand the situation of HDV infection in Shandong province and to explore the relationship between HDV and HBV infections, 2681 samples infected with HBV were tested for serum anti-HDV with enzyme linked immunosorbent assay, in Shandong. Among them, 207 (7.72%) were found to be anti-HDV positive. Anti-HDV positive rates in HBsAg carriers and in hepatitis B patients were 3.16% and 13.15%, respectively. A significant difference (P < 0.001) was noticed between the two groups. In the second group, anti-HDV positive rates in patients with chronic active hepatitis and severe hepatitis were both higher than those in patients with acute hepatitis and chronic persistent hepatitis and the difference was significant (P < 0.001). The longer the history and the more severe the condition the higher of the anti-HDV positive rate was seen. The results indicated that the combined and over-lapping HDV infections were both responsible for the chronic status of hepatitis B and its deterioration.

Typing of murine cell-surface antigens by cellular radioimmunoassay.

A cellular radioimmunoassay utilizing 125I-labelled Protein A was used for detecting antigen-antibody complexes on glutaraldehyde fixed cells attached to microtiter plates. This method is rapid, sensitive and specific for revealing H-2 private and public specificities as well as Ia and Lyt antigens. As plates may be kept for months, several reactivities can be tested in one step on a large panel rendering a regular supply of animals unnecessary.