Immunohistochemical examination of intracerebral aquaporin-4 expression and its application for differential diagnosis between freshwater and saltwater drowning.

Human brain samples were collected from 70 autopsy cases including 22 freshwater drowning (FWD), 26 saltwater drowning (SWD), and 22 non-drowning cases as controls. Then, immunohistochemical study combined with morphometry was carried out in order to examine the differential expression of AQP1 and AQP4 in the brain samples. Immunohistochemically, star-shaped cells bearing highly branched processes, often surrounding blood vessels, showed positive reactions for AQP1 and AQP4 in FWD, SWD, as well as control groups. Additionally, with double-color immunofluorescence analysis, AQP1- or AQP4-positive cells could be identified as GFAP-positive astrocytes. Moreover, AQP1-positive reaction was also observed in blood vessels. Morphometrically, there were no significant differences in AQP1 expression in astrocytes or in blood vessels among the three groups. In contrast, the average value of AQP4-positive astrocytes was significantly higher in FWD cases than in SWD and control groups. Moreover, AQP4 expression was significantly lower in SWD than in the control group (p < 0.05). Moreover, there was no significant correlation between post-submerged interval and AQP expression in drowning cases. Therefore, immunohistochemical analysis of intracerebral AQP4 expression would be forensically useful for differentiation between FWD and SWD.

Forensic application of intrarenal aquaporin-2 expression for differential diagnosis between freshwater and saltwater drowning.

Aquaporins (AQPs) are a family of homologous water channel proteins. In this study, the expressions of AQP1, 2, and 4 were immunohistochemically examined in kidney samples to evaluate their forensic applicability to differentiate between freshwater drowning (FWD) and saltwater drowning (SWD). Kidney samples were obtained from 51 drowning cases (23 FWD and 28 SWD) and 19 non-drowning cases. AQP1 was expressed in the proximal tubules and glomeruli, and AQP4 was localized in the collecting ducts. However, there were no significant differences in AQP1 and AQP4 expressions among FWD, SWD, and control groups. Immunohistochemically, AQP2 was predominantly expressed in the apical plasma membrane of the collecting duct principal cells in all kidney samples of FWD and SWD. Morphometrically, AQP2 expression at the apical plasma membrane of collecting ducts was significantly enhanced in SWD group, compared with FWD and control groups. On the other hand, AQP-2 expression was significantly lower in FWD group than in control group. Moreover, in drowning cases, there was no correlation between post-submersion intervals and AQP expression. From a forensic aspect, immunohistochemical detection of AQP2 in the kidney can be considered a valuable marker to differentiate between FWD and SWD.

Bioinspired Drug Delivery Carrier for Enhanced Tumor-Targeting in Melanoma Mice Model.

As a widely used first-line chemotherapy drug for tumor, Doxorubicin (DOX) can induce various side effects on normal tissues because of its non-specific distribution in the body. Emerging evidence has shown that platelets have the capability to recognize and interact with tumor cells. Inspired by this, the platelet-based drug delivery system was constructed by loading of DOX in platelet cytoplasm and modification of transferrin on the surface of platelet (Tf-P-DOX). The encapsulation efficiency of DOX in platelet was the highest at the DOX concentration of 0.05 mM, and reached to 64.9%. Fluorescence microscopy showed that the Tf-P-DOX facilitated cell uptakes and enhanced intracellular drug accumulation in B16F10 cells. Compared with free DOX, Tf-P-DOX exhibited an enhanced effect on cell apoptosis at the same concentration of DOX. In vivo imaging system showed that the near-infrared fluorescence of B16F10 tumor-bearing mice was mainly accumulated in the tumor site, which caused the inhibition of tumor growth in mice. The morphological changes of tumor tissue in Tf-P-DOX group was significant in comparison with those of the control group, including the small nucleus, the insufficiency of cancerous nest, and the infiltration of inflammatory cells, while Tf-P-DOX did not show significant adverse effects on normal tissues. Compared with the control group, the levels of caspase 9 and caspase 3 protein expressions were increased significantly in Tf-P-DOX group. Our studies suggest platelets can be repurposed as promising carriers for efficient targeting and treatment of solid tumors.

Clinicopathological analysis of CD8-positive lymphocytes in the tumor parenchyma and stroma of hepatocellular carcinoma.

Tumor-infiltrating lymphocytes may be a manifestation of antitumor immunity. In the present study, hepatocellular carcinoma (HCC) and pericancerous non-tumor liver tissues samples were obtained from 86 surgical patients who had not received preoperative treatment. The cellular expression levels of CD4 and CD8 were immunohistochemically examined in the two tissue groups using tissue microarrays, to evaluate their clinicopathological relevance. Immunohistochemically, CD4 and CD8 T cells were observed in the tumor parenchyma and tumor stroma, and the intensity of CD4 and CD8 immunoreactivity was homogeneous in all HCC samples examined. Morphometrically, the average numbers of CD4- and CD8-positive T cells were significantly increased in the tumor stroma, compared with those in the tumor parenchyma (tumor stroma versus tumor parenchyma: 22±3.6 versus 7.4±0.9 in CD4, 32.8±4.2 versus 16±2.5 in CD8; both P<0.01). Furthermore, the average numbers of CD8-positive T cells in the tumor parenchyma and stroma were significantly increased, compared with the average numbers of CD4-positive cells (P<0.05). In addition, in the tumor parenchyma and stroma, the average numbers of CD8 T cells were significantly higher in patients with tumor diameters ≤5 cm compared with those in patients with tumor diameters >5 cm (diameter ≤5 cm versus diameter >5 cm: 18.1±3.3 versus 12.2±3.8 in tumor parenchyma, 36.5±4.8 versus 21.9±8.9 in tumor stroma; both P<0.05). In addition, CD8 expression was significantly enhanced in patients with chronic hepatitis and cirrhosis, compared with paired tumor parenchymal tissues (P<0.01). Furthermore, a significant positive correlation was observed between CD4 and CD8 expression in the tumor parenchyma and stroma (both P<0.001). These observations suggest that tumor parenchyma- or stroma-infiltrating CD8 T cells may be involved in HCC tumor diameter control.

Immunohistochemical detection of CCR2 and CX3CR1 in sepsis-induced lung injury.

Sepsis is a systemic inflammatory disease with high mortality. In the present study, we immunohistochemically examined CCR2 and CX3CR1 expression in sepsis-induced lung injury, and discussed its availability for the postmortem diagnosis of sepsis. Lung samples were obtained from different lung lobes of nine sepsis and eight control cases with postmortem intervals between 12 and 48h. Immunohistochemically, mononuclear cells recruited into the lungs expressed CCR2 and CX3CR1 in both sepsis and non-septic groups. In double-color immunofluorescence analysis, CCR2- or CX3CR1-positive cells could be identified as CD68-positive macrophages. Moreover, most of CD68-positive macrophages expressed both CCR2 and CX3CR1. Morphometrically, the average of CCR2- and CX3CR1-positive macrophages was significantly increased in sepsis group, compared with control group (sepsis vs. control: 41.6+/-1.3% vs. 8.0+/-0.4% in CCR2; 36.2+/-1.3% vs. 9.2+/-0.4% in CX3CR1). These observations implied that CCR2- or CX3CR1-positive macrophages were recruited into the lungs under several pathological conditions. In particular, their recruitment might be more evident in sepsis. Moreover, from the forensic aspects, immunohistochemical detection of CCR2 and CX3CR1 expression in the lungs can be considered as valuable diagnostic tools for the postmortem diagnosis of sepsis.

Monosodium glutamate (MSG): a villain and promoter of liver inflammation and dysplasia.

Chronic inflammation is a common theme in a variety of disease pathways, including autoimmune diseases. The pathways of chronic inflammation are well illustrated by nonalcoholic steatohepatitis (NASH), which is of a serious concern due to its increasing prevalence in the westernized world and its direct correlation with lifestyle factors, particularly diet. Importantly, NASH may ultimately lead to the development of hepatocellular carcinoma. We previously reported that injection of monosodium glutamate (MSG) in ICR mice leads to the development of significant inflammation, central obesity, and type 2 diabetes. To directly address the long-term consequences of MSG on inflammation, we have performed serial analysis of MSG-injected mice and focused in particular on liver pathology. By 6 and 12 months of age, all MSG-treated mice developed NAFLD and NASH-like histology, respectively. In particular, the murine steatohepatitis at 12 months was virtually undistinguishable from human NASH. Further, dysplastic nodular lesions were detected in some cases within the fibrotic liver parenchyma. We submit that MSG treatment of mice induces obesity and diabetes with steatosis and steatohepatitis resembling human NAFLD and NASH with pre-neoplastic lesions. These results take on considerable significance in light of the widespread usage of dietary MSG and we suggest that MSG should have its safety profile re-examined and be potentially withdrawn from the food chain.

[Expression of PTEN, Cx43, and VEGF in hepatocellular carcinoma].

BACKGROUND & OBJECTIVE: Previous studies showed that expression of phosphatase and tensin homology deleted on chromosome ten (PTEN), gap junction protein connexin 43 (Cx43) and vascular endothelial growth factor (VEGF) may play an important role in tumor occurrence and progression. This study was designed to investigate the relationship among the protein expression of PTEN, Cx43, and VEGF in carcinogenesis and progression of hepatocellular carcinoma. METHODS: The expression of PTEN, Cx43 and VEGF were determined in 47 cases of HCC by streptavidin peroxidase immunohistochemistry. RESULTS: In the hepatocellular carcinoma, the total positive rates of PTEN, Cx43 and VEGF were 76.4%, 42.6%, and 70.2%, respectively. With progression of tumor course, PTEN and Cx43 positive rates decreased (P< 0.05), but the VEGF positive rate significantly increased (P=0.001). In comparison with the group of intrahepatic vascular embolism, PTEN positive rate increased apparently in the group without intrahepatic vascular embolism (P=0.006). Cx43 positive rate was lower in the group with cirrhosis background than that in the group without cirrhosis background (Chi(2)=4.713 5, P=0.03). No significant difference of PTEN positive rate was observed in tumor size, differentiation degree in cancerous tissue and cirrhosis background, Cx43 positive rate in tumor size, differentiation degree in cancerous tissue and intrahepatic vascular embolism, VEGF positive rate in tumor size, differentiation degree in cancerous tissue, cirrhosis background and intrahepatic vascular embolism. Correlation analysis revealed that PTEN positive rate was positively correlated with Cx43 positive rate (Cramer's V=0.394 9, P=0.023), negatively correlated with VEGF positive rate (Cramer's V=0.393 7,P=0.024), but no significant association was observed between Cx43 positive rate and VEGF positive rate (Cramer's V=0.307 2, P=0.064). CONCLUSION: The aberrant expression of PTEN, Cx43, and VEGF may play a role in occurrence, progression, and intrahepatic metastasis of the hepatocellular carcinoma. Determining the expression of PTEN, Cx43, and VEGF may contribute to synthetically evaluating the biological behavior of hepatocellular carcinoma.