Total mesopancreas excision for the treatment of pancreatic head cancer.

Mesopancreas is a controversial structure. This study aimed to explore the anatomical characteristics of the mesopancreas, define the range of the total mesopancreas excision (TMpE), and evaluate the feasibility, safety and effectivity of TMpE in the treatment of pancreatic head cancer. The clinical and pathological data of 58 consecutive patients undergoing TMpE for pancreatic head carcinoma from January 2013 to December 2015 were analyzed prospectively. The perioperative morbidity, mortality and clinical outcomes of patients undergoing TMpE were compared with the patients undergoing conventional pancreaticoduodenectomy. The mesopancreas was located in the retropancreatic area, extending from the head, neck, and uncinated process of pancreas to the aorto-caval groove, in which there were loose areolar tissue, adipose tissue, nerve plexus, lymphatic and capillaries. We observed significantly higher R0 rate (94.8% vs. 81.4%, P=0.035), more lymph nodes (16.2 vs. 11.4, P=0.000), lower total and local recurrence rate (half-year local recurrence rate 7.8% vs. 23.7%, P=0.036, one-year 18.2% vs. 39.5%, P=0.018) and longer disease-free survival (16.9 vs. 13.4 months, P=0.044) in TMpE group than in control group. In conclusion, mesopancreas is different from mesorectum because there is no fascial envelop or anatomical boundary in this area. TMpE could be safely and feasibly performed for the treatment of pancreatic head cancer to increase the R0 resection rate and improve the clinical outcomes.

[Expression, roles and therapy target values of CD24 in oral squamous cell carcinoma].

OBJECTIVE: To determine the expression profile and potential roles of CD24 in oral squamous cell carcinoma and explore the values of CD24 function as a potential target of clinical therapy. METHODS: Semi-quantitative immunohistochemistry was used to construct the expression profile of CD24 in 78 human oral tissues and 59 Hamster buccal pouch tissues. Real-time RT-PCR and Western blot were used to analyze the CD24 expression levels in oral DOK4 cells, oral cancer CAL-27 and WSU-HN6 cells. Then these two cancer cell lines were selected to evaluate the effect of all-trans retinoic acid (ATRA) and CD24 antibody on CD24 expression, and the proliferation and tumorsphere formation capacity of these two cell lines. RESULTS: CD24 expression was found significantly elevated in both human and animal tissues compared with normal and benign tissues (P<0.05), as well as in oral cancer CAL-27 and WSU-HN6 cells compared with DOK cells (P<0.05). CAL-27 and WSU-HN6 cells possess increased proliferative and specific tumorsphere formation capability compared with DOK cells (P<0.05). Both ATRA and CD24 antibody were able to effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells (P<0.05). Among them ATRA at least involved partially in the proliferation by down-regulating the CD24 expression (P<0.05), while CD24 antibody blocking had no effect on the CD24 expression. CONCLUSION: CD24 was upregulated in oral cancer and functioned as a potential factor that promoted the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells. Both ATRA and CD24 antibody might effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells and function as a potential therapy target.

[Corrigendum] MicroRNA-320a inhibits tumor invasion by targeting neuropilin 1 and is associated with liver metastasis in colorectal cancer.

After carefully checking the original data of migration and invasion experiment, we find we had selected the wrong picture to show the effect of microRNA320a on the migration of LoVo cells. The corrected version of Fig. 6A shown below contains appropriate representative images. We apologize for the error and appreciate the opportunity to correct the scientific record. All authors agree with this correction. [the original article was published in the Oncology Reports 27: 685-694, 2012 DOI: 10.3892/or.2011.1561]

Overexpression of SIRT1 is a poor prognostic factor for advanced colorectal cancer.

BACKGROUND: Sirtuin 1 (SIRT1) has been reported to have diverse roles in various biological processes through deacetylation of histone and nonhistone proteins. However, the correlations among SIRT1 protein expression, clinicopathological parameters, and survival of colorectal cancer patients remain unclear. METHODS: SIRT1 protein expression was measured by immunohistochemistry in a paraffin-embedded tissue microarray, including 120 paired colorectal cancer and normal mucosa tissues. The correlations among SIRT1 protein expression, clinicopathological features, and prognosis were analyzed. RESULTS: All samples (100%) were positive for SIRT1, with variable staining in the cytoplasm rather than in the nucleus. There was significant difference in SIRT1 overexpression between adenocarcinomas and normal mucosal tissue (P < 0.01, χ(2) test). SIRT1 overexpression was more frequently observed in advanced-stage tumors (P = 0.046, 0.002, χ(2) test). SIRT1 overexpression was significantly correlated with poor overall survival (P = 0.013, log-rank test) and diseasefree survival (P = 0.012, log-rank test). CONCLUSIONS: SIRT1 overexpression correlated with advanced stage and poor prognosis. SIRT1 may play an important role in the progression of colorectal cancer.

Clinicopathological significance of SIRT1 expression in colorectal adenocarcinoma.

Sirtuin 1 (SIRT1) has been reported to have diverse roles in various biological processes through deacetylation of histone and nonhistone proteins. However, the correlations between SIRT1 protein expression, clinicopathological parameters, and survival of colorectal cancer patients remain unclear. SIRT1 protein expression in a paraffin-embedded tissue microarray, including 13 benign adenomas, nine liver metastasis tissues, and 120 paired colorectal cancer and normal mucosa tissues, was measured by immunohistochemistry. SIRT1 mRNA and protein expression in colon cancer cell lines with different metastatic potential and normal colon cells were detected by real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. The correlations between SIRT1 protein expression, clinicopathological features, and prognosis were analyzed. All samples (100 %) were positive for SIRT1, with variable staining in the cytoplasm rather than the nucleus. There was significant difference in SIRT1 overexpression between adenocarcinomas and normal mucosal tissues (P < 0.01, χ(2) test). SIRT1 overexpression was more frequently observed in advanced-stage tumors and lymph node or liver metastases (P = 0.046, 0.002, and 0.004, respectively, χ(2) test). SIRT1 expression was also significantly elevated in the more aggressive colon cancer cell line SW620. SIRT1 overexpression was significantly correlated with poor overall survival (P = 0.013, log-rank test) and disease-free survival (P = 0.012, log-rank test). SIRT1 overexpression was correlated with advanced-stage and poor prognosis. SIRT1 may play an important role in the progression of colorectal cancer.

[Overexpression of high mobility group A2 and its correlation with microRNA let-7 family in serous ovarian cancers].

OBJECTIVE: To examine the expression of high mobility group A2 (HMGA2), P53 and let-7 family microRNA, to investigate the correlation of HMGA2 and let-7, and to compare the HMGA2 and P53 expressions in human serous ovarian cancer. METHODS: Immunohistochemistry assay was used to examine the expressions of HMGA2 and P53 in 50 paraffin-embedded tissue specimens of human serous ovarian cancer and 4 normal fallopian tube tissues. HMGA2 mRNA and let-7 family microRNA were detected by real time fluorescent quantitative reverse transcription polymerase chain reaction in the corresponding frozen tissues. RESULTS: HMGA2 and P53 were immuno-positive in 70% (35/50) and 78% (39/50) of the ovarian cancer tissues, respectively. HMGA2 was weakly expressed in the ciliated cells, but negative in the secretary cells of the fallopian tube. There was a tendency that the expression of HMGA2 increased with higher pathological grade of the ovarian cancer, but no correlation was observed between the HMGA2 overexpression and clinical stages. HMGA2 mRNA was detected in all the ovarian cancer samples, and its expression level was higher than that of the normal fallopian tube tissues in 72% (36/50) of the ovarian cancer samples. The expression of HMGA2 mRNA was much higher in more malignant SKOV3.ipl cells than in its corresponding SKOV3 cells. All let-7 family members were detectable in all ovarian cancer samples, and their expression were inversely correlated with HMGA2 mRNA expression (r=-0.305,P<0.05). CONCLUSION: HMGA2 can be a biomarker complement to P53, and its high expression has an inclination of more malignancy. The downregulation of let-7 is, but not the only mechanism of HMGA2 overexpression in serous ovarian cancer.

Genistein inhibit cytokines or growth factor-induced proliferation and transformation phenotype in fibroblast-like synoviocytes of rheumatoid arthritis.

The purpose of this research is to study the effect of genistein on cytokines or growth factor-induced proliferation and transformation phenotype of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). RA-FLS were primarily cultured. With respective stimulation of IL-1ß, TNF-α, and EGF, genistein was applied to elucidate its effect on synoviocytes' growth number, cell proliferation assay, cell cycle using cell counts, (3)H-TdR incorporation and flow cytometry, the colony numbers under anchorage-independent condition, and the expression of MMP-2 and MMP-9 in synovial fibroblasts. EGF, IL-1ß, and TNF-α increased (3)H incorporation in RA-FLS, respectively. EGF augmented clone numbers of RA-FLS under anchorage-independent condition and not IL-1ß and TNF-α. Genistein had an inhibitory role on cell number and (3)H-TdR incorporation of RA-FLS stimulated with IL-1ß, TNF-α and EGF; genistein arrested the cell cycle at G(1) restriction point; genistein decreased colony numbers under anchorage-independent condition stimulated by EGF in serum condition. IL-1ß or TNF-α increased expression of MMP-9 and MMP-2 in rheumatoid synoviocytes; EGF stimulated expression of MMP-9 but not of MMP-2; genistein suppressed production of MMP-9 more than MMP-2 induced by IL-1ß or TNF-α; rMMP-9, rMMP-2, or their inhibitors had no effect on the (3)H-TdR incorporation of synovial cells. Erk1/2 inhibitor (PD098 059) had obvious inhibitory effect on the (3)H incorporation induced by TNF-α or IL-1ß; inhibitors of JNK (SP600 125) had no significant effect on the (3)H incorporation. While pretreatment with PD098059 had no marked inhibitory effect on MMP-9 expression induced by TNF-α or IL-1ß, SP600125 decreased significantly the MMP-9 expression induced by TNF-α or IL-1ß. Neither PD098059 nor SP600 125 could inhibit the MMP-2 expression induced by TNF-α or IL-1ß. Genistein inhibited IL-1ß, TNF-α or EGF-induced proliferation and MMP-9 expression in fibroblast-like synoviocytes of rheumatoid arthritis; the proliferation of RA-FLS was mediated by Erk1/2 but not JNK activation, while JNK activation was involved in the signal transduction pathway leading to MMP-9 expression in rheumatoid synoviocytes.

microRNA-320a inhibits tumor invasion by targeting neuropilin 1 and is associated with liver metastasis in colorectal cancer.

MicroRNAs (miRNAs) have been implicated in regulating diverse cellular pathways. Although there is emerging evidence that various miRNAs function as oncogenes or tumor suppressors in colorectal cancer (CRC), the role of miRNAs in mediating liver metastasis remains unexplored. The expression profile of miRNAs in liver metastasis and primary CRC tissues was analyzed by miRNA microarrays and verified by real-time polymerase chain reaction (PCR). In 62 CRC patients, the expression levels of miR-320a were determined by real-time PCR, and the effects on migration and invasion of miR-320a were determined using a transwell assay. miR-320a target genes were confirmed by luciferase assay, real-time PCR and western blot analysis. A set of miRNAs was found to be dysregulated in the liver metastasis tissues compared to matched primary CRC tissues, and the expression levels of miR-320a were significantly decreased in the liver metastasis tissues examined. miR-320a was correlated with tumor progression in CRC. miR-320a was downregulated in liver metastatic colon cancer cells and inhibited liver metastatic colon cancer cell migration and invasion. miR-320a directly binds to the 3'UTR of neuropilin 1 (NRP-1), a protein that functions as a co-receptor of vascular epithelial growth factor. miR-320a downregulated the expression of NRP-1 at both the mRNA and protein levels. These data demonstrated that miR-320a may be useful for identifying CRC patients that are at an elevated risk for developing liver metastasis. Our findings suggest that miR-320a may be a novel therapeutic candidate for the treatment of colorectal cancer.

Efficacy and reliability of long-term implantation of multi-channel microelectrode arrays in the optical nerve sheath of rabbit eyes.

In addition to epiretinal and subretinal areas, the optic nerve (ON) is also a candidate location for implanting visual prosthesis to restore vision of patients with retinitis pigmentosa (RP). Since the ON receives all the signals from the retina, stimulating the ON may potentially evoke phosphenes over a wider range of visual field. In this study, we designed a 9-channel microelectrode array and implanted it between the dura mater and pia mater of rabbit ONs by lateral orbitotomy. We recorded the current thresholds and evaluated the efficacy of the array using electrically evoked potentials (EEPs). Spatial discrimination of approximately 20° was verified by EEP maps over visual cortex. A large area of the visual field (over 130° along horizontal meridian) could be activated by this microelectrode array. Visual evoked potentials (VEPs) and different pathological examinations were used to examine potential damage of ONs. One year post implantation, we did not notice significant damages to either the ONs or the microelectrode arrays. EEPs were successfully recorded up to 6months post implantations. However, further studies are still needed to reduce fibrous encapsulation of the microelectrode array, which resulted in a gradual elevation of current thresholds to elicit EEPs.

Effect of Robo1 on retinal pigment epithelial cells and experimental proliferative vitreoretinopathy.

PURPOSE: The Roundabout (Robo) family of proteins is related to the transmembrane receptors and plays a major role in neurogenesis. However, the role of the Robo proteins in proliferative retinopathy has not yet been defined. This study was conducted to determine whether Robo1 is expressed in the retina of patients with proliferative retinal disease and whether it has a pathobiological role in the disease. METHODS: Immunohistochemistry was used to determine the presence and distribution of Robo1 in the pathologic membranes in proliferative retinopathy. Small interfering (si)RNA technology was used to knockdown Robo1 expression and to study its effects on retinal pigment epithelial (RPE) cells in vitro. The impact on PVR development of blocking Robo1 expression was determined by applying specific siRNA in a PVR rabbit model. The prevalences of PVR and retinal detachment were determined by indirect ophthalmoscope on days 1, 3, 7, 14, 21, and 28 after the injection of RPE cells into the vitreous. RESULTS: Immunohistochemistry showed that Robo1 expression was detected in GFAP-labeled glial cells and cytokeratin-labeled RPE cells in proliferative membranes. Robo1 expression was also detected in CD31-labeled vascular endothelial cells. Knockdown of Robo1 expression not only reduced human RPE cell proliferation in vitro but also effectively suppressed the development of PVR in a rabbit model. CONCLUSIONS: Robo1 is present in the extracellular matrix of proliferative membranes and may be derived from dedifferentiated RPE cells. Silencing the expression of Robo1 in RPE cells inhibited cell proliferation and suppressed the development of PVR in an animal model, indicating a potential therapeutic usefulness in treating PVR.

Downregulation of ABI1 expression affects the progression and prognosis of human gastric carcinoma.

Abelson interactor protein-1 (ABI1) is a promising candidate tumor suppressor, and plays critical roles both in the pathogenesis of BCR-Abl-induced leukemia and in the spread of several solid tumors. The expression of ABI1 and its role in cancer progression and prognosis are largely unknown in the majority of solid tumors, including gastric cancer. In this study, we analyzed the correlation between ABI1 expression and the clinicopathological characteristics, tumor progression, and prognosis of patients with gastric carcinoma. Tissue specimens were from 103 gastric cancer patients who underwent gastrectomy in our hospital between January 2000 and December 2007. Among them 59 tumor tissue samples were matched with normal tissue samples. The expression of ABI1 protein was measured using immunohistochemical staining of paraffin-embedded tissue specimens. Meanwhile, quantitative real-time RT-PCR and Western blotting were used to identify the expression of ABI1 in human gastric normal mucosal cell line (GES-1) and gastric cancer cell lines (N87, AGS). We performed a statistical analysis of the potential correlation between ABI1 expression and the patients' clinicopathological characteristics, 5-year survival, and median survival time. The immunohistochemical staining results of 59 patients showed that ABI1 was expressed in 28.8% (17/59) of gastric cancer tissues, compared to 91.5% (54/59) of normal samples. ABI1 expression in 103 patients was strongly correlated with tumor differentiation, clinical stage, and lymph node status (P < 0.01). The 5-year survival rate was 15.3% in the ABI1-negative group and 63.7% in the ABI1-positive group. Median survival time in the ABI1-negative and ABI1-positive groups was 25.0 months (95% CI: 19.7-30.3) and 74.0 months (95% CI: 54.6-93.3), respectively. There was a significant difference between the two groups (chi(2) = 10.888, P = 0.001). Furthermore, we found that ABI1 expressed lowly in poor differentiated AGS, whereas highly in GES-1 and well-differentiated N87. Downregulation of ABI1 expression in human gastric carcinoma may play a critical role in tumor progression and in determining patient prognosis. ABI1 may be a useful diagnostic or prognostic molecular biomarker, and might be a potential target for therapeutic intervention.

Expression of Robo4 in the fibrovascular membranes from patients with proliferative diabetic retinopathy and its role in RF/6A and RPE cells.

PURPOSE: Robo4, a member of the roundabout (Robo) family, acts as a neuronal guidance receptor and plays some role in vasculogenesis and angiogenesis. This study investigated the effect of Robo4 on the formation of fibrovascular membranes (FVMs) from patients with proliferative diabetic retinopathy and its roles in choroid-retina endothelial (RF/6A) and human retinal pigment epithelial (RPE) cells. METHODS: RT-PCR and immunohistochemistry were used to determine the levels of mRNA and the presence and distribution of Robo4 in FVMs. Small interfering RNA (siRNA) technology was used to knock down Robo4 expression and to study its effects on RF/6A and RPE cells in vitro. Cell proliferation, migration, spreading, cycling, and apoptosis were assessed with MTT assay, Boyden chamber assay, immunocytochemistry, and flow cytometry. Tube formation by RF/6A on Matrigel was also analyzed. RESULTS: The level of Robo4 mRNA was high in FVMs. Robo4 was expressed in the vessels and fibrous-like tissue co-immunostained for CD31 and GFAP, respectively. Robo4 siRNA knockdown inhibited cell proliferation and migration. Tube formation by RF/6A cells was also disturbed. Under hypoxic conditions, more apoptotic cells were evident among the knockdown cells than among the control cells (p<0.01). CONCLUSIONS: Robo4 may play a role in the formation of FVMs. Silencing the expression of Robo4 in RF/6A and RPE cells inhibited their proliferation and reduced their tolerance of hypoxic conditions, suggesting physiologic functions of Robo4 in the cells of the retina.

[Effect of Abl-interacting protein 1 overexpression upon human gastric cancer cell proliferation in vitro].

OBJECTIVE: To investigate the expression of Abl-interacting protein 1 (ABI1) in normal gastric mucosal cell line GES-1 and gastric cancer cell line AGS, and the effects of ABI1 gene overexpression upon the proliferation of human gastric cancer cell AGS in vitro. METHODS: Firstly the ABI1 expression in GES-1 and AGS cells were identified by immunohistochemistry, immunofluorescence, RT-PCR, real-time PCR and Western blot. Secondly human gastric cancer cell line AGS was cultured and transfected with recombinant MSCV-GFP-ABI1 plasmid or blank plasmid MSCV-GPF. Real-time PCR and Western blot were used to detect the mRNA and protein expression of ABI1. And lastly the cell proliferation was detected by CCK-8 assay. RESULTS: ABI1 was expressed both in normal gastric mucosal cell line GES-1 and in gastric cancer cell line AGS. Compared to GES-1 cells, the ABI1 expression in AGS cells was lowered significantly. There were no significant differences in the ABI1 mRNA and protein expression between the AGS and AGS-MSCV-GFP groups. Compared to those of the AGS group, the ABI1 mRNA expression levels of the AGS-MSCV-GFP-ABI1 group increased by 1.87 times (P = 0.002). The protein expression levels of the AGS-MSCV-GFP-ABI1 group were remarkably higher than those of the AGS and AGS-MSCV-GFP groups (P = 0.002). CCK-8 assay showed that there were no significant differences in the proliferation rates at different time points between the AGS and AGS-MSCV-GFP groups. However, the proliferation rates at the time points of 24, 48, 72 and 96 hours of the AGS-MSCV-GFP-ABI1 were 1.46 +/- 0.31, 4.75 +/- 0.12, 6.62 +/- 0.32 and 8.96 +/- 0.27 respectively. And they were significantly lower than the proliferation rates of the AGS and AGS-MSCV-GFP groups (P < 0.01). CONCLUSION: ABI1 gene is down-regulated in gastric cancer cells. The ABI1 overexpression effectively inhibits the proliferation in human gastric cancer cell lines. It suggests that ABI1 may be involved in gastric cancer pathogenesis by regulating the proliferation of gastric carcinomas cells.

[Acute injury of inhalable particle matters on nasal mucous membrane of rat].

OBJECTIVE: To study the mechanism of the nasal mucous membrane inflammation induced by the inhalable particle matter (PM10). METHOD: Three dosage PM10 were instilled in rat nasal cavity of different groups for one week. The morphology of nasal mucosa and the numbers of inflammatory cell were observed in each samples. RESULT: The total numbers of inflammatory cells in PM10-treated groups were increased in a dose-respondent manner and significantly different from that in control group. The results of histopathological and scanning electron microscope (SEM) analysis indicated that PM10 caused nasal mucosa injury and pathological changes, such as the damage of cilia and nasal mucosa epithelium in a dose dependent way. The infiltration of inflammatory cells in nasal mucosa epithelium matrix, especially eosinophilia were observed. CONCLUSION: PM10 can cause rat's nasal mucosa inflammation and epithelial injury.

Effects of the phosphatidylinositol 3-kinase inhibitor in a mouse model of retinal neovascularization.

PURPOSE: To determine the effects of LY294002, a phosphatidylinositol 3-kinase inhibitor, on suppressing experimental retinal neovascularization in an animal model of ischemic retinopathy. METHODS: The effect of LY294002 on the survival of RF/6A cells stimulated by vascular endothelial growth factor (VEGF) was investigated colorimetrically. The inhibitory activity of LY294002 on the migration of cells stimulated with VEGF was measured by cell counting. C57BL/6N mice at postnatal day (P) 7 were exposed to 75 +/- 2% oxygen for 5 days (P7-P11) and then returned to room air for 5 days (P12-P17) to induce retinal neovascularization. Beginning on P12, mice received daily intraperitoneal injections of LY294002 or dimethyl sulfoxide and phosphate-buffered saline (control) through P17. Retinal neovascularization was examined by adenosine diphosphatase staining after 5 days in room air and was quantitated histologically by counting the neovascular endothelial cell nuclei anterior to the inner limiting membrane. RESULTS: LY294002 significantly inhibited VEGF-induced survival and migration. LY294002-treated and control animals demonstrated no perfusion regions in the posterior retina. Retinas from control mice at P17 contained neovascular tufts at the junction between the perfused and nonperfused retina. The tufts contained numerous neovascular nuclei. Retinas from mice treated with LY294002 demonstrated a significant reduction in neovascular cell nuclei compared with control mice. CONCLUSIONS: LY294002 significantly inhibits retinal neovascularization in a mouse model of retinal neovascularization.

Influential factors of thresholds for electrically evoked potentials elicited by intraorbital electrical stimulation of the optic nerve in rabbit eyes.

Influential factors of electrical thresholds for electrically evoked potentials elicited by intraorbital stimulation of the optic nerve, including stimulation positions of the optic nerve, stimulating electrodes, frequency and duration of electrical pulses and pathological status of the optic nerve, were evaluated in 48 pigmented rabbit eyes. Intravenous injection of sodium iodate was used to induce transneuronal degeneration of the retinal ganglion layer subsequent to photoreceptor death. Two equations were derived to predict electrical thresholds needed to elicit cortical responses.

Relationship between hepatitis B virus DNA levels and liver histology in patients with chronic hepatitis B.

AIM: To study the relationship between hepatitis B virus (HBV) DNA levels and liver histology in patients with chronic hepatitis B (CHB) and to determine the prevalence and characteristics of hepatitis B e antigen (HBeAg) negative patients. METHODS: A total of 213 patients with CHB were studied, and serum HBV DNA levels were measured by the COBAS Amplicor HBV Monitor test. All patients were divided into two groups according to the HBeAg status. The correlation between serum HBV DNA levels and liver damage (liver histology and biochemistry) was explored. RESULTS: Of the 213 patients with serum HBV DNA levels higher than 10(5) copies/mL, 178 (83.6%) were HBeAg positive, 35 (16.4%) were HBeAg negative. The serum HBV DNA levels were not correlated to the age, history of CHB, histological grade and stage of liver disease in either HBeAg negative or HBeAg positive patients. There was no correlation between serum levels of HBV DNA and alanine aminotransferanse (ALT), aspartate aminotransferase (AST) in HBeAg positive patients. In HBeAg negative patients, there was no correlation between serum levels of HBV DNA and AST, while serum DNA levels correlated with ALT (r=0.351, P=0.042). The grade (G) of liver disease correlated with ALT and AST (P<0.05, r=0.205, 0.327 respectively) in HBeAg positive patients. In HBeAg negative patients, correlations were shown between ALT, AST and the G (P<0.01, and r=0.862, 0.802 respectively). HBeAg negative patients were older (35 +/- 9 years vs 30 +/- 9 years, P<0.05 ) and had a longer history of HBV infection (8 +/- 4 years vs 6 +/- 4 years, P<0.05) and a lower HBV DNA level than HBeAg positive patients (8.4 +/- 1.7 Log HBV DNA vs 9.8 +/- 1.3 Log HBV DNA, P<0.001). There were no significant differences in sex ratio, ALT and AST levels and liver histology between the two groups. CONCLUSION: Serum HBV DNA level is not correlated to histological grade or stage of liver disease in CHB patients with HBV DNA more than 10(5) copies/mL. Compared to HBeAg positive patients, HBeAg negative patients are older and have a lower HBV DNA level and a longer HBV infection history. There is no significant difference in sex ratio, ALT and AST levels and liver histology between the two groups.

Diffusion of macromolecule through retina after experimental branch retinal vein occlusion and estimate of intraretinal barrier.

The disposition and diffusion knowledge of intravitreally injected macromolecule drugs through retina in pathological condition is crucial but the related studies are absent. Retinal edema is a common pathological change of fundus diseases and retinal vein occlusion (RVO) pig model were established to emulate it. FITC-dextrans of various molecular weights were dissolved in RPMI-1640 solutions and the rate of transretinal diffusion was determined with a spectrophotometer. Theoretical maximum size of molecule (MSM) was calculated by extrapolating the trend-linear relationship with the diffusion rate. In separate experiments to determine the sites of barrier to diffusion, FITC-dextrans were applied to either the inner or outer retinal surface, processed as frozen sections, and viewed with a fluorescence microscope. Paired-Samples T test was used to compared the diffusion rate of dextrans of the both eyes of one pig. The MSM in RVO tissues and normal tissue was 6.5+/-0.39 nm and 6.18+/-0.54 nm respectively (t=4.143, P=0.0001). FITC-dextrans applying to inner retinal surface, 4.4 kDa dextran were largely arrested at inner nuclear layer (INL). The INL of the 19.6-71.2 kDa dextran diffusion retina section became dark and the nerve fiber layer (NFL) and inner plexiform layer got brighter. As for 150 kDa dextran, the NFL was bright and the other layers were dark. FITC-dextrans applying to outer retinal surface, most dextrans were blocked before outer nuclear layer (ONL). In summary, ONL and INL may act as bottle-neck barriers to diffusion of macromolecules. Compared with normal neuroretina, the MSM of fresh edema retina after RVO increased limitedly.

Effects of subretinal implant materials on the viability, apoptosis and barrier function of cultured RPE cells.

BACKGROUND: Subretinal microphotodiode array (MPDA) is a type of visual prosthesis used for the implantation in the subretinal space of patients with progressive photoreceptor cell loss. The present study aimed to evaluate the effect of materials for MPDA on the viability, apoptosis and barrier function of cultured pig retinal pigment epithelium (RPE) cells. METHODS: Primary culture of pig RPE cells was performed and 24 pig eyes were used to start RPE culture. The third passage of the cultures was plated on different materials for MPDA and MPDAs. The tetrazolium dye-reduction assay (MTT) was used to determine RPE cell viability. Flow cytometry was measured to indicate the apoptosis rates of RPE cells on different materials. RPE cells were also cultured on microporous filters, and the transepithelial resistance and permeability of the experimental molecule were measured to determine the barrier function. RESULTS: The data from all the methods indicated no significant difference between the materials groups and the control group, and the materials tested showed good biocompatibility. CONCLUSIONS: The materials for MPDA used in the present study had no direct toxicity to the RPE cells and did not release harmful soluble factors that affected the barrier function of RPE in vitro.

[The effects of hematopoietic stem cells on mice vascular endothelial cells of diabetic retinopathy].

OBJECTIVE: The purpose of the study was to determine whether hematopoietic stem cell (HSCs) mobilization can regulate early diabetic retinopathy in mice. METHODS: Mice were divided into four groups: control group, normal mice HSC-mobilized group, diabetic mice control group and diabetic mice HSC-mobilized group. Murine stem cell growth factor (SCF) and recombined human granulocyte colony stimulating factor (rhG-csf) were administered to the mice with diabetes and without diabetes for continuous 5 days to induce autologous HSCs mobilization, and subcutaneous injection of physiological saline was used as control. The changes associated with autologous HSCs mobilization were characterized using flow cytometry, Immunohistochemistry and semiquantitative RT-PCR. Evans blue quantitative test was used to measure the breakdown level of blood-retina barrier. RESULTS: HSCs were marked by CD34-/low and Sca1+ in this study. The acceleration of endothelial cell regeneration was observed. A decrease of VEGF expression due to autologous stem cell mobilization was found. HSCs could increase the content of VEGFR-2 in mouse retina and significantly downregulated the expression of VEGF and ang-2 in diabetic mice. CONCLUSIONS: The experiment suggest that autologous HSCs mobilization can be approach of therapeutic vascular reconstruction and functional restoration of blood-retina barrier in mice.