Objective This study analyzes the expression and clinical significance of Gli1 and Gli3 proteins in hepatocellular carcinoma.Methods 36 patients with hepatocellular carcinoma were studied.The expressions of Gli1 and Gli3 in the carcinoma and adjacent non-tumor tissues were detected with immunohistochemical assay,and their correlations with clinicopathological factors were statistically analyzed.Results Expression rates of Gli1 in hepatocellular carcinoma and adjacent nontumor tissues were 75 ％ and 36.1％,respectively.Expression rates of Gli3 in hepatocellular carcinoma and adjacent non-tumor tissues were 58.3％ and 30.6％,respectively.Expression rates of Gli1 and Gli3 in hepatocellular carcinoma were significantly higher than in adjacent non-tumor tissues (P＜0.05),and a positive correlation was found between the expression of Gli1 and Gli3 (r=0.423,P＜0.05).There was no association between the expression of Gli3 and clinicopathological factors such as age,tumor size,tumor differentiation and lymphatic metastasis.The expression of Gll1 was not related witha patient's age and tumor size,hut there were significant associations with tumor differentiation and lymphatic metastasis.Conclusions Therefore,the expression rate of Gli1 was positively correlated with tumor malignancy,which makes the detection of Gli1 and Gli3 valuable for the diagnosis and prognosis of hepatocellular carcinoma.

Objective To identify highly sensitive and specific antigens in Aeromonas hydrophila strain ATCC7966 by using immunoproteomics.Methods Outer membrane proteins were extracted from the Aeromonas hydrophila strain by using two-dimension electrophoresis and identified by LC-LTQ-XL-MS ( liq-uid chromatography coupled with linear ion trap mass spectrometry).Western blot assay was performed to screen out the immunogenic proteins.Results A total of 43 peptides representing 39 proteins were identi-fied by LC-LTQ-XL-MS.Among the 39 proteins, 69% were outer membrane proteins and 12% were inner membrane proteins.They were involved in the process of transportation, cell motility, biosynthesis, etc. One candidate vaccine antigen was identified by using two-dimensional Western blot analysis .Conclusion The immunoproteomics approach could be used to identify immunogenic proteins of Aeromonas hydrophila. This study provided references for further investigation on candidate vaccine antigens.

Objective To compare the clinical value of three visual rating scales (VRS) for white matter lesions (WML) in Alzheimer's disease (AD) and mild cognitive impairment (MCI).Methods Totally 184 subjects (including 107 AD patients,47 MCI patients and 30 normal controls)were recruited.All subjects underwent comprehensive neuropsychological tests and were examined with a standard protocol of MR imaging.WML burden was rated with the Age-Related White Matter Changes (ARWMC) rating scale,Cholinergic Pathways Hyperintensities Scale (CHIPS) and Fazekas scale.Consistence of the three rating scales were analyzed,and detection results of WML in AD/ MCI/NC group with the three rating scales were compared.The relationship between WML and cognitive function in AD and MCI groups were explored.Results ARWMC rating scale,CHIPS and Fazekas scales were moderately to highly correlated with WML (r=0.61-0.78,all P＜0.01).ARWMC showed that WML were in the left and right frontal lobes in AD and MCI groups and in the right frontal lobe and left temporal lobe in NC group,which had significant differences (F=4.20,4.69,4.69,3.36,respectively,all P＜0.05).WML in the left temporal lobe had a significant difference between MCI group and NC group (F=3.36,P＜0.05).CHIPS showed that WML in the front left side of centrum semiovale in AD and MCI groups had significant difference as compared with that in NC group (F=4.88,P＜0.05),and WML in the rear right side of low external capsule had a significant difference between AD and NC groups (F=3.04,P＜0.05).Fazekas scale showed that there was no difference in WML among AD,MCI and NC groups.ARWMC showed that scores of WML in right temporal lobe were positively correlated with concentration and visual construction,and bilateral basal ganglia were negatively correlated with the concentration.Fazekas scale showed that scores of WML in periventricular were negatively correlated with concentration,MMSE and orientation,scores of WML in deep white matter were negatively correlated with concentration,abstraction and judgment,and the whole scores were negatively correlated with concentration.CHIPS showed that the scores of WML in the rear left side of low external capsule were negatively correlated with short-term memory in MCI group.Conclusions The three rating scales well consistent with each other.ARWMC rating scale and CHIPS can detect the differences in WML between AD,MCI and NC groups.The scores of CHIPS in WML are related with cognition in MCI group,while scores of ARWMC and Fazekas scale in WML are correlated with many aspects in cognitive function in AD group.In general speaking,CHIPS in MCI patients and ARWMC rating scale in AD patients show more advantages in the detection of WML and have better correlation with cognitive function.

Objective To investigate the incidence of occult pancreatobiliary reflux and to evaluate its relation to gallbladder epithelial dysplasia and cancer. Methods From July 2006 to Feb 2008,956 cases underwent selective biliary procedure or preoperative endoscopic retrograde cholangiopanereatography (ERCP), bile was collected and amylase was measured. All removed gallbladders were pathologically examined for dysplasia and cancer. Results Occult pancreatobihary reflux was found in 75 of 754 patients in this study, with an incidence of 9. 9%. The biliary amylase values in the patients with occult pancreatobiliary reflux and in controls were 7701±20 378 IU/L and 16±51 IU/L, respectively ( P <0. 01 ).Gallbladder dysplasia and cancer were found in 31.0% and 3.4% of the patients with occult pancreatobiliaryreflux, respectively, and both were higher than those in the patients without pancreatobiliary reflux ( P <0. 05). In the patients with occult pancreatobiliary reflux, the biliary amylase level with gallbladder dysplasia or cancer was 2388 ± 2745 IU/L and was higher than those without gallbladder dysplasia or cancer (P < 0. 01 ). Conclusions With an incidence of 9.9% in patients of normal pancreatobiliary junction,the occult pancreatobihary reflux may contribute to the pathogenesis of gallbladder epithelial dysplasia and cancer.

Objective To study the clinicopathologic characteristics of triple-negative breast cancer (TNBC) and its value in the prediction of prognosis. Method In this study,500 cases of female breast cancers were examined immunohistochcmically for the TNBC. The clinicopathologic characteristics of the 243 TNBC cases were inspected. Results TNBC accounted for 17.6% (88/500) of the 500 breast cancers. The histological types of the TNBC included mainly infihrative ductal carcinoma, metaplastic carcinoma and medullar carcinoma. Among those, histological grade Ⅲ accounted for 72.7% (64/88) of all the TNBC and was more common than that in hormone receptor positive breast cancers (HR+ group ) and Her-2 overexpression breast cancers (Her-2 group)(P=0.000). The positive rates of CK5/6 and EGFR in the TNBC were 30.7% (27/88) and 34.1% (30/88), respectively. The positive rates of ERCC1 and KIT in the TNBC were 28.4% (25/88) and 34.1% (30/88), respectively, Both of which were higher than those in the HR + group and Her-2 group, respectively (P=0.032 and P=0.026). 3-year survival rate of the TNBC was 71.5% and it was lower than that of HR group (P=0.021) and not significantly different from that of Her-2 group (P=0.474). Conclusions TNBC is the breast cancer with high aggressive pathologic futures and poor prognosis. EGFR and ERCC1 expression were positive in a portion of TNBC cases.

OBJECTIVETo investigate the role of R-Spondinl in the activation of hepatic stellate cells (HSCs).

METHODSTwenty-four healthy male Kunming mice were randomly divided into the following two groups:fibrosis model group (n=16) and control group (n=8). Hepatic fibrosis was induced by subcutaneous injections of CC14 (20% in olive oil) at a dose of 5 ml/kg twice per week. After 10 weeks, the animals were sacrificed by CO(2) over-exposure and liver tissues were harvested.The protein and mRNA levels of R-Spondin1, alphat-SMA,and collagen I were examined by Western blot assay and real-time PCR respectively. Additionally,HSCs were isolated from the mice liver tissues to examine the time-series expression changes of R-Spondinl, alpha-SMA, and nuclear beta-catenin.TCF activity was analyzed by luciferase reporter assay.Moreover,HSCs were cocultured with recombinant R-Spondin1 and DKK1 to evaluate dose-response.

RESULTSR-Spondinl expression was significantly higher in the fibrosis model group than in the control group (protein level:3.16 ± 0.18 vs. 0.99 ± 0.16, t =13.31, P < 0.01; mRNA level:4.36 ± 0.26 vs. 0.98 ± 0.12, t =21.46, P < 0.01).The culture-activated mouse HSCs showed up-regulated TCF activity (5.33 ± 0.34 vs. non-activated: 1.03 ± 0.09, t =20.93, P < 0.01), nuclear beta-catenin expression (4.47 ± 0.21 vs. 0.97 ± 0.14, t =25.25, P < 0.01), and R-Spondin1 expression (protein level: 4.54 ± 0.18 vs. 1.04 ± 0.12, t =31.17, P < 0.01; mRNA level:5.13 ± 0.15 vs. 1.01 ± 0.16, t=38.06, P < 0.01). Exogenous stimulation of freshly isolated mouse HSCs with recombinant R-Spondin1 induced a dose-dependent increase in both TCF activity and the expression of nuclear beta-catenin and alphat-SMA. DKK1 down-regulated activities of factors in the WNT signaling pathway and repressed activation of HSCs. Conclusion R-Spondin1 may promote HSC activation by enhancing the canonical WNT signaling pathway.

Animals ; Down-Regulation ; Extracellular Matrix Proteins ; Hepatic Stellate Cells ; Liver Cirrhosis ; Male ; Mice ; RNA, Messenger ; Wnt Signaling Pathway ; beta Catenin

Objective: To investigate the relationship between the recurrence of benign paroxysmal positional vertigo(BPPV) and the levels of bone mineral density(BMD) and estrogen in postmenopausal women. Methods: A total of 38 postmenopausal women with recurrent BPPV were recruited as study group, in the First Affiliated Hospital of Soochow University from December 2013 to June 2017. Meanwhile, 49 normal menopausal women were included as control. All patients were natural menopausal for over one year.The patients were diagnosed as BPPV based on results of Dix-Hallpike test and Roll-test, with at least two episodes of recurrent onset. In the subjects, BMD was measured by dual X-ray absorptiometry of lumbar vertebrae. Estrogen levels were obtained by testing serum estradiol (E2) levels in early morning fasting venous blood. In the present study, we compared the level of E2 and the value of BMD in two groups by SPSS 21.0. In the study group, patients with decreased BMD were divided into two groups: treatment and untreated group. The recurrence rate of BPPV was compared between the two groups within 12 months. Results: ①The averagel levels of E2 and BMD in the study group were (16.21±11.00)ng/L and -1.68±0.98) respectively, which were significantly lower than those in the control group (t value was 7.03 and 8.05 respectively, both P<0.05). The averagel levels of E2 and BMD incontrol group were(28.52±6.34)ng/L and -0.18±0.77 respectively. ②The number of patients with decreased BMD in the study group (30 cases) was more than that in control group (6 cases), and the difference was statistically significant (P<0.05). ③ The recurrence rate of BPPV in treatment group [17.6%(3/17)] was significantly lower than that of untreated group [61.5%(8/13)], and the difference was statistically significant (P<0.05). Conclusion Recurrent BPPV in postmenopausal women usually accompany with low levels of estrogen and BMD. Active treatment is helpful for their recurrence of BPPV.

Objective: Establish a murine model for hyperuricemia (HU) and periodontitis to explore whether there is correlation between them and provide a basis for periodontal treatment. Methods: Fourteen male KM mice were divided into 2 groups; the HU group (n=7) was fed food supplemented with potassium oxonate and uric acid, the NC group (n=7) was fed standard food, and the induction period was 35 days. On the 25th day, the molars on one side were ligated to induce periodontitis (P side), while the opposite was true for the control (C side). Baseline and terminal serum uric acid (UA) levels were detected, and alveolar bone resorption was analyzed by micro-CT. Results: The serum UA level of HU mice was (112.94 ± 26.82 )mol/L, that of the NC group was (72.21 ± 19.95) μmol/L, and the difference in UA level was statistically significant (P < 0.05). The P side bone volume fractions of the HU and NC groups were( 29.01 ± 11.09)% and (29.56 ± 15.27)%, respectively, which were not significantly different (t=-0.072, P=0.944). The P side bone mineral densities of the HU and NC groups were(0.53 ± 0.16) g/cm3 and (0.52 ± 0.14) g/cm3, respectively, which were not significantly different (t=0.038, P=0.970). Additionally, there was no correlation between HU or serum UA and alveolar bone resorption (P > 0.05). Conclusion This research established a murine model for HU and periodontitis, but based on micro-CT analysis of alveolar bone, no relationship between HU or UA levels and periodontitis was found.

BACKGROUND: Pancreatic β-cells elevate insulin production and secretion through a compensatory mechanism to override insulin resistance under metabolic stress conditions. Deficits in β-cell compensatory capacity result in hyperglycemia and type 2 diabetes (T2D). However, the mechanism in the regulation of β-cell compensative capacity remains elusive. Nuclear factor-Y (NF-Y) is critical for pancreatic islets' homeostasis under physiological conditions, but its role in β-cell compensatory response to insulin resistance in obesity is unclear. METHODS: In this study, using obese ( ob/ob ) mice with an absence of NF-Y subunit A (NF-YA) in β-cells ( ob , Nf-ya βKO) as well as rat insulinoma cell line (INS1)-based models, we determined whether NF-Y-mediated apoptosis makes an essential contribution to β-cell compensation upon metabolic stress. RESULTS: Obese animals had markedly augmented NF-Y expression in pancreatic islets. Deletion of β-cell Nf-ya in obese mice worsened glucose intolerance and resulted in β-cell dysfunction, which was attributable to augmented β-cell apoptosis and reactive oxygen species (ROS). Furthermore, primary pancreatic islets from Nf-ya βKO mice were sensitive to palmitate-induced β-cell apoptosis due to mitochondrial impairment and the attenuated antioxidant response, which resulted in the aggravation of phosphorylated c-Jun N-terminal kinase (JNK) and cleaved caspase-3. These detrimental effects were completely relieved by ROS scavenger. Ultimately, forced overexpression of NF-Y in INS1 β-cell line could rescue palmitate-induced β-cell apoptosis, dysfunction, and mitochondrial impairment. CONCLUSION Pancreatic NF-Y might be an essential regulator of β-cell compensation under metabolic stress.
Rats ; Mice ; Animals ; Reactive Oxygen Species/metabolism* ; Diabetes Mellitus, Type 2/metabolism* ; Insulin Resistance ; Insulin ; Insulin-Secreting Cells/metabolism* ; Apoptosis ; Stress, Physiological ; Transcription Factors/metabolism* ; Palmitates/pharmacology* ; Obesity/metabolism*