OBJECTIVETo compare the Golgi proteome of hepatocellular carcinoma (HCC) with that of the adjacent non-tumor tissues.

METHODSHepatocellular carcinoma and adjacent non-tumor tissues were obtained from HCC patients. The protein expression maps in Golgi were obtained by two-dimensional gel electrophoresis (2-DE), and the differentially expressed protein spots were analyzed by PD-Quest software. Peptide mass fingerprint (PMF) of differential protein spots was obtained with MALD-TOT-MS.

RESULTSAccording to 2-DE maps, the average numbers of protein spots were (1153+/-49) and (1086+/-37) in hepatocellular carcinoma and the adjacent non-tumor tissues. Compared to the adjacent non-tumor tissues, 27 proteins were upregulated, and 20 proteins were downregulated in HCC Golgi.

CONCLUSIONSThe Golgi proteome in HCC tissues is different from that in the adjacent non-tumor tissues, and the differential expression proteins are involved in energy metabolism, tumor metastasis, and cell cycle regulation.

Annexin A5 ; analysis ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; methods ; Golgi Apparatus ; metabolism ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Proteins ; analysis ; metabolism ; Proteome ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Objective To analyze the changes of clinical and pathological features in the patients of IgA nephropathy with anemia.Methods Four hundred and nine patients of IgA nephropathy diagnosed by renal biopsy were classified into two groups:IgA nephropathy with nonanemia (group 1) and IgA nephropathy with anemia (group 2).Changes were studied retrospectively between the groups.Results Serum hemoglobin level was correlated with the clinical parameters of IgA nephropathy.Companed to group 1,changes in group 2 were as followed:serum creatinine increased,eGFR decreased,proteinuria increased; the global sclerosis,segmental sclerosis,crescents and tubulointerstitial lesions worsened.The glomerular and tubulointerstitial lesions were negatively correlated with serum hemoglobin and eGFR,but positively correlated with serum uric acid and proteinuria (P＜0.05).Multivariate Logistic regression analysis revealed that anemia was an independent risk factor for the tubulointerstitial lesion.Conclusion Clinical feature and pathological damages in the patients of IgA nephropathy with anemia are more serious than those with non-anemia.

Objective To investigate the influences of UVA on the secretion and expression of chemokine CXCL11/I-TAC by HaCaT cells induced by interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α). Methods HaCaT cells were cultured in the presence of IFN-7 and TNF-a and irradiated with UVA of 2, 4 and 8 J/cm~2, respectively; those cells receiving neither treatment with IFN-γ or TNF-α nor UVA irradiation served as the negative control, and those receiving only cytokine treatment but no irradiation as the positive control. After another 24-hour culture, enzyme-linked immunosorbent assay (ELISA) was performed to detect the protein levels of CXCL11/I-TAC in the supernatant of HaCaT celb, real time PCR to measure the mRNA expression of CXCL11/I-TAC in these HaCaT cells. Results As far as the negative control HaCaT cells were concerned, there was a minor secretion of CXCL11/I-TAC protein and expression of CXCL11/I-TAC mRNA. After treatment with IFN-7 and TNF-a of 10 μg/L, the protein and mRNA expressions of CXCL11/ I-TAC were synergistically upregulated, whereas the induced secretion and expression of CXCL11/I-TAC by HaCaT cells were dose-dependently inhibited by UVA irradiation. Conclusions UVA irradiation inhibits the secretion and expression of CXCL11/I-TAC by HaCaT cells, which in turn suppresses the chemotaxis of Th1/ Tel cells in some degree.

Objective To investigate the effects of baicalein and acitretin on the apoptosis in a human cutaneous squamous cell carcinoma cell line, SCL-12. Methods Cultured SCL-12 cells were treated with different concentrations of baicalein (3.125, 6.25, 12.5 μmol/L) and acitretin (2.5, 5.0, 10.0 μ mol/L), alone or in combination, for 48 hours. Subsequently, cell proliferation was detected by MTT assay, and cell apoptosis by ELISA as well as annexin V-FITC and propidium iodide double staining. Real-time quantitative RT-PCR was used to detect the expression of Fas mRNA in SCL-12 cells. Results The cell proliferation of SCL-12 cells was inhibited by baicalein and acitretin alone or in combination. The combination of baicalein and acitretin at the three tested concentrations, except for that of baicalein at 3.125 μmol/L and acitretin at 2.5 μmol/L, more strongly inhibited the proliferation of SCL-12 cells compared with baicalein or acitretin alone, and the inhibitory effect was in a dose-dependent manner. The early apoptosis rate was 9.39% ± 1.52%, 20.86% ± 2.16%,36.85% ± 3.26% in SCL-12 cells treated with baicalein of 3.125 μmol/L, acitretin of 5.0 μmol/L alone and their combination, respectively, significantly higher than that in untreated cells (4.39% ± 0.64%, all P <0.05); the induction of apoptosis in SCL-12 cells by the combination of baicalein and acitretin was stronger than that by baicalein or acitretin alone (F = 138.44, P < 0.05). Baicalein and acitretin alone or in combination significantly increased the mRNA expression of Fas in SCL-12 cells, and the effect of their combination was stronger than that of baicalein or acitretin alone. Conclusions Baicalein and aeitretin could inhibit the growth of and induce the apoptosis in SCL-12 cells, and the effect is enhanced by the combination of baicalein and acitretin, which may be associated with the upregulation of Fas expression in SCL-12 cells.

Objective To investigate the expression of survivin and bcl-2 in human squamous cell carcinoma (SCC) lesions and cell line SCL-1. Methods Tissue samples from 60 patients with SCC and 10 normal human controls were immunohistochemically stained to detect the expressions of survivin and bcl-2.Western blot was used to measure the expressions of bcl-2 and survivin proteins in HaCaT human keratinocytes and SCL-1 human squamous cell carcinoma cells. Results In normal control tissues, there was no expressions of survivin or bcl-2, while in SCC, the expression rates of bcl-2 and survivin were 70% and 60%, respectively,and there was no statistical correlation between the expressions of bcl-2 and survivin (P >0.05). Neither the expression of survivin nor that of bcl-2 was correlated to patients' age, gender or lesional site (all P >0.05). A statistical correlation was observed between the pathological stage in patients and expression of bcl-2 as well as between lymph node metastasis and expression of survivin (both P < 0.05). Western blot analysis revealed a significant increase in the expression of survivin and bcl-2 in SCL-1 cells compared with HaCaT cells. Con-clusion In SCC, survivin and bcl-2 seem to play their roles via different anti-apoptotic pathways.

Objective To investigate the feasibility and clinical value of ultrasound elastography in assessment of sternocleidomastoid fibrosis in patients after radiotherapy for nasopharyngeal carcinoma . Methods Forty‐five patients with nasopharyngeal carcinoma after radiotherapy ( study group) and 52 healthy volunteers ( control group ) underwent ultrasound elastography . Bilateral sternocleidomastoid elasticity score , thickness , width , resistance index ( RI ) , and pulsatility index ( PI ) were analyzed and compared . Results The sternocleidomastoid elastography in study group was mainly blue ,and that in control group was mainly green . The difference of sternocleidomastoid elasticity score ,width ,and thickness was significant between study group and control group ( P <0 .05) ,reseparately . There was no significiant difference in the RI ,PI between study group and control group ( P > 0 .05) . Conclusions Ultrasound elastography could be effectively and noninvasively used for evaluating the sternocleidomastoid fibrosis in patients after radiotherapy for nasopharyngeal carcinoma . It has a potential clinical value in the grading and classification of fibrosis .

OBJECTIVETo analyze hepatotoxicity of Polygonum multiflorum and clinical character- istics of drug-induced liver injury (DILI) caused by Polygonum multiflorum and its preparations.

METHODSA retrospective study was performed in 158 patients treated at 302 Military Hospital between January 2009 and January 2014. All of them had used Polygonum multiflorum and its preparations before the onset of DILI, and their clinical characteristics and prognoses were analyzed.

RESULTSOf the 158 DILI patients who used Polygonum multiflorum or its preparations, 92 (58.2%) combined with Western medicine or Chinese herbal preparations without Polygonum multiflorum; 66 patients (41.8%) used Polygonum mult florum and its preparations alone. In 66 DILI patients induced by Polygonum multiflorum or its preparations alone, 51 cases (77.3%) were induced by Polygonum multiflorum compounds and 22.7% by single Po- lygonum multiflorum; 4 cases (6.1%) were caused by crude Polygonum multiflorum and 62 (93.9%) by processed Polygonum multiflorum and its preparations. Clinical injury patterns were hepatocellular 92.4% (61 cases), cholestatic 1.5% (1 case), and mixed 6.1% (4 cases). Pathological examination was per- formed by liver biopsy in 32 cases (48.15%), manifested as hepatocellular degeneration and necrosis, fibroplasia, Kupffer cells with pigment granule, and a large number of eosinophil infiltration, were ob- served. Four patients were developed into liver failure, 4 into cirrhosis, and 1 died.

CONCLUSIONPolygo- num multiflorum and its preparations could induce DILI, but clinical diagnosis of Polygonum multiflorum induced hepatotoxicity should be cautious.

Asian Continental Ancestry Group ; Chemical and Drug Induced Liver Injury ; diagnosis ; Cholestasis ; Drugs, Chinese Herbal ; adverse effects ; Fallopia multiflora ; Humans ; Liver Cirrhosis ; Liver Failure ; Plant Preparations ; adverse effects ; Polygonum ; Retrospective Studies

OBJECTIVETo investigate the association between the expression of galectin- 3 protein and peritoneal metastasis in gastric cancer.

METHODThe expressions of galectin- 3 was detected in matching- samples including primary gastric cancer lesions,lymph node metastases,peritoneal metastases and paratumor normal tissues by immunohistochemistry. All specimens were gained from 35 patients who had synchronous peritoneal metastasis from gastric cancer.

RESULTSThe over- expression of galectin- 3 was observed in 97% (34/35) of the gastric cancer lesions, the peritoneal metastases and the lymph node metastases,whereas in 14% (5/35) of paratumor normal tissues. There were significant differences in the expression of galectin- 3 between paratumor normal tissues and the gastric carcinoma lesions,peritoneal metastases and lymph node metastases (P< 0.05),but there were no significant differences among the gastric cancer lesions,the peritoneal metastases,and the lymph node metastases (P> 0.05).

CONCLUSIONThe expression of galectin- 3 in gastric cancer lesions can be used as a biological marker of peritoneal metastasis from gastric cancer before operation and as a prognostic factor of gastric cancer.

Galectin 3 ; metabolism ; Humans ; Immunohistochemistry ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Neoplasm Staging ; Peritoneal Neoplasms ; metabolism ; pathology ; secondary ; Stomach Neoplasms ; metabolism ; pathology

Objective To evaluate the role of vascular endothelial growth factor-D(VEGF-D)in lymphangiogenesis of breast cancer,and investigate its relationship with some clinicopathological parameters and prognosis. Methods VEGF-D expression was detected in 85 cases with primary breast carcinoma by immunohistochemistry,and VEGF-D mRNA was detected by RT-PCR.Podoplanin monoclonal antibody was used to mark lymphatic endothelial cell and lymphatic vessel density(LVD) was counted.The relationship among the aboved parameters,clinicopathological parameters and prognosis was analysed. Results It was revealed by immunohistochemistry that VEGF-D expression was ranked by 5 levels: negative,n=5(5.88%);"+",n=17(20%);"++",n=34(40%);"+++",n=22(25.88%),and "++++",n=7(8.24%).VEGF-D expression was associated with tumor lymph node metastasis and TNM clinical stage(P

Objective To investigate the inhibitory effect of metformin on high glucose-induced lipolysis and further elucidate the underlying mechanisms,for better understanding of metformin.Methods Adipocytes were isolated from epididymal fat pads of Sprague-Dawley rats.After isolation and digestion,packed adipocytes were divided into four groups according to the glucose and metformin in the media,control A group containing 5 mmol · L-1 glucose,control B group containing 25 mmol · L-1 glucose,test A group containing 500 μmol · L-1 metformin with 5 mmol · L-1 glucose and test B group containing 25 mmol · L-1 glucose with 500 μmol · L-1 metformin or at the concentrations as planned.After incubation,glycerol accumulated or released into the media in 30 minutes was determined by the use of a colorimetric assay and served as an index of lipolysis.The expressions of phosphorylated and total perilipin as well as adipose triglyceride lipase (ATGL) were examined by Western Blot.Adipose lipases activity was assayed by using an enzymatic method.Results The glycerol release in the control B and test B groups were (1.61 ± 0.08) and (0.50 ± 0.06) μmol · mL-1 packed cell volume,respectively,suggesting that metformin significantly inhibited the lipolytic action of high glucose (P ＜0.001).Such an inhibitory effect lasted from 16 h to 24 h after incubation.The adipose lipases activity in the control B and test B groups were (344.28 ± 65.98) and (200.44 ± 64.25)μmol glycerol · mg protein-1 · h-1,respectively,and thus metformin caused a 41.78％ (P ＜0.05) reduction of adipose lipases activity elevated by high glucose.Compared with control B group,metformin in test B group significantly attenuated the phosphorylation of perilipin induced by high glucose (P ＜ 0.01).However,the protein amount of total ATGL was not changed in control B group by high glucose or in test group (A and B) by metformin compared with that of control A group (all P ＞ 0.05).Conclusion Metformin inhibits high glucose-induced lipolysis through eliminating perilipin phosphorylation and suppressing lipolytic lipases activity.We suggest that such antilipolytic effect of metformin in hyperglycemia could be a molecular basis by which metformin reduces the release of free fatty acids from adipose tissue to bloodstream and thus ameliorates insulin resistance in diabetes.