Rhubarb anthraquinone derivatives (AQs) have been documented to have both therapeutic and toxic effect on liver and kidney, leading to a complex puzzle to assess their benefits and risks. In this study, the tissue distributions of AQs in SD rats after orally administrated extracts of raw and prepared rhubarb were examined whether they undergo different uptake. The total rhubarb extract (14.49 g x kg(-1) of body weight per day od, counted on the quantity of crude material) was administrated orally for 12 weeks. The concentrations of the AQs in different tissues were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The five major AQs, aloe-emodin, rhein, emodin, chrysophynol and physcion, could all be detected in the liver, kidney and spleen, while only rhein, aloe-emodin and emodin reached the quantitative limit. The tissue concentrations of AQs in raw rhubarb group were higher than that in steamed rhubarb group with rhein > emodin > aloe-emodin in the same tissue. On the whole, the tissue distribution of rhein was higher than that of emodin and aloe-emodin in liver, spleen and kidney. AQs could not be detected in those tissues after drug withdraw for 4 weeks, which suggested scarcely any accumulative toxicity of rhubarb. The result indicated that raw rhubarb had more tissue toxicity than steamed rhubarb and rhein may be one of the major poisonous ingredients. The results were concordant with the traditional Chinese medicine theory of toxicity-attenuating effect of processing.

OBJECTIVETo study the clinicopathologic features, immunophenotype and prognosis of primary cutaneous anaplastic large cell lymphoma (CALCL).

METHODSHistopathologic evaluation and immunohistochemical study by Envision method were carried out in 44 archival cases of CALCL. The clinical information and follow-up data were analyzed.

RESULTSThe patients presented with skin nodules, masses or plaques, sometimes associated with ulceration. The commonest sites of involvement were the extremities. Follow-up data were available in 39 patients. The overall survival rate was 87.2% (34/39). Disease relapses were detected in 46.2% (18/39) of the patients. Statistical analysis indicated that patients older than 50 years of age or with no less than two involved anatomic sites were more likely to have disease relapses (P < 0.05). Histologically, 31 cases were classified as common variant, 6 cases as small cell variant and 7 cases as neutrophil/eosinophil-rich variant. Immunohistochemical study showed that the rates of expression of CD30, CD45, CD45RO, CD43, CD3, cytotoxic protein and epithelial membrane antigen were 100% (44/44), 91.2% (31/34), 82.6% (19/23), 94.7% (18/19), 70.0% (28/40), 73.3% (22/30) and 31.8% (7/22), respectively. The CD4(+)/CD8(-), CD4(-)/CD8(+) and CD4(-)/CD8(-) immunophenotypes were found in 58.3% (21/36), 22.2% (8/36) and 19.4% (7/36) of the CALCL cases, respectively. Only one case (3.7%) expressed CD56.

CONCLUSIONSCALCL is a form of low-grade primary cutaneous T-cell lymphoma with a wide spectrum of clinicopathologic pattern. Special variants of CALCL should not be confused with other types of cutaneous lymphomas and inflammatory lesions. CALCL patients older than 50 years of age or with no less than two involved anatomic sites are more likely to have disease relapses.

Adult ; Age Factors ; Aged ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Immunophenotyping ; Ki-1 Antigen ; metabolism ; Lymphoma, Large-Cell, Anaplastic ; metabolism ; pathology ; Lymphoma, Primary Cutaneous Anaplastic Large Cell ; drug therapy ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Proportional Hazards Models ; Skin Neoplasms ; drug therapy ; metabolism ; pathology ; Survival Rate ; Young Adult

OBJECTIVETo investigate the clinicopathologic feature, immunophenotype and differential diagnosis of cutaneous Rosai-Dorfman disease (CRDD).

METHODSClinical manifestation, morphologic features and immunohistochemical staining were studied in 8 cases of CRDD.

RESULTSAll 8 patients presented with multiple papules, nodules and/or coalescent patches or plaques distributing over the extremities or trunk, without lymphadenopathy or other systemic abnormalities. Microscopically, the lesions were located intradermally and/or subcutaneously. CRDD was characterized by the presence of S-100 positive histiocytic cells exhibiting emperipolesis, accompanying with infiltration of mixed inflammatory cells. Fibrosis, somewhere in vague storiform pattern due to stromal responses, with distribution of individual neutrophil microabscess was seen in cases with a long course of illness. Dilated vascular spaces in dermis containing numerous large typical histiocytes were seen in 2 cases.

CONCLUSIONSCRDD is a benign, persistent proliferative disease of histiocytes. Systemic involvement is rare, outcome favorable. It should be differentiated from other types of histiocytosis, dermatofibrosarcoma protuberans, xanthoma and lymphoproliferative disorders. Immunohistochemical staining for S-100 protein and CD68 is helpful in making a correct diagnosis.

Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Diagnosis, Differential ; Female ; Histiocytosis, Sinus ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Prognosis ; S100 Proteins ; metabolism ; Skin Diseases ; metabolism ; pathology ; surgery

OBJECTIVETo study the clinicopathologic features and differential diagnosis of cutaneous regressing/regressed melanoma.

METHODSHistopathologic evaluation and immunohistochemical study by EnVision method were performed in 8 cases of cutaneous regressing/regressed melanoma. The clinical presentation, treatment and follow-up data were analyzed.

RESULTSThe age of the patients ranged from 40 to 69 years (mean 58 years). The male-to-female ratio was 3: 1. Tumors were located on the back (4 cases), sole of the foot (2 cases), ventral aspect of the toes (1 case), and the forearm (1 case). Clinically, 6 patients presented with progressive black mole of the skin, followed by subsequent focal hypopigmentation, even scarring. Two patients presented with multiple foci of dark-brown pigmentation. Microscopically, 3 cases were completely regressed malignant melanoma. Tumoral melanosis was found in 1 of 3 cases. The other 5 cases were melanoma with severe regression. The extent of regression ranged from 75% to 90%. The Breslow depth of the tumors ranged from 0.5 to 1.0 mm. Immunohistochemically, both metastatic and primary tumor cells were diffusely positive for S-100, HMB45 and Melan A, while melanophages were positive for CD68. Follow-up data were available in 8 patients, ranging from 8 to 27 months. Five patients were alive with no evidence of disease, 1 patient was alive with stable disease and 2 patients died of metastatic melanoma.

CONCLUSIONSCorrelation between clinical presentation and pathologic features is important for diagnosis of cutaneous regressing/regressed melanoma. Thin melanoma with extensive regression ( ≥ 75%) should not been regarded as low metastatic risk and wide excision combined with sentinel lymph node biopsy is recommended.

Adult ; Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Back ; pathology ; Female ; Follow-Up Studies ; Foot ; pathology ; Humans ; Lymphatic Metastasis ; MART-1 Antigen ; metabolism ; Male ; Melanoma ; metabolism ; pathology ; surgery ; Melanoma-Specific Antigens ; metabolism ; Melanosis ; pathology ; Middle Aged ; S100 Proteins ; metabolism ; Sentinel Lymph Node Biopsy ; Skin Neoplasms ; metabolism ; pathology ; surgery ; Toes ; pathology ; Treatment Outcome

OBJECTIVETo establish a drug-resistance cell line of human glioma mediated by MGMT.

METHODSSimulated the clinical usage of BCNU to establish a BCNU-resistant human glioma subline by cyclic exposing the U251 parent cells to a constant concentration of BCNU. The resistance index and the expression of MGMT mRNA of U251/BCNU were detected and compared the difference of in vitro proliferation between U251 and U251/BCNU.

RESULTSA subline--U251/BCNU was successfully established in about 4-month culture, which had a stable resistance to BCNU. U251/BCNU cells showed 17-fold higher resistance to BCNU than did U251 cells by MTT assay, while U251/BCNU cells expressed MGMT mRNA. The doubling time of U251 and U251/BCNU had no statistical difference.

CONCLUSIONA drug-resistance cell line of human glioma mediated by MGMT is established, which could provide experimental basis for further studies on the resistance mechanism and reversal methods of glioma chemotherapy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; Brain Neoplasms ; pathology ; Cell Line, Tumor ; DNA Modification Methylases ; biosynthesis ; genetics ; Drug Resistance, Neoplasm ; genetics ; Glioma ; pathology ; Humans ; O(6)-Methylguanine-DNA Methyltransferase ; metabolism ; physiology

OBJECTIVETo evaluate the effect of adenovirus-mediated double suicide gene (CD/TK) for selective killing of breast cancer cells.

METHODSVascular endothelial growth factor (VEGF)-expressing MCF-7 cells and normal human mammary epithelial cells that did not express VEGF were infected with the adenovirus containing VEGFP-CD/TK-GFP genes. CD/TK gene expression in the infected cells was detected by RT-PCR. After treatment of the infected cells with GCV and/or 5-FC, the cell growth status was evaluated using MTT assay, and the cell cycle changes were detected with flow cytometry. In nude mice bearing human breast cancer, the recombinant adenovirus vector was injected directly into the tumor followed by intraperitoneal injection of the prodrugs GCV and/or 5-FC, and the subsequent tumor growth was observed.

RESULTSThe recombinant adenovirus achieved similar infection rates in MCF-7 and human mammary epithelial cells, and the rates increased gradually with the multiplicity of infection (MOI) of the virus. RT-PCR demonstrated the presence of CD/TK gene product in infected MCF-7 cells, but not in the infected mammary epithelial cells. The infected MCF-7 cells, but not the mammary epithelial cells, were highly sensitive to the pro-drugs. The CD/TK fusion gene system showed significantly greater efficiency than either of the single suicide gene in killing the target cells (P<0.01). At the MOI of 100, treatment of the infected cells with the pro-drugs resulted in increased cell percentage in G(0)-G(1) phase and decreased percentage in S phase. In nude mice bearing MCF-7 cell-derived subcutaneous tumor, treatment with the double suicide gene system significantly inhibited the tumor growth, showing much stronger effect than either of the single suicide gene (P<0.01).

CONCLUSIONThe adenovirus-mediated CD/TK double suicide gene driven by VEGF promoter combined with GCV and 5-FC treatment can be an effective therapy against experimental breast cancer, and produces much greater efficacy than the single suicide gene CD/TK combined with GCV or 5-FC.

Adenoviridae ; genetics ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cytosine Deaminase ; genetics ; metabolism ; Female ; Flow Cytometry ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Kinase ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the role of C-type natriuretic peptide receptor (NPR-C) and large-conductance calcium-activated potassium channels (BK(Ca)) in brain natriuretic peptide (BNP) induced porcine coronary artery dilation.

METHODSPorcine coronary artery rings were obtained and treated with BNP (10(-6) mol/L), BNP + NPR-C antagonist cANF4-28 (10(-6) mol/L) and BNP + BK(Ca) blocker tetraethylammonium (TEA, 1 mmol/L). The vascular tone experiments were observed on 10 vessel segments. BK(Ca) current density was measured by the whole-cell patch clamp technique.

RESULTSThe maximum diastolic rate was similar between BNP group (68.51% ± 11.50%) and cANF4-28 + BNP group (65.67% ± 11.90%, P > 0.05) while significantly reduced in TEA + BNP group (28.87% ± 4.55%, all P < 0.05). When the holding potential was set at +60 mV, the BK(Ca) current density of BNP group was (78.48 ± 5.86) pA/pF, which was significantly higher than control group [(53.84 ± 4.55) pA/pF, P < 0.05], which was equally reduced in the TEA group and TEA + BNP group [(28.80 ± 2.76) pA/pF and (30.60 ± 3.88) pA/pF respectively, all P < 0.05 vs. control group].

CONCLUSIONBNP could relax the porcine coronary arterial smooth muscles by increasing BK(Ca) current, and this effect is not mediated by NPR-C.

Animals ; Coronary Vessels ; drug effects ; physiology ; Large-Conductance Calcium-Activated Potassium Channels ; physiology ; Natriuretic Peptide, Brain ; pharmacology ; Patch-Clamp Techniques ; Receptors, Atrial Natriuretic Factor ; physiology ; Swine

[ ABSTRACT] AIM:To investigate the effects of Chaihu Shugan powder ( CSP) on lipid metabolism and the pro-teins involved in adenosine 5’-monophosphate-activated protein kinase (AMPK)/sirtuin 1 (SIRT1) pathway in the liver tissues of the rats with non-alcoholic fatty liver disease (NAFLD).METHODS: Sprague-Dawley rats were randomly di-vided into normal control ( NC) group, with HFD ( HFD) group and CSP group.The NAFLD models were established by feeding with HFD for 16 weeks in the rats.The rats in CSP group were intragastrically administered with CSP extracts (9.6 g· kg-1 · d-1 ) , and blood and liver samples were collected 16 weeks later.Serum and liver levels of total cholesterol ( TC) and triglyceride ( TG) , and serum levels of alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) were measured using an automatic biochemical analyzer.The histological changes of liver tissues were observed with HE staining, while the lipid deposition was observed with Oil Red O staining.The ultrastructural changes of the liver tissues were observed under transmission electron microscope.Moreover, the protein levels of AMPK, phosphorylated AMPK (pAMPK), SIRT1 and uncoupling protein 2 (UCP2) in the liver were detected by Western blot.RESULTS:The results of HE staining, Oil Red O staining and electron microscopy demonstrated that NAFLD rat model was successfully estab-lished.Compared with NC group, the serum and liver levels of TC and TG, and serum level of AST in model group were markedly elevated ( P<0.01) .Moreover, the protein levels of pAMPK and SIRT1 in HFD group were markedly reduced (P<0.01), whereas UCP2 level was elevated (P<0.01).Furthermore, liver levels of TC and TG, and serum level of AST in GSP group were markedly reduced as compared with HFD group ( P<0.05 ) .The protein levels of pAMPK and SIRT1 were elevated ( P<0.05 ) , whereas the UCP2 level was reduced as compared with HFD group ( P<0.01 ) .The protein level of AMPK between the 3 groups had no significant difference.CONCLUSION: CSP attenuates hepatic lipid disorder and hepatic lipid deposition in NAFLD rats induced by feeding with HFD for 16 weeks, which is associated with the activation of AMPK/SIRT1 pathway.

Objective To explore the effect of atorvastatin on the proliferation and apoptosis of K562 cells andto investigate its mechanisms.Methods The cells were treated by different concentrations of atorvastatin.The CCK-8 assay was employed to detect the cell proliferation.The cell apoptosis was detected by AnnexinV-FITC/PI dual staining;the flow cytometry was used to detect the cellular cycle;the activities of caspase-3,-8,-9 were detected by the colorimetric method;qRT-PCR was employed to measure the mRNA expression levels of Bcl-2 and PDCD5 in K562 cells.Results Atorvastatin could inhibit the proliferation of K562 cells in a time-and dose-dependent manner(P＜0.05);and induced the apoptosis of K562 cells,the percentage of G0/G1 phase cells was increased after atorvastatin treating k562 cells(P＜0.01),while the percentage of S phase cells was decreased(P＜0.01),moreover which showing the concentration dependence(P＜0.01);atorvastatin activated the caspase-3,-8,-9 (P＜0.01);down-regulated Bcl-2 mRNA expression and up-regulated PDCD5 mRNA expressionin a concentration dependence(P＜0.01).Conclusion Atorvastatin can inhibit the proliferation and induce apoptosis in K562 cells.

Objective:This study aims to explore the expression of miR-143 in gallbladder cancer tissues and its correlation with tumor angiogenesis, clinicopathologic features and clinical prognosis.Methods:The clinical and pathological data of 78 patients with gallbladder cancer who were surgically resected and pathologically confirmed from January 2008 to June 2016 in the Department of General Surgery, Xuancheng Hospital Affiliated to Wannan Medical College were collected, including 20 males and 58 females, with an average age of (68.0±1.4) years. 78 cases of gallbladder cancer tissue were used as the tumor group, and 32 cases of normal gallbladder tissue adjacent to cancer were randomly selected as the control group. The expression level of miR-143 in the tumor group and the control group was detected by in situ hybridization. Microvessel density (MVD) were evaluated with immunohistochemistry to detect the expression of CD34 in tumor group. The expression of miR-143 and its correlation with tumor MVD, pathological characteristics and prognosis were analyzed by Cox regression analysis.Results:The results of in situ hybridization showed that the positive rate of miR-143 in gallbladder cancer tissues and average expression score were significantly lower than those in adjacent normal gallbladder tissues [29.49% (23/78) vs 62.5% (20/32), χ 2=10.39, P=0.001; (2.705±0.172) vs (4.688±0.405), t=5.33, P<0.001]. Correlation analysis showed that the expression of miR-143 was associated with lymph node metastasis of gallbladder cancer ( P=0.036). The MVD counts in the miR-143 positive group were significantly lower than those in the miR-143 negative group [(126.2±8.483) vs (157.4±6.412), t=2.75, P=0.007)]. The median survival time of gallbladder cancer patients with miR-143 positive and negative was 16 and 9 months, respectively. Cox regression analysis showed that the low expression of miR-143 was an independent risk factor affecting the clinical prognosis of gallbladder cancer. Conclusion:The expression of miR-143 is down-regulated in gallbladder cancer and participates in angiogenesis and clinical prognosis of gallbladder cancer.