Kaposi sarcoma-associated herpesvirus (KSHV),also known as human herpesvirus 8 (HHV-8),is discovered in 1994 from Kaposi's sarcoma (KS) lesion of an acquired immunodeficiency syndrome (AIDS)patient.In addition to its association with KS,KSHV has also been implicated as the causative agent of two other AIDS-associated malignancies:primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD).KSHV is a complex DNA virus that not only has the ability to promote cellular growth and survival for tumor development,but also can provoke deregulated angiogenesis,inflammation,and modulate the patient's immune system in favor of tumor growth.As KSHV is a necessary but not sufficient etiological factor for KS,human immunodeficiency virus (HIV) is a very important cofactor.Here we review the basic information about the biology of KSHV,development of pathogenesis and interaction between KSHV and HIV.

Animals ; Bone Marrow Cells ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Graft vs Host Disease ; Male ; Mesenchymal Stromal Cells ; Mice ; Mice, Inbred C57BL ; Pregnancy

Objective To package the recombinant lentivirus containing HIV-1 Vpr gene and detect the effect of Vpr protein expression on the latent infection and lyric replication of KSHV.Methods The fragment of Vpr gene from expression plasmid pCI-neo-Vpr was cloned into the lentivirus vector pHAGE-CMV-MCS-IzsGreen,then the recombinant plasmid pHAGE-Vpr,package vector psPAX2 and envelope vector pMD2.G were cotransfected into 293T cells.GFP expression was observed by fluorescent microscopy.Culture media of 293T cells were harvested and filtered through 0.45 μm filter.After 293T cells were infected by a series of diluted lentivirus,the virus titer was checked by observing GFP expression.Vpr mRNA transcripts in 293T cells were detected by RT-PCR.Then BCBL-1 cells were infected by the recombinant lentivirus with 1 MOI,GFP expression was observed by fluorescent microscopy,and the mRNA transcripts and protein expression of Vpr in BCBL-1 cells were detected by RT-PCR and Western blot.Meanwhile,the mRNA transcripts and protein expression of KSHV lytic cycle gene Rta were detected by RT-PCR and Western blot,respectively.Results The recombinant lentivirus carrying HIV-1 Vpr was packaged successfully with the virus titer of 4 × 107 TU/ml.After infected with lentivirus,BCBL-1 cells could express GFP,and the exact band of Vpr was detectable by RT-PCR and Western blot.Moreover,the expression of KSHV Rta mRNA and protein were downregulated by Vpr protein.Conclusion Overexpression of HIV-1 Vpr mediated by the recombinant lentivirus could inhibit KSHV lytic replication and enhance KSHV latent infection.

Adult ; Aged ; Angiography, Digital Subtraction ; Carcinoma, Hepatocellular ; diagnostic imaging ; therapy ; Chemoembolization, Therapeutic ; Female ; Hemangioma ; diagnostic imaging ; therapy ; Humans ; Liver Neoplasms ; diagnostic imaging ; therapy ; Male ; Middle Aged ; Radiography, Interventional ; methods ; Tomography, X-Ray Computed ; methods

Objective To explore the role of glycogen synthase kinase (GSK)-3β/β-catenin signaling pathway on human immunodeficiency virus 1 (HIV-1) negative factor (Nef) protein promoting of human herpesvirus-8 (HHV-8) viral interleukin-6 (vIL-6)-induced angiogenesis.Methods GSK-3β mutant plasmid GSK-3β-S9A,dominant negative (DN) form GSK-3β-DN and the control vector pcDNA3.1+ were transfected into endothelial cells which stably expressed HHV-8 vIL-6 or HIV-1 Nef,or co-expressed vIL-6 and Nef protein.Microtubule formation assay was performed to explore microtubule formation ability.A chick embryo chorioallantoic membrane (CAM) model was used to detect angiogenesis.The expression of GSK-3β/β-catenin signaling pathway-related kinases in transfected cells and CAM tissue were further detected by Western blot.The measurement data were compared by t test.Results The activity of GSK-3β was decreased and the ability of HIV-1 Nef protein was enhanced by transfection with GSK-3β-DN in promoting vIL-6 induced microtubule formation (3.42 vs 2.51,t =3.67,P＜0.01) and angiogenesis (6.25 vs 3.97,t=4.06,P＜0.01).In contrast,the activity of GSK-3β was significantly increased and these functions of HIV-1 Nef protein mentioned above were inhibited by transfection with GSK-3β-S9A (0.62 vs2.51,t=8.48,P＜0.01; 0.39 vs 3.97,t=8.59,P＜0.01).The results of Western blot showed that with the elevated level of,β-catenin (in cells:3.53 vs 2.07,t=6.60,P＜0.05; in tissues:2.76 vs 1.74,t=17.40,P＜0.01) and vascular endothelial growth factor (VEGF,in cells:2.68 vs 1.87,t=4.28,P＜0.01; in tissues:2.20 vs 1.39,t=7.08,P＜0.01) were increased in the GSK-3β-DN transfected cells or tissues,while the opposite results were achieved in the GSK-3β-S9A-transfected cells (GSK-3β phosphorylation:0.50 vs 1.47,t=7.33,P＜0.01; β-catenin:1.05 vs 2.62,t=29.50,P＜0.01; VEGF:0.74 vs 2.16,t=20.95,P＜0.01) or tissues (GSK-3β phosphorylation:0.35 vs 1.97,t=10.72,P＜0.01; β-catenin:0.79 vs 1.77,t=5.72,P＜0.01; VEGF:0.43 vs 1.65,t=11.89,P＜ 0.01).Conclusion GSK-3β/β-catenin signaling pathway is involved in vIL-6-induced angiogenesis promoted by HIV-1 Nef protein,which would be valuable for the therapy of Kaposi's sarcoma,an acquired immunodeficiency syndrome,as a potential molecular target.

Background Stromal-derived factor 1α /chemokine receptor 4(SDF-1α/CXCR4) axis is one of the important signals which mediates several different activities such as chemotaxis,adhesion,proliferation and survival resulting in recruitment to sites of immune and inflammatory reactions.Considerable evidence suggests that CXCR4/SDF-1α axis is involved in tumor angiogenesis and plays a key role in the development of ocular neovascularization.Objective The purpose of this study was to explore the effect of CXCR4 antagonist on the development of cxperimental corneal neovascularization(CNV).Methods CNV model was established in the left eye of 8-weekold clean BALB/c mouse by putting the filter with 1 mol/L NaOH at the central cornea for 40 seconds.The animals were randomizcd into hyaluronate group and CXCR4 antagonist group,and the edydrops was topically administered respectively on the day of modeling 4 times per day for 14 days.CNV was examined under the slit lamp at the fourteenth day,and then the corneal suspension and section were made.Expressions of CXCR4 mRNA and protein in corneas were detected using RT-PCR and Western blot.The CD31 level in cornea was assayed by flowcytometry and immunochemistry.The expression of VEGF in burned corneas and suspension from mouse peritoneal macrophages stimulated with CXCR4 antagonist in vitro was detected by ELISA.The use of the animal followed Ragulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results Two weeks after corneal alkali burn,the growth of CNV peaked under the slit lamp.Compared with hyaluronate group,CNV was obviously decreased in the CXCR4 antagonist group.Immunochemistry showed that intensity of positive staining for CD31 in cornea in the CXCR4 antagonist group was weaker than the hyaluronate group.Flowcytometry clarified that CD31 positive cells rate was 9.50％ ±2.34％ in the CXCR4 antagonist group and 17.50％ ±3.16％ in the hyaluronate group,showing a significant difference between them (t=-7.312,P＜0.05).In 2,4,7 days after cornea alkali burn,the expressions of CXCR4 mRNA and protein were significantly enhanced in burn corneas compared with normal corneas(P＜0.01 ;P＜0.05).ELISA showed that the VEGF expression level in corneal tissue and supernatant of mouse peritoneal macrophages in vitro were significantly lower in the CXCR4 antagonist group than that of hyaluronate group(t =10.927,5.151,P＜0.05).The expression level of VEGF in corneal suspension was lower in the GM-CSH+CXCR4 antogonist group than that in the GM-CSH group (P＜0.05).Conclusions CXCR4 antagonist can reduce experimental CNV by down-regulating VEGF expression in cornea.

The contents of adenosine, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, parishin and sulfur dioxide residue were compared in differently-processed Gastrodiae Rhizoma to provide the basis for a reasonable processing method of Gastrodiae Rhizoma. The analysis was performed on a Merck Purospher STAR column (4.6 mm x 250 mm, 5 μm) with a mobile phase consisting of methanol and water (containing 0.1% formic acid) under gradient elution at a flow rate of 1.0 mL x min(-1). The eluates were detected at 270 nm, and the column temperature was 35°C. The content of adenosin, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxy-benzaldehyde and parishin in processing of boiling or sulfur-fumigated were lower than that of in processing of steaming. Furthermore, the sulfur dioxide residue of sulphur-fumigated groups exceed 400 mg x kg(-1). This stable and reliable method will contribute to the quality control of different processed Gastrodiae Rhizoma.
Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Gastrodia ; chemistry ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods

Drugs are exogenous compounds for human bodies, and will be metabolized by many enzymes after administration. CYP450 enzyme, as a major metabolic enzyme, is an important phase I drug metabolizing enzyme. In human bodies, about 75% of drug metabolism is conducted by CYP450 enzymes, and CYP450 enzymes is the key factor for drug interactions between traditional Chinese medicine( TCM) -TCM, TCM-medicine and other drug combination. In order to make clear the interaction between metabolic enzymes and TCM metabolism, we generally chose the enzymatic activity as an evaluation index. That is to say, the enhancement or reduction of CYP450 enzyme activity was used to infer the inducing or inhibitory effect of active ingredients and extracts of traditional Chinese medicine on enzymes. At present, the common method for measuring metabolic enzyme activity is Cocktail probe drugs, and it is the key to select the suitable probe substrates. This is of great significance for study drug's absorption, distribution, metabolism and excretion (ADME) process in organisms. The study focuses on the interaction between TCMs, active ingredients, herbal extracts, cocktail probe substrates as well as CYP450 enzymes, in order to guide future studies.
Animals ; Cytochrome P-450 Enzyme Inhibitors ; metabolism ; pharmacology ; Cytochrome P-450 Enzyme System ; chemistry ; metabolism ; Drugs, Chinese Herbal ; metabolism ; pharmacology ; Enzyme Activation ; drug effects ; Enzyme Activators ; metabolism ; pharmacology ; Humans

Objectives To analyze the diagnosis and management of steroid-resistant nephrotic syndrome (SRNS) accompanied with irreversible leukoencephalopathy in children. Methods The clinical, laboratory and imaging data were retrospectively analyzed in a SRNS child accompanied with irreversible leukoencephalopathy. A literature review was performed. Results After clinical diagnosis of SRNS, glucocorticoid, immunosuppressant, and hemodialysis were administrated for 10 months. During the course of treatment, the seizures, visual problems, and hypertension were repeatedly occured. The cranial MRI showed bilateral occipital parietal lobe hyperintensity and right frontotemporal lobe hyperintensity on T2-weighted imaging and bilateral occipital parietal lobe hypointensity on T2-Flair imaging, which indicated that encephalomalacia was accompanied with gliosis. Conclusions A variety of reasons may induce leukoencephalopathy in children. The accompanied irreversible leu-koencephalopathy should be strongly considered in management of SRNS.

Objective:To test the expression of Minichromosome maintenance complex component 7(MCM7) protein in hepato-cellular carcinoma(HCC) of different species including human, rat and tree shrew (tupaia) by cross-species oncogenomics approach, and to investigate the relationship between the expression of MCM7 and the development of hepatocellular carcinoma and its clinical significance. Methods:Western blot and Immunohistochemistry were applied to detect the expression levels of MCM7 protein in HCC tissues,corresponding HCC-adjacent liver tissues and normal liver tissues collected from different species including human, rat and tree shrew, respectively. The clinicopathologic factors were also analyzed with the results of Immunohistochemistry. Results:Western blot analysis showed that the expression of MCM7 protein in HCC tissues of human and rat were higher than that in corresponding HCC-ad-jacent liver tissues and normal liver tissues, respectively and significantly (P<0.05). However, the expression of MCM7 protein in HCC tissues of tree shrew were also higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, but no significant difference was found among three types of tissues (P>0.05).There was also no significant difference between HCC-adjacent liver tis-sues and normal liver tissues in three species (P>0.05). Immunohistochemical analysis showed that MCM7 protein was mainly ex-pressed in nucleus of HCC cells, and the positive rate of MCM7 protein in HCC tissues of human, rat and tree shrew were significantly higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, respectively (P<0.05). However, no significant difference was found between HCC-adjacent liver tissues and normal liver tissues (P>0.05). Moreover, the protein level of MCM7 was intimately related to patient's HCC stage, extrahepatic metastases and postoperative recurrence (P<0.05). Conclusion:MCM7 protein might play a pivotal role in hepatocarcinogenesis. In addition, it was probably related to patient's HCC stage, extrahepatic metastases and postoperative recurrence. It seems very likely that MCM7 may be applied as a new molecular target in HCC prevention and treat-ment.