Objective To investigate the expression of Clq complement receptor(ClqRp and gClqR)in the peripheral blood mononuclear cells(PBMC)of systemic lupus erythematosus(SLE), and the corre|ation between serum levels of complement lq(Clq)and anti-Clq autoantibodies(ClqAb)with ClqRp and gClqR is analyzed. The probable mechanism of ClqRp and gClqR in the development of SLE is explored. Methods The peripherial blood monouclear cells of 58 SLE patients and 30 healthy subjects were collected for the detection of the expression of ClqRp and gClqR by flow cytometry. The serum levels of Clq and ClqAb were detected by single radial immunodiffusion and enzyme-linked immunosorbent assay(ELISA)re- spectively. The correlation between ClqRp and gClqR and other disease activity parameters, such as Clq, ClqAb, SLEDAI score, anti-dsDNA antibody, C3, C4 levels were analyzed. Results The expression of ClqRp on mononuclear and neutrophiles of SLE was(7.2?2.3)% and(3.4?2.1)%, lower than that in healthy individuals [(10.6?2.1)% and(9.0?8.7)% ], P

Humans ; Lung Neoplasms ; therapy ; Medicine, Chinese Traditional ; methods ; trends

OBJECTIVE: To construct the expressing vector of siRNA which can inhibit the Smad3 activity. METHODS: Sixty-four bases of 2 pair oligos for hairp in RNA expression which targeted Smad3 gene were chemically synthesized and annealed. pSUPER vector was linearized with BgL II and Hin d III treated with alkaline phosphatase (CIP). Anneled oligos were inserted into the downstream of the treated pSUPER's pol III H1 promoter to construct RNAi plasmid (pSUPER Smad3). Oligos with a scrambled sequence were used as a negative control. pSUPER Smad3 was transfected into human renal tubular epithelial cells (HKC). RESULTS: Recombinant pSUPER Smad3 vector was identified by the digestion with Eco R I and Hin d III, and confirmed by the sequencing analysis with T3 primer. Sixty-four bases had been inserted into the expected site. Furthermore, the insertion sequence was exactly corrected. The activity evaluation indicated that mRNA and protein of Smad3 but not Smad2 were inhibited by pSUPER Smad3 in HKC. CONCLUSION The pSUPER Smad3 system has been constructed successfully, and has high inhibition and specificity in vitro.
Epithelial Cells ; metabolism ; Humans ; Kidney Tubules ; cytology ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Smad3 Protein ; antagonists & inhibitors ; genetics ; Transfection

OBJECTIVETo study the change in ultrastructure of C6 glioma cells after photodynamic therapy (PDT), to compare morphological differences in necrosis and apoptosis before and after PDT treatment, and to evaluate the effect of photodynamic therapy on the blood brain tumor barrier (BTB) of C6 glioma.

METHODSThe model was produced by transplanting C6 glioma cells cultured in vitro using Peterson method into the caudate nuclei of Wister rats. The experiment group received PDT for two weeks after the operation. The sub-cellular structure, blood-brain-barrier (BBB) and BTB in both groups were observed under electron microscope.

RESULTSApoptosis in different phases and necrosis could be observed in some C6 glioma cells. Swelling occurred on the ultrastructure of cellular organs such as mitochondria and endoplasmic reticulum in most of the cells. Damage to the BTB, reduction of the number of cellular organs in endothelial cells of the capillary blood vessels, stretch of the tight junction, and enlargement of the gaps between endothelial cells were also seen in the experiment group. Meanwhile, limited impact on the normal sub-cellular structures and BBB was observed.

CONCLUSIONPDT could induce apoptosis and necrosis of C6 glioma cells due to the damage to the ultrastructure of mitochondria and endoplasmic reticulum. The weakened function of C6 glioma BTB initiated by PDT makes it possible to perform a combined therapy of PDT and chemotherapy for glioma.

Animals ; Blood-Brain Barrier ; Brain Neoplasms ; drug therapy ; ultrastructure ; Cell Line, Tumor ; Glioma ; drug therapy ; ultrastructure ; Photochemotherapy ; Rats

The purpose of this study was to investigate the effect of bisphosphonate combined with chemotherapy on bone mineral density (BMD) of patients with multiple myeloma (MM) and analyse its value of BMD detection in clinic of these patients. 53 MM cases were enrolled in this study, including 33 newly diagnosed, 10 refractory/relapsed and 10 stable cases. They were divided randomly into two groups, 33 MM cases were treated with bisphosphonates combined with chemotherapy and 20 MM cases were treated with chemotherapy alone. The chemotherapy schedules for all patients were same. BMD was tested using the dual energy X-ray absorptiometry at 2 time points, i.e. pretreatment (basal level) and 12 months after treatment with chemotherapy and bisphosphonates. Comparisons was performed with t tests using SPSS 11.0 software. The results indicated that there was minor difference between 2 groups for BMD scores of whole body and lumbar vertebra (L1-L4), but no difference for scores of the near-end of left femur. After treatment for 12 months, all BMD scores (whole body, lumbar vertebra and the near-end of left femur) increased significantly in the bisphosphonate combined with chemotherapy group (P < 0.05). In contrast, only minor changes were seen in chemotherapy alone group. It is concluded that the bisphosphonate combined with chemotherapy has displayed promotive effect on BMD of MM patients, the detection of BMD is sensitive and special method for monitoring therapeutic effect of bisphosphonate in MM patients, thus has useful value in clinic of these patients.
Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Bone Density ; drug effects ; Diphosphonates ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; metabolism ; pathology

This study was purposed to explore the expression of p57kip2 in the bone marrow of patients with de novo myelodysplastic syndrome (MDS) and its role in MDS pathogenesis, as well as the relationship between the expression of p57kip2 and SDF-1/CXCR4 signal. The expression of p57kip2 and CXCR4 in 67 de novo MDS patients was measured by real-time quantitative PCR. The percentage of CD34(+) cells in the bone marrow from MDS patients was measured by flow cytometry. 18 healthy volunteers were recruited for control. The effect of SDF-1 on p57kip2 expression in bone marrow mononuclear cell (BMMNC) from MDS or normal controls was investigated in vitro, and difference between them was compared. The results showed that low-risk MDS and high-risk MDS displayed a significant reduction of p57kip2 mRNA expression in BMMNC compared with that in control group (P < 0.001) and there was a negative correlation between p57kip2 expression and percentage of CD34(+) (r = -0.458, P < 0.001); the patients with abnormal karyotype showed lower expression of p57kip2 gene, compared to patients with normal karyotype (P = 0.045). Although the expression of CXCR4 had no difference between MDS patients and normal controls, a positive correlation between p57kip2 and CXCR4 in MDS patients was still found (r = 0.609, P < 0.001). Moreover, SDF-1 increased p57kip2 expression in normal BMMNC in dose-dependent manner, but BMMNC from MDS patients showed no response to SDF-1. SDF-1-induced p57 expression was blocked by AMD3100. It is concluded that the low expression of p57 gene in MDS may play a role in the pathogenesis of MDS. Furthermore, SDF-1-induced p57kip2 expression in BMMNC, and the decreasing response of BMMNC to SDF-1 may contribute to the low expression of p57kip2 in MDS patients.
Case-Control Studies ; Chemokine CXCL12 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p57 ; genetics ; metabolism ; Flow Cytometry ; Humans ; Myelodysplastic Syndromes ; genetics ; metabolism ; Receptors, CXCR4 ; metabolism

AMD3100 (Plerixafor) is an antagonist of CXCR4, receptor for stromal cell-derived factor-1 (SDF-1).It disrupts binding of SDF-1 to CXCR4 by competing binding site, thus blocking the physiological function of SDF-1/CXCR4 axis. SDF-1/CXCR4 axis has been shown to play critical roles in stem cell mobilization, migration and homing, and in immunoregulation, inflammatory disease, autoimmune disorder, embryonic development, and tumor cell proliferation, migration and location. AMD3100 has been confined effective for the mobilization of HSC and MSC, inhibition of carcinoma growth and metastasis, suppression of some inflammatory and autoimmune disorder. Therefore, further research on AMD3100 will be helpful to understand the effects of bone marrow microenvironment on the pathogenesis of neoplasm, and to restore the traumatic tissues by mobilizing HSC effectively, that might provide a new idea and measure for the treatment of certain neoplasms. Some research progress of basic research and application on AMD3100 are summarized in this review.
Heterocyclic Compounds ; pharmacology ; Receptors, CXCR4 ; antagonists & inhibitors

The purpose of this study is to evaluate the effect of oligomeric grape seed proanthocyanidins (GSP) on isoproterenol (ISO)-induced cardiac remodeling in rats. ISO was given subcutaneously (5 mg x kg(-1), sc, 7 days) to induce cardiac remodeling in rats. Therapeutic groups were given GSP (50, 100, and 150 mg x kg(-1)) after ISO treatment. After 2 weeks intervention, heart rate (HR), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), rate of rise of left ventricular pressure (+/- dp/dt(max)) were examined. The myocardial hypertrophy index was expressed as heart weight/body weight (HW/BW) and left ventricle weight/body weight (LVW/BW), the histological changes were investigated by HE and Van Gieson stain. SOD activity and MDA content in serum, contents of hydroxyproline (Hyp) in the left ventricular tissue were assayed by xanthinoxidase method, thiobarbituric acid (TBA) method and alkaline hydrolysis method, respectively. After the onset of ISO-treatment, GSP therapy potently improved cardiac function, inhibited myocardial hypertrophy, improved cardiac pathology change, decreased the myocardial cross-section area (CSA), collagen volume fraction (CVF) and perivascular circumferential collagen area (PVCA), reduced the content of Hyp in the left ventricular tissue, inhibited the decrease of SOD activity and increase of MDA content in serum. GSP possess protective effect against ISO induced cardiac remodeling in rats, this may be related to reducing the oxidative stress and improving antioxidant capacity.
Animals ; Antioxidants ; isolation & purification ; pharmacology ; Grape Seed Extract ; isolation & purification ; pharmacology ; Heart Rate ; drug effects ; Hydroxyproline ; metabolism ; Isoproterenol ; Male ; Malondialdehyde ; blood ; Myocardium ; metabolism ; pathology ; Organ Size ; drug effects ; Plants, Medicinal ; chemistry ; Proanthocyanidins ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood ; Ventricular Function, Left ; drug effects ; Ventricular Pressure ; drug effects ; Ventricular Remodeling ; drug effects ; Vitis ; chemistry

OBJECTIVETo investigate whether murine soluble Fas gene transfected marrow graft could block the immune escape of leukemia cells, so as to eliminate the residual leukemia cells and reduce relapse after bone marrow transplantation (BMT).

METHODSThe murine leukemia/lymphoma models were established by inoculating female C57BL/6 mice (H-2b) with 10(5) EL4 cells/mouse through caudal vein. Donors of BM grafts were C57BL/6 male mice. Bone marrow mononuclear cells (BMMCs) were transfected with sFas or EGFP by adenovirus (adsFas or adEGFP) 24 hours before BMT (group D or E). The following three groups were set simultaneously: group A, no BMMCs transplanted; group B, BMMCs transplanted with no adenoviruses transfection; group C, EL4 cells transfusion only. Hematopoietic reconstitution, generation of leukemia/lymphoma and the survival rate were observed in all the groups after BMT.

RESULTSThe spleen indices examined 11 days after BMT were not obviously different among group B, D and E (P > 0.05), but in group A were significantly lower than those in the groups B, D, E (P < 0.01). The leukocyte and platelet counts on day 30 after BMT were recovered in group B and D, but were very low in group C and E. The Y-chromosomes appeared 2 months after BMT. Bone marrow pictures in group B and D were almost normal, but in group C and E had plenty of lymphoblast-like tumor cells. Tumors were obviously revealed in the mice of group C and E by histopathology examination, but did not in group B and D. The survival rate was 0 in group A, 100% in group B and D, 12.5% in group C and 6.25% in group E. Compared with that in group E, the survival was significantly increased in the sFas group (P < 0.01).

CONCLUSIONSGraft transfected with sFas gene prolonged the post-BMT survival of leukemia/lymphoma mice. The transfection of sFas might block the effect of the immune escape of EL4 cells through FasL.

Animals ; Bone Marrow Transplantation ; immunology ; Combined Modality Therapy ; Female ; Genetic Therapy ; methods ; Leukemia, Experimental ; immunology ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Transduction, Genetic ; Transfection ; Transplantation, Homologous ; Tumor Escape ; fas Receptor ; genetics

OBJECTIVETo investigate the expression of heparanase mRNA and its relation with the clinicopathological features and angiogenesis in primary hepatocellular carcinoma (HCC).

METHODSExpression of heparanase mRNA was detected by RT-PCR in 51 HCC lesions, and microvessel density (MVD) was detected by immunohistochemical stain with a factor VIII-related monoclonal antibody.

RESULTSExpression of heparanase mRNA was shown in 49.0% (25/51) HCC lesions. The positive rate of heparanase expression in tumors larger than 3 cm (63.6%, 21/33) was significantly higher than those in smaller tumors (22.2%, 4/18; P < 0.01). Heparanase expression was more frequent in highly invasive tumors (70.0%, 14/20) compared with moderately invasive tumors (46.7%, 7/15) and low invasive ones (25.0%, 4/16; P < 0.05). Moreover, heparanase expression in tumors with high MVD (62.5%, 20/32) was significantly higher than those in tumors with low MVD (26.3%, 5/19; P < 0.05).

CONCLUSIONHeparanase mRNA expression may be important for the growth, invasion and angiogenesis of hepatocellular carcinoma.

Adult ; Carcinoma, Hepatocellular ; blood supply ; enzymology ; pathology ; Female ; Glucuronidase ; biosynthesis ; genetics ; Humans ; Liver ; enzymology ; Liver Neoplasms ; blood supply ; enzymology ; pathology ; Male ; Microcirculation ; pathology ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Burden