With the introduction and development of evidence-based medicine in China, it has been spread rapidly in the area of integrative medicine (IM) and has become a new unique discipline. During almost 20 years, as one of the most important parts of evidence-based IM, systematic review (SR)/meta-analysis (MA) of IM have shown a good development momentum in the aspects of quantity, depth, breadth and influence, but also face the harsh situation of the uncontrolled quantity and quality, especially for SRs in Chinese. Therefore, how to supervise and standardize this area effectively becomes a problem to be solved. Based on the experience both at home and abroad, the authors put forward several kinds of solutions for laying the foundation for further development such as promoting the registration system of SR/MA of IM, effectively setting up the regulatory platform of quality and quantity, launching professional training for SR/MA reviewers, forming qualification registration, developing the data transfer and sharing platform to realize the transparency of evidence process.

Objective To compare the pharmacokinetic profiles between a new generic and a branded reference formulation of isosorbide -5 -mononitrate sustained release capsules , and to assess the bioequivalence of the two products in healthy Chinese male subjects.Methods Fifty subjects participated in the open -label, randomized -sequence, 2-way crossover study.Twenty-four subjects and 26 subjects were ran-domly assigned in a 1:1 ratio to receive single dose (50 mg) or multiple dose (50 mg, qd, 6 days) of the test or reference formulation , followed by a one -week washout period and administration of the alternate formulation , respectively.Serial blood samples were collected , and isosorbide -5 -mononitrate concentration in plasma was determined by LC-MS/MS.The relative bioavailability and related parameters of pharmacokinetics were calculated. Results The pharmacokinetic parameters of test formulation and the reference formulation after a single dose were as follows: Cmax were ( 554.18 ± 117.84 ) and (526.29 ±91.58 )μg· L-1; AUC0-t were ( 7834.21 ±1227.70 ) and (7658.86 ±927.74) h· μg· L-1 , respectively.The 90% confidential interval of Cmax and AUC0-t of test formulation were 99.82%-113.03%and 99.13%-106.43%of reference formulation , respectively.The pharmacokinetic parame-ters of test formulation and the reference formulation after multiple doses were as follows :Cmax were (612.96 ±171.32) and (527.12 ±114.36 ) μg · L-1; AUC0-t were ( 8408.71 ±1321.91 ) and ( 7781.88 ±1325.12 ) h · μg · L-1 , respectively.The 90% confidential interval of Cmax and AUC0-t of test formulation were 108.44% -122.17% and 105.35%-111.57% of reference formulation , respectively.The 90% confidence interval of Cmax and AUC0-t of isosorbide-5-mononitrate for the test formulation after single and multiple oral doses were fall within 70%-143%and 80%-125%of reference formulation.Conclusion The test formulation was considered bioequivalent to the reference formulation.

OBJECTIVETo study the phenotype and tumorigenicity of SHG-44 glioma stem cell spheres and the pathological characteristics of their xenograft tumors.

METHODSSHG-44 glioma cells were cultured under neural stem cell medium and glioma stem cell spheres were collected. Immunocytochemistry was used to dectet the expression of CD133, nestin, A2B5, vimentin, VEGFR-2 and IDH R132H. Cell spheres were induced using serum-containing medium, and the expression of CD133, nestin, vimentin, GFAP, β-III tubulin and GalC in the cell spheres were detected. The expression of CD133, nestin, VEGFR-2, GFAP, S-100 and CD34 in the intracranial xenograft tumor tissues was detected using immunohistochemistry. The pathological characteristics of orthotopic xenograft tumors generated from the SHG-44 glioma cells and SHG-44 glioma stem cell spheres were compared.

RESULTSSHG-44 glioma stem cell spheres were collected successfully after cultured under neural stem cell medium. The ratio of CD133(+) cells in the passage 10 SHG-44 glioma stem cell spheres was (71.63 ± 5.92)%, significantly higher than that in the SHG-44 glioma cells [(1.95 ± 1.45)%]. Immunocytochemistry showed that in the SHG-44 glioma cell spheres, the ratio of nestin(+) cells was (84.06 ± 7.58)%, vimentin(+) cells (29.11 ± 3.44)%, VEGFR 2(+) cells (64.44 ± 3.69)%, and A2B5(+) cells (14.08 ± 2.19)%. A subpopulation of cells with mutation of IDH R132H was detected harboring in the SHG-44 glioma cell spheres. After induction of differentiation with serum-containing medium, the ratio of CD133(+) cells was (1.89 ± 1.27)%, nestin(+) cells (6.67 ± 2.75)%, vimentin(+) cells (93.75 ± 2.95)%, GFAP (+) cells (91.33 ± 4.75)%, β-III tubulin(+) cells (82.36 ± 4.02)%, and GalC(+) cells (8.92 ± 3.19)%. Immunohistochemistry showed positive expression of GFAP, S-100, VEGFR-2, and negative of CD133 and nestin in the orthotopic xenograft tumors. A very small amount of human-specific CD34 cells formed a tubular structure. Compared with the SHG-44 glioma cell-formed xenograft tumor, the SHG-44 glioma stem cell-formed xenograft tumor exhibited a higher local invasiveness.

CONCLUSIONSSHG-44 glioma cell spheres are successfully collected after cultured under neural stem cell medium. They belong to the CD133(+)A2B5(-) GSC subpopulation, highly expressing VEGFR-2, possess the ability of both self-renewal and multi-directional differentiation, and may participate in the formation of vasculogenic mimicry.

Animals ; Brain Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cells, Cultured ; Glial Fibrillary Acidic Protein ; metabolism ; Glioma ; metabolism ; pathology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; pathology ; S100 Proteins ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

[Objective]To establish stable glioma stem cells with a high expression level ofred fluorescent protein (RFP) in vitro.[Methods]The glioma stem cells SU2 were transfected with lentivirus vector containing RFP gene;cell expression of RFP was observed by fluorescent microscopy;RFP-positive glioma stem cells were sorted out by fluorescence-activated cell sort.The transfection efficiency of SU2 cell before transfection and 20-passaged RFP-SU2 cells after transfection were assayed by flow cytometry.Dynamic viewing was performed to observe the cell division and cloning of RFP-SU2 single cell;RFP -positive cells were collected and immunostained with antibodies against CD133 and nestin.[Results] PFP-SU2 cells grew as levitated sphere with high RFP expression;the transfection efficiency of SU2 cell before transfection was only 1.5％,while that of20-passaged RFP-SU2 cells after transfection reached to 75％.RFP-SU2 single cell could proliferate into brain tumor stem cell spheres,having the abilities of self-renewing and clonal proliferation.Immunofluorescence showed positiveCDt 33 and nestin expressions in RFP-SU2 cells.[Conclusion] A SU2/RFP cell line marked by RFP is established;and it can serve as a promising tool for further basic research.

BACKGROUNDIt is generally accepted that gliomas are the most common primary brain tumors with poor prognosis. We aimed to explore the relationship of the immunity of the central nervous system and the genesis and development of glioma.

METHODSG422 glioma was implanted in the brain of BALB/c mice (immuno-competent mice), nude mice (T cell related immuno-deficient) and complement C3 knock-out mice (complement C3 related immunodeficient). The survival time of the host, growth and histopathology of the tumor, and concentrations of tumor necrosis factor-α (TNF-α) and interferon-γ (INF-γ) in tumor tissues were assessed.

RESULTSTumor spheres were formed in all mice after injection, and glial fibrillary acidic protein (GFAP) positive staining of the cells declared their glioma origin. The longest median survival time of (44.3 ± 6.0) days was found in BALB/c mice, followed by (24.8 ± 5.2) days in nude mice and the shortest (18.6 ± 5.8) days in complement C3 knock-out mice. Accordingly, the growth of the tumor was fastest in complement C3 knock-out mice, followed by the nude mice and slowest in the BALB/c mice. Although the proportions of infiltrating CD68(+) lymphocytes in tumor tissues showed no significant difference (P > 0.05), TNF-α level in the nude and C3 knock-out mice, (28.11 ± 4.86) µmol/L and (22.87 ± 6.36) µmol/L respectively, were significantly lower (P < 0.01) than that in the BALB/c mice, which was (230.21 ± 39.17) µmol/L. The INF-γ level was highest in the BALB/c mice ((180.76 ± 29.19) µmol/L), followed by the nude mice ((113.46 ± 23.76) µmol/L) and then the C3 knock-out mice ((16.84 ± 4.45) µmol/L).

CONCLUSIONSThe G422 glioma implanted in the brains of mice with different immune ability would be a useful model for studying the relationship of the immune system and tumor in the central nervous system. Furthermore, the T cells and complement C3 compartments of the immune response may affect the growth of implanted tumors and inflammatory factors such as TNF-α and INF-γ.

Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; Complement C3 ; genetics ; metabolism ; Glioma ; metabolism ; pathology ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Knockout ; Mice, Nude ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the effect of cyclooxygenase-2 (COX-2) on lymphangiogenesis in breast cancer.

METHODSBy the means of immunohistochemistry, COX-2, vascular endothelial growth factor-C (VEGF-C) and D2-40 were examined in the tissue samples of primary tumors from 94 patients underwent surgical resections for breast cancer from November 1998 to March 2002. Eighty-three patients were followed-up. The expressions of VEGF-C mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot in MDA-MB-231 cell lines by the treatment of selective COX-2 inhibitor Nimesulide at different doses. The expressions of VEGF-C protein were evaluated in MDA-MB-231 cells treated by PGE2 (1 microg/ml) and Trastuzumab (1 microg/ml), respectively.

RESULTSCOX-2 over-expression was observed in 46.8% of surgical specimens (44/94), while VEGF-C overexpression occurred in 51.1% of tumor samples (48/94). COX-2 was strongly correlated with VEGF-C expression (P < 0.01), micro-lymphatic vessels (P = 0.032) and metastatic lymph nodes (P = 0. 035). Patients with COX-2 positive tumors had a significant shorter survival time than those with negative tumors did, including disease-free survival (P = 0.010) and overall survival (P = 0.040). Nimesulide could down-regulate the expressions of VEGF-C mRNA and protein in a does-dependent manner, while PGE2 could up-regulate the expressions. The expression of VEGF-C protein up-regulated by PGE2 treatment was decreased by Trastuzumab.

CONCLUSIONSCOX-2 over-expression can up-regulate the expression of VEGF-C. VEGF-C might promote lymph node metastasis by a lymph-angiogenic pathway, then affect the prognosis of the patients with breast cancer.

Adolescent ; Adult ; Aged ; Breast Neoplasms ; metabolism ; pathology ; Cyclooxygenase 2 ; metabolism ; physiology ; Female ; Follow-Up Studies ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Middle Aged ; Prognosis ; Vascular Endothelial Growth Factor C ; genetics ; metabolism

OBJECTIVETo research on the protective effect of 3'-methoxy-puerarin (3'-mo-pue) on rat brain suffering from ischemia and to offer the experimental basis for widening the applied areas of Radix Puerariae.

METHODAll rats were randomly divided into sham operation group, control group, Pue group and 3'-mo-pue group. The influence of different dosage on relevant indexes were measured through cerebral artery occlusion (MCAO) model rat covered by Fecl3, and cerebral ischemia-reperfusion model rat after 2VO.

RESULTThe result showed that 3'-mo-Pue could reduce the scores of neurological deficit, cerebral infarcted zone to varying degrees and the water content of brain tissue in MCAO model. It could obviously increase the activity of CAT and GPx in the hippocampus and also increase the activity of CAT and GPx in the cortex in the ischemia field. Moreover, 3'-mo-Pue could decrease the content of LPO, LD in the brain tissue of cerebral ischemia-reperfusion model rat.

CONCLUSION3'-mo-Pue has favorable protective effect on brain suffering from ischemia. The mechanism is possibly related to its effect of anti-oxidation.

Animals ; Brain ; drug effects ; enzymology ; metabolism ; Brain Ischemia ; drug therapy ; metabolism ; prevention & control ; Catalase ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Enzyme Activation ; drug effects ; Glutathione Peroxidase ; metabolism ; Isoflavones ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar

OBJECTIVETo discuss the treatment and mechanisim of acetabulum fracture combined with ipsilateral lower extremity fracture.

METHODSFourteen cases(9 males and 5 females) of acetabulum fracture were involved in the study with the mean age of 35 years (range, from 18 to 65 years). According to Letournel classification, 11 cases were the fracture of posterior wall and column, 3 cases were only the fracture of posterior column. All of them were treated with titanium plate internal fixation. Of three cases with ipsilateral femoral intertrochanteric fracture, one was treated with external fixiation device, other two were treated with DHS fixation; Three cases with ipsilateral femoral neck fracture were treated with compressive cannulated screw fixation; Among six cases with ipsilateral femoral shaft fracture,one was fixed with steel plate,the rest underwent intramedullary nailing fixation; Two cases with ipsilateral tibial plateau fracture were treated with anatomic plate internal fixation.

RESULTSAll patients except the died old patient were followed up from 18 months to 5 years, with mean of 30 months. According to standard of American institute of orthopaedics, 9 cases obtained excellent results, 3 good and 2 poor and the excellent and good rate was 92.3%.

CONCLUSIONMost cases of acetabulum fracture combined with ipsilateral lower extremity fracture are due to great violence, their mechanisms are complex, and easily lead to missed diagnosis, therefore the early correct diagnosis, and reasonable internal fixation should be considered.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Aged ; External Fixators ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged

OBJECTIVETo compare the long-term effect and safety of tacrolimus (FK506) and cyclosporine (CsA) in kidney transplant (KT) recipients carrying hepatitis B Virus(HBV).

METHODSA total of 109 patients with HBV were randomized into FK506 group (52 cases) and CsA group (57 cases) after KT, and a 2-year-long follow-up of the patients was conducted to record the patient and graft survival, incidence of acute graft rejection and postoperative liver function.

RESULTSThe 2-year patient/graft survival was 86.0%/73.7% and 94.2%/90.3% in CsA and FK506 groups, respectively (P<0.05), with incidence of acute rejection of 10.5% and 9.6% (P>0.05), and rate of abnormal liver function of 26.3% and 15.4% (P<0.05), respectively. Eight patients (14.4%) in CsA group required a drug conversion but none in FK506 group. The drug conversion resulted in significant reduction of ALT/AST level from 255.13+/-31.38/201.88+/-21.25 U/L to 31.25+/-11.50/25.13+/-9.68 U/L (P<0.01).

CONCLUSIONFor HBV-carrying renal transplant recipients, FK506 as the primary choice of immunosuppressant can be more effective and safer than CsA.

Adolescent ; Adult ; Carrier State ; physiopathology ; Cyclosporine ; administration & dosage ; adverse effects ; pharmacology ; Drug-Related Side Effects and Adverse Reactions ; Female ; Graft Rejection ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B virus ; Humans ; Kidney Transplantation ; adverse effects ; Liver ; drug effects ; physiology ; Male ; Middle Aged ; Tacrolimus ; administration & dosage ; adverse effects ; pharmacology ; Young Adult

OBJECTIVETo investigate the intracellular localization of S100A4 in gastric carcinoma cells and the relationship between S100A4 expression status and lymph node metastasis of gastric carcinoma.

METHODSWestern blotting analysis was performed to locate the expression of S100A4 protein in sub-fraction components of frozen tissues. S100A4 protein expression was also determined by immunohistochemical method in 131 samples of gastric cancer and 20 samples of matched metastatic lymph nodes.

RESULTSThirty-two of 131 (24.4%) gastric carcinoma showed positive S100A4 nuclear expression and 50/131 (38.2%) carcinoma showed positive cytoplasmic expression. In 32 samples with positive S100A4 nuclear expression, 30 (93.8%) carcinomas had positive lymph node metastases. S100A4 nuclear expression level was higher in gastric carcinoma with lymph node metastasis (29.1%) than that without lymph node metastasis (7.1%) (P=0.016).

CONCLUSIONNuclear expression of S100A4 is associated with lymph node metastasis of gastric carcinoma.

Cell Nucleus ; metabolism ; Humans ; Lymphatic Metastasis ; Neoplasm Staging ; S100 Calcium-Binding Protein A4 ; S100 Proteins ; metabolism ; Stomach Neoplasms ; metabolism ; pathology