Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estrogen Antagonists ; pharmacology ; Glioma ; pathology ; Humans ; Protein Kinase C ; antagonists & inhibitors ; Tamoxifen ; pharmacology

Objective: To observe the tropism of bone marrow stromal cells (BMSCs) to intracranial glioma and their differentiation in the brain of rats bearing glioma, and to investigate the corresponding mechanism. Methods: The in vitro tropism of cultured BMSCs to glioma cells, vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), basic fibroblast growth factor(FGF), platelet derived growth factor (PDGF) were observed under microscope and detected by performing Transwell experiment. The in vivo tropism of BMSCs to intracranial glioma was observed by immunofluorescence method. The differentiation of BMSCs was induced in vitro and observed. After BMSCs were transplanted in the brain of glioma-bearing rats for 15 days, their in vivo differentiation was observed by immunofluorescence staining. Results: BMSCs displayed obvious in vitro tropism to glioma, PDGF, and EGF and in vivo tropism to intracranial glioma. They could migrate to satellite lesions of glioma in vivo. BMSCs were induced to differentiate into neural progenitor cells (8.4% ± 3.5%), neurons (53.7% ± 7.4%), and astrocytes (22.3% ± 5.2%) in vitro. After being transplanted into the brain of glioma-bearing rats, they also differentiated into neural progenitor cells (8.3% ± 3.6%), neurons (15.7% ± 4.3%) and astrocytes (32.5% ± 7.2%). There was significant difference in the differentiation ratio to neurons between in vitro and in vivo experiments (P< 0.05), but the differentiation ratios to neural progenitor cells and astrocytes had no significant difference (P>0.05). Conclusion: BMSCs display extensive tropism to glioma. The direction of the differentiation of BMSCs may be related with local microenvironment.

AIMTo explore the effect of tamoxifen on voltage-dependent sodium channels in SHG-44 glioma cell line.

METHODSWhole-cell patch clamp technique was used to record the Na currents in SHG-44 cell line and to investigate the effect of tamoxifen of different concentration on this channel currents.

RESULTSThis channel activated and inactivated quickly. Tamoxifen could significantly decrease the amplitude of Na currents of SHG-44 cell line. This block effect was dose dependent and voltage dependent. When the holding potential was 0 mV, 8 micromol/L tamoxifen could block this currents 69%. The half inhibition concentration (IC50) was 5.54 micromol/L.

CONCLUSIONTamoxifen could significantly block the voltage dependent sodium channel in malignant glioma cell line SHG-44. It might be one of the mechanisms that tamoxifen inhibit glioma proliferation. clamp technique was used to record the Na currents in SHG-44 cell line and to investigate the effect of tamoxifen of different concentration on this channel currents.

RESULTSThis channel activated and inactivated quickly. Tamoxifen could significantly decrease the amplitude of Na currents of SHG-44 cell line. This block effect was dose dependent and voltage dependent. When the holding potential was 0 mV, 8 micromol/L tamoxifen could block this currents 69%. The half inhibition concentration (IC50) was 5.54 micromol/L.

CONCLUSIONTamoxifen could signifi-cantly block the voltage dependent sodium channel in malignant glioma cell line SHG-44. It might be one of the mechanisms that tamoxifen inhibit glioma proliferation.

Antineoplastic Agents, Hormonal ; pharmacology ; Brain Neoplasms ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Glioma ; pathology ; Humans ; Patch-Clamp Techniques ; Sodium Channel Blockers ; pharmacology ; Tamoxifen ; pharmacology

Three osmotolerant yeasts were isolated from three batches of "swollen can" soy sauce produced by a Guangdong condiment plant. These strains grew faster in the media containing 50%～60% glucose or 15% NaCl than in common yeast media. The three yeasts were identified as Pichia etchellsii by using morphological characteristics, physiological and biochemical tests.

Objective To develop a candidate reference method for the measurement of progesterone in human serum.Methods The serum sample is mixed with the internal standard [3,4-~(13)C_2] progesterone.After extraction with n-hexane and purified by a aqueous solution of 2-Hydroxypropyl-?- cyclodextrin (HP-?-CD),the serum progesterone and labeled progesterone are converted to the 3-enol heptafluorobutyrate and analyzed by gas chromatography mass spectrometry (GC/MS) with selected ion monitoring.The concentration of serum progesterone is calculated by bracketing method.Results The results gave coefficients of variation (CVs) of 0.69% to 2.12%.The analytical recoveries ranged from 98.3% to 100.1%.The results of measuring certified reference materials of serum progesterone are agree with the target value.Conclusion The procedure for measuring progesterone in serum is a highly accurate and precise method and may be used as a candidate reference method for serum progesterone assays.

OBJECTIVETo study the anti-glioma activity of treatment by bone marrow stromal cells (BMSCs) transfected with AdCMV-tk containing HSV-tk gene in rats.

METHODSPrimary cultured BMSCs were obtained and transfected with HSV-tk (BMSCs/tk) and were injected into contralateral brain of glioma-bearing rats to observe their tropism for glioma cells. RT-PCR was performed to examine the transduct of tk gene after it was transduced into BMSCs. C6 glioma cells were co-cultured with BMSCs transfected with HSV-tk. MTT test was performed to examine its antitumor activity. BMSCs, after being transfected with HSV-tk, were injected into contralateral brain tissue of glioma-bearing rats to show their in vivo antitumor activity. Dynamic MRI was performed to monitor the development of intracranial glioma.

RESULTSPurified BMSCs were obtained by primary cultured bone marrow cells. After being transfected with HSV-TK, the cells still stably displayed extensive tropism for intracranial glioma and transcripted tk gene. RT-PCR showed that BMSCs/tk were transduced tk gene obviously at 21 days after AdCMV-tk transfection. BMSCs/tk showed a clear bystander effect after being co-cultured with C6 glioma cells in vitro. TUNEL assay showed that BMSCs/tk could obviously show bystander effect and induce apoptosis of glioma cells in vivo with an apoptosis positive ratio of 20.38% +/- 2.57%, showing a statistically significant difference in comparison with BMSCs group (2.56% +/- 0.52%, P = 0.023) and control group (2.74% +/- 0.38%, P = 0.025). Compared with the control group (21.40 +/- 1.63 days), BMSCs/tk transplantation significantly prolonged the survival time of glioma-bearing rats (52.60 +/- 13.11 days, P = 0.000). MRI detection showed that the least volume of intracranial glioma in BMSCs/tk group (8.28 +/- 2.64 mm3), significantly smaller than that in BMSCs group (134.51 +/- 16.37 mm3, P = 0.001) and control group (147.22 +/- 31.05 mm3, P = 0.001). Some of the intracranial gliomaa disappeared after transplantation of BMSCs/tk.

CONCLUSIONBMSCs transfected with AdCMV-tk may become an effective therapy method in the treatment for glioma.

Animals ; Apoptosis ; Bone Marrow Cells ; cytology ; Brain ; pathology ; Bystander Effect ; Cell Line, Tumor ; Coculture Techniques ; Genetic Therapy ; methods ; Glioma ; pathology ; therapy ; Magnetic Resonance Imaging ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Simplexvirus ; enzymology ; Stromal Cells ; cytology ; enzymology ; transplantation ; Thymidine Kinase ; genetics ; metabolism ; Transfection

OBJECTIVETo observe the effect of long-term application of Shengmai Capsule (SMC) on recovery of patients after myocardial infarction.

METHODSA total of 120 myocardial infarction patients were: assigned into two groups. Changes of angina pectoris, electrocardiogram (ECG), living capacity and heart function in patients were observed after 6-month treatment.

RESULTSThe total effective rate in alleviating angina: pectoris was 90.0% and that in improving ECG figure was 93.3% in the treatment group, both were significantly higher than those in the control group, 73.4% and 70.0% respectively (P<0.05). The Karnofsky Performance Status scores of heart function were increased and the Activity of Daily Living scores in living capacity decreased in both groups, but the improvements were better in the treatment group (P<0.01 and P<0.05). The parameters of cardiac function, including cardiac output, stroke volume, cardiac index and ejection fraction, were increased in both groups, but the increments in the treatment group were more significant (P<0.01 or P<0.05).

CONCLUSIONLong-term application of SMC could effectively prevent and treat angina pectoris, improve the living capacity and accelerate the recovery of heart function in patients after myocardial infarction.

Electrocardiography ; Humans ; Karnofsky Performance Status ; Medicine, Chinese Traditional ; Myocardial Infarction ; physiopathology ; therapy ; Quality of Life

Adolescent ; Adult ; Aged ; Apoptosis ; Cell Cycle ; Cell Division ; Child ; Female ; Glioma ; chemistry ; pathology ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; analysis ; Middle Aged ; Neoplasm Proteins ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; analysis

Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.

Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.