Objective To demonstrate the superiority and feasibility of using bioluminescent image to monitor the islet graft after islet transplantation.Methods Diabetic models were established by intraperitoneal injection of streptomycin into mature male C57BL/6 mice.Islets were harvested from the pancreas of C57BL/6 and Bclb/c mice by digestion and purification,and transfected with Lueiferase gene.The mouse diabetic models were divided into iso-transplantaion group (n=20) and allo-transplantation group (n=7).The islets of C57BL/6 were transplanted into iso-transplantaion mice with different doses (10,50,110 and 200,n=5 in every dose),and Bclb/c mouse islets were transplanted into allo-transplantation group.The islets were transplanted into the subcutaneous fat tis- sue near left scapula.The receptor mice were scanned with CCD camera to get bioluminescent images at different scheduled time points,and the changes in random blood glucose of allo-transplantation group were observed.Results On day 6 after transplantation,the scanning image showed that the pi- xel intensity from the region of interest (ROI) was increased with the increased number of islet grafts and they had a positive correlation.The random blood glucose was reduced to the normal level in the first 2 days,and then increased again to the diabetic level on 11 days averagely,while pixel intensity from the ROI reached the peak on day 6-7,and then reduced rapidly after islet transplantation in allo- transplantation group.The beginning of pixel intensity reduction occurred on day (6.14?0.90), while that of the random blood glucose raise occurred on day (10.00?0.82) after transplantation,and the former alteration occurred obviously earlier than the latter (P

Objective To estimate the exposure level of PM2.5,CO and O_3 in the elders in a community in Beijing.Methods The concentrations of PM2.5,CO and O_3 in 10 main activity sites of the elders were measured and 24 h time-activity data of 30 elders was collected by recording of activity log paper,Nov.28,2007—Jan.17,2008.Results The 24 h average exposure concentrations of PM2.5,CO and O_3 for the elders were(146.54?6.60)?g/m~3,(2.67?0.18)mg/m~3 and(32.30?2.79)?g/m~3, respectively,there was no significant difference in the exposure level of PM2.5 and O_3 between male and female elders except CO (P

OBJECTIVETo assess the ability of tetrandrine (Tet) to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcinoma p53-mutant HT-29 as well as in C26 colorectal carcinoma-bearing BALB/c mice.

METHODSMCF-7/ADR, HT-29 and MCF-7 cells were exposed to irradiation in the absence or presence of tetrandrine. The effect of Tet on the cytotoxicity of X-irradiation in these three cells was determined and the effect of tetrandrine on cell cycle arrest induced by irradiation in its absence or presence was studied by flow cytometry. Moreover, mitotic index measurement determined mitosis of cells to enter mitosis. Western blotting was employed to detect cyclin B 1 and Cdc2 proteins in extracts from irradiated or non-irradiated cells of MCF-7/ADR, HT-29 and MCF-7 treated with tetrandrine at various concentrations. Tumor growth delay assay was conducted to determine the radio-sensitization of tetrandrine in vivo.

RESULTSClonogenic assay showed that tetrandrine markedly enhanced the lethal effect of X-rays on p53-mutant MCF-7/ADR and HT-29 cells and the sensitization enhancement ratio (SER) of tetrandrine was 1.51 and 1.63, but its SER was only 1.1 in p53-wt MCF-7 cells. Irradiated p53-mutant MCF-7/ADR and HT-29 cells were only arrested in G2/M phase while MCF-7 cells were arrested in G1 and G2/M phases. Radiation-induced G2 phase arrests were abrogated by tetrandrine in a concentration-dependent manner in MCF-7/ADR and HT-29 cells, whereas redistribution within MCF-7 cell cycle changed slightly. The proportion of cells in M phase increased from 1.3% to 14.7% in MCF-7/ADR cells, and from 1.5% to 13.2% in HT-29 cells, but 2.4% to 7.1% in MCF-7 cells. Furthermore, the levels of cyclin B 1 and Cdc2 expression decreased after X-irradiation in MCF-7/ADR and HT-29 cells, and the mitotic index was also lower. Tet could reverse the decrease and induce the irradiated cells to enter mitosis (M phase). Endosomatic experiment showed that tetrandrine caused tumor growth delay in irradiated mice.

CONCLUSIONTetrandrine boosts the cell killing activity of irradiation both in vitro and in vivo. Tetrandrine is a potent abrogator for G2 checkpoint control and can sensitize the cells to radiation.

Animals ; Benzylisoquinolines ; pharmacology ; CDC2-CDC28 Kinases ; metabolism ; Cell Line, Tumor ; Cyclin B ; metabolism ; Cyclin B1 ; Drug Screening Assays, Antitumor ; G2 Phase ; drug effects ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Radiation Tolerance

OBJECTIVETo investigate the effect of transurethral resection of ejaculatory ducts (TURED) for azoospermia with ejaculatory duct obstruction (EDO).

METHODSFrom June 2003 to December 2004, 20 azoospermia with EDO were diagnosed, diagnostic criteria included a history, physical examination, semen analyses, semen fructose measurement, endocrine assessment, testicular biopsy and transrectal ultrasonography (TRUS); All 20 cases were treated by TURED. Fifteen of them were followed up more than 3 months after the treatment. The semen samples of them were analysed at 3-month intervals in post-therapy.

RESULTSSemen analyses in all 20 cases showed the typical characteristics of EDO, low semen volume (0.4-1.6 ml), azoospermia, low pH, absent or low semen fructose. TRUS showed the main etiology factor of EDO was a midline cyst in 11, lateral cystic lesions in 2, the remaining 7 cases had dilated ejaculatory duct with or without dilated seminal vesicles. Among 15 cases followed up more than 3 months after TURED, 10/15 (67%) had an improvement in semen parameters and 3/15 (20%) had pregnancies. Semen analyses had not been done in anther 5 cases.

CONCLUSIONTransurethral resection of ejaculatory ducts may be a safe and effective method for the treatment of azoospermia with EDO.

Adult ; Azoospermia ; diagnosis ; surgery ; Ejaculatory Ducts ; diagnostic imaging ; pathology ; surgery ; Electrosurgery ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Oligospermia ; diagnosis ; Ultrasonography

Homo sapiens longevity assurance homologue 2 (LASS2) is a novel gene isolated from a human liver cDNA library by our laboratory, and it is a human homologue of the yeast longevity assurance gene LAG1 (Saccharomyces cerevisiae longevity assurance gene). According to our previous results, LASS2 could interact with subunit c of vacuolar type H(+)-ATPase (V-ATPase), and the overexpression of LASS2 could inhibit the cell growth of a human hepatocellular carcinoma (HCC) cell line, SMMC-7721. In order to understand the role of the interaction between LASS2 and V-ATPase in HCC cell growth, we transiently transfected plasmid pCMV-HA2-LASS2 into HCCLM3, a HCC cell line without the significant expression of endogenous LASS2. The pH-sensitive fluorescence probes, BCECF and BCECF-AM, were used to measure the intracellular and extracellular H(+) concentrations of HCCLM3 cells respectively. The effect of LASS2 gene on apoptosis was evaluated with Annexin-V/FITC and propidium iodide (PI) by flow cytometry. Western blot was used to detect cytochrome c (Cyt c) in the cytosol and mitochondria, as well as pro-caspase-3 in cytosol. The results showed that the cell growth of LASS2-transfected HCCLM3 cells was significantly inhibited compared with that of the mock control. LASS2 transfection increased intracellular H(+) concentration of HCCLM3 cells, while decreased extracellular H(+) concentration. Moreover, LASS2 transfection significantly enhanced the apoptosis of HCCLM3 cells. In LASS2-transfected cells, the amounts of Cyt c increased in the cytosol, while decreased in the mitochondria. Meanwhile, the expression of pro-caspase-3 in the cytosolic extracts was decreased. These results implicate that LASS2 gene might increase intracellular H(+) of HCC cells via the interaction with V-ATPase, thereby inducing cell apoptosis through mitochondrial pathway.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Membrane Proteins ; metabolism ; RNA, Small Interfering ; Sphingosine N-Acyltransferase ; metabolism ; Transfection ; Tumor Suppressor Proteins ; metabolism ; Vacuolar Proton-Translocating ATPases ; metabolism

OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

BACKGROUND: The immediate implantation in the anterior maxillary region is in a high risk of aesthetic complications. OBJECTIVE: To assess the prognosis of the labial bone plate after anterior maxillary repair with immediate implant combined with immediate restoration. METHODS: Thirty-two patients with single failed tooth in the anterior maxillary region were subjected to implantation of ZIMMER implants immediately after minimally invasive extraction. Good primary stability was achieved and immediate restoration was carried out. Final restoration was finished after 6-12 months of osteosynthesis and gingival shaping. The loading situation of the labial bone plate was recorded at 6 months post operation. RESULTS AND CONCLUSION: Final restoration was finished with normal loading in all the patients. No bleeding and swelling of the gingiva was recorded. The horizontal absorption of the labial bone plate at the upper margin, 5 mm and 10 mm below the upper margin was (-2.12±0.05), (-1.54±0.04), and (-1.01±0.06) mm, respectively. Therefore, absorption of the labial bone plate with varying degrees exists after anterior maxillary repair with immediate implant combined with immediate restoration.

Matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF-IMS) has been a classical technique for studying proteomics in present and a tool for analyzing the distribution of proteins and small molecules within biological tissue sections. MALDI-TOF-IMS can analyze multiple unknown compounds in biological tissue sections simultaneously through a single measurement which can obtain molecule imaging of the tissue while maintaining the integrity of cellular and molecules in tissue. In recent years, imaging mass spectrometry technique develops relatively quickly in all biomedical domain. This paper based on the relevant data and reviews the present developing level of MALDI-TOF-IMS, the principle of imaging mass spectrometry, methology and the prospect in forensic pathology.
Diagnostic Imaging ; Forensic Sciences/methods* ; Humans ; Male ; Proteins ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

AIM To investigate the effects of drying temperature,growing area and plucking time on hederacoside C,α-hederin from leaves of Hedera helix L..METHODS Three batches of H.helix leaves plucked in different time from two growing areas were dried in a vacuum oven to the constant weight at 60 ℃,70 ℃,80 ℃,90 ℃ and 105 ℃,respectively.Two saponins in the processed leaves were determined by HPLC.The powders of the processed H.helix leaves of different batches were mixed with proper ratios,which were determined by least squares optimization method with constraints.RESULTS The content of hederacoside C in the processed H.helix leaves of the three batches increased while that of α-hederin decreased with increasing temperature.The relative error between measured value and desired contents of hederacoside C and α-hederin in the mixed H.helix leaves was less than 5.5％.CONCLUSION The effects of three factors on the content of two saponins in the H.helix leaves are in the order of drying temperature,growing area and plucking time.Mixing processed H.helix leaves of different quality statues reasonably can control the contents of two saponins in a certain range.

OBJECTIVECastrated rats exhibit significant shrinkage of the ventral prostate and apoptosis of prostatic cells, which can be attributed to the reduced blood supply to the prostate. But what causes the blood decrease in the prostate remains unknown. This study aims to explore the molecular mechanism of the changes in the microcirculation of the ventral prostate of rats following castration.

METHODSWe randomized 24 male adult rats into 6 groups of equal number, and collected their ventral prostates at 0, 1/2, 1, 2, 3 and 7 d, respectively, after castration. Then we observed the changes of the microvessels under the transmission electron microscope, detected the apoptosis of endothelial cells by TUNEL, and determined the expressions of VEGF, endostatin, angiostatin and angiopoietin-2 by Western blot.

RESULTSThe castrated rats showed dramatic changes in the microvessels of the ventral prostate, obvious apoptosis of the endothelial cells, down-regulated expression of VEGF, and up-regulated expressions of endostatin and angiostatin, while angiopoietin-2 remained unchanged.

CONCLUSIONThe decreased level of VEGF and increased levels of endostatin and angiostatin might underlie the mechanism of the changes in the microcirculation of the ventral prostate of rats following castration.

Angiopoietin-2 ; metabolism ; Angiostatins ; metabolism ; Animals ; Endostatins ; metabolism ; In Situ Nick-End Labeling ; Male ; Microcirculation ; Orchiectomy ; Prostate ; blood supply ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism