Objective To analyze the effect of combined detection of serum tumor markers, Ki-67 and P-glycoprotein (PGP) in lymph node metastasis and postoperative recurrence of colorectal cancerpatients. Methods The clinical data of 517 colorectal cancer patients who had underwent surgicaltreatment were retrospectively analyzed. Among them, lymph node metastasis was found in 165 cases(lymph metastasis group), and 352 cases did not have lymph node metastasis (non lymphoid metastasis);postoperative recurrence was found in 224 cases (postoperative recurrence group), and 293 cases did havepostoperative recurrence (non postoperative recurrence group). The serum tumor markers levels of cancerantigen 19- 9 (CA19- 9), carcinoembryonic antigen (CEA), cancer antigen 125 (CA125) and theexpressions of Ki-67, PGP were compared. The correlation between the serum tumor markers levels andthe expressions of Ki-67, PGP was analyzed. Results The serum levels of CEA, CA19-9 and CA125 and the positive expression rates of Ki-67 and PGP in lymph metastasis group were significantly higher than those in non lymph metastasis group:(21.39 ± 3.15)μg/L vs. (10.12 ± 2.48)μg/L, (68.48 ± 5.82) U/L vs. (35.26 ± 3.51) U/L, (82.16 ± 7.53) U/L vs. (32.46 ± 6.24) U/L, 69.70% (115/165) vs. 13.64% (48/352) and 72.73%(120/165) vs. 14.77%(52/352), and there were statistical differences (P<0.01). The serum levels of CEA, CA19-9 and CA125 and the positive expression rates of Ki-67 and PGP in postoperative recurrence group were significantly higher than those in non postoperative recurrence group:(18.26 ± 2.34)μg/L vs. (9.18 ± 1.26)μg/L, (47.52 ± 4.85) U/L vs. (21.43 ± 2.18) U/L, (59.16 ± 4.25) U/L vs. (33.17 ± 3.46) U/L, 60.27% (135/224) vs. 9.56% (28/293) and 6.70% (15/224) vs. 7.51% (22/293), and there were statistical differences (P<0.01). The correlation analysis results showed that the positive expression rates of Ki-67 and PGP were positive correlated with the serum levels of CEA, CA19-9 and CA125 (P<0.05). Conclusions The levels of serum tumor markers and the positive expression rates of PGP and Ki-67 in colorectal cancer patients with lymph metastasis and postoperative recurrence are high. The levels of tumor markers are closely related to the positive expression rates of Ki-67 and PGP, which indicates that the combined detection of these indexes has a good effect on evaluating lymph metastasis and postoperative recurrence of colorectal cancer patients.

This study is to develop a sensitive method by using reversed-phase high performance liquid chromatography coupled with UV detector (HPLC-UV) to simultaneously determine four bioactive compounds, iriflophenone 3-C-beta-D-glucoside, iriflophenone 3,5-C-beta-D-diglucoside, mangiferin, and iriflophenone 2-O-alpha-L-rhamnoside in the leaves of Aquilaria sinensis. An Agilent Zorbax SB-C, column (4, 6 mm x 250 mm, 5 microm) was used, and the gradient elution was performed with mobile phase of 0.1% aqueous phosphoric acid and acetonitrile at a flow rate of 1 mL x min(-1). The detection wavelength was 280 nm, and the column temperature was 25 degrees C. The four marker compounds were well separated with good linearity (R2 > 0.9990), precision, stability and repeatabili y. The-recovery rates were in the range of 98.80%-101.39%. For 15 branch of the leaves, the contents of iriflophenone 3-C-beta-D-gluoside, iriflophenone 3,5-C-beta-D-diglucoside, mangiferin, and iriflophenone 2-O-alpha-L-rhamnoside were between 0.41-14.48, 0.72-3.85, 4.30-29.07, 0.24-5.06 mg, respectivley. This method is precise, accurate and reliable, which provides an efficient way for the quality control of the leaves of A. sinensis. This will promote the comprehensive usage of this plant.
Benzophenones ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry ; Spectrophotometry, Ultraviolet ; methods ; Thymelaeaceae ; chemistry ; Xanthones ; analysis

AIM: To study the expression of endostatin in ischemic myocardium of myocardial infarction(MI) rats in various periods and the correlation with VEGF expression and microvascular density(MVD).METHODS: Thirty-two male Sprague-Dawley rats after myocardial infarction were randomly divided into 7,14,21 and 28 days group.The sham group was normal control group(eight rats in each group).The expression of endostatin,VEGF and MVD in ischemic myocardium were observed by immunohistochemistry.RESULTS: The expression of endostatin significantly increased in the ischemic myocardium after MI,peaked at 7 days,then gradually decreased at 14,21 and 28 days.The endostatin level at 28 days was the same as the shams.The changing trends of expression of endostatin in ischemic myocardium after MI were similar to that of VEGF and were significantly correlated with the MVD.CONCLUSION: The expression of endostatin increased in ischemic myocardium of myocardial infarction rats.The changing trends of endostatin were similar to that of VEGF and positively correlated with the MVD.These data suggest that endostatin may modulate ischemic myocardium angiogenesis after myocardial infarction.

0.05). H/R injury significantly decreased the GSH level (P

BACKGROUND: The dynamic characteristics of electroencephalogram (EEG) include a decrease in the chaotic dimension, the correlation dimen sion, the Lyapunov exponent, the chaotic complexity, the freedom of EEG and an enhanced synchronization and periodicity of the EEG from several minutes to tens of minutes before epileptic seizures. All these characteristics prefigure the forthcoming seizures. Some studies have proven that the non linear dynamical system can be used as a feasible approach to explore the potential variables for describing the chaos portrait of EEG. OBJECTIVE: To analyze the electric characteristics of EEG signal in the epileptic seizures in rat model by investigating the nonlinear dynamical variables, such as the approximate entropy (ApEn) and correlation dimen sion. DESIGN: Observational and experimental study based on animals. SETTING: Department of Medical Engineering, Department of Gastroen terology, Second Artilleryman General Hospital of Chinese PLA; Department of Physics, Faulty of Biomedical Engineering, Fourth Military Medical Uni versity of Chinese PLA. MATERIALS: From September 2001 to January 2002, this study was conducted at the Complexity Laboratory of the Biomedical Department of the Fourth Military Medical University of Chinese PLA. Six male SD rats,weighing 150- 200 g, were selected.INTERVENTIONS: After intraperitoneal injection of chloral hydrate (0. 5 mL), the male SD rats were deeply anesthetized. When their EEG signal became stable, bemegride injection was diluted at 1:1 with saline and was given on a volume of 0.5 mL to the rats intraperitoneally. After a while,the epileptic seizures started marked by a spasm with a deep roar. The entire epileptic seizures were recorded. According to the shape of EEG waves and the corresponding symptoms of the rats during their seizures, data of the four phases, referring to normal condition, preictal phase, ictal phase and postictal phases of epileptic seizures, were selected for nonlinear analysis. The variations of the ApEn and the correlation dimension were calculated.MAIN OUTCOME MEASURES: In the four phases of the seizures, before seizures, preictal phase, ictal phase and postictal phases, the changes in the ApEn and correlation dimension were observed.RESULTS: All the 6 rats entered the statistical procedure. During epilepsy, the ApEn and correlation dimension of the EEG signal in ictal phases (0. 447 ±0. 126, 2. 166 ±0. 377) decreased significantly while those in preictal phases(0. 807 ±0. 182, 4. 773 ±0. 319) and postictal phases (1. 241 ±0. 125, 6. 042 ±0. 373) (t = -3. 984to 17. 902, P ＜0. 01). The ApEn and the correlation dimension of the EEG signal in preictal and ictal phases had significant difference with those observed under normal conditions (1.313 ± 0. 090, 6. 405 ± 0. 694) (t = -5. 228 to 12. 740, P ＜ 0. 01 ).CONCLUSION: The changes in ApEn and correlation dimension showed by nonlinear dynamical approach in this study reflect the characteristics of EEG signals in preictal time, ictal time and postictal timeof the epileptic seizures and the differences among them. Additionally, they also reveal the laws in the changes of the complex ictal EEG signal.

OBJECTIVETo investigate the levels of secretions from the prostate and seminal vesicles and their association with the expressions of aquaporins (AQP) in the prostatic tissue and seminal vesicles of castrated rats.

METHODSWe randomly divided 18 eight-week-old male SD rats into a control, a castration, and a testosterone (T) replacement group. Four weeks after surgical castration, we detected the plasma T level and measured the volumes of the secretions and the expressions of AQPs 3, 7, and 10 - 12 in the prostate and seminal vesicles of the rats.

RESULTSThe plasma T level was significantly lower in the castrated models ([30. 98 ± 28. 84] ng/dl) than in the rats of the control ([700.78 ± 123.8] ng/dl) and T replacement groups ([688.08 ± 132. 47] ng/dl) (P <0. 05). The castration group, in comparison with the control and T replacement groups, showed remarkably reduced ratios of prostatic secretion volume / prostate weight ([11.1 ± 0.30] vs [2.32 ± 0.61] and [2.13 ± 0.56] %, P <0. 05) and seminal vesicle secretion volume / seminal vesicle weight ( [4. 78 ± 1. 97 ] vs [57. 36 ± 11. 86] and [55. 74 ± 7. 21] %, P < 0. 05). Immunohistochemistry revealed the expressions of AQPs 3 and 7 in the epithelial envelop and cytoplasm and that of AQP 11 the in endothelial envelop and cytoplasm of the prostate and seminal vesicles. Western blot exhibited significantly lower expressions of AQPs 3, 7, and 10 - 12 in the prostate and seminal vesicles of the castrated rats than in the animals of the control and T replacement groups (P <0. 05).

CONCLUSIONSignificant decreases of the secretions from the prostate and seminal vesicles may be related to the reduced expressions of AQPs 3, 7, and 10 - 12 in the prostatic tissue and seminal vesicles in castrated rats.

Animals ; Aquaporins ; metabolism ; Humans ; Male ; Orchiectomy ; Prostate ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminal Vesicles ; metabolism ; Testosterone ; blood

Purpose To investigate whether FoxM1 participate in inhibitory effect of cisplatin (CDP) in resistant osteosarcoma cell lines by down-regulation of Rad51.Methods The resistant osteosarcoma cell lines were induced by gradually increasing dose intermittent action,and were named MG-63/R and HOS-MNNG/R respectively.The mRNA and protein level of FoxMl and Rad51 were detected by qRT-PCR and Western blot analysis in resistant cells and parental cells.The mRNA and protein level of FoxM1 and Rad51 were detected by qRT-PCR and Western blot analysis in resistant cells after treatment of 4 μmol/L Thiostrepton.The effect of single or combined treated of 4 tμmol/L Thiostrepton or 2 tμg/mL CDP on the rate of cell proliferation in resistant cells was examed by cell counting.Results Resistant osteosarcoma cell lines MG-63/R and HOSMNNG/R were established and stablely growthed in the concentration of 2 μg/mL CDP,and the resistance index was 30.52 and 37.87 respectively (severe CDP resistance).The mRNA and protein level of FoxM1 and Rad51 were significantly increaced in resistant cells compared with parental cells.The proliferation rate of resistant cells in conbination of 4 μmol/L Thiostrepton and 2 μg/mL CDP treated group was significantly lower than these two drugs single treated group.The level of mRNA and protein of FoxM1 and Rad51 were significantly decreased after 4 μmol/L Thiostrepton treatment in CDP resistant cells.Conclusion The results suggest that FoxM1 and Rad51 may participate in the resistant osteosarcoma cells to CDP.FoxM1 inhibitor Thiostrepton may strengthen the inhibitory effect of CDP in the resistant cells by down-regulation of Rad51.

OBJECTIVE To establish an in vitro cell model based on patient-specific human neural stem cells to study the pathomechanism of sporadic AD as well as screen candidate drugs.METHODS The peripheral blood cells from sporadic AD patients and cognitive normal controls were repro-grammed into inducedpluripotent stem cells(iPSCs),which were further induced into neural stem cells and neurons. The cell growth curve during the differentiation process was recorded by the IncuCyte ZOOM, and neural stem cells and neurons were identified by immunofluorescence. The apoptosis of neural stem cells and neurons was detected by Click-iT?Plus TUNEL Assay. RESULTS Neural stem cells derived from AD patients and cognitive normal controls can express neural stem cell markers Nes-tin,Sox1,Sox2 and Ki67.TUNEL assay results showed that the number of TUNEL-positive cells in neu-ral stem cells derived from AD patients was significantly higher than that of cognitive normal controls (P<0.01). When neural stem cells were differentiated into neurons, the percentage of MAP2 positive cells in the neural stem cell-derived culture dish of AD patients was significantly higher than the cogni-tive normal controls at day 16 of neuronal differentiation (P<0.01); the TUNEL assay showed that the number of TUNEL-positive cells in AD-derived neurons was significantly greater than that in cognitive normal controls (P<0.01) at day 16 of neuronal differentiation. CONCLUSION Our study revealed that AD-iPSC-derived neural stem cells exhibit premature neuronal differentiation and increased neural apoptosis,which might be relevant to the neuronal loss of AD,thus may provide valuable new tools to screen candidate drugs for the disease and to discover the mechanisms underlying AD pathogenesis.

Objective To investigate the correlation between apolipoprotein E (protein:apoE;gene:APOE) polymorphisms and intracellular Ca2 + concentration in the early stage after astrocyte injury.Methods ( 1 ) The CDS region of three APOE alleles was obtained by using reverse transcription polymerase chain reaction (RT-PCR). Then, the recombinant plasmid pEGFP-N1-APOE was constructed and identified by sequencing. (2) Astrocytes were separated from APOE gene-knockout mice for immunocytochemical identification. The recombinant plasmid was transfected into the astrocytes with liposome-mediated method to screen the cell lines that could stably express APOE information. (3) Cell injury models were set up by scarification. Laser scanning confocal microscope (LCSM) was used to detect the dynamic changes of intracellular Ca2+ at 12, 24, 48 and 72 hours postinjury. Results Compared with the control group ( before injury ), every allele showed significant changes of fluorescence intensity of Ca2 + ( P ＜0.05). At 12 hours after injury, the fluorescence intensity of Ca2+ was weak, with no statistical difference between three groups ( P ＞ 0. 05 ). At 24,48 and 72 hours postinjury, the fluorescence intensity was increased progressively, with significant higher intensity in ε4 group than the other two groups (P ＜0.05 ). Conclusions The concentration of intracellular Ca2+ in the astrocytes carrying APOEε4 allele is higher than that of those carrying APOEε2 and ε3 alleles, indicating that APOEε4 carriers may activate Ca2+ channel and lead to aggravation and poor prognosis of acute injury.

· AIM: To investigate the role of intercellular adhesion molecule-1 (ICAM-1) in the graft rejection of rabbit penetrating keratoplasty (PKP).· METHODS: Sutures were used to produce corneal neovascularization (CNV) on New Zealand white rabbits to make PKP model. Graft mean survival time and rejection index were determined. The levels of sICAM-1 in serum and aqueous humor were dynamically determined by enzyme-linked immunosorbent assay (ELISA).Immunohistochemical technique was used on cornea to get the evidence of expression and distribution of ICAM-1.· RESULTS: Graft mean survival time is (12.4±1.3) days.The mean sICAM-1 levels in aqueous humor and serum in the control animals that did not undergo surgery were (16.6±3.6)ng/L and (95.2±6.3) ng/L respectively. Levels in aqueous humor and serum for the graft rejection group increased postoperatively, continued to increase to (53.9±19.2)ng/Land (378.8±30.6)ng/L, compared to those in the other group (P ＜0.01). ICAM-1 positive cells were found in the graft rejection group.· CONCLUSION: ICAM-1 plays a critical role in initiating and maintaining corneal graft rejection. Sequential determination of serum sICAM-1 after operation may be of value in the prediction and diagnosis of acute rejection.