Objective To determinted whether GM1 had a protective effect on injury induced by serum-deprivation and the possible mechanism in PC12 cells. Methods The viability of PC12 cells was quantified by MTT after serum-deprivation.The number of apoptotic cells and necrotic cells were determined by Hoechst 33258/PI staining.And the change of PKC protein expression on PC12 cells' membrane and cytosols was detected by Western blotting. Results 1.The viability of PC12 cells decreased after serum-deprivation and the serum-deprivation for 24 hours was chosen as an injury model in this research.Most of the PC12 cells presented apoptosis 24 hours after serum-deprivation.In addition,the PC12 cells' cytosols PKC protein decreased,while the PC12 cells' membrane PKC protein increased significantly,and this result suggested PKC's translocation to membrane and its activation.2.The viability of PC12 cells preincubated with GM1 in high concentrations(10,1,0.1?mol/L) increased significantly and GM1 protected PC12 cells from apoptosis after serum-deprived injury.GM1 reduced the damage of serum-deprivation on PC12 cells and inhibited PKC protein translocation after injury.3.The repair function of GM1 was effective to neuronal resume after serum-deprived injury.Conclusion Neuroprotective effects of GM1 on serum-deprived injury may be partly mediated through the regulation of PKC pathways and it is helpful for the recovery after injury.

ObjectiveA HPLC-FLD method was developed to determine the levels of serum AAA in CRIpatients, and to studythe variationof serum AAAinCRI patientsanditsclinical significances. MethodsSerumsampleswerecollected from100healthcontrolsand 80CRI patients. According to 2002 National Kidney Foundation (NKF) staging diagnosis, CRI patients included 4 of stage 2, 12 of stage 3, 12 of stage 4, 52 of stage 5. According to pathogenesis, CRI patients were also divided into 3 groups :chronic nephritis group ( n = 32), DM group ( n = 36), hypertension group ( n = 12 ).Serums were deproteinized by equal volume of 5％ (v/v) PCA and supernate were analyzed direcdy. External standard method was used as quantitative method. The analytical column was Megres C18. 10％ acetonitrile in water was used as mobile phase. Flow rote was 1.0 ml/min. The wavelengths of fluorescence excitation and emission were changed with specific time. The levels of Tyr, Phe and Trp in CRI groups, different CKD stages and different pathogenesis were compared with healthy control groups to evaluated the sensitivity and specificity of serum AAA for CRI diagnosis. ResultsThe linear ranges of the method were 0. 550 -275.000, 3. 050 - 1220. 000 and 0. 049 -49. 000 pμmol/L for Tyr, Phe and Trp, respectively. The limit of detection (LOD) was 0.014 μmol/L for Tyr, 0.500μmol/L for Phe, and 0.005 μmol/L for Trp. The average recovery was 100. 9％, 101.3％ and 98. 5％ for Tyr, Phe and Trp, respectively. Intra-day CVwas 3. 18％ -4. 20％ ( mean was 3. 13％ )and inter-day CV was 3. 18％ -4. 20％ ( mean was 3. 58％ ). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in CRI patients were( 135.74 ±23.23 )μmol/L, (52.27 +8.25) μmol/L, (21.49 ±4.25) μmol/L and[0.87(0.68 - 1.05)]μmol/L. which were lower than that in healthy groups (t value was -14. 709, 4.452, 22. 100, U value was 266.000,respectively, P＜0. 05). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in healthy groups were ( 174. 47 ± 11.57 ) μmol/L, ( 63.53 ± 4. 68 ) μmol/L, (44. 22 ± 3. 67 ) μmol/L and[0. 97(0. 94 - 1.00)]μmoL/L. There were no statistically significant difference between the different stage of CRI. Compared with the concentration of Tyr, Phe and Trp among chronic nephritis group, DM group,hypertension group, the concentration of Tyr had no significant changes among these three kinds of diseases (P ＞ 0. 05 ). The concentration of Phe had significant changes between Chronic nephritis group and DM group, Chronic nephritis group and hypertension group ( U = 395.00, 114. 00, P ＜ 0. 05 ) ; the concentration of Trp haad significant changes between Chronic nephritis group and DM group ( U = 349.00, P ＜ 0. 05 ).The diagnostic sensitivity and specificity of serum AAA for CRI were 90％ (72/80) and 100. 0％ (100/100).ConclusionsThe method of high-performance liquid chromatography with fluorescence detection ( HPLC-FLD) is simple, rapid, sensitive and specific. Simultaneous determination of serum AAA was benefit to the diagnosis and evaluation of CRI patients.

OBJECTIVETo quantitatively evaluate the metabolic changes of benign prostatic hyperplasia (BPH) and prostate cancer in the transitional zone using magnetic resonance spectroscopic imaging (MRSI), and to analyze the characteristics and differences of the spectra in this zonal area.

METHODSEighteen patients with prostate cancer in the transitional zone underwent magnetic resonance imaging (MRI)/MRSI examinations. The (Choline + Creatine)/Citrate (CC/Ci) ratio and the Choline/Creatine (Cho/Cr) ratio were evaluated in each voxel with cancer or BPH confirmed by pathological results. Discriminant analysis was used to determine the power of the two ratios in differentiation between cancer and BPH.

RESULTSThe CC/Ci ratio and Cho/Cr ratio for cancer voxels were significantly higher than those in the voxels with BPH in the transitional zone (CC/Ci: 2.36 +/- 1.31 vs. 0.85 +/- 0.29, P < 0.01; Cho/Cr: 4.14 +/- 1.79 vs. 1.26 +/- 0.45, P < 0.01). As for the discriminant function with the CC/Ci ratio and the Cho/Cr ratio, the specificity, sensitivity, and accuracy were 98.6%, 85.7%, 92.9% respectively for the differentiation between cancer and BPH.

CONCLUSIONSThe prostate cancer is characterized by higher CC/Ci ratio and Cho/Cr ratio compared to BPH in the transitional zone. Both CC/Ci ratio and Cho/Cr ratio have high specificity, sensitivity, and accuracy in their discriminative power between cancer and BPH in this zonal area.

Adult ; Diagnosis, Differential ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Prostatic Hyperplasia ; diagnosis ; pathology ; Prostatic Neoplasms ; diagnosis ; pathology

OBJECTIVETo explore the ability of interictal diffusion-weighted imaging (DWI) to localize the temporal lobe of seizure origin and to predict postoperative seizure control in patients with temporal lobe epilepsy (TLE).

METHODSTwenty-seven patients with intractable TLE considered for surgery and 19 healthy volunteers were studied with conventional magnetic resonance imaging (MRI) and DWI. Apparent diffusion coefficients (ADCs) of bilateral hippocampi in both TLE patients and control subjects were obtained. Lateralization to either temporal lobe with hippocampal ADC was based on the threshold values derived from +/- SD of right/left ratios in normal subjects. And the postoperative pathology was reviewed.

RESULTSHippocampal ADCs were higher on the side of surgery compared with those on the contralateral side as well as the ipsilateral side in control subjects [resected side (109.8 +/- 7.3) x 10(-5) cm2/s, contralateral side (91.7 +/- 4.7) x 10(-5) cm2/s, control subjects (81.6 +/- 5.2) x 10(-5) cm2/s, all P < 0.01]. Right/left hippocampal ADC ratio and conventional MRI lateralized to the operated temporal lobe in 21 of 27 (77.8%) and in 18 of 27 (66.7%) patients. Lateralization to the surgical side was not associated with postoperative seizure control with right/left hippocampal ADC ratio (P > 0.05).

CONCLUSIONSConventional MRI is a sensitive method to detect hippocampal sclerosis. Accuracy of the right/left hippocampal ADC ratio for lateralizing to the side of surgery is very high, but it isn't a better predictor of surgical outcome.

Adolescent ; Adult ; Brain ; anatomy & histology ; metabolism ; Child ; Diffusion Magnetic Resonance Imaging ; methods ; Epilepsy, Temporal Lobe ; diagnosis ; physiopathology ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Prognosis ; Seizures ; physiopathology ; Treatment Outcome ; Young Adult

Objective To explore the effect of in utero exposure to inlfammation on innate immune response in preterm infants. Methods Forty-seven premature infants with gestational age<35 weeks were recruited in this study. According to his-tological evidence of placental infection, all neonates were divided into intrauterine inlfammation positive group and negative group. Mononuclear cells and monocytes were isolated from umbilical cord blood, and were cultured in vitro in the presence or absence of LPS (100 ng/ml). The levels of interleukin 1β(IL-1β), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α) and interleukin 10 (IL-10) in cord blood plasma and monocyte cultural supernatants were measured by ELISA respectively. The level of IL-1β, TNF-α, IL-6 and IL-10 mRNA were detected by Real-time PCR. Expression of HLA-DR on surface of CD14+monocytes and ratio of CD3+CD4+/CD3+CD8+T was analyzed by lfow cytometry. Results (1) The level of cord plasma IL-6 in intrauterine inlfammation positive group was signiifcantly higher than in negative group. (P=0.02). (2) After stimulation of LPS, levels of IL-1β, IL-6, TNF-α, IL-10 in supernatants were increased signiifcantly, in consistence with their mRNA expression (P<0.05) in both groups. (3) Expression of HLA-DR on surface of monocytes was signiifcantly decreased after stimulation with LPS in intrauterine inlfammation positive group (P=0.012), but was signiifcantly increased in negative group (P=0.0305). Con-clusions In utero exposure to inlfammation does not suppress the response of monocytes to LPS in preterm neonates, but impairs the antigen presenting function in monocytes.

AIM:To compare the biological characteristics , surface markers and multi-differentiation potential of the mesenchymal stem cells (MSCs) derived from the umbilical cord and bone marrow in the green fluorescent protein (GFP) transgenic mice.METHODS:Umbilical cord MSCs (UCMSCs) were isolated by collagen type II enzymatic diges-tion and bone marrow MSCs ( BMSCs) were isolated by density gradient centrifugation .The growth of the 2 types of MSCs was observed under inverted microscope .The cell proliferation was detected by determining the growth curve and MTT as-say.The Trypan blue method was performed to analyze the cell viability rate .The cell cycle and cell surface markers were measured by flow cytometry .The differentiation potentials of the 2 types of MSCs were tested by the differentiation kits to-ward adipocytes and osteoblasts .RESULTS:The UCMSCs attached to the culture surface 1 d after the isolation , and the cells showed spiral shape with notable growth and proliferation after 2 d of culture.After 3 d, the cell arrived sub-confluent and was ready for passage .BMSCs still showed circular shape and started to attach to the surface 4 d after culture .They formed the small colony shape only after 5 d with obvious proliferative potential .The cells became confluent 7 d after the culture.The original generation of cultivating UCMSCs growth curve was shown typically an “S” shape.But the BMSCs growth was slower than the UCMSCs .The cell proliferation was obvious for UC-MSCs in 3~5 d.BMSCs proliferated signif-icantly only after 7 d.The viability rate arrived more than 96%for both types of MSCs .The cell cycle of both MSCs did not show significant difference (G0/G1 phases were above 85%, P>0.05).Both MSCs positively expressed CD44, CD90 and CD105 (60.7%±2.3%) but the expression of CD45, CD19, CD14 and CD79 was negative (less than 25.6%±4.8%, P>0.05).More than 90%of the MSCs from the umbilical cord and bone marrow differentiated towards the adipocytes and osteoblasts without significant difference (P<0.05).CONCLUSION:UCMSCs have stronger ability of proliferation and multi-directional differentiation potentials .UCMSCs in GFP transgenic mice as a high-quality tracer can serve for tracking the stem cells in vivo.

Acupuncture Therapy ; Altitude ; Animals ; Brain Edema ; prevention & control ; Female ; Hypoxia, Brain ; therapy ; Male ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Adenosine A1 ; metabolism

Objective:To conduct a systematic study of the immunologic response of rats to transplanted glutaraldehyde ( GA)-treated porcine blood vessels in vivo.Methods: The experiment was divided into two groups:fresh group and glutaraldehyde-treated group.Twenty cases of fresh and glutaraldehyde-treated porcine pulmonary arteries were subcutaneously embedded in rats.We compared the changes using HE staining and immunohistochemistry.Results:HE staining showed that there were stronger expression on day 12 and day 30 in the fresh group than that in the glutaraldehyde group.There were similar results in morphology in CD68,C3,IgG.The results of integral optical density ( IOD) in immunohistochemistry showed that IOD started rising from day 4 and got the peak on day 12 or day 30 and or fell on day 60.Conclusion: Innate immunity played an important role in the research on xenogenic immunological rejection mechanism.The immunogenicity of glutaraldehyde-treated xenogenic blood vessels is lower than that in fresh blood vessels.However there is still immunogenicity in glutaraldehyde-treated xenogenic blood vessels.We will explore better ways to obviously weaken the rejection.

BACKGROUNDAmide proton transfer (APT) imaging has recently emerged as an important contrast mechanism for magnetic resonance imaging (MRI) in the field of molecular and cellular imaging. The aim of this study was to evaluate the feasibility of APT imaging to detect cerebral abnormality in patients with Alzheimer's disease (AD) at 3.0 Tesla.

METHODSTwenty AD patients (9 men and 11 women; age range, 67-83 years) and 20 age-matched normal controls (11 men and 9 women; age range, 63-82 years) underwent APT and traditional MRI examination on a 3.0 Tesla MRI system. The magnetic resonance ratio asymmetry (MTR asym ) values at 3.5 ppm of bilateral hippocampi (Hc), temporal white matter regions, occipital white matter regions, and cerebral peduncles were measured on oblique axial APT images. MTR asym (3.5 ppm) values of the cerebral structures between AD patients and control subjects were compared with independent samples t-test. Controlling for age, partial correlation analysis was used to investigate the associations between mini-mental state examination (MMSE) and the various MRI measures among AD patients.

RESULTSCompared with normal controls, MTR asym (3.5 ppm) values of bilateral Hc were significantly increased in AD patients (right 1.24% ± 0.21% vs. 0.83% ± 0.19%, left 1.18% ± 0.18% vs. 0.80%± 0.17%, t = 3.039, 3.328, P = 0.004, 0.002, respectively). MTR asym (3.5 ppm) values of bilateral Hc were significantly negatively correlated with MMSE (right r = -0.559, P = 0.013; left r = -0.461, P = 0.047).

CONCLUSIONSIncreased MTR asym (3.5 ppm) values of bilateral Hc in AD patients and its strong correlations with MMSE suggest that APT imaging could potentially provide imaging biomarkers for the noninvasive molecular diagnosis of AD.

Aged ; Alzheimer Disease ; diagnosis ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male

Anti-Müllerian tube hormone(AMH), also known as Müllerian inhibitory substances, is a member of the transformation system. In males, AMH is secreted by immature Sertoli cell which promotes the degradation of male fetal Müllerian tubes,and participates in testicular development and spermatogenesis.AMH can regulate gona-dotropin-releasing hormone (GnRH), pituitary secretes follicle stimulating hormone (FSH) and luteinizing hormone (LH),testicular stromal cells secrete testosterone(T) and inhibin B causes male reproductive endocrine related diseases through HPG axis.