OBJECTIVETo establish a model of hypoxic postconditioning of dermal microvascular endothelial cells.

METHODSDuring reoxygenation after hypoxia, cells received three times of hypoxic/ reoxygenation alternate treatment for a certain time. The cells were seeded on 6-well plates, with one plate for one group. They were divided into 6 groups as group 1 ( Control), group 2 (8h hypoxia + 24h reoxygenation), group 3 (8h hypoxia + 2 min x 3 times post-hypoxia treatment) , group 4 (8h hypoxia + 5 min x 3 times post-hypoxia treatment), group 5 (8h hypoxia + 10 min x 3 times post-hypoxia treatment), 6 group (8h hypoxia + 20 min x 3 times post-hypoxia treatment). Each group underwent 8 h hypoxia + 24 h hypoxia Buffer and reoxygenation. Lactate dehydrogenase (LDH) was detected during the process. Apoptosis rate was calculated by staining Tunel method. Bcl-2, Bax and activated caspase-3 protein were detected by Western Blot.

RESULTSIn the continuous hypoxia process, the LDH was (1563 ± 83.35) IU/L at 8h and (582.85 ± 58.25 ) IU/L at 0h, showing a statistical difference (P = 0.0001). Western blotting results showed that the expression of Bax/Bcl-2 in group 2 was 0.38 ± 0.02, showing a significant difference when compared with that in group 3 (0.23 ± 0.01) and group 4 (0.22 ± 0.02) (P = 0.012, P = 0.005), while not when compared with that in group 5 (0.33 ± 0.02) and 6 groups (0.34 ± 0.01) P > 0.05). The ratio of Activated caspase-3/caspase-3 in group 2 (6.30 ± 1.50) was significantly higher than that in group 3 (2.17 ± 0.26) and group 4 (2.63 ± 0.31) (P = 0.008, P = 0.019); while not in group 5 (4.36 ± 0.29) and group 6 (4.97 ± 0.51) (P > 0.05).

CONCLUSIONSThe model of hypoxic postconditioning of human dermal microvascular endothelial cells is successfully established.

Apoptosis ; Blotting, Western ; Caspase 3 ; analysis ; Cell Hypoxia ; Endothelial Cells ; metabolism ; Humans ; In Situ Nick-End Labeling ; Ischemic Postconditioning ; methods ; L-Lactate Dehydrogenase ; analysis ; Oxygen Consumption ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Time Factors ; bcl-2-Associated X Protein ; analysis

Objective To investigate the diagnostic values of DCE-MRI quantitative evaluation of the activity of sacroiliac joints in ankylosing spondylitis.Methods Forty two patients (36 male and 6 female) with ankylosing spondylitis in our hospital were enrolled prospectively according to the standard of diagnostic criteria for ankylosing spondylitis revised in 1984.All of them were evaluated with the blood sedimentation,C-reactive protein and ankylosing spondylitis activity index (BASDAI).We divided the patients with ankylosing spondylitis into stationary phase group and active phase group.We analyzed each of the MRI images of the two patients groups using the extended Tofts model to determine the quantitative parameters of DCE-MRI,such as contrast transfer coefficient (Ktrans),reflux constant (Kep),extravascular extracellular volume fraction (Ve),plasma volume fraction (Vp).And then the differences of quantitative data between the two groups were compared.Spearman correlation analysis was performed between the parameters with statistically significant difference and BASDAI scores.Results Among the 42 patients,22 patients (18 male and 4 female) were in active phase group and the other 20 patients (18 male and 2 female) were in stationary phase group.Ktrans,Kep,Ve were (0.750± 1.130)/min,(1.008±0.732)/min,0.460± 1.735;(0.163±0.401)/min,(0.505 ± 0.902)/min,0.345 ±3.460 for active phase group and the stationary phase group,respectively.The results of the active phase group was significantly higher than the stationary phase group (Z=3.727,2.317,3.696,respectively;all P＜0.05).The Vp had no statistically significance for the active phase group (0.125 ± 0.310) and the stationary phase group (0.160 ± 0.329) (Z=1.209,P＞0.05).Strong correlations existed between Ktrans,Kep,Ve and BASDAI score,the correlation coefficients were 0.714,0.430,and 0.676 (P＜0.05).Conclusions Quantitative DCE-MRI parameters can evaluate the activity of the ankylosing spondylitis.Strong correlations exist between Ktrans,Kep,Ve and BASDAI score.

BACKGROUND:Vertebral column resection is the frequently mentioned spinal orthopaedic concept. Due to the high requirement of the operation skil , difficulty and more complications, the previous studies have reported from different aspects, and many researchers have focused on the analysis of complications, that may be related with the procedure and manner. OBJECTIVE:To analyze the efficacy and complications of posterior vertebral column resection combined with titanium screw rod fixation for the treatment of severe spinal deformity. METHODS:We retrospectively analyzed 48 patients with severe spinal deformity who treated with posterior vertebral column resection and titanium screw rod fixation, with an average removal of 1.6 vertebral. The patients were fol owed-up for 15-64 months. The Cobb angle (coronal plane and sagittal plane) of the patients before treatment, after treatment and in the final fol ow-up was analyzed, and the relative complications of the surgery were analyzed. RESULTS AND CONCLUSION:The patients with spinal deformity were divided into five categories, included kyphoscoliosis (n=11), severe scoliosis (n=20), congenital spinal deformity (n=4), spherical kyphosis (n=3), and angular kyphosis (n=10). The average coronal plane deformity angle of the patients was corrected from 84° preoperation to 35° postoperation, with the total correction rate of 54%. The average sagittal plane deformity angle was corrected from 90° preoperation to 42° postoperation, and the sagittal plane Cobb angle was decreased for 48°. The mean operation time was 545 minutes (204-1 355 minutes), the intraoperative blood loss was 1 610 mL (50-8 244 mL), and the average blood loss was 65%. After treatment, 31 cases had complications, including 13 cases of intraoperative neurological dysfunction (observed through electrophysiological monitoring and wake), permanent neurological dysfunction did not occur after timely treatment. The posterior vertebral column resection and titanium screw rod fixation can obtain better effect in the treatment of severe spinal deformity, but the procedure has high complications and is difficult for operation.

Objective To investigate the experience of laparoscopic complete mesocolic excision ( CME) for colon cancer. Methods There were102 patients,of which 68 cases with colon cancer were performed laparoscopic CME,34 cases were treated by traditional surgery. The 2 groups were reviewed retrospectively. Results As compared with the traditional group,the operation time,time of first flatus,hospital stay in the CME group increased. The postoperative suction drainage was decreased in CME group. The CME group had less blood loss and more mean lymph nodes clearance than the traditional group. The complication incidences had no significant differences between 2 groups. Conclusion Laparoscopic CME for colon cancer,with the advantages of less tumor spreading and more thoroughly lymph node dissection,is worthy of clinical application.

Objective To evaluate the clinical effects of combined therapy of Fuyuan Huoxue decoction and transcatheter hepatic arterial chemoembolization in the treatment of primary hepatic carcinoma. Methods 80 patients with primary hepatic carcinoma were randomly divided into a control group, treated by transcatheter hepatic arterial chemoembolization, and a treatment group, additionally treated by Fuyuan Huoxue decoction on the basis of the control group. By observing the change of gross tumor volume、tumor markers、clinical symptoms、Karnofsky Performance Status(KPS) score、quality of life and so on,compare the clinical effects and quality of life between the two groups. Results The effective rate of solid tumor was 47.50%and 35%in the treatment and the control group respectively, with no significant difference(χ2=-1.229, P>0.05);The total effect rate was 87.50%and 32.50%in the treatment and the control group respectively, with significant difference(χ2=-5.633, P<0.05);The rate of patients merged with portal vein tumor thrombus whose cancer embolus narrowed more than 1/2 after the treatment was 78.95%and 33.33%in the treatment and the control group respectively, with significant difference(χ2=7.836, P<0.05);The rate of alpha fetoprotein(AFP) decreasing or turning negative was 78.95%and 37.83%after the treatment in the treatment and the control group respectively, with significant difference(χ2=-3.857, P<0.05);Both groups have improvement in Karnofsky Performance Status(KPS) score after the treatment, the ratios was 80% and 72.50% in the treatment and the control group respectively, with no significant difference(χ2=-1.203, P>0.05);The accumulated scores change of quality of life(QOL) has asignificant difference(χ2=-3.025, P<0.05) between the two groups after the treatment. Conclusion The combined therapy of Fuyuan Huoxue decoction and transcatheter hepatic arterial chemoembolization can alleviate the clinical symptoms, improve treatment effects and quality of life of patients with primary hepatic carcinoma.

Objective To investigate the effects of Gualou Guizhi Granules on excitatory toxic damage of PC12 induced by glutamate; To primarily explore the involved protective mechanism of Gualou Guizhi Granules. Methods Excitatory toxic damage of PC12 induced by glutamate was used to establish models. Cells were divided into normal group, glutamate group, and Gualou Guizhi Granules low- (200 μg/mL), medium- (400 μg/mL) and high-dose (800 μg/mL) groups. MTT and LDH assay methods were used to detect PC12 activity; Caspase-3 activity detection method and Annexine V/PI double staining method were used to detect cell apoptosis; Western blot and RT-PCR were used to detect Bcl-2, Bax protein and mRNA expression. Results Compared with glutamate group, MTT showed that all Gualou Guizhi Granules groups could improve PC12 activity, and LDH showed that cell activity in all Gualou Guizhi Granules groups decreased; Annexine V/PI double staining method showed that all Gualou Guizhi Granules groups could decrease the PC12 apoptosis; Caspase-3 activity detection method showed that all Gualou Guizhi Granules groups could decrease the activity of Caspase-3; Western blot and RT-PCR showed that all Gualou Guizhi Granules groups could reduce Bax expression and increase Bcl-2 expression. Conclusion Gualou Guizhi Granules have certain protective effects on excitatory toxic damage of PC12 induced by glutamate, which may be related to its anti-apoptotic activity.

Objective To compare and study species homology of two kinds of Antrodia Cinnamomea; To conduct the study on quality control. Methods HPLC fingerprint of two kinds of Antrodia Cinnamomea was applied. The HPLC fingerprints were determined on an Diamonsil C18 (2) column (250 mm × 4.6 mm, 5 μm) eluted with the mobile phase consisting of 1% formic acid solution and acetonitrile in gradient mode at a flow rate of 0.8 mL/min; the detection wavelength was set at 254 nm; the column temperature was 30 ℃. Results There were 9 common peaks in the HPLC fingerprints of Antrodia Cinnamomea. The similarities varied from 0.906 to 0.995 and from 0.956 to 0.998 in 11 batches and 10 batches of two kinds of Antrodia Cinnamomea, respectively. Conclusion The method is simple and with good reproducibility, which shows that the HPLC fingerprint and infrared spectrum share similarity for two kindsof Antrodia Cinnamomea.

In order to explore the differences of chemical constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra, a qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) was developed for identification of multi-constituents and an HPLC-DAD analytical method was developed for simultaneously determining 14 major compounds (gallic acid, protocatechuic acid, paeoniflorin sulfonate, protocatechuic aldehyde, methyl gallate, oxypaeoniflorin, catechin, albiflorin, and paeoniflorin, ethyl gallate, benzoic acid, pentagaloylglucose, benzoyl-paeoniflorin, and paeonol) in Paeoniae Radix Alba and Paeoniae Radix Rubra. Q-TOF/MS qualitative analysis was performed under negative ion mode and inferred 38 components of Paeoniae Radix Alba and 30 components of Paeoniae Radix Rubra. HPLC-DAD quantitative method result showed the contents of 8 ingredients were different between Paeoniae Radix Alba and Paeoniae Radix Rubra. The results indicated that the new approach was applicable in qualitative and quantitative quality control of Paeoniae Radix Alba and Paeoniae Radix Rubra.
Bridged-Ring Compounds ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Molecular Structure ; Monoterpenes ; chemistry ; Paeonia ; chemistry ; classification ; Tandem Mass Spectrometry ; methods

Objective:To investigate polymorphism of human leukocyte antigen (HLA)-DRB1 and -DQB1 genes in Bai ethnic group in Dali,Yunnan province.Methods:Polymerase chain reaction-sequence specific primers (PCR-SSP) were used to determine HLA-DRB1 and -DQB1 alleles in 124 unrelated healthy Bai ethnic individuals living in Eryuan County of the Dali Bai autonomous prefecture,Yunnan province.Results:Among all the 21 DRB1 alleles and 15 DQB1 alleles were identified,the predominant alleles were DRB1*1202(26.61%),DRB1*0901(13.89%) and DRB1*0803(9.92%) on DRB1 locus and DQB1*0301(31.45%),DQB1*0601(10.08%),DQB1*0401(8.06%)and DQB1*0502(8.06%)on DQB1 locus.The most common haplotypes were DRB1*1202-DQB1*0301(20.08%)and DRB1*0803-DQB1*0601(7.19%).Conclusion:The phylogenetic tree constructed according to the HLA-DRB1,-DQB1 allele frequencies of Bais with those of other 10 populations suggests that the Bai ethnic group belongs to the southern group of China,but it keeps genetic distance from others and the HLA genes exhibits a unique profile.This study would provide HLA polymorphism information of Bai for the future investigation on the disease related to the genetic polymorphism.

[ ABSTRACT] AIM:To investigate the protective effect of naringin ( Nar) on the injury of human umbilical vein endothelial cells ( HUVECs) induced by 33 mmol/L high glucose ( HG) and to explore its possible mechanisms.METH-ODS:The injury model was established by treating HUVECs with HG medium for the indicated time (6, 12, 24, 48 and 72 h) , and then the levels of NO, eNOS and p-eNOS were detected, respectively.The effects of Nar on high glucose-in-duced endothelial cell injury were observed.HUVECs were treated with Nar at concentrations of 5, 10, 25, 50 and 100 mg/L for 6 h, 12 h, 24 h, 36 h and 48 h.The levels of NO in the supernatants were measured.The effects of Nar on HG-injured HUVECs were explored by treating the cells with 10 μmol/L of LY294002, a PI3K inhibitor, or 0.5 μmol/L of AKT inhibitorⅣ, an AKT inhibitor, and then the levels of NO, PI3K, AKT, eNOS and their phosphorylated proteins were determined by Western blot.RESULTS:Nar at concentration of 50 mg/L significantly attenuated the injury of endothelial cells induced by high glucose ( P<0.01) , and the protective effects of Nar were abolished by pretreating with the inhibitor of PI3K or AKT (P<0.01).CONCLUSION: Nar protects endothelial cells against the injury induced by high glucose through PI3K/AKT/eNOS pathway.