Objective To study the expression of ARHI mRNA,an imprinted suppressor gene,in ovarian serous carcinoma and the methylation status of its three CpG islands,and to determine the possible role of aberrant methylation of ARHI gene in pathogenesis of ovarian serous carcinoma.Methods Twenty normal ovarian specimens and 20 ovarian serous carcinoma specimens were analyzed.Total RNA was extracted and semi-quantitative RT-PCR was employed to detect the mRNA expression of ARHI gene.Methylation status of three CpG islands were examined by DNA bisulfite treatment,PCR amplification and methylation specific restriction enzymes,TagI for CpG I and III,BstUI for CpG II.Results The average of ARHI/G3PDH was 0.73?0.09 in normal ovarian specimens,and it was 0.43?0.37 in ovarian carcinoma specimens(P

Objective To perfect gene profile expressed in pre-implantation embryos. Methods Using nested RT-PCR to investigate the expression of seven imprinted genes: P57~ KIP2, LIT1, TSSC3, GRB10, PEG3, ARHI, and ZAC1 in human oocytes and pre-implantation embryos. Results Transcripts of P57~ KIP2 and ZAC1 were detected in human oocytes and at all stages of pre-implantation; LIT1 was expressed only in stages of 8-cell and blastocyst; transcripts of TSSC3 could not be detected; GRB10 mRNA could be detected in oocytes and pre-implantation embryos except for 2-cell embryo; ARHI was expressed in oocytes and 2,8-cell embryos and blastocyst; Peg3 mRNA existed in 4,8-cell embryos and blastocyst. Conclusion Except for TSSC3, transcripts of the other six imprinted genes are detected in human pre-implantation development, which are helpful for pre-implantation diagnosis of imprinted diseases, and provide the theoretical basis for understanding the correlation among assisted reproductive technology, genetic imprinted diseases and tumor.

Objective To study the impact of vaginal mesh exposure on quality of life in patients undergoing transvaginal reconstructive pelvic surgery (RPS) with polypropylene mesh.Methods From May 2004 to March 2011,114 patients with severe pelvic organ prolapse(POP) undergoing transvaginal RPS with polypropylene mesh were enrolled in this study,which were divided into exposure and non-exposure group according to appearing vaginal mesh exposure at 2 months,6 months and 1 year after operation.At the same time,pelvic floor distress inventory short form 20 ( PFD1-20 ) and pelvic floor impact questionnaire short form 7 ( PFIQ-7 ) were completed in those patients.Results At 2 months after operation,96 patients were followed up,including 19 patients in exposure group and 77 patients in non-exposure group,and the rate of exposure was 19.8c (19/96); At 6 months after operation,85 patients were followed up,including 13 patients in exposure group and 72 patients in non-exposure group,and the rate of exposure was 15.3％( 13/85 ) ; At 1 vear after operation,77 patients were followed up,including 6 patients in exposure group and 71 patients in non-exposure group,and the rate of exposure was 7.8％ (6/77).Mean score of PFDI-20 and PFIQ-7 in exposure group before operation was 39.6 and 57.1,which was statistically improved to 8.3 and 9.5 at 2 months after operation,8.3 and 9.5 at 6 months after operation,2.1 and 0 in I year after operation (P ＜0.01 ). Mean score of PFDI-20 and PFIQ-7 of non-exposure group before operation was 54.2 and 66.7,which was improved to 8.3 and 4.8 at 2 months after operation,0 at 6 months and 1 vear after operation,but there was no significant difference in mean score of PFDI-20 and PFIQ-7 between the two groups (P ＞ 0.05 ).Conclusion Vaginal mesh exposure was common after transvaginal RPS with polypropylene mesh,however,most of them were moderate,and there was no significant impact on patients'qualifies of life.

BACKGROUND: The spinal instability would accelerate the degeneration of normal disk. The injuries of anterior longitudinal ligament (ALL) and intervertebral disk were usually caused by cervical trauma, which leaded to spinal instability. Currently, there were few animal researches about the degeneration of injured intervertebral disk affected by ALL destruction.OBJECTIVE: To investigate the degeneration of injured intervertebral disk after spinal instability in the rabbit model of different degrees of ALL and disc destruction.METHODS: A total of 24 New-Zealand rabbits were randomly divided into intervertebral disk injury group (Group A, n=6), partial injury of ALL with disc injury group (Group B, n=6), injury of bony attachment point of ALL with disc injury group (Group C, n=6) and entirely injury of ALL with disc injury group (Group D, n=6). The L2-L3 intervertebral disk and ALL were injured through abdominal cavity. Different groups received different treatments. Computed tomography (CT) scans of the injured disc were performed at the postoperative 4 and 8 weeks, and the middle high of injured discs was calculated on CT sagittal reconstruction. Three rabbits were selected from each group. Hematoxylin-eosin staining of injured disc was performed after the animals were killed. Results were examined under the light microscope.RESULTS AND CONCLUSION: (1) At postoperative 4 weeks, the middle height of injured discs in Group D was decreased significantly compared with Group A (P < 0.05). There were hyperosteogeny and calcification in Group C and Group D. There were nucleus pulposus cells reduction and inflammatory reaction in Group C and Group D on histological staining. (2) At postoperative 8 weeks, the middle height of injured discs respective was decreased significantly compared with Group A (P < 0.05). The hyperosteogeny and calcification became clearer in Group C and Group D than before. There were morphologic changes of nucleus pulposus cells and fibrillation by hematoxylin-eosin staining, and the degree of disc degeneration was Group D > Group C > Group B > Group A. (3) In conclusion, the injury of ALL would accelerate the degeneration of correspondingly injured disk, and the degree of injury of ALL was positively correlated with the degeneration of disk.

BACKGROUND:Under hypoxic environment, hypoxia-inducible factor 1α plays a dualregulatory role in cel apoptosis. Severity of hypoxia is the key to determine whether cels appear to have apoptosis or adapt to survive. When the cels are exposed to chronic or extreme hypoxia, a lack of protection mechanisms from hypoxia-inducible factor-1α can induce cel apoptosis. OBJECTIVE: To research the expression of hypoxia-inducible factor 1α in human lumbar nucleus pulposus of different herniated types and its relationships with cel apoptosis. METHODS: The nucleus pulposus was harvested from 60 cases of herniation of lumbar intervertebral discs, L4-5 in 41 cases and L5-S1 in 19 cases. The nucleus pulposus tissues were equaly divided into protruded and sequestered groups. Meanwhile, the nucleus pulposus tissues from another 10 cases of lumbar spine fracture were taken as control group. Expression of hypoxia-inducible factor 1α and apoptosis of lumbar nucleus pulposus cels were observed and detected with immunohistochemical technique and TUNEL method. Correlation of hypoxia-inducible factor 1α and apoptosis in human lumbar nucleus pulposus of different herniated types was analyzed. RESULTS AND CONCLUSION: The expression of hypoxia-inducible factor 1α was visualized in each case, but it was significantly higher in the sequestered group than in the protruded group and control group (P < 0.01). Apoptosis of nucleus pulposus cels were found in al the three groups, but the apoptotic rate was also higher in the sequestered group than in the protruded group and control group (P < 0.01). Expression of hypoxia-inducible factor 1α was positively correlated with cel apoptosis in human lumbar nucleus pulposus (P < 0.01). Overal, the expression of hypoxia-inducible factor1α in degenerative human lumbar nucleus pulposus is associated with herniated types, which is the highest in the sequestered type. The relationship between hypoxia-inducible factor 1α and apoptosis is positive.

BACKGROUND:Under hypoxic environment, hypoxia inducible factor-1 plays an important role in regulation of hypoxia-induced gene expression in the intervertebral disc. Hypoxia-inducible factor-1 consists of α and βsubunits, and which hypoxia inducible factor-1α determines the stability and activity of hypoxia-inducible factor-1. OBJECTIVE: To observe the expression of hypoxia inducible factor-1α in the human lumbar nucleus pulposus of different herniated types and to judge their relationships. METHODS:A total of 60 nucleus pulposus samples were harvested from the lumbar vertebra, including 41 from L4-5 and 19 from L5-S1, and then divided into protruded group and sequestered group, with 30 cases in each group. Meanwhile, another 10 samples of lumbar nucleus pulposus served as controls. Hematoxylin-eosin staining and streptavidin-biotin peroxidase complex immunohistochemical technique were used to observe the expression of hypoxia inducible factor-1α in the human lumbar nucleus pulposus in different groups. RESULTS AND CONCLUSION:The expression level of hypoxia inducible factor-1α was (58.2±7.5)% in the sequestered group, (27.3±2.3)% in the protruded group, and (10.5±4.7)% in the control group, which was significantly higher in the sequestered group than the other two groups (P < 0.01). These findings indicate that the expression of hypoxia inducible factor-1α in the lumbarnucleus pulposus is associated with the herniated types, which is the highest in the prolapse sequestered type.

By combining frequency modulation Chirp Z transform ion mobility spectrometers (IMS) and multi nozzle electrospray array ionization source, a method of NanoESI-Chirp Z transform ion mobility spectrometry-high performance liquid chromatography was developed for the determination of n-alkyl ammonium bromide compounds.The parameters of NanoESI-Chirp Z transform IMS such as electric field intensity, solvent composition, and solution flow rate were investigated and optimized.Subsequently, four kinds of n-alkyl ammonium bromide compounds were respectively detected by this developed Chirp Z transform method and Fourier transform method, and the obtained results were compared.The result indicated that the optimum conditions were electric field intensity of 4.5 kV, and ESI solution flow rate of 8 μL/min.Then a test mixture containing tetrabutylammonium bromide, tetrapentylammoniumbromide, tetrahexylammonium bromide, tetraheptylammonium bromide, tetranoctylammoniumbromideandtetrakis(decyl) ammonium bromide was successfully separated and determined by the HPLC-nanoESI-Chirp Z IMS method.Chirp Z transform method provided higher signal to noise ratio compared to conventional signal averaging method, and was superior to FT method in the determination of drift time.

Objective: To evaluate the influence of metabolic syndrome (MS) on coronary lesions in elder patients with hypertension.
Methods: A cohort of 210 hypertensive patients at the age of 60 years or elder who received coronary angiography in our hospital from 2012-06 to 2013-01 were studied. The patients were divided into 2 groups, MS group,n=85 and Non-MS group,n=125 patients without MS. The coronary lesion distribution, number and Gensini score were analyzed and compared between 2 groups.
Results: Compared with Non-MS group, MS group showed increased BMI, TG, HDL-C and fasting blood glucose, allP<0.001. There were no real differences for lesion distribution in left main, left anterior descending branch and the number of patients with normal coronary artery between 2 groups, allP>0.05. MS group had higher Gensini score, more lesions at left circumlfex and right coronary artery, more patients with coronary lesions and more patients with 3-branch disease,P<0.05. Logistic regression analysis indicated that MS is the independent risk factor for coronary lesions and the risk is higher than any single components of MS.
Conclusion: MS is related to the severity of coronary lesions in elder patients with hypertension.

Objective To observe the effects of eluting stents coated with arsenic trioxide(As2O3)and suspended in poly-L-lactic acid(PLLA)on expression of monocyte chemoattractant protein-1 (MCP-1)and interleukin-6(IL-6)and to assess the effects of As2O3 eluting stents on local inflammatory reaction in injured coronary arteries in pigs. Methods Bare metal stents,rapamycin eluting stents and As2O3-eluting stents were randomly and double-blindly implanted into the anterior descending branches,circumflex branches and right coronary arteries in eight pigs.Animals were sacrificed and coronary arteries were isolated 7 days after stents implantation.The expression levels of protein and mRNA of MCP-1 and IL-6 were determined by Western blot analysis and reverse transcription polymerase chain reaction(RT-PCR),and the inflammatory cell infiltration was observed by HE staining and immunohistochemistry. Results Compared to bare metal stents,As2O3-eluting stents and rapamycin-eluting stents identically and markedly inhibited the protein expression level of MCP-1(0.421±0.055 and 0.406±0.042 vs.0.857±0.053,P＜0.01)and IL-6(0.151±0.032 and 0.146±0.051 vs.0.551±0.032,P＜0.01)and correspondingly lowered the mRNA expression level of MCP-1(0.338±0.047 and 0.327±0.051 vs.0.724±0.027,P＜0.01)and IL-6(0.531±0.052 and 0.523±0.061 vs.1.015±0.041,P＜0.01),and significantly reduced the inflammatory cell infiltration of injured coronary arteries in pigs. Conclusions As2O3-eluting stents can effectively inhibit the expressions of MCp-1 and IL-6 and reduce the inflammatory cell infiltration of injured coronary arteries in pigs.

BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P ＜ 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.