Objective To investigate the inhibitory effects of RNA interference(RNAi)on the expression of matrix metalloproteinase-9(MMP-9)gene and invasiveness and adhesion of ovarian cancer cells.Methods Four groups of different specific target sequence in coding region of MMP-9 and one non- specific sequence were chosen,which were Sitel,Site2,Site3,Site4 and Site5.Small interference RNA (siRNA)expression cassettes(SEC)were constructed by PCR and transfected into ovarian cancer HO- 8910PM cells.RT-PCR and western blot were used to detect mRNA and protein expression of MMP-9 gene; the abilities of invasion and adhesion were detected by Matrigel invasion assay and cell adhesion assay. Results The expression of MMP-9 was inhibited and the inhibitory effects of different sequence were varied.The mRNA expression was 0.64?0.06,0.47?0.07,0.55?0.10 in Sitel,Site2,Site3 group, and protein expression was 0.30?0.09,0.27?0.08,0.37?0.12,respectively.Site2 group had the most efficient inhibitory effect,followed by Sitel and Site3 groups.Cell growth curve revealed that cell growth was significantly inhibited in Site2 group.Invasiveness and adhesion were significantly reduced,the inhibitory rate on invasion in Site1,Site2,Site3 groups were 50.0%,50.0% and 37.5%,respectively;the inhibitory rate on adhesion in Site1,Site2,Site3,Site4 groups were 43.8%,48.8%,33.9%,24.2% at 60 min and 41.6%,40.2%,35.1%,16.0% at 90 min,respectively.Conclusions RNAi exists in ovarian HO-8910PM cells.MMP-9 siRNA can specifically down-regulate MMP-9 expression and lead to the inhibition of invasiveness and adhesion in ovarian cancer cells.

OBJECTIVETo investigate the changes of CD4 + CD25 + Foxp3 + regulatory T cells in the peripheral blood mononuclear cells (PBMC) and their association with insulin resistance in different stages of prostate cancer (PCa).

METHODSUsing flow cytometry, we counted the CD4+ CD25 + Foxp3 + regulatory T cells in the PBMCs of 62 PCa patients (5 cases of TNM stage I, 16 cases of stage II, 21 cases of stage III, and 20 cases of stage IV) and 42 normal healthy controls, and calculated their proportion in the CD4+ T-lymphocytes. We determined the levels of fast blood glucose (FBG) and fast insulin (FINS) for the insulin resistance index (HOMA-IR), obtained the serum IGF-1 level by ELISA, and analyzed the relationship of the count and proportion of CD4+ CD25+ Foxp3+ regulatory T cells with insulin resistance by comparison between the PCa patients and normal healthy controls.

RESULTSCompared with the control group, the PCa patients showed significantly increased HOMA-IR (3.68 ± 1.42 vs 6.68 ± 1.66), decreased level of serum IGF-1 ([164.56 ± 30.58] vs [96.39 ± 21.21] ng/ml), and elevated count ([1.99 ± 0.78 ] x 10(7) vs [3.55 ± 0.29] x 10(7)) and proportion ([5.33 ± 0.65] vs [13.88 ± 0.96]%) of CD4 + CD25 + Foxp3 regulatory T cells in the PBMCs. The TNM stage was correlated positively with the count and percentage of CD4 + CD25+ Foxp3 + regulatory T cells and HOMA-IR, but negatively with the level of serum IGF-1. Meanwhile, the count and percentage of CD4 + CD25 + Foxp3 + regulatory T cells were found to have a positive correlation with HOMA-IR (r = 0.722 and 0.689, P < 0.01) but a negative correlation with the level of serum IGF-1 (r = -0.747 and -0.896, P < 0.01).

CONCLUSIONThe count and proportion of CD4+ CD25 + Foxp3 + regulatory T cells in the peripheral blood and insulin resistance increase with the elevated stage of PCa. CD4 + CD25 + Foxp3 + regulatory T cells may be involved in the occurrence and progression of PCa by regulating insulin resistance.

Aged ; CD8-Positive T-Lymphocytes ; Case-Control Studies ; Disease Progression ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Hyperinsulinism ; Insulin ; blood ; Insulin Resistance ; Insulin-Like Growth Factor I ; metabolism ; Leukocytes, Mononuclear ; Lymphocyte Count ; Male ; Prostatic Neoplasms ; blood ; immunology ; pathology ; T-Lymphocytes, Regulatory

OBJECTIVETo investigate the changes in cerebral blood flow in patients with maxillary defect treated with prosthesis insertion.

METHODSThirty patients with maxillary defect receiving obturator prosthesis insertion were enrolled with another 30 subjects without dentition defect as the control. The cerebral blood flow rate was recorded before and at 5 and 10 min during mastication, and the results were analyzed statistically.

RESULTSThere was no significant difference in Vs, Vd or Vm between the two groups at the time points for measurement.

CONCLUSIONThe blood supply by the middle cerebral artery is similar between the patients receiving obturator prosthesis insertion for maxillary defect and the subjects with full denture.

Adult ; Aged ; Case-Control Studies ; Cerebrovascular Circulation ; Denture, Complete ; Female ; Humans ; Male ; Mastication ; physiology ; Maxilla ; injuries ; Maxillofacial Prosthesis ; Middle Aged ; Middle Cerebral Artery

0.05).Conclusion MK mRNA overexpresses in the breast cancer tissues.It might be considered to be a reference indicator for deter- mining the angiogenesis and invasion of breast carcinoma.

OBJECTIVETo observe the inhibitory effects of antisense MMP-9 oligodeoxynucleotides on invasiveness and adhesion ability in vitro of ovarian cancer cells, and to investigate the mechanisms of action.

METHODSMMP-9 antisense oligonucleotides were transfected by lipofectinmin into ovarian cancer cell line HO-8910PM cells expressing MMP-9 induced with fibronectin. RT-PCR, Western blot and gelatin zymography were used to detected MMP-9 expression of mRNA and protein and enzymatic activity. The ability of invasion and migration of ovarian cancer cells was assayed in Transwell cell culture chamber. Cell adhersion assay was carried out in a microculture well pre-coated with fibronectin.

RESULTSMMP-9 expressions of mRNA and protein were significantly decreased in the antisense-transfected cells. Comparing with the control group, the inhibition rate was 34. 8% and 42. 5% , respectively (P <0. 05). Its gelatin enzymatic activity was inhibited. Matrigel invasion assay and Transwell migration assay revealed markedly reduction in invasion and migration for the antisense group. The inhibition rates were 22. 4% and 24. 8% , respectively. The adhesion ability was also reduced. The inhibition rates were 49. 8% and 38. 3% at 60 min and 90 min, respectively.

CONCLUSIONMMP-9 down-regulation can significantly inhibit the ability of invasion and attachment of ovarian cells in vitro. MMP-9 may play an important role in invasion and metastasis of ovarian cells and potentially be a molecular target of blocking invasion and metastasis of ovarian cancer.

Blotting, Western ; Cell Adhesion ; genetics ; physiology ; Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Down-Regulation ; Female ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Oligodeoxyribonucleotides, Antisense ; genetics ; Ovarian Neoplasms ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo retrospectively study medium-term follow-up outcomes of total hip arthroplasty (THA) for patients with ankylosing spondylitis (AS).

METHODSFrom January 2000 to December 2008, 67 patients (88 hip joints) with AS were treated with all ceramic interface THA. And 55 patients (74 hips) were finally followed up. Among them, there were 30 males and 25 females, with an average age of 32.6 (ranged 19 to 58) years old. Sixty-one hips were treated with biological prosthesis and 13 hips were treated with hybrid prosthesis. Fifty-five patients were followed up at least 5 years, with an average of (75.2 +/- 8.6) months. Clinical symptoms and radiography information were evaluated after follow-up.

RESULTSHarris hip score were significantly improved from 30.8 +/- 7.0 preoperatively to 85.2 +/- 5.5 at the last follow-up (P<0.01). The hip movement range increased from (21.2 +/- 8.5) degrees preoperatively to (142.0 +/- 10.2) degrees postoperatively (P<0.01). The 5-year survival of prosthesis was 95.9%. One patient were renovated because of internal wall broken caused by injury, 1 was renovated for infection, 1 was renovated for fracture arround femoral stem prostheses, and 1 was treated with conservative treatment by dislocation. Three cases with abnormal sound were cured with non-operation. 7 cases with heterotopic ossification were not treated, 2 cases with thigh pain received conservative treatment. Bone dissolve around prosthesis, loose and sink of femur and acetabulum prosthesis were occurred in other cases.

CONCLUSIONTHA for the treatment of AS is a reliable method, which has a satisfied medium-term follow-up outcomes.

Adult ; Arthroplasty, Replacement, Hip ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spondylitis, Ankylosing ; surgery ; Treatment Outcome ; Young Adult

The structure analysis of porcine hemoglobin alphabeta dimer and the calculation of solvent accessible surface of the amino acids showed the epsilon-amino groups of the lysine are suitable for modification by polyethylene glycol (PEG). The modification of the lysine residues will not affect the carring oxygen capacity of Hb. Three types of linker have been designed to connect PEG and porcine hemoglobin. The lysines between porcine and bovine hemoglobin (pHb and bHb) are highly conserved, but the solvent accessible surface of conserved lysines are different. These suggested that the properties of homologous proteins are similar in pHb and bHb, but the characteristic derived from the homology analysis will be deviated from the actual status. The results of molecular dynamics simulation suggested that the chemical modified porcine hemoglobin would be no immunogenicity.

In this study, we observed the levels of adrenomedullin (ADM) and proadrenomedullin N-terminal 20 peptide (PAMP) in myocardium and aorta of spontaneously hypertensive rats (SHRs) in comparison with Wistar-kyoto (WKY) rats. Contents of ADM and PAMP were measured by radioimmunoassay (RIA) in plasma, myocardium and aorta. The amount of Pro-ADM mRNA of myocardium and aorta was determined by competitive quantitative reverse transcription polymerase chain reaction (RT-PCR). In SHRs the amounts of Pro-ADM mRNA of myocardium and aorta were 66.7% (P<0.01) and 73% (P<0.01) higher than those in WKY rat, respectively. In SHRs, the levels of ADM in plasma, myocardium and aorta were 29%, 76.7% and 79% (all P<0.01) higher than those in WKY rats, respectively. The level of PAMP in SHRs was increased by 42.5% in plasma (P<0.01), 47.2% in myocardium (P<0.0.1) and 27.3% in aorta (P<0.05) compared to WKY rats, respectively. In addition, the ratio of ADM content to PAMP content in SHRs group was increased compared with that in WKY group (2.0+/-0.25 vs 1.64+/-0.3 and 2.2+/-0.18 vs 1.56+/-0.28, in myocardium and aorta, respectively, P<0.01). These results suggest that ProADM gene expression is up-regulated and the increase in ADM and PAMP is different in SHRs. The significance of inconsistency of increase in ADM and PAMP in SHRs needs to be further investigated.
Adrenomedullin ; genetics ; metabolism ; Animals ; Aorta ; metabolism ; Female ; Male ; Myocardium ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Up-Regulation

Virus inactivation of plasma can be achieved by methylene blue/photochemical method. To investigate the effect of this method on immunological properties and biochemical functions of plasma components, the virus-inactivation method was performed on single-donor plasma that was exposed to visible light (40,000 lux) at room temperature for 1 h in the presence of 1 micro mol/L methylene blue. The results showed that activities of the factor VIII, PT and APTT were decreased to a certain degree while most of other plasma proteins were not affected significantly. Human plasma components including albumin, glucose and minerals as well as plasma pH were also not affected. By using different electrophoreses and immunochemical techniques, no neoantigens were found in photodynamically treated plasma and electrophoretic mobility revealed identical patterns for untreated and treated plasma. In conclusion, methylene blue/photochemical method dose not considerably influence the properties of major of plasma components.
Blood Coagulation Factors ; metabolism ; Complement C3 ; metabolism ; Electrophoresis ; methods ; Factor VIII ; metabolism ; Humans ; Hydrogen-Ion Concentration ; Light ; Methylene Blue ; pharmacology ; Partial Thromboplastin Time ; Plasma ; drug effects ; metabolism ; radiation effects ; Thrombin Time ; Time Factors ; Virus Inactivation ; drug effects ; radiation effects

OBJECTIVETo study MMP-2, MMP-9, TIMP-2 and TIMP-1 expression, and their association to invasion and metastasis of neuroblastoma (NB).

METHODSThe staining status was compared of MMP-2, MMP-9, TIMP-2 and TIMP-1 in cryostat sections of tumor tissue in 35 NB patients by immunohistochemistry.

RESULTSStrong expression of MMP-2 was detected only in 2 patients with early stage NB (group A without metastasis), but in 9 and 10 respectively with advanced stage NB (group B with local metastasis and group C with distant metastasis) (compared to group A, P < 0.01). Strong MMP-9 staining was found in 4, 8 and 11 patients for group A, B and C patients (group A vs group C, P < 0.05). The expression of TIMP-2 was the strongest in 4 group A patients, but it decreased with progression of the disease. There was no statistical difference in TIMP-1 expression among the three groups of patients.

CONCLUSIONMMP-2, MMP-9 expression may be related to metastasis and progression of neuroblastoma, while TIMP-2 may have an inhibitory effect.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Lymphatic Metastasis ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Neuroblastoma ; enzymology ; metabolism ; secondary ; Retroperitoneal Neoplasms ; enzymology ; metabolism ; pathology ; Testicular Neoplasms ; enzymology ; metabolism ; secondary ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism