Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence found within several glycoprotein ligands and also in snake-venom disintegrins. Adinbitor, a protein with 73 amino acid residues including 12 cysteins and an RGD motif, was cloned from Gloydius blomhoffi brevicaudus in my laboratory. As a new member of disintegrin family, adinbitor can inhibit both human platelet aggregation induced by ADP and angiogenensis in vivo and in vitro, the typical characters of disintegrin family. To separate the effect of inhibiting platelet aggregation from that of inhibiting angiogenensis, the motif KGD was introduced into adinbitor cDNA to replace RGD by site-directed and PCR-based mutagenesis. The recombinant protein (recombinant adinbitor (KGD)) was expressed in E. coli BL21 and purified through the His· Bind affinity chromatography. Recombinant adinbitor (KGD) could inhibit ADP-induced platelet aggregation with IC50 value of 85 nmol/L. Considerably, it was a more effective inhibitor on platelet aggregation than recombinant adinbitor (RGD), which has an IC50 of 150 nmol/L. Interestingly, recombinant adinbitor (KGD) has no potency in inhibiting angiogenesis in vivo compared with recombinant adinbitor (RGD). These findings showed that KGD containing adinbitor was more suitable for inhibiting ADP-induced human platelet aggregation as a potential and specific inhibitor of human platelet aggregation, which might have promising therapeutic potential as an antithrombotic agent.

OBJECTIVETo study the role of lateral collateral ligament complex on the posterolateral rotatory instability and the relationship between the radiocapitellar ratio (RCR) and the injury of lateral collateral ligament complex on X-ray images.

METHODSTwenty elbow joints from fresh-frozen adult cadavers were used to make osteo-ligamentous elbow specimens. The specimens were fixed with a self-made device to maintain posterolateral rotatory instability of the elbow joint. All the specimens were divided into two groups: group A and group B. Surgical procedures were carried out as follows in the lateral structures of group A: A1, intact specimen; A2, transection of radial ulnar collateral ligament firstly; A3, transection of annular ligament secondly; A4, final transection of the radial collateral ligament. The procedures in group B were carried out as follows: B1, intact specimen; B2, transection of the radial collateral ligament firstly; B3, transection of the annular ligament secondly; B4, final transection of the radial ulnar collateral ligament. Lateral X-ray films of elbow joint were taken, and the radiocapitellar ratio (RCR) was measured by using PACS. All analysis was performed with SPSS 17.0 software.

RESULTSGroup A: the increases in RCR had statistical differences among A1, A2, A3, and A4 groups. Group B: the increases in RCR had no statistical differences among B1, B2 and B3 groups; but the increase in RCR in group B4 was more than that in B1, B2 and B3 groups.

CONCLUSIONThe radial ulnar collateral ligament is a key structure to maintain posterolateral rotatory stability;the radial collateral ligament and the annular ligament are the secondary important structures. There are 4 grades of the posterolateral rotatory instability of the elbow, according to the X-ray imaging classification.

Collateral Ligaments ; injuries ; Elbow Joint ; diagnostic imaging ; physiopathology ; Female ; Humans ; Joint Instability ; diagnostic imaging ; etiology ; physiopathology ; surgery ; Magnetic Resonance Imaging ; Male ; Radiography

OBJECTIVETo explore the correlation of males'age with sperm apoptosis, sperm DNA integrity and other seminal parameters.

METHODSWe collected 104 semen samples and divided them into three groups according to the males' age: <35 yr (n = 43), 35 -39 yr (n = 31), and > or = 40 yr (n = 30). Based on the WHO Laboratory Manual (4th ed), we detected the seminal parameters, calculated the percentage of apoptotic sperm by flow cytometry (FCM), determined sperm DNA integrity by Acridine orange staining, and compared the results among the three groups.

RESULTSThere were no statistically significant differences among the < 35 yr, 35 -39 yr and > or = 40 yr groups in semen volume ([2.87 +/- 0.89] ml vs [2.98 +/- 1.09] ml vs [2.65 +/- 0.95] ml), sperm concentration ([60.40 +/- 25.43] x 10(6)/ml vs [69.74 +/- 28.33] x 10(6)/ml vs [55.97 +/- 27.22] x 10(6)/ml) (P>0.05). The percentage of progressively motile sperm was significantly lower in the > or = 40 yr ([39.00 +/- 8.35 %) than in the <35 and 35 -39 yr groups ([48.73 +/- 9.89]% and [45.65 +/- 10.55]%) (P<.0.1), and so was the percentage of morphologically normal sperm in the > or = 40 yr than in the < 35 yr group ([11.11 +/- 8.26]% vs [16.43 +/- 8.75 ]%, P<0.01). The percentage of apoptotic sperm was markedly higher in the > or = 40 yr than in the <35 yr group ([11.82 +/- 5.77]% vs [7.04 +/- 3.50]%, P<0.01), while the sperm DNA integrity significantly reduced in the > or = 40 yr group ([75.52 +/- 10.60]%) as compared with the <35 yr ([86.55 +/- 5.60])% and 35 -39 yr group ( [81.39 +/- 8.94]%) (P<0.01). The males' age was correlated positively with the rate of sperm apoptosis (P<0.01), and negatively with sperm DNA integrity and the percentage of progressively motile sperm (P<0.01).

CONCLUSIONThe advance in males' age increases sperm apoptosis and reduces sperm progressive motility, normal morphology and DNA integrity.

Adult ; Age Factors ; Apoptosis ; genetics ; DNA ; Flow Cytometry ; Humans ; Infertility, Male ; genetics ; Male ; Sperm Count ; Sperm Motility ; Spermatozoa ; cytology

OBJECTIVETo study the influence of male age on the outcome of conventional IVF-ET.

METHODSBased on male age, 170 couples undergoing conventional IVF-ET were divided into three groups: <35 yr (n = 60), 35 -39 yr (n = 77) and > or = 40 yr (n = 33). We observed the rates of fertilization, cleavage, good quality embryo, implantation, clinical pregnancy and abortion in different groups.

RESULTSThere were no significant differences among the three groups in semen volume ([3.10 +/- 1.22] ml vs [2.84 +/- 1.05] ml vs [2.80 +/- 0.79] ml), sperm concentration ([54.23 +/- 26.07] x 10(6)/ml vs [60.27 +/- 24.80] x 10(6)/ml vs [60.21 +/- 27.42] x 10(6)/ml) and sperm viability ([53.93 +/- 13.25]% vs [56.10 +/- 16.58]% vs [51.82 +/- 15.45]%) (P>0.05). The men of the > or = 40 yr group showed a significantly lower percentage of grade a + b sperm ([40.97 +/- 11.91]%) than those of the <35 and 35 - 39 yr groups ([48.47 +/- 11.78]% and [46.84 +/- 13.51]%) (P<0.05), and morphologically normal sperm ([11.76 +/- 5.97]%) than those of the <35 yr group ([15.25 +/- 6.94]% (P<0.05). The rates of fertilization, cleavage, good quality embryo, implantation, clinical pregnancy were 81.52%, 82.61%, 52.33%, 18.06% and 33.33% in the > or = 40 yr group, with no significant differences from those of the <35 yr group (83.18%, 82.68%, 56.99%, 22.40% and 40.00%) and the 35 - 39 yr group (78.78%, 80.66%, 55.01%, 21.74% and 38.96%) (P>0.05), while the abortion rate was markedly increased in the > or = 40 yr group as compared with the <35 yr group (36.36% vs 8.33%, P>0.05).

CONCLUSIONIncreasing male age is related with decreasing percentages of progressively motile sperm and morphologically normal sperm, but not obviously with the rates of fertilization, good quality embryo, implantation, pregnancy and abortion.

Adult ; Age Factors ; Female ; Fertilization in Vitro ; Humans ; Male ; Middle Aged ; Paternal Age ; Pregnancy ; Pregnancy Outcome

Objective To evaluate the short-term efficacy and safety of percutaneous liver puncture for local management of portal vein thrombosis(PVT).Methods Variations in thrombus,blood flow,and laboratory examination results were observed before and after percutaneous liver puncture in 197 patients with PVT,and the occurrence of comorbidities was recorded and followed up for one year after treatment.Results After treatment,the thrombus in the main portal vein vessels almostly disappeared in 119 patients(60.41％)with PVT,the thrombus had a significant reduction in 57 patients(28.93％),and the thrombus had a smaller change or an increase in 21 patients(10.66％);146 patients(74.11％)had smooth blood flow in the main portal vein vessels,29 patients(14.72％)showed significant improvement in blood flow,and 22 patients(11.17％)showed no significant improvement or worsening of blockage.The mean portal venous pressure was significantly lower than that before treatment(P＜0.001);thrombin time,activated partial thromboplastin time,and prothrombin time were prolonged compared to those before thrombolysis(P＜0.001),and fibrinogen were reduced compared to those before thrombolysis(P＜0.001).A total of 35 patients(17.77％)occured comorbidities during treatment.One year after treatment,196 patients(99.49％)with PVT survived,of which thrombus essentially disappeared in 141(71.94％),thrombus stabilized(or decreased)in 42(21.43％),and thrombus increased in 13(6.63％).Conclusion percutaneous liver puncture for local management of PVT is effective and reliable in the short-term and requires standardized management of the entire process.

Objctive To investigate the protective effect of resveratrol on oxidative stress damage of follicular granulosa cells induced by hydrogen peroxide. Methods Granulosa cells were collected from the follicular fluid of in vitro fertilization(IVF) patients after oocyte retrieval and cultured. The cultured granulosa cells were divided into four groups: control group, injury model group, 10 μmol / L resveratrol group and 50 μmol / L resveratrol group. Cell viability was determined by CCK-8 assay, malondialdehyde(MDA) content by thiobarbituric(TBA) assay, superoxide dismutase(SOD) level by water soluble tetrazdium-1(WST-1) assay, apoptosis by AnnexinV-FITC / PI double-staining flow cytometry, Bcl-2 and Caspase-3 protein expression by Western blotting, and progesterone secretion by competitive ELISA. Resutls Compared with the control group, the cell viability, SOD level, Bcl-2 protein expression and progesterone secretion were significantly decreased in the injury model group, while MDA content, apoptosis rate and Caspase-3 apoptotic protein expression were significantly increased (P<0. 05) . Compared with the injury model group, the 10 μmol / L resveratrol group showed no statistically significant differences in various parameters; however, the cell viability, SOD level, progesterone secretion, and Bcl-2 and silent information regulator factor 2 related enzyme 1(SIRT1) protein expression were significantly increased, and the MDA content, apoptosis rate, and Caspase-3 apoptotic protein expression were significantly decreased in the 50 μmol / L resveratrol group (P < 0. 05) . Conclusion 50 μmol / L resveratrol can increase the activity of SIRT1, enhance the anti-oxidation and anti-apoptosis ability of granulosa cells and improve the function of granulosa cells.