OBJECTIVETo investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures.

METHODSUsing alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method.

RESULTSThree agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05).

CONCLUSIONThe optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.

Asthenozoospermia ; genetics ; Comet Assay ; standards ; DNA Damage ; DNA Fragmentation ; Humans ; Hydrogen Peroxide ; toxicity ; Male ; Oligospermia ; genetics ; Oxidants ; toxicity ; Reproducibility of Results ; Sperm Count ; Spermatozoa ; drug effects ; enzymology ; Time Factors

Objective Delayed xenograft rejection (DXR) is a major barrier to the long-term xenograft survial.This study evaluated the interaction between human peripheral blood mononuelear cells (PBMC) and porcine endothelial cells (PEC),and the effects of new generation of rabbit antihuman leukocyte polyclonal antibody (newRALG) inhibiting xenogeneic cell-mediated immune responses.Methods newRALG was obtained from rabbits after immunization with activated lymphocytes and monoeytes.PEC were isolated from aorta,and human PBMC were isolated from peripheral blood.Co-cultures of PKH-26 labeled PEC with PBMC were established,newRALG,thymoglobulin,isotype Ig and scavenger receptor (SR) ligand poly G were added into the co-cultures.Cells were collected,then FACS analysis was carried out to detect the up-take of PEC membrane by monocytes and the expression of costimulatory molecules.Lymphocyte proliferative responses to PEC with or without antibody were evaluated by a xenogeneie mixed lymphocyte-endothelial cell reaction (xMLER).Results FACS analysis revealed that monocytes from PBMC-PEC co-cultures became positive for PKH-26 following their interaction with PKH-26 labeled PEC,indicating that they engulfed PEC membranes during activation.PKH-26 positive monocytes up-regulated the CD40 and CD80 expression.Furthermore,SR blockade with poly-G prevented PEC membrane up-take by monocytes,newRALG greatly reduced SR-mediated PEC membrane up-take.The effects of thymoglobulin in inhibiting PEC membrane uptake were limited.xMLER demonstrated strong lymphocyte proliferation in response to PEC,and lymphocyte proliferation was dramatically inhibited by newRALG but not isotype Ig at a dosmdependent manner.Conclusions Monocytes play an important role in xenogeneic immune responses.SR ligand poly G inhibits PEC membrane up-take.newRALG inhibits PEC membrane up-take by monocytes,suggesting that newRALG blocks SR.Additionally,newRALG inhibits lymphocyte proliferation in response to PEC.These results suggest that this new polyclonal preparation may thus impair the initiation of xeno-specific immune responses and prevent xenograft rejection.

Aim To investigate the expression and im-plication of HIF-1α, ROCK-2 , FoxM1 in PC12 cell in-jury induced by lead acetate. Methods PC12 cells were treated with lead acetate at the doses of 100 , 200 and 400 μmol·L-1 . The cell viability was determined by MTT reduction assay and LDH assay, the intracellu-lar production of oxygen species was measured by as-sessing SOD and MDA levels, cell apoptosis was deter-mined by Hoechst 33342 staining, the expressions of HIF-1α, ROCK-2 , FoxM1 , Bcl-2 and Bax were deter-mined by immunoblotting analysis. Results Lead ac-etate induced cell injury in PC12 cells in a dose-de-pendent manner, and it potentiated oxygen radical pro-duction and cell apoptosis. In addition, lead acetate enhanced HIF-1α and ROCK-2 expressions, increased Bax/Bcl-2 ratio and decreased FoxM1 expression. Conclusion Lead acetate can induce PC12 cell apop-tosis, which may be related with the expressions of HIF-1α, ROCK-2 and FoxM1 . Cellular oxidative stress may contribute to the injury as well.

OBJECTIVETo study the effects of rhizoma sparganii and radices zedoariae on hepatic fibrosis.

METHODThe rat immunohepatic fibrosis model was made by intraperitoneal injection of porcine serum and treated with rhizoma sparganii and radices zedoariae. The ALT, GGT, TP, ALb, A/G, IVC, LN, HA and the pathological change of the liver were observed.

RESULTRhizoma sparganii and radices zedoariae could increase TP, ALb, A/G, decrease ALT, GGT, IVC, LN, HA and improve the pathological change.

CONCLUSIONRhizoma sparganii and radices zedoariae can protect hepatic cells, alleviate degeneration and necrosis, recover structure and function, and reduce the proliferation of fibrous tissue.

Animals ; Curcuma ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Liver Cirrhosis ; drug therapy ; pathology ; Magnoliopsida ; chemistry ; Male ; Phytotherapy ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Rhizome ; chemistry

Objective To discuss the semilateral supine position for retroperitoneoscopic adrenalectomy. Methods From Jan. 2006 to Dec. 2008, 36 patients (20 males and 16 females with mean age of 43 years) underwent retroperitoneoscopic adrenalectomy in 60° -70° semilateral supine position. There were adrenal cortex adenomas in 18 cases, pheochromocytoma in 6 cases, adrenal cysts in 3 cases, myelolipoma in 2 cases, gangliocytoma in 1 case, lymphangioma in 1 case, metastatic tumor in 1 case and corticohyporplasia in 4 cases. The mean diameter of the tumors was 2.6 cm( 0.5 - 7.7 cm ). The tumors were superior to the renal pole in 5 cases, anteromedial in 10 cases and superomedial in 17 cases. The three ports that were usually used in lateral position and were placed anteriorly to create retroperitoneal place: the first port was placed 2 -4 cm superior to the iliac crest along the anterior axillary line, the other two were placed just below the costal margin along the midaxillary line and at the same level along the midclavicular line, and dissected along the anterior surface of kidney to its superomedial aspect, so as to avoid the hampering of the kidney in the exposing of the diseased adrenal gland. Results The procedure was completed successfully in all of the cases with the operating time of 37 - 145 min ( mean 69 min) and intraoperative blood loss of 30 - 100 ml (mean 48 ml). Six cases had rupture of peritoneum, which were sutured and the procedure was continued to completion. The postoperative hospital stay was 3 -8 d (mean 5 d ). Thirty-five patients were available for follow-up of 3 - 28 months ( mean 14 months). The case of metastatic tumor died of the primary diseases in the 12th month postoperatively. No other complication was found. Conclusion With this alternative position and ports' location, the procedure of retroperitoneoscopic adrenalectomy could be easier and safer than the conventional position.

Thermodynamic parameters of complexation of naphto-15-crown-5 with four alkaline earth ions in aqueous media was determined using titration microcalorimetry at 298.15 K. The stability of the complexes, thermal effect and entropy effect of the complexation is discussed on the basis of the guest ions structure and the solvent effect. The stability constants tendency to vary with ion radius was interpreted. Complex of naphtha-15-crown-5 with calcium ion is very stable due to the synergism of static electric interaction and size selectivity between the host and the guest.
Calorimetry ; methods ; Crown Ethers ; chemistry ; Ions ; chemistry ; Macromolecular Substances ; analysis ; chemistry ; Metals, Alkaline Earth ; chemistry ; Microchemistry ; methods

To evaluate the clinical outcomes of loupe-assisted intussusception vasoepididymostomy (VE) in the treatment of epididymal obstructive azoospermia (EOA), we retrospectively analyzed data from 49 patients with EOA who underwent two-suture longitudinal intussusception vasoepididymostomy (LIVE) between 2000 and 2007. The data included the surgical method, postoperative motile sperm count per ejaculation, percentage of progressive motile sperm and patency and pregnancy outcomes. There were a total of 49 men undergoing scrotal exploration, and epididymal obstruction was found in all cases. Bilateral or unilateral anastomoses were performed in 40 and 6 men, respectively. The postoperative courses of 42 patients were followed up for more than 6 months, and the courses of 38 patients were followed up for more than 1 year. The overall patency and pregnancy rates were 71.4% and 26.3%, respectively. Moreover, progressive motile sperm was more frequently present in those patients who had undergone anastomosis at cauda than at corpus or caput. Pregnancy was achieved only in those patients who had undergone anastomosis at least on one side of the cauda epididymis. We think that the loupe-assisted method, with a lower overall cost and a simplified surgical procedure, can achieve satisfactory patency outcomes and pregnancy results. Data from this paper also suggest that paternity outcomes occur more frequently after anastomoses at cauda than at corpus or caput.
Adult ; Anastomosis, Surgical ; Azoospermia ; etiology ; pathology ; surgery ; Constriction, Pathologic ; complications ; pathology ; surgery ; Ejaculatory Ducts ; pathology ; surgery ; Epididymis ; pathology ; surgery ; Humans ; Male ; Microsurgery ; methods ; Retrospective Studies ; Sperm Motility ; Treatment Outcome ; Vasovasostomy ; methods

AIM:To investigate the inhibitory effect of sinapine thiocyanate(ST)on hyperglycemia,hyper-lipemia,atherosclerosis and hepatocellular steatosis of ApoE-/-mice with insulin resistance(IR)and the possible mecha-nisms.METHODS:ApoE-/-male mice(n=60)were assigned randomly into control group ,saline group,rosiglitazone group and ST treatment groups(at low,middle and high doses )with 10 mice in each group.The mice in control group were fed with fundamental diet ,while the mice in other groups were fed with high-fat diet for 12 weeks.The mice in ST groups were given gavage with different doses of ST(10,30 and 90 mg· kg-1· d-1)simultaneously,while the mice in rosiglitazone group received gavage with rosigltazone(1.33 mg· kg-1 · d-1 ).In the last 3 weeks,the mice in control group received daily intrape-ritoneal injection of physiological saline ,and IR was induced in other groups by daily intrape-ritoneal injection of dexamethasone(0.8 mg/kg).The blood sample was collected and fasting plasma glucose was tested weekly through tail vein.After all animals fasted for 12 h at the end of the 12th week,they were sacrificed and the levels of fasting insulin,tumor necrosis factor-α(TNF-α),triglyceride,total cholesterol and liver lipids were measured.The li-ver tissue and aortic immobilized sections were detected by HE staining.The expression of the proteins related to liver lipid metabolism and skeletal muscle MAPK signaling pathway was determined by Western blot.RESULTS:ST showed dose-dependently reduced serum lipids ,plasma glucose and TNF-α(P<0.05),delayed hepatocellular steatosis and atheroscle-rosis,and dose-dependently regulated hepatic lipid metabolism signaling molecules(HMGR and SREBP-2)and MAPK signaling molecules(ERK and p38)(P<0.05).CONCLUSION:ST has the biological potential of reducing blood li-pids and relieving IR.The mechanism may be related to the regulation of liver lipid metabolism and skeletal muscle MAPK signaling pathway.

Objective To observe the protective effects of a mixture for protecting liver and supplementing stomach (保肝益胃合剂) on rat acute liver damage induced by carbon tetrachloride (CCl_4). Methods The model of rat acute liver damage was established by injection of CCl_4 2 ml/kg into the abdominal cavity.The rat models were treated respectively by the mixture for protecting liver and supplementing stomach 30 g?kg~(-1)?d~(-1),the polyene phosphatide acid radical choline capsule [Yi Shanfu (易善复), 180 mg?kg~(-1)?d~(-1)],the glycyrrhizic acid diaminogen capsule [Gan Lixin (甘利欣),30 mg?kg~(-1)?d~(-1)] infused into the stomach.The activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) were detected.In the mean time,the liver pathological changes were observed,the degree of liver cell necrosis was evaluated,and the rat mortality was noted in various groups of treatment.Results The values of ALT,AST and the score of liver cell necrosis in the group treated with the mixture for protecting liver and supplementing stomach [(1.168?1.066) kU/L,(1.845?2.212) kU/L,(0.56?0.53) score] were significantly lower than those in the model group [(4.982?3.502) kU/L,(7.030?3.616) kU/L, (1.38?0.92) scores],and all the differences being statistically significant (all P

OBJECTIVETo study the changes of serum interleukin-2 (IL-2), interleukin-8 (IL-8) and immunoglobulin (IgG, IgA, IgM) in patients with esophageal cancer, and to probe the relationship between the levels of IL-2, IL-8, IgG, IgA and IgM and the progress of cancer.

METHODSThe serum levels of IL-2 and IL-8 were detected by enzyme-linked immunosorbent assay for 72 case of primary esophageal cancer, 68 advanced esophageal cancer and 120 healthy controls, and the level of immunoglobulin (IgG, IgA, IgM) in patients with esophageal cancer was dynamically observed.

RESULTSThe IL-2 level in patients with early esophageal cancer [(1.69 +/- 0.53) ng/ml] or late esophageal cancer [(1.11 +/- 0.60) ng/ml] was lower than the control group [(2.78 +/- 0.51) ng/ml] (P < 0.01), the late esophageal cancer group was lower than early esophageal cancer group (P < 0.05). The level of IL-8 in patients with early esophageal cancer [(85.48 +/- 6.14) ng/L] or late esophageal cancer [(121.41 +/- 6.22) ng/L] was much higher than the control group [(54.48 +/- 12.20) ng/L] (P < 0.01), the late esophageal cancer group was much higher than early esophageal cancer group (P < 0.01); There was correlation between the levels of IL-2 and IL-8 and the worsen-extent of the tumour in patients with early esophageal cancer or late esophageal cancer. But the level of IgG [(12.23 +/- 2.50) g/L], IgM [(1.60 +/- 0.80) g/L] in the patients with esophageal cancer compared with the level of IgG [(11.65 +/- 3.70) g/L], IgM [(1.46 +/- 0.71) g/L] in the health control group have no significant difference (P > 0.05), the level of IgA [(3.50 +/- 1.10) g/L] in patients with esophageal cancer Compared with the control group [(1.88 +/- 1.08) g/L] has significant difference (P < 0.01), and along with the worsen-extent of the tumor in patients the level of IgA has the increased tendency.

CONCLUSIONThe IL-8 might accelerate the pathogenesis of esophageal cancer, and the IL-2 might restrain. The positive correlation between the level of IgA and the patients with esophageal cancer is observed in this study; the immune maladjustment of IL-2, IL-8 and IgA might be correlative to esophageal cancer, and the IL-2, IL-8 and IgA levels might be an available index for the severity of esophageal cancer, Which may be of some help for clinic practitioners to judge the progress, curative effect and prognosis of the cancer.

Adult ; Aged ; Case-Control Studies ; Esophageal Neoplasms ; blood ; pathology ; Female ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Interleukin-2 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged ; Neoplasm Staging