BACKGROUND: Preliminary study has proven that adult human bone marrow-derived mesenchymal stem cells (MSCs) suppress peripheral blood lymphocytes proliferation.But the mechanism was still to be investigated.OBJECTIVE: To study the negatively regulatory effect of adult human MSCs on allogeneic lymphocyte proliferation by cell-free condition.DESIGN,TIME AND SETTING: Cytological observation in vitro,which was performed in the Lanzhou General Hospital,Lanzhou Military Area Command of Chinese PLA between October 2005 and December 2007.MATERIALS: The bone marrow sample was provided by the allo-transplantation donor.The peripheral blood lymphocytes were provided by the healthy volunteer.METHODS: Adult human MSCs were separated with Percoll + adherence method.Allogeneic peripheral blood lymphocytes were obtained from healthy donors with Ficoll solution and the cell concentration was adjusted as 2×109/L for use.100μ L MSCs culture supernatant was taken out in 96-well plates.The groups were following: A superuatant + 3-day MSCs culture media (100 μ L/well); B superuatant + phytohemagglutini (PHA; 1 g/L,5 μ L); C medium + LG-DMEM culture media containing 10% fetal bovine serum (100 μ L); D medium + PHA (1 g/L,5 μ L).The cells were incubated at 37 ℃ with 5% CO2 in a fully humidified atmosphere for three days.MAIN OUTCOME MEASURES: Effect of MSCs supematant on proliferation and transformation of variant lymphocytes.RESULTS: Peripheral blood lymphocytes proliferation was suppressed as compared with the blank control group and PHA group after MSCs culture,and the inhibition ratio was 9.00% (P ＜ 0.05).When lymphocytes were stimulated by PHA,the suppression effects were even stronger and the inhibition ratio was 20.91% (P ＜ 0.01).CONCLUSION: Adult human MSCs supernatant can suppress peripheral blood lymphocytes proliferation and transformation; furthermore,PHA can enhance the inhibitory effect,suggesting the negative regulation is at least in part due to indirectly inhibiting lymphocytes via soluble cytokines.

AIM:ToinvestigatetheeffectofP21-activatedkinase1(PAK1)andlymphoidenhancer-binding factor 1(LEF1) on the proliferation of esophagus cancer cells .METHODS:Real-time PCR was applied to detect the mR-NA expression of PAK1 and LEF1 in the esophagus cancer tissues .MTT assay were used to measure the proliferation of hu-man esophagus cancer cell line KYSE transfected with PAK 1 and LEF1.RESULTS: The mRNA expression of PAK1 in the esophagus cancer tissues was lower than that in control group (P<0.05).The mRNA expression of LEF1 and tran-scription factor 4 (TCF4) in the esophagus cancer tissues was higher than that in control group (P<0.05).The prolifera-tion of KYSE cells with over-expression of PAK1 and LEF1 was higher than that in control group .No significant change of apoptosis between the KYSE cells with over-expression of PAK1 and LEF1 and control group was observed .CONCLU-SION:The expression of PAK1 decreases and the expression of LEF 1 increases in esophagus cancer tissues .LEF1 domi-nantly regulates the proliferation of esophagus cancer cells .

Objective:To study the chemical constituents of Buddleja lindleyana.Method:Separation by chromatographic methods and identification by spectral analysis.Result:Seven compounds vanillic acid,octacosanoic acid，β-sitosterol-3-O-β-D-glucopyranoside,stigmasterol-3-O-β-D-glucopyranoside,α-spinasterol-3-O-β-D-glucopyranoside,betulin acid were isolated.Conclusion:All the compounds were obtained from this plant for the first time.

OBJECTIVESTo analysis the correlation between parameters of sperm chromatin structure assay(SCSA) and semen routine analysis, and to discuss the reliable methods of semen quality evaluation.

METHODSFive hundred and eleven semen samples were detected to analyse the mutiple-parameter correlation between results of SCSA (COMP alpha t) and semen routine analysis.

RESULTSThe parameters that have low-level positive correlation(r: 0.10-0.30) with denatured sperm percentage(COMP alpha t) were viscosity, ejaculation interval, abnormal sperm ratio, concentration of grade c sperm; those having low-level negative correlation(r: -0.30(-)-0.10) were VDL, VSL and VAP; those having mid-level positive correlation (r: 0.30-0.70) were sperm concentration, percentage of grade d sperm; those having mid-level negative correlation (r: -0.70(-)-0.30) were MAD, percentage of grade a sperm and survival rate.

CONCLUSIONSFlow cytometry can be used to evaluate the percentage of denatured or injured sperm rapidly, correctly and simply. The result (COMP alpha t) correlates partly with semen parameters, and it is not a conclusively parameter compared with routine semen analysis. It is important to use SCSA to evaluate productivity under the above situation.

Adult ; Chromatin ; chemistry ; DNA Damage ; Flow Cytometry ; Humans ; Hydrogen-Ion Concentration ; Male ; Semen ; Sperm Count ; Spermatozoa ; ultrastructure ; Viscosity

OBJECTIVESTo detect the sperm DNA damage and to evaluate its significance in male reproductive using single cell gel electrophoresis (SCGE).

METHODSFour hundred and eighteen sperm samples were analysed using the computer assisted analysis system and SCGE. The sperms samples were divided into five grades according to the extent of the sperm nuclear DNA damage.

RESULTS1. When the sperm density is less than 20 x 10(6)/ml, the occurence of grade II and III are increased significantly; 2. In the unmotile grade d sperm the occurence of grade I comet amounts was 5.39%, the occurence of grade II and III was remarkably increased. There was a evidently variance between the grade d and grade a + b sperm.

CONCLUSIONSSCGE can be used to detect the sperm DNA breakage and to evaluate the sperm quality and damage.

Adolescent ; Adult ; Comet Assay ; methods ; DNA Damage ; Humans ; Male ; Sensitivity and Specificity ; Sperm Count ; Sperm Motility ; genetics ; Spermatozoa ; cytology ; metabolism

OBJECTIVETo study the influence of varicocele on sperm chromatin structure and sperm motility.

METHODSRoutine semen analysis and sperm chromatin structure assay (SCSA) were performed in a varicocele group (n=74) and a control group (n=89).

RESULTSSperm concentration (41.4 +/- 38.7] x 10(6)/ml) grade a+b sperm percentage ([31.7 +/- 16.9]% and sperm viability ([62.8 +/- 22.2]%) in the varicocele group were evidently lower than those ([80.9 +/- 63.1] x 10(6)/ml, [46.8 +/- 20.5]%, [77.2 +/- 17.5])% in the control group (P < 0.05) and so were VCL, VSL and VAP ([37.4 +/- 12.5 microm/s, [23.4 +/- 7.8] microm/s, [26.5 +/- 8.2] microm/s) in the varicocele group than those ([42.4 +/- 10.7] microm/s, [27.3 +/- 7.3] microm/s, [30.7 +/- 7.8] microm/s) in the control (P < 0.05). MAD was increased (P < 0.01), and the COMP alphat of SCSA (23.2 +/-16.2) was obviously higher in the former than in the latter (14.1 +/- 11.8) (P < 0.05).

CONCLUSIONVaricocele causes damage to sperm DNA and changes sperm motility, which may result in male infertility.

Adolescent ; Adult ; Chromatin ; genetics ; metabolism ; DNA Damage ; Humans ; Male ; Sperm Count ; Sperm Motility ; genetics ; physiology ; Spermatozoa ; cytology ; metabolism ; Varicocele ; genetics ; physiopathology

OBJECTIVETo evaluate the inflicted by radar electromagnetic radiation to the sperm DNA of radar operators.

METHODSSperm concentration, viability, motility, sperm abnormality were determined by routine sperm analysis and sperm chromatin structure assay (SCSA) in the highly exposed group(n = 88), lowly exposed group(n = 143) and control group(n = 39).

RESULTSSperm motility, viability of the highly exposed group reduced compared with that of the lowly exposed group and control group, while sperm abnormality increased. The COMP alpha reduction of the highly exposed group indicated that the highly exposed group had a medium fertility potential. The multifactor variable analysis showed that daily working time was a dangerous factor in sperm abnormality and abstinence time was a dangerous factor in the parameter of SCSA.

CONCLUSIONSRadar radiation inflicts damage to male reproduction system and it is important to take protective measures.

Adolescent ; Adult ; Chromatin ; ultrastructure ; DNA Damage ; Dose-Response Relationship, Radiation ; Electromagnetic Phenomena ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Radar ; Spermatozoa ; radiation effects ; ultrastructure

OBJECTIVETo identify the effects of overtraining on human sperm DNA.

METHODSMolecular epidemiological investigation of 249 men from different groups (training and non-training) was carried out by using flow cytometer to detect the integrity and damage of in situ DNA of sperm nucleus, and sperm chromatin structure assay was performed.

RESULTSThe average COMPalpha(t) in training group was 11.02% while that in control group was 5.90% (P < 0.01). COMPalpha(t) was significantly correlated with sperm activity (r = 0.41, P < 0.05).

CONCLUSIONOvertraining could induce sperm DNA injury and affect sperm activity, thus to decrease the potentiality of reproduction.

Adult ; Chromatin ; genetics ; metabolism ; DNA Fragmentation ; Exercise ; physiology ; Humans ; Male ; Sperm Motility ; physiology ; Spermatozoa ; cytology ; metabolism

A Kinase Anchor Proteins ; metabolism ; Animals ; Cell Cycle Proteins ; metabolism ; Lipopolysaccharides ; pharmacology ; Liver ; metabolism ; pathology ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Objective To investigate the clinical efficacy of finger reconstruction in child with second toe transplantation,and evaluate the postoperative appearance and function regarding the reconstructed donor feet.Methods From June 2002 to May 2011,sixteen cases were reconstructed in sub-emergency with second toe transplantation.Two thumbs,eight index fingers,and 6 middle fingers were reconstructed.All patients were followed-up from 12 to 24 months.The functions of reconstructed fingers were analysed.Results All the reconstructed fingers survived.Vascular crisis occurred in 1 patient,and survived after re-anastomosis.Necrosis of skin grafts at the domon site with exposed tedons was seen in 1 ease,and healed after changing dressings.All the reconstructed fingers showed good in growth and development,and performed good functions as grabbing,grasping and nipping.Two-point discrimination was between 6 mm and 10 mm.The donor site of the foot had normal gait,without obvious influence on walking.Also,no pain was complained.Conclusion The method of transplanting the second toe can reconstruct the appearance and function of the finger defects in child,and has little effect on the appearance and motion of feet.It is an effective treatment method.