Objective　To establish the method of 2-dimensional electrophoresis(2-DE) for proteins of human spermatozoa and to construct a protein map of human spermatozoa. Methods　The sperm pellet was prepared with simple Percoll layer protocol. We studied the effects of various sample preparation methods, loading quantities and isoelectric-focusing protocols on the quality of silver-stained 2-DE map, and constructed a primary protein map of human spermatozoa. Result　Up to 703 protein spots were acquired with sample preparation Method Ⅰwhile only 194～210 spots with Method Ⅱ.With immobilized pH gradients and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(IPG-DALT) we could acquire over 700 spots while only 280～300 with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(ISO-DALT). Conclusion　It is satisfactory to lyse sperm with sample preparation Method Ⅰ and to separate sperm proteins by IPG-DALT for establishing 2-D map of human sperm.

Objective To study the effect of Naotongliting capsule (NLC) on hemorheology and platelet aggregation in blood stasis rats. Method The rat model of blood stasis was established with adrenaline subcutaneous injection in combination with ice-water socking. Aspirin and Yangjiao capsule were selected as contrast medicine to observe the effect of NLC on whole blood viscosity, plasma viscosity, haematocrit, fibrin and platelet aggregation in blood stasis rats. Result NLC could reduce hemorheology and platelet aggregation in blood stasis rats. Conclusion NLC is helpful to adjust hemorheology and plateiet aggregation in blood stasis rats, indicating its good effect of activating blood circulation to remove blood stasis.

This article explains the purpose of setting up the sperm bank, the role of the sperm bank in auxiliary reproduction technology, and exhaustively clarifies the ethical problems in the various aspects of the social life that are caused by the sperm bank, and finally it also proposes some relevant suggestions to these problems.

Objective To analyze the effect of sperm factor on recurrent pregnancy lass.Methods Accordingto WHO criteria,we routinely analyzed the sperm of 21 unexplained recurrent pregnancy loss patients as study group.and 20 proven fertility men as control group.Meanwhile,we evaluated sperm chromosome aneuploidy for chromosomes 18,X and Y using standard FISH procedure,and sperm DNA ingegrity using sperm chromatin structure assay(SCSA)procedure.Results The percentage of normal morphology sperm is(10.8±2.8)%,and DFI value is(21.7±6.7)%,and the percentage of sperm aneuploidy for sex chromosome and chromosome 18 is(0.80±0.14)%and(0.23 ±0.09)%respectivelyin study group.The differences between the two groups were statistically significant(P＜0.01).Conclusion Sperm morphology,DNA integrity and chromosome aneuploidy are the correlative factors to recurrent pregnancy loss.

BACKGROUND:Spermatogonial stem cel s with abilities of differentiation, self renewal and proliferation are a kind of adult stem cel s that can transfer genetic information into offspring, which have great application prospects in medicine, genetics and zoology. OBJECTIVE:To review the source, biological characteristics, and application of spermatogonial stem cel s as wel as self-renewal and molecular regulation underlining these differentiations. METHODS:PubMed and CNKI databases were searched by the first author using key words of“spermatogonial stem cel , biological characteristics, self-renewal, differentiation”in English and in Chinese, respectively, to retrieve relevant articles published from 1990 to 2015. Literatures addressing spermatogonial stem cel s were included, and 46 articles were chosen for further analysis eventual y. RESULTS AND CONCLUSION:Spermatogonial stem cel s can be cultured in vitro, cryopreserved, and genetical y modified as wel as used for al ogeneic or xenogeneic transplantation, al of which contribute to understanding the mechanisms of spermatogenesis, thereby providing new means for treatment of male sterile disease and genetic diseases and providing new hopes for chemotherapy-induced germ cel damage in young cancer patients. Microenvironment and Plzf, GDNF, SCF/c-Kit signaling pathways can play an important role in the regulation of spermatogonial stem cel self-renewal and differentiation. As a cel model, spermatogonial stem cel s become an important tool for the researches on spermatogenesis mechanism, regeneration of spermatogenesis in sterile individuals and reproduction of transgenic animals.

Objective To identify the effects of androgen on mouse model of autoimmune ovary failure.MethodsFifty mice were randomly divided into five groups as control group,early therapeutic group and its model group,late therapeutic group and its model group.Autoimmune POF was induced in forty mice by Pzp3.Then,the animals were randomly divided into four groups: two were treated with androgen from the 15th and 20th day after immune.The control group was treated with distilled water.The therapeutic efficacy of each treatment was evaluated by the changes in sexual cycles of mice,the serum level of AzpAb,infiltration of cells positively expressing CD45 in the ovary as well as the percentage of growing follicles of the ovary.Results The sexual cycles,serum level of AzpAb,infiltration of CD45 cells in the ovary of each model group were more significant than those in control group;early therapeutic group was less than model group;and serum level of AzpAb in early therapeutic group was lower than that of late therapeutic group.The percentage of growing follicles in the ovary of each model group was lower than that of control group;early therapeutic group was higher than model group and late therapeutic group.Conclusion The use of androgen in mice with autoimmune POF may notably ameliorate the pathogenetic conditions of the disorder,and the treatment during prophase was apparently more effective than that during the advanced stage.

Objective To evaluate the correlation between homologous recombination repair protein RAD51 and methotrexate-enhanced radiosensitivity.Methods Western blot and RT-PCR assays were used to detect RAD51 expression in HOS osteosarcoma cells exposed to γ-ray irradiation alone and in combination with methotrexate.Colony formation assay was used to test the survival fraction of HOS cells exposed to γ-rays and methotrexate.Results Methotrexate inhibited both protein and RNA expressions of RAD51,and the combination of radiation and methotrexate enhanced the inhibition of RAD51 expression.Moreover,transfection of cells with RAD51 gene decreased cellular sensitivity to methotrexate and γ-rays.The sensitizer enhancerment ratios after irradiation in combination with methotrexate were 1.51 and 0.99,respectively.Methotrenate was a preferred radiosensitizer to HOS cell.Conclusions RAD51 might be involved in the methotrexate-enhanced radiosensitivity.

Objective To investigate the effect of fetal hemoglobin( HbF) in improving anemia of women who were β‐thalasse‐mia carriers at childbearing age .Methods 289 cases of women at childbearing age diagnosed as β‐thalassemia carriers in Shenzhen City were enrolled in this study .The high performance liquid chromatography (HPLC) was uesed for quantitative analysis of HbF , and the red blood cell parameters were detected by using the LH750 Automatic blood cell analyzer .The differences of red blood cell parameters between the HbF normal group and the HbF increased group were compared ,and the relationship between the high HbF expression rates and gene mutations were also analyzed .Results Comapred with the HbF normal group ,the hemoglobin (HGB) levels ,mean corpuscular volume (MCV) and the mean corpuscular hemoglobin (MCH) increased significantly ,while the red blood cell (RBC) count reduced ,there were significant differences between the two groups (P<0 .05) .No significant differ‐ences of the mean corpuscular hemoglobin concentration (MCHC) and RDW coefficient of variation (RDW‐CV) between the two groups were found(P>0 .05) .There were no significant differnces of rates of HBF high expression between different types of β‐globin gene mutations and the overall rate of HBF high expression (P>0 .05) .Conclusion Compared with the HbF normal group , anemia may improve more significantly in cases of women diagnosed as β‐thalassemia carriers at childbearing age in the HbF in‐creased group ,and there may be no relationship between gene mutations and high expression of HbF .

Objective To study the curative effect of XRCC2 gene silencing mediated by shRNA combined with radiation on human colonic trans?planted carcinoma in nude mice. Methods Colonic carcinoma T84 cells were transfered into BALB/c nude mice to establish a tumor xenograft mod?el in vivo. Mice were divided into three groups:control,shRNA?SC and shRNA?XRCC2 and exposed to X?ray radiation. The change of volume and weight of the xenografts were examined after receiving radiotherapy and the pathological analysis of tumor tissues were conducted. Results Tumor xenografts transfected with shRNA?XRCC2 in nude mice grew slowly. The xenograft volume in the shRNA?XRCC2 group was decreased significant?ly from day 12 to day 28 after radiotherapy compared with the control group(P<0.01). The xenograft weight in the shRNA?XRCC2 group was small?er than in the control group,with statistically significant difference(t=18.843,P<0.01). The inhibited rate of xenografts in the shRNA?XRCC2 group(56.25%),was markedly higher than that in the shRNA?SC group(4.69%). Pathological analysis of colonic transplanted carcinoma showed that nuclear atypia was not obvious,karyokinesis was decreased and small areas of necrosis were present in tumor xenografts treated with shRNA?XRCC2 transfection. Conclusion XRCC2 gene silencing combined with radiation has significant inhibition effect on colonic transplanted carcino?ma in nude mice.

Objective To study effects of palm pressing peripherally inserted untral catheter(PICC) and mandible-pressing-on-shoulder on PICC′s misplacement in jugular veins during intubation. Methods Seventy-six patients undergoing intubation with PICC were randomized into observation group and control group equally by random digital table. The palm pressing PICC method was used in the observation group and the mandible-pressing-on-shoulder method was used in the control group. The two groups were compared in terms of the incidence of PICC misplacement. Result The misplacement rate of PICC in the experiment group was significantly lower than that in control group (P<0.01). Conclusion Palm pressing method is much effective in prevention of PICC misplacement in jugular veins and worth promoting especially in hospitals of basic level.