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 know the influence of using health belief model among chronic prosatitis patients on their ability of self-controL Methods Divided 96 patients with chronic prosatitis into the observation group and the control group randomly,there were 48 cases in each group. Routine health education was used in the control group,while the health education according to health belief model was used in the observation group. Compared the ability of self-control between the two groups. Results Before the health education,there was no significant difference about the ability of self-control between the two groups,while after 3-month follow-up,the difference was significantly. Conclusions Health education based on health belief model can effective promote the ability of self-control in patients with chronic prosatitis.

Objective To establish human embryonic stem cells (hESCs) feeder-independent and cell factor-free culture system. Methods Effect of high and low clone densities of hESCs culture was compared and impact of the clone densities to hESCs culture media was analyzed. Results HESCs could maintain their undifferentiated states at high clone density (34 clones/cm2) without cell factors. At the same time,the bone morphology protein (BMP)-like induction of N2 and B27 supplements (NB) medium could be modulated by the clone density,and high level of BMP-like induction was accompanied by high clone density. Conclusion High clone density of hESCs can change the environments by themselves to maintain the undifferentiated states,which provides a new clue to explore the mechanism of undifferentiated states of hESCs and simplify the culture medium.

Objective To investigate the effects of Rosa Laevigata Michx Flavoid( RLMF) and Rosa laevigata Michx Polysaccharose (RLMP) on expression of TRPV5 in IgA Nephropathy (IgAN) rat renal tissue. Methods Forty Sprague-Dawley rats were randomly assigned to four groups.The rat model of IgA nephropathy was induced by intragastric administration of bovine serumalbumin and injections of LPS and CC14.Eight weeks later,the rats with IgAN were treated with RLMF or RLMP (4 weeks), or normal saline.Rats was sacrificed at thirteenth weeks, and RNA was extracted from the kidney.Expression of TRPV5 in tubulointerstitial tissues were analyzed by fluorescent quantitative RT-PCR. Results After RFLP intervention,the expression levels of TRPV5 were markedly increased (P<0.01) than model control group,while decreased (P<0.05) than normal control group but had no significance with model control group after RFLF intervention. Conclusion TRPV5 expression is decreased in IgAN,and RLMP can adjust TRPV5 expression and improve renal function of IgAN.

OBJECTIVETo construct the two-dimensional electrophoretic (2-DE) protein map of the human sperm head.

METHODSProtein extracts of the normal human sperm and sperm head were loaded on an 18 cm immobilized pH gradients (IPG) strip holder and separated with isoelectric focusing electrophoresis as the first dimension, and then with upright SDS-PAGE as the second. Different protein spots between them were compared with Image Master 3.0, and so were the maps of method I with Urea/Thiourea/Guanidine HCl and method II with Kit/Guanidine HCl.

RESULTSOf the whole-sperm proteome, 802 protein spots were obtained by method I, and 797 by method II. Among them, distribution patterns of 492 spots were the same. A sperm protein map was constructed with 1107 protein spots after the interaction of the spots obtained by both the methods. Of the sperm-head proteome, 428 protein spots were obtained, all found in the whole-sperm protein map after comparison.

CONCLUSIONBoth the methods could be complementarily used to construct a sperm protein map, and the map could serve as a model for the establishment of the sperm protein profile.

Electrophoresis, Gel, Two-Dimensional ; methods ; Humans ; Hydrogen-Ion Concentration ; Male ; Proteins ; analysis ; Proteomics ; methods ; Sperm Head ; chemistry

OBJECTIVE: To summarize the genetic characteristics of 671 Chinese pedigrees affected with Duchenne/Becker muscular dystrophy (DMD/BMD). METHODS: Clinical data of the pedigrees were collected. Multiplex PCR, multiple ligation dependent probe amplification (MLPA), next generation sequencing (NGS), Sanger sequencing and long read sequencing were used to detect the variant of DMD gene in the probands and their mothers, and prenatal diagnosis was provided for high risk pregnant women. RESULTS: Among 178 pedigrees analyzed by multiplex PCR, 44 variants of the DMD gene were detected, with the genetic diagnosis attained in 110 pedigrees. Among 493 pedigrees analyzed by MLPA in combination with NGS or Sanger sequencing, 294 pathogenic/possible pathogenic variants were identified, among which 45 were unreported previously, and the genetic diagnosis attained in 484 pedigrees. Structural variants of the DMD gene were identified in two pedigrees by long-read sequencing. Among 444 probands, 341 have inherited the DMD gene variant from their mothers (76.8%). Among 390 women with a high-risk, 339 have opted to have natural pregnancy and 51 chose preimplantation genetic testing for monogenetic disease (PGT-M). The detection rate of neonatal patients and carriers following natural pregnancy was significantly higher than that for PGT-M. CONCLUSION Combined application of MLPA, NGS, Sanger sequencing and long-read sequencing is an effective strategy to detect DMD/BMD. PGT-M can effectively reduce the risk of fetuses. Above finding has expanded the spectrum of DMD gene variants and provided a basis for reproductive intervention for pregnancies with a high risk for DMD/BMD.
China ; Dystrophin/genetics* ; Exons ; Female ; Genetic Testing ; Humans ; Infant, Newborn ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne/genetics* ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis