Spermatogenesis is a complex process. Current knowledge about human spermatogenesis is mainly based on the mouse model while little is known about the initial stage of this fundamental process in humans. The establishment of the model of spermatogenesis in vitro may contribute to an overall understanding of male germ cell development, an insight into the mechanisms of infertility, and clinical management of male infertility. This review summarizes current knowledge about the generation of germ cell-like cells from induced pluripotent stem cells (iPSCs) in vitro and discusses the potential application of iPSCs in the treatment of male infertility.
Animals ; Cell Differentiation ; Germ Cells ; Humans ; Induced Pluripotent Stem Cells ; Infertility, Male ; therapy ; Male ; Mice ; Spermatogenesis

OBJECTIVETo evaluate the effects of Attractin (Atrn) silence on the anti-oxidative and anti-apoptotic abilities of TM4 Sertoli cells and its influence on the expressions of superoxide dismutase (SOD) and caspase6 in the cells.

METHODSWe observed the apoptotic indexes of TM4 Sertoli cells with normal expression (control), partial deletion, and complete deletion of the Atrn gene (psiRNA-TM4, psiAtrn-TM4, and mu-SC). We determined the mRNA and protein expressions of SOD and caspase6 by Q-PCR and Western blot, measured the SOD activity and malondialdehyde (MDA) contentby spectrophotometry, and detected the apoptotic index of the cells by TUNEL.

RESULTSCompared with psiRNA-TM4, after inhibition of the Atrn expression, the Sertoli cells in the psiAtrn-TM4 and mu-SCgroups showed significantly decreased expressions ofSOD mRNA (70.76% and 92.58%) and protein (65.11% and 71.0%) (both P < 0.05). The levels of caspase 6 mRNA and protein were increased 5.28 and 3.40 times in the psiAtrn-TM4 and 2.97 and 2.50 times in the mu-SCgroup as compared with the normal control (both P < 0.05). Atrn deletion markedly increased the apoptotic indexes of the cells in the psiAtrn-TM4 and mu-SC groups by 16.22% and 22.03% (P < 0.05) and reduced the activity of SOD by 23.00% and 39.37% (P < 0.05); it also elevated the level of MDA by 155.22% (P < 0.05).

CONCLUSIONThe Atrn gene exerts influence on the function of Sertoli cells in multiple ways, in which antioxidative stress and apoptosis regulation may play an important role.

Animals ; Apoptosis ; Caspase 6 ; metabolism ; Cells, Cultured ; Gene Deletion ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Oxidative Stress ; Sertoli Cells ; metabolism ; pathology ; Superoxide Dismutase ; metabolism

This paper is aim to investigate the pharmacokinetics and absolute bioavailability of neoline in Beagle dogs, and provide a theoretical basis for further study. Ethyl acetate was used for liquid-liquid extracting after 10% ammonia alkalizing. The method of UPLC-Q-TOF-MS was established for the determination of neoline plasma concentrations. Beagle dogs were orally or intravenously administered with neoline for pharmacokinetic and absolute bioavailability study. Good linear relationship of neoline was found over the range of 0.1-4 mg x L(-1) (R2 = 0.9982) and 2-100 microg x L(-1) (R2 = 0.9945). Intra-and inter-day precision, expressed as the relativestandard (RSD) were less than 5.0%. Accuracy, expressed as the relative error (RE) was within 90.0%-115%. The recovery of neoline in dog plasma was more than 80%. After 6 mg x kg(-1) for ig and 1 mg x kg(-1) for iv administration of neoline, the main pharmacokinetic parameters were analyzed with Winnonlin software. t(1/2) were (313.88 +/- 63.18), (236.33 +/- 229.84) min, and AUC(0-infinity) were (58,027.40 +/- 14,132.69), (473,578.02 +/- 82,333.08) min x microg x L(-1) for ig and iv administration respectively. The absolute bioavail ability was (73.15 +/- 10.29) %. The method of UPLC-Q-TOF-MS described in the report was sensitive, reliable and specific, and suitable for pharmacokinetic study of neoline in Beagle dog. The high absolute bioavailability of neoline in dog suggested good absorption of neline which was worth of further investigation.
Aconitine ; analogs & derivatives ; chemistry ; pharmacokinetics ; Animals ; Biological Availability ; Dogs ; Drug Stability ; Female ; Male

In recent years, the problem of bacterial resistance has become more and more serious, which has brought troubles to global public health and medical care. The time and money required to develop new antibiotics is even greater than before. Bacteriophage is a kind of virus that can specifically infect bacteria, fungi, actinomycetes and other microorganisms. Relying on host bacteria to replicate in large numbers, rich species, low research and development cost, the value of anti-infection therapy is very considerable. It is a new generation of biological antimicrobial agents with great potential. This paper briefly describes the sterilization mechanism, progress of research on anti-infection aspect and clinical application of phage, in order to provide reference for phage anti-infection treatment and clinical application.

The expression of Attractin mRNA and protein in testis and semen of human and male mice was investigated. Human testis and semen samples were all collected from Reproductive Center of Renmin Hospital, Wuhan University in December, 2012. Testis samples were collected from 7 cases of obstructive azoospermias when they were subjected to diagnosed testis biopsy, and 30 normal human semen samples were obtained from those cases of semen analysis. Adult mice testis tissues were obtained from 10 2-month-old male BALB/c mice, and 60 male mice at different ages were classified into 10 groups (day 1, 5, 10, 15, 21, 28, 35, 42, 56, and 120 respectively, n=6 each). The expression of Attractin mRNA and protein in testis was detected by RT-PCR and Western blotting respectively. Human semen samples were centrifuged into sperm plasma (SP) and sperm extract (SE), and mice sperm samples were collected from the epididymis of 10 adult male BALB/c mice. Western blotting was used to determine the Attractin protein expression level. Attractin mRNA and protein were expressed in the testis of both patients with obstructive azoospermias and adult Bcl/B mice. Quantitative RT-PCR revealed that no Attractin mRNA was detectable in day 1 male BALB/c mice group. The Attractin mRNA and protein levels were low on the day 10, and increased with age until day 56. On the day 120, the expression levels of Attractin were decreased. As for human semen samples, Attractin protein was expressed in both SP and SE, but didn't exist in samples from the epididymis of male BALB/c mice. It was suggested that Attractin acted as a novel active substance and was involved in male reproduction in both human and BALB/c mice, but it exerted a different expression profile in different mammal species.

OBJECTIVETo explore the effects of testis murine cytomegalovirus (MCMV) infection on mature sperm viability in mice.

METHODSBALB/c mice without MCMV infection, screened by ELISA, were randomly divided into two groups: an experimental group (n = 64) and a control group (n = 40). The former were directly inoculated with MCMV into the testis, while the latter treated by inoculation of DMEM without MCMV. The mice in both of the groups were sacrificed respectively at 1, 2, 4, 6, 9, 14, 21, 38 d postinoculation (D1, 2, 4, 6, 9, 14, 21, 38 PI), the testis was examined histopathologically, and meanwhile the viability of mature sperms in the epididymis cauda was measured.

RESULTSMCMV basophil inclusion bodies were found in the Leydig cells in the experimental group, and spermatogenic cells were vacuolated and arranged disorderly. Compared with the control group, the sperm viability in the experimental group was decreased significantly by 71.42% to 56.04% (P < 0.05) on D1 PI.

CONCLUSIONThe sperm viability in mice might be descended significantly by MCMV infection in the early period, but restored to normal with time. This shows that MCMV infection might influence procreation transiently.

Animals ; Cell Line ; Cell Survival ; physiology ; Cytomegalovirus Infections ; pathology ; physiopathology ; Male ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Spermatozoa ; physiology ; Testicular Diseases ; pathology ; physiopathology ; virology ; Testis ; pathology

OBJECTIVETo identify the proteins that could improve the resistance of human sperm to cryopreservation using comparative proteomics.

METHODSA total of 31 semen samples from 10 donors were divided into a high recovery and a low recovery group. Differentially expressed proteins in sperm and seminal plasma were detected and compared between the two groups by two-dimensional differential gel electrophoresis and mass spectrometry.

RESULTSTotally, 22 differentially expressed proteins were found in the two groups, 12 seminal plasma proteins, 9 sperm proteins, and 1 belonging to both. These identified proteins were involved in the maturation, movement, energy metabolism, DNA repair and other activities of spermatozoa.

CONCLUSIONMany proteins were identified in sperm and seminal plasma that might influence the resistance of human sperm to cryopreservation.

Adult ; Cryopreservation ; Humans ; Male ; Proteomics ; Semen ; metabolism ; Seminal Plasma Proteins ; metabolism ; Sperm Motility ; Spermatozoa ; metabolism ; Young Adult

OBJECTIVETo study the influence of root of medicated serum with the root of Crataegus cuneata on human sperm motility in vitro.

METHODSperm specimens of 16 asthenospermia patients were co-incubated with the medicated serum in vitro. The sperm motility characteristics were evaluated by computer-assisted semen analysis (CASA) at 5, 15, 30, 60 min and 120 min.

RESULTCompared with the control group, the medicated serum significantly increased the sperm progressive motility in 5 and 15 min (P < 0.05), and the motility and progressive motility were both increased significantly in 60 and 120 min (P < 0.01). The excitatory effect of the medicated serum on the sperm motility exhibited in a time-dependent manner.

CONCLUSIONRoot of C. cuneata medicated serum can improve sperm motility of asthenospermia patients in vitro.

Adult ; Animals ; Coculture Techniques ; Crataegus ; chemistry ; Culture Media ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Male ; Oligospermia ; physiopathology ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Sperm Motility ; drug effects

Chemical constituents of Leonurus japonicus were isolated and purified by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, MCI, and Rp C18. Structures of the isolates were determined by spectroscopic analysis as 10 coumarins: bergapten (1), xanthotoxin (2), isopimpinellin (3), isogosferal (4), imperatorin (5), meransin hydrate(6), isomeranzin(7), murrayone(8) , auraptenol(9), and osthol(10). In addition to compound 9, the others were isolated from the genus Leonurus for the first time. In the in vitro assay, compounds 4 and 8 significantly inhibited the abnormal increase of platelet aggregation induced by ADP.
Blood Platelets ; drug effects ; Coumarins ; chemistry ; pharmacology ; Leonurus ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; chemistry ; pharmacology

The chemical constituents of 95% ethanol extract of Melastoma dodecandrum were isolated and purified by chromatography on silica gel, Sephadex LH-20, and HPLC, to obtain thirteen compounds eventually. On the basis of their physico-chemical properties and spectroscopic data, these compounds were identified as quercetin (1), quercetin-3-O-β-D-glucopyranoside (2), quercetin-3-O-(6"-O-p-coumaroyl) -β-D-glucopyranoside (3), kaempferol (4), kaempferol-3-O-β-D-glucopyranoside (5), kaempferol-3-O- [2",6"-di-O-(E)-coumaroyl]-β-D-glucopyra-noside (6), luteolin (7), luteolin-7-O-(6"-p-coumaroyl) -β-D-glucopyranoside (8), apigenin (9), apigenin-7-(6"-acetyl-glucopyranoside) (10) , naringenin (11), isovitexin (12), and epicatechin-[8,7-e] -4β-(4-hydroxyphenyl)-3,4-dyhydroxyl-2(3H)-pyranone (13). Eight compounds(3,5,6,8-11 and 13) were obtained from M. dodecandrum for the first time.
Apigenin ; analysis ; Chromatography ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Flavanones ; analysis ; Flavonoids ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Kaempferols ; analysis ; Luteolin ; analysis ; Magnoliopsida ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analysis ; Silica Gel