Micro RNA differential expression in primary retroperitoneal liposarcoma / 中国现代普通外科进展

Objective:To explore the miRNA (micro RNA)differential expression profile between primary retroperitoneal liposarcoma tissues and normal fat tissues,and to provide the evidence that miRNA were involved in the molecular pathways of primary retroperitoneal liposarcoma tissues' occurrence.Methods:Collecting retroperitoneal liposarcoma tissues and normal fat tissues from 4 patients after radical surgery of retroperitoneal lipsarcoma.Using microarray analysis.The tissues' miRNA hybridizated with human's LC Sciences microRNA Microarray-Single (miRBase 21.0) expression profile gene chips,and got the date.Analyzing the differential expressing of the siginal date by LOWESS.Results:Total 38 differential expressed miRNA were found (P＜0.05),including 23 over-expression and 15 low-expression miRNAs.10 of them(38 differential miRNAs) was significant deviation (P＜0.01),including 4 over-expression and 6 low-expression.Date analysis revealed that some miRNAs were associated some different tumors,Conclusion:The number of over-expression were more than the low-expression in primary retroperitoneal liposarcoma compared with the normol fat tissue,which indicate that the genes expression are less abundant in primary retroperitoneal li-posarcoma;some of the miRNAs might involved in the molecular pa-thways of primary retroperitoneal liposarcoma tissues' occurrence and recurrence,they might become the target point of the targetedtherapy of the primary retroperitoneal liposarcoma,some of the over-expressed miRNAs can become new biomarkers in the following diagnosis of the primary retroperitoneal liposarcoma.

Protective effects of puerarin on high glucose-induced oxidative injury in HUVEC-12 cells / 中成药

AIM To explore the action mechanism of puerarin's protective effects against oxidative stress of HUVEC-12 cells induced by high glucose.METHODS HUVEC-12 cells cultured with 100 mmol/L glucose medium and 10,25,50 μmoL/L puerarin for 36 h had the cell proliferation,the levels of lactate dehydrogenase (LDH),intracellular reactive oxygen species (ROS),the activities of caspase-3,superoxide dismutase (SOD)and catalase (CAT),and the contents of malondialdehyde (MDA) and glutathione (GSH) measured.The mRNA expressions of SIRT1 and PGC-1α were detected by real-time flu orescence quantitative PCR,and the protein contents of SIRT1 and PGC-1 α were determined by enzyme-linked immunosorbent assay.RESULTS The puerarin treatment to HUVEC-12 cells resulted in markedly lowered LDH level,caspase-3 activity,intracellular levels of MDA and ROS,and notable improvement of the cell viability,the activities of SOD and CAT,GSH content,the mRNA expressions and the protein contents of SIRT1 and PGC-1α as well.CONCLUSION The protective effect of puerarin on high glucose-induced oxidative damage of HUVEC-12 cells may be attributed to the SIRT1/PGC-1α pathway activation.

Expression and synergistic function of ENHANCIN-like gene of Agrotis Segetum granulovirus / 病毒学报

ENHANCIN is an enhancing protein chiefly found in insect baculoviruses. One ENHANCIN homologue was identified, by blast method, in Agrotis Segetum granulovirus (AgseGV) genome, named enhancin-like. Sequence analysis indicated that this gene includes the conserved domains, conserved in other ENHANCIN, and it has no signal peptide or a-transmembrane helix. A proline-rich domain, which is similar to those of mammals, is present at its C-terminal. To analyze the synergistic function of AgseGV enhancin-like gene, prokaryotic expression vectors of its whole gene and the 5'-truncated fragment (1, 017bp) were constructed. Expression product of truncated fragment was purified by chromatography, and then it was used to prepare antibody. The expression product of whole gene was identified by Western blot with specific antibody and anti-His-Tag antibody. Bioassay proved that the expression product of whole gene can increase the mortality with 16.25% to 3th instar larvae of Helicoverpa armigera (HaNPV: 1.17 x 10(2) PIBS/mL), while the truncated fragment has no obvious synergistic function.

Effects of Di (2-ethylhexyl) phthalate on the testis and testicular gubernaculum of fetal KM mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of Di (2-ethylhexyl) phthalate (DEHP) on the testis and testicular gubernaculum of fetal KM mice in vivo and to investigate the mechanism of DEHP-induced cryptorchidism.</p><p><b>METHODS</b>Thirty healthy pregnant KM mice were randomly and equally divided into a blank control group, a corn oil control group and a DEHP group. The pregnant mice in the latter group were exposed to DEHP by gavage at the dose of 500 mg/kg body weight per day from gestation day 12 (GD12) through gestation day 19 (GD19). The effects of DEHP were observed on the number of fetuses per pregnancy, the ratio of male to female pups, the weight of the testis, the morphology and location of the testis and gubernaculum, the relative testis-bladder neck distance (TBD) and cranial suspensory ligament (CSL) residual. The expressions of the androgen receptor (AR), estrogen receptor (ER) and actin and proliferating cell nuclear antigen (PCNA) in the gubernaculum were detected by immunohistochemistry.</p><p><b>RESULTS</b>DEHP reduced the testis weight and TBD, induced different degrees of testis maldescent, but produced no obvious effect on the body weight, the number of fetuses per pregnancy, the sex ratio and the testis gubernacular morphology. Under the light microscope, hypotrophy was seen in all the testis seminiferous tubules, spermatogenic cells and Sertoli cells, marked Leydig cell hyperplasia was noted, and the positive expression of AR in the gubernaculum was decreased in the DEHP group (P < 0.01).</p><p><b>CONCLUSION</b>DEHP could cause dysfunction of the testis gubernaculum via its anti-androgen effect, induce cryptorchidism, and cause dysplasia and dysfunction of Sertoli cells, Leydig cells and spermatogenic cells in fetal mice.</p>

Di(2-ethylhexyl) phthalate affects the testes and leydig cells of neonatal KM mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of di(2-ethylhexyl)phthalate (DEHP) on neonatal mice's testes and Leydig cells in vivo.</p><p><b>METHODS</b>Pregnant mice were exposed to DEHP at the dose of 100 mg/kg, 200 mg/kg or 500 mg/kg (body weight) per day by gavage from gestation day 12 (GD 12) through postnatal day 3 (PND 3), respectively. The testis and body weights, testicular histopathology and the activity of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) of the neonatal mice were investigated.</p><p><b>RESULTS</b>The body and testis weights of the male mice's offspring were significantly reduced following DEHP exposure. Leydig cell morphology was affected significantly by DEHP as compared with the controls. Leydig cells obviously increased in the neonatal mice's testes on PND 15 and PND 30 when exposed to DEHP (500 mg/[kg x d]). Activities and positive area of the steroidogenic enzymes 3beta-HSD immunoexpression decreased markedly when exposed to DEHP (100 mg/[kg x d] or 200 mg/[kg x d]). Image analysis showed a decrease in the activities of 3beta-HSD in the animals exposed to DEHP (500 mg/[kg x d]), but an increase in the positive area of 3beta-HSD immunoexpression as compared with the control animals on PND 15 (P < 0.01).</p><p><b>CONCLUSION</b>DEHP affects the Leydig cell morphology, the activity of 3beta-HSD, the testis and body weights and the testicular histopathology of neonatal mice, and it may function as an antiandrogenic agent.</p>

Isolation, cultivation, identification and functional study of fetal mice Leydig cells in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the methods of isolation, cultivation, purification, identification of the fetal mice testis Leydig cell and to observe its biological characteristics in vitro.</p><p><b>METHODS</b>Leydig cells were isolated by 0.03% collagenase (type I) from fetal mice testis and cultured in DMEM/F12 medium. The identity and purity of Leydig cell were assessed by 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD). Cell viability was measured by trypan blue. Testosterone level in the medium of cultured Leydig cells was measured in various culture phases and cell density by radioimmunoassay.</p><p><b>RESULTS</b>The purity of Leydig cell was (45.10 +/- 1.66)% before culture, and (81.17 +/- 2. 32)% 72 h after culture. The level of testosterone secreted by Leydig cells could be detected in the medium and its level was associated with the density and time of cultured Leydig cells. The secretion capacity of testosterone by single Leydig cell decreased gradually during the culturing period.</p><p><b>CONCLUSION</b>The fetal Leydig cells isolated from fetal mice testis have high purity. It can be cultured and kept the secretion ability of testosterone for a few days in vitro. This system can provide a valuable model for further study on the cellular function of the Leydig cells of fetal mice.</p>