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

Objective To study the inhibitory effects of stachyose on the hyperacute rejection in pig-to-human heart xenotransplantation.Methods A pig-to-human xenogeneic heart transplantation model was established based on an in vitro free heart blood perfusion system.The pig hearts were di- vided into two groups:group A(pig hearts treated with human blood perfusion as control)and group B(pig hearts treated with human blood plus stachyose perfusion).After perfusion for 1h,the heart xenografts were examined for deposit of lgG and IgM by immunohistochemistry and pathological analy sis.Results The mean survival time of perfusion hearts in groups A and B was(9.5?2.5)min and (46.8?8.1)min respectively(P

Objective To observe the effect of transplantation of telomerase immortalized human neural progenitor cells to acute injured canine spinal cord by using MR diffusion tensor imaging (DTI).Methods Telomerase immortalized human neural progenitor cells with expression of green fluorescent protein were prepared for transplantation. Eight adult canines with left spinal cord hemisection at the level of T13 were examined by MR diffusion tensor imaging four times sequentially: prior to injury, one week after injury, one week after transplantation (two weeks after injury), and four weeks after transplantation. Results The ADC values of the injured spinal cord were (1.00?0.15)?10 -3 mm2/s, (1.65?0.45)?10 -3 mm2/s, (1.44?0.48)?10 -3 mm2/s, and (1.43?0.26) ?10 -3 mm2/s, respectively. There was statistically significant difference between the data obtained at different times (F=6.038, P=0.005). The FA values of the injured spinal cord were 0.59?0.11, 0.30?0.17, 0.36?0.25, and 0.34?0.11, respectively. There was also statistically significant difference between the data obtained at different times ( F=5.221,P=0.009). The ADC values of the intact spinal cord were (1.01?0.17)?10 -3 mm2/s, (1.32?0.06)?10 -3 mm2/s, (1.10?0.24)?10 -3 mm2/s, and (1.14?0.22) ?10 -3 mm2/s, respectively. There was no statistically significant difference between the data obtained at different times ( F=1.303,P=0.306). The FA values of the intact spinal cord were 0.60?0.09, 0.38?0.25, 0.46?0.15, and 0.50?0.21, respectively. There was also no statistically significant difference between the data obtained at different times (F=2.797,P=0.072).Conclusion DTI can provide useful information for spinal cord injury and regeneration in experimental spinal cord injury.

Objective:To study the interaction of the inflammatory cytokines expression between CD4+ T cells and primary human umbilical vein endothelial cells (HUVECs) infected by dengue virus (DENV-2).Methods:PBMC was extracted from white blood cells by density gradient centrifugation,CD4+ T cells were sorted by immunomagnetic beads.The expression of CD3 and CD4 molecules on the surface of cells was detected by flow cytometry to identify the purity of CD4+ T cells.First,HUVECs were pretreated by specific-S1 P1 receptor agonist CYM-5442 for 24 h,second,infected by DENV-2 on the titer of 103 TCID50,then co-culturing with CD4+ T cells,The relative expression of NS1 partial sequence and IL-6,L-8 mRNA of HUVECs,and IL-4,IL-17,TNF-α,IFN-γ of CD4+ T cells detected by Real-time RT-PCR.IL-6 and IL-8 secreted in cultured supematant analyzed by ELISA.Results:The purity of CD4+ T cells was (98.02±0.32) ％.The expression of NS1 gradually increased to 24 h (3.03±0.26,P＜0.001),decreased after reaching the peak.The relative expression of NS1 in the group of co-cultured with CD4+ T cells was lower than other groups.After infection,the expression of IL-6 and IL-8 were up-regulated,and the expression of IL-6 and IL-8 at each time point was significantly increased after co-culturing with CD4+ T (P＜0.01).IL-6 of CYM-5442 pretreatment group,in 24 h (28.91±2.34,P＜0.05),36 h(19.36±0.1,P＜ 0.05) and 72 h(13.84±0.82,P＜0.05) was significantly decreased,the expression of IL-8 also decreased significantly.The mRNA expression of IL-4,IL-17,TNF-α and IFN-γ in CD4+ T cells was significantly increased after co-culturing with HUVECs.After the treatment with CYM-5442 group,the expression was decreased.Conclusion:DENV-2 could infect the primary HUVECs,and the expression of NS1 was inhibited after co-culturing with CD4+ T cells.CD4+ T cells can not only enhance the activation of HUVECs infected by DENV-2,but also can be activated by the infected HUVECs infected with DENV-2.

Objective To study the effect of mechanical ventilation guided by esophageal pressure on hemodynamics and oxygen metabolism of severe acute pancreatitis (SAP) swine model with intra-abdominal hypertension (IAH) Methods By self-controlled study,SAP model was made by infusing sodium taurocholate (5％) into the pancreatic duct in 6 domestic swine.Mechanical ventilator mode was volumeassist control with tidal volume 10 ml/kg ; FiO2 40％ and PEEP 5 cm H2O (routine ventilation) given to SAP model swine.After 3 h ventilation,pneumo-peritoneum was made with N2 gas to increase the intraabdominal pressure (IAP) to 25 mm Hg in SAP swine.Three more hours later,PEEP was adjusted as the measurement of esophageal pressure (Pes guided ventilation) to such a level that trans-pulmonary pressure stayed above 0 cm H2O during end-expiratory occlusion.During the investigation period,heart rate (HR),cardiac output index (CI),central venous pressure (CVP),mean arterial pressure (MAP) and pulmonary arterial wedge pressure (PAWP) were continuously recorded with the aid of Swan-Ganz catheter and ECG monitor,and oxygen partial pressure of artery (PaO2) and carbon dioxide partial pressure of artery (PaCO2) were measured by blood-gas analysis.In addition,systemic oxygen delivery (DO2) and systemic oxygen consumption (VO2) were calculated by using the data of blood-gas analysis of arterial and central venous blood.Results No swine model was subjected to barotrauma.After routine mechanical ventilation,there were significant differences in HR,CI,MAP,CVP,PAWP,Ppeak,Pplat,Pes,pulmonary compliance (Cstat),PaO2,and DO2 between SAP and IAH in swine (all P ＜ 0.05).Compared with routine ventilation,however,PaO2 and Cstat improved significantly with lower CI and increased Pplat after PEEP adjusted according to measurements of esophageal pressure (all P ＜ 0.05).The lactate decreased significantly after esophageal pressure guided ventilation (all P ＜ 0.05).There were no significant changes in PaCO2,HR,MAP,CVP and PAWP in IAH swine after mechanical ventilation with routine parameters (all P ＞ 0.05).Conclusions There were remarkable effects on oxygen metabolism in response to mechanical ventilation guided by esophageal pressure.In case of clinical application of mechanical ventilation,the results of this study are in favor of setting transpulmonary pressure according to measurements of esophageal pressure in SAP patients with IAH in an early stage.

In order to reveal feasibility of different concentrations of Ca2+ and SA on Platycodon grandiflorum under high temperature stress, the effects of Ca2+ on physiological index and related photosynthetic parameters were studied. Pot cultured P. grandiflorum leaves under the same outdoor conditions were sprayed with CaCl2 and SA separately, and then placed in the high incubator [35 degrees C/25 degrees C (day/night), light intensity 3 600 lx], and sprayed with distilled water at 25 degrees C and under high temperature stress were set as the control. The related photosynthesis, relative conductivity, contents of proline, malondialdehyde, soluble protein, activities of SOD and CAT, ASA and GSH content were measured. The results show that the 6 mmol x L(-1) CaCl2 and 1.5 mmol x L(-1) SA enhanced the activities of SOD and CAT, the contents of proline and soluble protein, and effectively reduced the damage of heat stress on cell membrance. At the same time, the exogenous Ca2+ and SA increased the contents of chlorophyll and carotenoid, the efficiency of leaf photosynthesis and ASA and GSH content, and thus effectively resisted the oxidative stress caused by high temperatures, but with the increasing concentration of spraying, P. grandiflorum decreased the ability to resist high temperature stress. In conclusion, the foliage spraying CaCl2 and SA could reduce the damage of high temperature stress on P. grandiflorum leaves.
Calcium ; metabolism ; Hot Temperature ; Malondialdehyde ; metabolism ; Oxidative Stress ; Photosynthesis ; Plant Leaves ; metabolism ; Platycodon ; metabolism ; Salicylic Acid ; metabolism ; Stress, Physiological

Objective To investigate the effects of interaction between human umbilical vein endothelial cells (HUVECs) which were infected with dengue virus type 2 (DENV-2) and CD4+T cells on the expression of ICAM-1 (intercellular adhesion molecule 1),VCAM-1 (vascular cell adhesion molecule 1),IL-10 and TGF-β1 at mRNA level for further understanding the immunological mechanism of DENV infection.Methods HUVECs were treated with CYM-5442,a selective agonist for sphingosine-1-phosphate receptor 1 (S1P1),for 24 hours and then infected with 103 TCID50 (50% tissue culture infective dose) of DENV-2 before co-culturing with CD4+T cells.Changes in the expression of NS1 (DENV-2 nonstructural protein),SPHK1 (sphingosine kinase 1,phosphorylating sphingosine to S1P),ICAM-1,VCAM-1,IL-10 and TGF-β1 at mRNA level were detected by real-time PCR after 4,8,12,24,48 and 72 hours of co-culturing.Results There was a certain timeliness in the expression of NS1 at mRNA level after infecting HUVECs with DENV-2 and the expression reached a peak at 24 h.Treating HUVECs with or without CYM-5442 had no significant influence on the expression of DENV-2 NS1 at mRNA level.The expression of SPHK1 at mRNA level was significantly increased after treating HUVECs with CYM-5442 and DENV-2 (P<0.05).Compared with DENV-2-infected or untreated HUVECs,Co-culturing DENV-2-infected HUVECs with CD4+T cells increased the expression of ICAM-1 and VCAM-1 in HUVECs at mRNA level (P<0.01) as well as the expression of IL-10 in CD4+T cells at mRNA level (P<0.05),but had no significant influence on the expression of TGF-β1 in CD4+T cells at mRNA level.Conclusion This study shows that DENV-2 can replicate and proliferate in HUVECs,but CD4+T cells inhibit the replication and proliferation.CD4+T cells play an important role in promoting the expression of VCAM-1 and ICAM-1 in DENV-2-infected HUVECs at mRNA level,activating HUVECs and increasing inflammation,which may be associated with increased vascular permeability induced by DENV-2 infection.Co-culturing CD4+T cells with DENV-2-infected HUVECs promotes the expression of IL-10 in CD4+T cells at mRNA level,but has no significant effect on TGF-β1.

Background Dominant eye is one of the functional asymmetric organ,and the dfference between dominant eye and undominant eye is a researching hotspot.But the study about accommodation in adult myopia is less.Objective This study was to determine the association between ocular dominances and accommodative factors in the subjects with adult myopia.Methods This study used prospective descriptive research method.Thirty-five subjects aged from 18 to 35 years with the myopia ranged from-2.00 D to-10.00 D and anisometropia less than 1.5 D,BCVA≥ 1.0 were recruited consecutively in this study.Ocular dominance was determined using the hole-inthe-card test and thumb test.Refractive error was measured with objective and subjective optometry,and amplitude of accommodation was measured by push-up test.Fusion cross cylinder(FCC) was used to measure the accommodative lag,and flipper test was applied to determine the accommodative facility.Oral informed consent was obtained from each subject before any relevant examination.Results No significant differences were found in the amplitude of accommodation (D),accommodative facility (cpm) and accommodative lag (D) between the dominant eye and undominant eye (accommodative amplitude:9.69 D±2.30 D vs.9.60 D±2.37 D,P =0.294 ;accommodative facility: 11.08 D±4.20 D vs.10.63 D± 4.60 D,P=0.260;accommodative lag:P=0.141).In the patients with the right eyes as dominance eyes,the accommodative amplitude of both eyes were (9.48±2.29) cpm and (9.33 ± 2.49) cpm,and accommodative facility were (10.50 ± 4.70) cpm and (9.99 ± 4.90) cpm.There were no significant differences between the right and left eyes in the accommodative amplitude,accommodative facility and accommodative lag (P =0.319,0.116,0.590).In the patients with the left eyes as dominant eyes,the accommodative amplitude of both eyes were (9.91±2.35)D and (9.88±2.26) D,and accommodative facility were (10.70±3.77)cpm and (11.25 ±4.27) cpm.No significant differences were seen between the right eyes and left eyes in the accommodative amplitude,accommodative facility and accommodative lag (P =0.749,0.295,0.238).Conclusions The amplitude of accommodation of the dominant eye is not significantly enhanced,and less accommodative lag and better accommodative facility also are found in the demonstrate eye in myopia adults with low anisometropia.

Objective: To observe the effects of recombinant adenovirus TRAIL (AdS-TRAIL & Ad5F35-TRAIL) on apoptosis of non-small cell lung (NSCLC) cells, so as to assess the value of Ad-TRAIL in gene therapy of NSCLC. Meth-ods: CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells from samples of 10 NSCLC patients were assayed by flow cytometry analysis. The lung cancer cell lines and primary lung cancer cells were infected with Ad5-TRAIL & Ad5F35-TRAIL adenoviral vectors at MOI 10 or 50, re-spectively; the percentage of apoptosis cells labeled by Annexin V-FITC in different cells were measured by flow cytometry 48 h after transfection. Results: The expression of CD46 were higher than that of CAR in all the lung cancer lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5-TRAIL or Ad5F35-TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)% (Ad5-TRAIL), (15.96±2.89) % (Ad5F35-THAIL) and (6.05±1.58) % (Ad5-TRAIL), (10.11±1.26) % (Ad5F35-TRAIL), respectively, compared to no adenovirus-transfected cells ([2.33±0.37] % and [5.95±1.89]%, respectively, P < 0.05). Less than 10% of apoptosis cells were detected in NCI-H520 cells transfected with Ad5- or Ad5F35-TRAIL at MOI 50 ([12.89±3.2] % for AdS-TRAIL and [9.08±1.35]% for Ad5F35-TRAIL, respectively) compared to no adenovirus-transfected cells ([7.04±2.17] %, P > 0.05). Moreover, apoptosis induced by Ad5- or Ad5F35-TRAIL transfection in A549 cells was not detected both at MOI 10 and 50. About half of the primary lung cancer cells from 10 patients induced apoptosis after transfected with Ad5-TRAIL or Ad5F35-TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35-TRAIL group than those in Ad5-TRAIL and control groups. Conclusion: Ad5-TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35-TRAIL is more potent than Ad5-TRAIL, so Ad5F35-TRAIL is more suitable for gene therapy of NSCLC.

BACKGROUND: Experimental autoimmune encephalomyelitis has become the most classical animal model for multiple sclerosis. However, the experimental autoimmune encephalomyelitis model of China presented one-way course of disease. By using proteolipid protein 139-151 and proteolipid protein 178-191, relapse remitting experimental autoimmune encephalomyelitis models may be induced in SJL/J mice which were susceptible to immune, which have similar clinical situation, course and histologicallterations to multiple sclerosis.OBJECTIVE: To establish the relapse remitting experimental autoimmune encephalomyelitis mouse model induced by proteolipid protein, which has similar clinical situation, course and histological alterations to multiple sclerosis.DESIGN: Completely randomized controlled study.SETTING: The centre of Neuro-information, and Neurological Institute,General Hospital of Chinese PLA.MATERIALS: The study was carried out at the Laboratory of Neuro-pathology, General Hospital of Chinese PLA, from February to June 2004.Sixty female SJL/J mice with 8-12 weeks old were selected and randomly divided into proteolipid protein 139-151 group and proteolipid protein-178-191 group with 30 in each.INTERVENTIONS: After injected with proteolipid protein-139-151 or proteolipid protein-178-191, the models of relapse remitting experimental autoimmune encephalomyelitis were induced, and the body weight and neurological signs of each female SJL/J mouse were viewed. The tissue morphological changes of models were observed with hematoxylin and eosin and uxol fast blue stain.MAIN OUTCOME MEASURES: The neurological symptoms and signs,features of relapse and remitting and the perivascular inflammatory cell infiltration, demyelinated lesion of the model of experimental autoimmune encephalomyelitis mouse induced by two proteolipid protein peptides.RESULTS: All 60 mice entered the final analysis. ① Significant neurological symptoms, signs and features of relapse and remitting was manifested in the model of experimental autoimmune encephalomyelitis mouse induced by two proteolipid protein peptides. Obvious phenomena of perivascular inflammatory cuffing, satellitism, predominant perivascular inflammatory cell infiltration and demyelinated lesion were found in spinal and cerebral tissue. ②Changes of body mass: Before immunity, the body mass of mice in two groups was( 17. 84 ± 2.59) g and (17. 88 ± 0.52) g respectively. Onset of relapse of the mice in proteolipid protein-178-191 group was earlier and faster, their body mass had no distinctive change after immunization and the mean body mass was(23.52 ± 2.37) g till the 60th day. Meanwhile, Onset of relapse of the mice in proteolipid protein-139-151 group was later and slower. After the immunity, the body mass of mice was little decrease, and the body mass was (16. 70 ±0.46) g on the 60th day. ③ Neural functional scores: The highest functional scores in the two groups were not different(3.86 ± 1.10vs 3.71 ±1.05, t=0.49, P=0.628).CONCLUSION: The two different antigenic peptides of proteolipid protein can all cause the autoimmune response of central nervous system. Both models have the same characters of relapse and remitting and the severity has no significant difference. But compared with proteolipid protein 139- 151 group,onset and recover of the experimental autoimmune encephalomyelitis of the mice in proteolipid protein 178-191 group were earlier, as well as weight variance was larger, which maybe due to the different structure of two peptides.