OBJECTIVE: To observe monocyte chemoattractant protein 1 (MCP-1) changes in ischemic tissue during the process of angiogenesis induction in ischemic limbs by autologous mesenchymal stem cell (MSC) transplantation.METHODS: Twenty male Sprague-Dawley rats were randomized to 2 groups (n = 10): model and MSC transplantation. Femoral and tibial bone marrow was taken to isolate and culture MSCs by percoll density gradient method. Cells of the 3~(rd) or 4~(th) passage were used for transplantation. Severe bilateral hind limb ischemia was surgically created in each group rats. Two hours after model establishment, MSCs (1×10~(11)/L) were infused into the ischemic region of rats from the MSC transplantation group, and the model group received the same amount of phosphate buffered saline. Collateral artery formation was determined by angiographic analysis and histological assessment. CD68~+ macrophage infiltration was examined by immunohistochemistry. MCP-1 protein expression in the plasma and ischemic tissue was detected by enzyme-linked immunosorbent assay. MCP-1 mRNA expression in ischemic tissue was detected by reverse transcription-polymerase chain reaction.RESULTS: At postoperative 28 days, treatment with MSC transplantation lead to collateral vessel formation, and immunohistochemistry demonstrated that CD68~+ macrophage infiltration was lower compared with the model group. MCP-1 protein and mRNA expression in the plasma and ischemia tissue was significantly lower in the MSC transplantation group than in the model group (P < 0.05).CONCLUSION: Following MSC transplantation, MCP-1 may play an important role in ischemia-induced angiogenesis. This indicates that MCP-1 would become one possible target molecule for modulating inflammatory angiogenesis by MSC Transplantation.

Objective To investigate the role of AGE-RAGE system on intimal hyperplasia in autogenous vein graft of diabetic rats.Methods The experiments were performed on 40 male Sprague-Dawley rats which divided into two groups:diabetes group and control group.Autogenous vein graft model was established in all rats.Specimens were harvested 7 days and 14 days after surgery for histological examination,and RAGE,NF-?B p65 and VCAM-1mRNA were measured by semi-quantitative reverse transcription-PCR.Western Blotting and immunohistochemistry were used to detect the protein expression of RAGE,NF-?B p65 and VCAM-1.At the same time,the levels of serum AGE were tested.Results Compared with non-diabetic rats of control group,the intimal hyperplasia of vein graft in diabetic rats obviously increased(P

Objective To investigate the role of ribosomal S6 kinase (RSK) in the pathogenesis of rat hepatic fibrosis. Methods Intra-abdominal injection of dimethylnitrosamine was carried out to (establish) the model of rat experimental hepatic fibrosis. Immunofluorescent double labeling laser scanning (confocal) microscope was performed to detect the location of RSK, ?-smooth muscle actin(?-SMA) and the collagen type Ⅰ. Immunohistochemistry was used to determine the correlation of the expression of RSK and collagen type Ⅰ, Ⅲ in the tissue of hepatic fibrosis. Results In the tissue of hepatic fibrosis, RSK was coexpressed with ?-SMA, and the collagen type Ⅰ located around RSK. The expression of RSK was also correlated with that of collagen type Ⅰ and Ⅲ( P

Purpose To investigate the effects of intervention with Tanakan on anterior ocular segment in diabetic retinopathy (DR) after retinal photocoagulation. Methods Prospective random controlled study was performed on 72 patients (72 eyes) with ultrasound biomicroscopy (UBM),by obtaining and quantitatively analyzing the changes of anterior ocular segment including anterior chamber, anterior chamber angle, ciliary body and choroids before and the 3rd day and the 7th day after retinal photocoagulation. Results Three days after photocoagulation, significant elevated IOP and narrowed chamber angle were observed in control group and 4 eyes (11.11%) in Tanakan group (P

Objective To investigate the early influences of laser photocoagulation on retinal function in diabetic retinopathy(DR). Methods The multifocal electroretinograms (MERG) of 30 eyes with DR (phase Ⅲ～Ⅳ) were tested with visual evoked response image system IV before,and the 3 rd day and the 7 th day after laser photocoagulation. Results Three days after photocoagulation, the latency of N1 prolonged in the central macula 5? area and superionasal quadrant.The response densities of N1,P1 and N2 markedly reduced, and most significant changes occurred in the central macula 5? area and then in the central 10?area. There were also differences in the changes of the amplitude of N1 and P1 in different quadrants .The changes of visual acuity were positively related to the decrease of amplitudes of N1,P1 and N2 in the macula. Conclusion The reduction of response densities in MERG reveals functional damage in diabetic retina occurring early after photocoagulation.The functional damage in macula induced indirectly by photocoagulation may explain the reduction of visual acuity after panretinal photocoagulation in some degree.

Purpose To investigate the relationship between the changes of the thickness of retina in macula and the abnormalities in multifocal electroretinograms (mERG) in diabetic retinopathy. Methods mERG and optical coherence tomography (OCT) examination were performed in 38 patients (60 eyes) with DR (phase Ⅲ～Ⅳ). The data were processed with software SPSS and line relation analysis was done. Results The response densities of N 1, P 1 and N 2 in central 5? area was significantly negative related to the thickness of neuroretina in macular fovea (correlation efficient -0.252～-0.266, P

Objective To investigate the early influences of laser photocoagulation on macular retinal thickness in diabetic retinopathy(DR). Methods Optic coherence tomography examination was performed in 30 eyes with DR(phase Ⅲ~Ⅳ) before, and on the 3rd day and the 7th day after photocoagulation respectively. The thickness of neuroretina and pigment epithelium were measured in the areas of fovea macula and 750 ?m from fovea macula. Results Three days after photocoagulation, significant thickening of neuroretina was observed in the fovea macula, which is positively related with age, fasting blood sugar and duration of DR. There was no significant changes in the thickness of pigment epithelium in macula and in the thickness of neuroretina 750 ?m from fovea macula. Conclusion Significant thickening of neuroretina in fovea macula in DR early after photocoagulation reveals progressed macular edema induced by photocoagulation which is positively related with age, fasting blood sugar and duration of DR.

Objective To investigate the early effects of intervention with tanakan on retinal function in diabetic retinopathy(DR) after laser photocoagulation. Methods Prospective random controlled study was performed on 60 Patients (60 eyes) from 23 to 69 years old with DR(phase Ⅲ～Ⅳ). The multifocal electroretinograms (MERG) were tested with VERIS Ⅳ before , the 3rd day and the 7th day after photocoagulation. Results No significant differences were found in the latencies and response densities of N1,P1 and N2 between the two groups before photocoagulation. Compared with that before photocoagulation, three days after photocoagulation the latencies in tanakan group had no significant change. The response densities of N1,P1 and N2 reduced and the changes were much smaller than that in control. Three days after photocoagulation ,the response densities of P1 and N2 in the central macula 5? area were much higher and the latencies of P1 and N2 were significantly shorter than that in control group. There were no significant differences in the response densities in the 7th day and the differences in the latencies between two groups still existed. Conclusion Tanakan may be effective in preventing the retina from damage of retinal photocoagulation in some degree in DR.

OBJECTIVETo investigate the semen quality of cancer patients and search for a better way of sperm cryopreservation for them.

METHODSWe retrospectively analyzed the quality of the semen from 43 cancer patients under cryopreservation in the Sperm Bank of Zhejiang Province, and compared the semen parameters between the cancer patients and 248 normal donors as well as between the testicular cancer cases (n=22) and non-testicular cancer cases (n=21).

RESULTSThe cancer patients exhibited significantly lower semen quality than the normal donors as in sperm concentration (60.90 x 10(6)/ml vs 74.27 x 10(6)/ml), progressive motility (41.07% vs 51.79%), and recovery rate (49.98% vs 57.33%) (all P <0.05). Furthermore, the progressive sperm motility and sperm recovery rate after freezing were significantly decreased in the testicular cancer cases (15.68% and 42.81%) than in the non-testicular cancer cases (28.36% and 57.53%) (both P <0.05).

CONCLUSIONSemen quality declines in cancer patients, and therefore early sperm cryopreservation is essential for them. Due to the poor sperm motility and recovery rate of testicular cancer patients after freezing, further investigation is required on the improvement of sperm cryopreservation methods.

Adult ; Cryopreservation ; methods ; Humans ; Male ; Neoplasms, Germ Cell and Embryonal ; Semen ; Semen Analysis ; Semen Preservation ; methods ; Sperm Count ; Sperm Motility ; Testicular Neoplasms

[Objective]The oxidative injury of retinal pigment epithelium(RPE)plays a key role in the pathogenesis of age-related macular degeneration(ARMD). This study is to investigate the effects of endoplasmic reticulum stress and the vital transcriptional factor X-box binding protein 1(XBP1)in acrolein-induced oxidative damage of RPE.[Methods]RPE cells were treated with acrolein (75μmol/L)for 2~24 h,expression of glucose regulated protein 78(GRP78)and XBP1 was determined by Western blot analysis. After being transfected with XBP1 siRNA with 24 h,the expression of XBP1 was knocked-down in RPE cells. Protein level of Nrf2 and SOD2 was then determined by Western blot analysis and intracellular Reactive Oxygen Species(ROS)generation was determined by DCF staining. Acrolein was added for 8 h after being transfected with XBP1 siRNA or control siRNA for 24 h. Apoptosis was detected by TUNEL assay before and after the treatment. Subretinal injection of Cre or GFP adenovirus was performed in XBP 1flox mice. After 1 week the mice were sacrificed. Total RNA was extracted from mice eyecups using TRIzol and real-time RT-PCR was performed to determine the two XBP1 down-stream genes,ERdj4 and p58IPK. Cryosectioning and immunofluorescent staining were performed to look at the expression of XBP1,Nrf2 and SOD2 in mice RPE.[Results]Protein level of GRP78 was significantly un-regulated after exposure to acrolein for 2 and 4 h. XBP1 was activated after acrolein treatment for 6 h. Knock-down of XBP1 by siRNA down-regu?lates anti-oxidant genes expression and increased ROS generation in RPE cells. Loss of XBP1 exacerbates acrolein-induced cell apoptosis. XBP1 was knocked-down in the RPE of XBP1flox mice after subretinal injection of Cre adenovirus. Decreased mRNA level of ERdj4 and p58IPK,and decreased Nrf2 and SOD2 expression were seen in the Cre-injected group.[Conclusions]Acrolein induces ER stress and activates XBP1 in RPE cells. Knock-down of XBP1 down-regulates anti-oxidant genes expression ,increases ROS generation,and exacerbates acrolein-induced cell apoptosis. XBP1 plays a role in the anti-oxidant defense in the RPE cells.