Objective To explore the effect on effect of combined use of Y27632 (ROCK II inhibitor)and TDZD-8(GSK-3β inhibitor)on axonal regeneration of dorsal root ganglion (DRG) neurons in neogenetic rats. Methods All the thoracolumbar DRGs of two neogenetic Sprague-Dawley (SD)rats(<5 days)were harvested under the stereopsis raicrostat,and then the DRG neurons were cultured,purified and indentified.Fifteen adult female SD rats were randomly divided into three groups,ie,complete paraplegia group(5 rats),sham operation control group(5 rats)and normal group(5 rats)respectively.The T8-10 spinal cord extracts (SCEs) were harvested in the complete paraplegia group,sham operation control group and normal group respectively at day 7 after spinal cord injury.The experiment was divided into group A(DRG neurons + PBS),group B(DRG neurons + complete paralysis SCE),group C(DRG neurons + complete paralysis SCE + different concentration Y27632),group D(DRG neurons + complete paralysis SCE + different concentration TDZD-8)and group E(DRG neurons + complete paralysis SCE + proper concentration Y27632 and TDZD-8).The average axonal length and expression intensity of Tubulin βⅢ at distal end of neuronal axons were observed after two days of co-culture respectively in intro. Results (1)The average axonal length and expression intensity of Tubulin βⅢ at axon shaft and growth cone in the group B were significantly shorter and weaker than that in the group A,with statistical difference.(2)In the group C,the average axonal length and expression intensity of Tubulln βⅢ at axon shaft and growth cone in 5-10 μmol/L Y27632 treatment groups were more than that in the group B but lower than that in the group A.The average axonal length and expression intensity of Tubuhn βⅢ at axon shaft and growth cone in 20-50 μmol/L Y27632 treatment group were longer and stronger than that in the group A and the group B,especially the group B.Among different concentration Y27632 treatment groups,there was a longest average axonal length and strongest expression intensity of Tubulin βⅢ in 30 μmol/L treatment group.(3) In the group D,there was a longer average axonal length in 0.5-3 μmol/L TDZD-8 treatment group than that in the group A and the group B,with the longeat average axonal length in l μmool/L TDZD-8 treatment group.In 5-25 μmol/L TDZD-8 treatment groups,the average axonal length showed no difference compared with the group B but wns shorter than that in the group A.In all different concentration TDZD-8 treatment groups,the expression intensity of Tubulin βⅢ at axon shaft and growth cone was significantly stronger than that in the groups A and B.(4) In the group E,although the average axonal length was increased in the group E,there was no statisilcal difference compared with the group A,30 μmol/L Y27632 treatment group and l μmol/L TDZD-8treatment group.There was a significantly longer average axonal length in the group E than it in the group B and the expression intensity of Tubulin βⅢ at axon shaft and growth cone was stronger in the group E compared to the group A,30 μmol/L Y27632 treatment group and l μmol/L TDZD-8 treatment group.Conclusion The complete paralysis SCEs obviously inhibits DRG axonal growth,induces axonal retraction and growth cone collapse.High concentration of Y27632 can more obviously promote the axon growth compared with the low concentration,while the low concentration of TDZD-8 can obviously promote the axon growth.Combined use of appropriate concentration of TDZD-8 and Y27632 can promote the axon growth and induce the axons branching,as facilitates the formation of the axon circuit.

Objective To observe expression of human beta-defensin-3 (HBD-3) in tissues around the infected artificial prostheses and investigate its value in treatment and diagnosis of periprosthetic joint infection (PJI).Methods According to clinical diagnosis,periprosthetic tissues and normal synovial membrane excised in operation were collected and divided into the following four groups:PJI group (n =13),aseptic loosening group (loosening group,n =9),spacer treatment group (treatment group,n =12),and normal group (n =15).HE staining was used to observe infiltration of inflammatory cells.Immunofluorescence staining was used to detect positive cells number and fluorescence intensity.Image-pro plus (IPP) 7.0C software was used to measure the average value of absorbance.Preoperative peripheral white blood cell count,erythrocyte sedimentation rate (ESR),and C-reactive protein (CRP) results were documented.Then,differences of those parameters were analyzed and compared among groups.Results HE staining revealed that all groups had different degree of inflammatory cell infiltration except for normal group.Immunofluorescence staining revealed that the most number of positive cells and highest fluorescence intensity existed in PJI group.Value of absorbance in PJI group was 0.430 ± 0.013,followed by 0.308 ±0.005 in loosening group,0.234 ± 0.009 in treatment group,and 0.089 ± 0.019 in normal group.Preoperative peripheral white blood cell count,ESR and CRP were the highest in PFI group,but were not significantly different among the remaining three groups.Conclusion HBD-3 is highly expressed in tissues around the prostheses which had infection or aseptic loosening,but its expression in response to infection and loosening has difference.

Objective To explore the pathological changes of lung, expression of the relevant inflammatory factors and oxidative stress markers of Sprague-Dawley (SD) rats undergoing autologous orthotopic liver transplantation (AOLT). Methods Thirty SD rats were randomized into sham group and AOLT group. The pathological changes of lung, expression of the relevant inflammatory mediators and oxidative stress markers were detected . Results ( 1 ) Compared with the sham group , the pathological scores of lung tissue in AOLT group increased significantly and reached its peak at 8 h after surgery. Then the pathological scores decreased to the level of sham within 24 h to 48 h after surgery; (2)The relative expression of inflammatory mediators including TNF-α, IL-1β, IL-6 and IL-8 increased significantly and reached its peak at 8 h after surgery in AOLT group. Then decreased to the level of sham group within 24 h to 48 h after surgery; (3)The change trends of MDA and H2O2 were similar to inflammatory mediators.The relative SOD expression decreased significantly and touched the nadir at 8h after surgery and then increased. Conclusion The pathological changes of lung expression, the relevant inflammatory mediators and oxidative stress markers of rats underwent AOLT were consistent.

Fura—2 was used as a Ca~(2+) indicator to determine the intracellular calcium ion concentra-tion(〔Ca~(2+)〕i)of rat peritoneal macrophages(RPM?s),and APAAP enzyme immnoassay was ap-plied to detect the expression of Ia antigens on RPM?s.The results showed that norepinephrine(NE,10~(-9)mol/L)could markedly increase the〔Ca~(2+)〕i of the RPM?s(p

This study investigated the protective effect of ATP on skeletal muscle satellite cells damaged by H2O2 in neonatal rats and the possible mechanism. The skeletal muscle satellite cells were randomly divided into four groups: normal group, model group (cells treated with 0.1 mmol/L H2O2 for 50 s), protection group (cells treated with 16, 8, 4, 2, 1, 0.5, or 0.25 mmol/L ATP for 24 h, and then with 0.1 mmol/L H2O2 for 50 s), proliferation group (cells treated with 16, 8, 4, 2, 1, 0.5, or 0.25 mmol/L ATP for 24 h). MTT assay, FITC+PI+DAPI fluorescent staining, Giemsa staining and immunofluorescence were performed to examine cell viability and apoptosis, and apoptosis-related proteins. The results showed that the survival rate of skeletal muscle satellite cells was decreased and the apoptosis rate was increased after H2O2 treatment (P<0.01). Different doses of ATP had different effects on skeletal muscle satellite cells damaged by H2O2: the survival rate of muscle satellite cells treated with ATP at 4, 2, or 1 mmol/L was increased. The protective effect was most profound on cells treated with 2 mmol/L ATP. Immunofluorescence showed that ATP could increase the number of Bcl-2-positive cells (P<0.01) and decrease the number of the Bax-positive cells (P<0.01). It was concluded that ATP could protect skeletal muscle satellite cells against H2O2 damage in neonatal rats, which may be attributed to the up-regulation of the expression of Bcl-2 and down-regulation of Bax, resulting in the suppression of apoptosis.

Objective To study the changes of serum level of high mobility group box-1(HMGB- 1)in patients with multiple trauma in order to forecast organ dysfunction(OD)and deaths.Methods The optical densities of HMGB-1 in serum of 35 patients with multiple trauma were determined on 1st,3rd, and 7th days after trauma,and the incidence of organ dysfunction and deaths were evaluated,then analyzed statistically to learn the relation between the serum levels of HMGB-1 and deaths with an attempt of predic- ting the incident of organ dysfunction and deaths.Results (1)As OD was concerned,there was a statis- tically significant difference in optical density of HMGB-1 on 1st and 3rd days between the two groups of multiple injury patients(t=4.411,P

20(Z= -2.117, P=0.034), and between the patients with OD and without OD (Z=-3.089, P=0.002), but PCT was not so between the non-surviror and survivor (Z=-1.307, P=0.191). The serum PCT level correlated with the incidence of organ dysfunction (x~2=14.82, P=0.033) and APACHEII (x~2=12.83, P

Objective To observe the different expression of somatomedin-receptor in cell membrane of prostate epithelial cells at anoxic or normoxic condition.Methods Human prostate epithelial cells line RWPE-1 were cultured in vitro.At 4,8,12,24,48 h after cells had been seeded,the gene and protein expression of epidermal growth factor receptor (EGFR),fibroblast growth factor receptor (FGFR),transforming growth factor β1 receptor (TGF- β 1R),insulin-like growth factor-1 receptor (IGF-1 R) and vascular endothelial growth factor receptor (VEGFR) in prostate epithelial cells were tested by RT-PCR and immunohistochem-istry methods,respectively.Results The expression of mRNA and protein of EGFR,FGFR,IGF-1R,TGF- β1R,VEGFR were significantly increased in anoxic and normoxic prostate epithelial cells (P ＜ 0.01 ).At different time point,the expression of mRNA and protein of EGFR,FGFR,IGF-1R,TGF- β1R,VEGFR significantly higher in anoxic than those in normoxic prostate epithelial cells (P＜ 0.01 )besides 4 h EGFR mRNA,12 h EGFR protein,4 h IGF-1R mRNA,4 and 8 h IGF-1R protein,4 and 8 h TGF-β 1R mRNA,4 and 8 h TGF-β 1R protein,4 h VEGFR mRNA (P ＞ 0.05).Conclusion Anoxic prostate epithelial cell can up-regulate the expression of somatomedin-receptor.

Objective To study the influence of donor GFR on the early renal function in recipients undergoing living donor transplantation.Methods A total of 172 living donor transplant recipients in our kidney transplantation center from 2006 to 2011 were enrolled into this study.Among them,166 were genetically related (96.5％),while 6 were genetically unrelated (spouses in 5 and other in 1).The predonation GFR was measured by isotope clearance (99mTC-DTPA with few exceptions).The range of donor GFR was 62 to 148 ml/min.The recipients were classified into two groups according to donor graft GFR level (GFR≤45 ml/min,n=76; GFR＞45 ml/min,n =96).The predonation dialysis,cold and warm ischemia time,antibody induction,immunosuppressive regimens and HLA mismatch were not significantly different between two groups.Results There were no significant differences in the incidence of postoperative acute rejection and delay graft function (DGF).The postoperative Scr of GFR＞45 ml/min group in 1 week,1 month,3 months and 1 year was lower compared with the GFR ≤45 ml/min group,and only the difference of Scr in 1 week was significantly different (P＜0.05).A repeated-measure ANOVA revealed no significant differences were found in Scr variation of two groups during the first year after transplantation.Conclusions Predonation GFR of the donor has effect on the Scr of postoperative Ⅰ week of recipients,not on the Scr within a year.Recipients with graft GFR＞45 ml/min have lower Scr levels.

Objective To explore the effects of lactoferrin on T cells ( the levels of CD4 + T and CD8 +T lymphocytes) and the development of intestinal mucous membrane (villus heights, crypt depths, villus circumferences, and villus areas) in neonatal SD rats. Methods Totally 96 neonatal (one week old) SD rats were equally and randomly divided into twelve groups, in which animals were fed with lactoferrin at a dose of 1.0 g/( kg · d) (dose Ⅰ group), 3.0 g/(kg · d) (dose Ⅱ group), or 5.0 g/(kg · d) (dose Ⅲ group) for 2, 3, or4 weeks,with corresponding blank control groups. Rats in the dosage groups were killed at the set time points and the levels of venous blood CD4 + and CD8 + T lymphocytes were detected using immunofluorescence method. Jejunum ( 1 cm)and ileum (1 cm) specimens were obtained for pathological sectioning, and the villus height, crypt depth, villus circumferences, and villus areas were measured through image analysis system. Results The CD4 + T lymphocyte levels at two weeks were significantly different among dose I group, dose Ⅱ group, and control groups ( all P ＜0. 05).The CD8 + T lymphocyte levels at two weeks were significantly different among dose Ⅱ group, dose Ⅲ group,and control groups ( all P ＜ 0. 05 ). The villus heights, crypt depths, villus circumferences, and villus areas of jejunum at two weeks between feeding groups and control groups were not significantly different ( all P ＞ 0. 05 ), while the condition in ileum was on the contrary. The CD4 + T lymphocyte levels at three weeks were significantly different between feeding groups and control groups ( P ＜ 0. 05 ). The CD8 + T lymphocyte levels at three weeks between dose Ⅲ group and control groups were significantly different ( P ＜ 0. 05 ). The villus heights, crypt depths, villus circumferences, and villus areas of jejunum and ileum at three weeks were significantly different between feeding groups and control groups ( all P ＜ 0. 05 ). The CD4 + T lymphocyte levels at four weeks between feeding groups and control groups were significantly different (P ＜0. 05). The CD8 + T lymphocyte levels at four weeks were significantly different among dose Ⅱ group, dose Ⅲ group, and control groups ( all P ＜ 0. 05 ). Except villus areas of ileum, the villus heights, crypt depths, villus circumferences of jejunum and ileum, and villus areas of jejunum at four weeks were significantly different between feeding groups and control groups ( all P ＜ 0.05 ). Conclusions Lactoferrin can promote the levels of CD4 + and CD8 + T lymphocytes in venous blood and facilitate the development of the mucous membranes of jejunum and ileum. However, such effects are affected by the dose and timing of lactoferrin feeding.