Objective To study the effect of sensory nerves on the expression of epidermal growth factor(EGF), basic fibroblast growth factor(bFGF) by fibroblasts in the granulation tissues in skin wound healing. Methods In vivo and in vitro studies were conducted. Fifty wistar rats were randomly divided into capsaicin group and control group. The rats in the capsaicin group were subcutaneously injected with neurotoxin dose of capsaicin(50 mg/kg for 3 days) to chemically destroy the sensory nerves. Full thickness skin incisive wound was made on the rat′s back on 8th day after last injection. The contents of substance P(SP), which was mainly released from afferent sensory nerve terminal, and EGF, bFGF in granulation tissue of skin lesion were determined using immunohistochemistry combined with computerized imaging analysis on 3rd, 6th, 9th, 12th days after the incision. In the control group no capsaicin was injected. In the in vitro study the fibroblasts from granulation tissue of rats were cultured with different doses of SP(10-9 ~ 10-5 mol/L), and the protein expression of EGF, bFGF by cultured fibroblasts was determined using Western blot. Results Immunoreactive SP was not only found around the small blood vessels and in the intercellular space, but also in the cytoplasm of fibroblasts in granulation tissue. SP content and the protein expression of EGF, bFGF in granulation tissue decreased in the capsaicin group, compared with those in the control group, which indicated that SP(10-7 mol/L) added into the culture medium for fibroblasts could upregulate the levels of EGF, bFGF expression. Conclusions Protein expression of EGF and bFGF by fibroblasts in granulation tissue in skin wound healing might be related with the SP released from sensory nerves.

Objective To study autocrine characteristics of substance P(SP)in epidermal stem cells(ESCs). Methods In vitro cultured ESCs was stimulated by exogenous SP.Then,the expressions of SP gene,protein and exogenous SP protein in ESCs and the supernatant fluid were detected by immunofluorescence,ELISA,Western blot and real-time PCR. Results After stimulation by exogenous SP,the expression of SP protein in ESCs and supernatant fluid was up-regulated and reached peak at 24th hour,with higher concentration than the added SP.The gene expression of SP was up-regulated to the peak 6 hours after stimulation by SP.Both expression of gene and protein remained unchanged after the natural killer cell receptor Was blocked even by stimulation with exogenous SP. Conclusion Anticrine of SP of ESCs can be induced by exogenous SP.

Objective To investigate the Laparoscopic management for Nutcraker Syndrome(NCS) with resection of fibrous ring and placing extravascular stent.Methods This was a retrospective analysis of clinical data and treatment process of cases from March 2010 to February 2015 in urology department,affiliated hospital of Guizhou medical university.Five cases with NCS,4 males and 1 female;age were 28-40 years,mean age was 35 years,all cases were afflicted with gross hematuria and flank pain,the history of gross hematuria were 6-72 months.3 cases were afflicted with proteinuria.Duplex ultrasound scanning before surgery revealed the compressed left renal vein (LRV) between the aorta and the superior mesenteric artery(SMA),with peak velocity 110-132 cm/s,an average of 121 cm/s.The flow velocity of LRV in the renal hilum were 18-25 cm/s,an average of 21 cm/s.CT scanning showed that the stricture segment diameter of LRV were 1.2-2.5 mm,an average of 1.8 mm;and the max diameter of proximal dilatation of the LRV in renal hilum were 8.3-15.2 mm,an average of 10.1 mm.The ratio between the dilated segment inner diameter and the stricture segment were 3.4-9.5.Bleeding from the left ureteral orifice was detected by cystoscopy in 3 cases.5 cases were treated by resection of fibrous ring and placing extravascular stent with Laparoscopic management,and the average length of extravascular stent was 4.0 cm.Results The operation was successful in the 5 cases.The average operation time was 83 min.The average blood loss was 65 ml.Hematuria gradually reduce 5-6 days and resolved 7-20 days after surgery in 5 patients.Proteinuria was disappeared successful 2 weeks after surgery in 3 patients.There was no recurrence at 8-24 months' follow-up.3 days after surgery Doppler ultrasound showed the stricture segment diameter of LRV were 3.8-5.6 mm,an average of 4.9 mm;the ratio between the dilated segment inner diameter and the stricture segment decreased were 1.1-2.0,an average of 1.6;the peak velocity of compressed LRV were 25-45 cm/s,an average of 34 cm/s.6 months after surgery,CTA result showed no LRV compression in the aortomesenteric region;the max diameter of LRV in renal hilum were 7.9-9.8 mm and 6.0-8.8 mm in the aortomesenteric region of LRV.Conclusion Etiology of NCS exist a fibrous ring around the left renal vein outflow of the inferior vena cava besides the commonly anatomic extrinsic compression on the LRV as it crosses between the superior mesenteric artery and the aorta.The Laparoscopic management for NCS with resection of fibrous ring and placing extravascular stent is an effective minimally invasive treatment.

OBJECTIVETo investigate the effects of reducing APE/Ref1 expression in the cultures of rat spiral ganglion cells with oxidative damage induced by H(2)O(2).

METHODSPrimary cultured rat spiral ganglion cells were infected with small interfering RNA to APE/Ref1 (Ape1siRNA) for 72 h, followed by treating with H(2)O(2) (0, 10, 25, 50, 100 and 300 µmol/L) for 1 h , and then cultured in normal medium for 24 h. Western blot were used to detect the level of APE/Ref1 protein and phosphorylation of histone protein H2AX in the infected cells. The caspase3 activation was tested by spectrophotometric method . The cell viability was determined by MTT and the apoptosis of spiral ganglion cells was determined by terminal-deoxynucleotidyl transferase mediated nick and labeling (TUNEL).

RESULTSWestern blot showed that infection with Ape1siRNA resulted in APE/Ref1 reduced expression in the spiral ganglion cells. Exposing spiral ganglion cultures with reduced expression of APE/Ref1 to H(2)O(2) (50, 100, 300 µmol/L) for 1 h resulted in increasing in the phosphorylation of histone protein H2AX. The reduction in APE/Ref1 significantly reduced cell viability in cultures 24 h after 1 h expression to 50-300 µmol/L H(2)O(2). The apoptosis of cells and caspase 3 activity was detected significantly improved.

CONCLUSIONSThe induced of APE/Ref1 results in significantly decrease in spiral ganglion cells viability in oxidative stress. The repairing function of APE/Ref1 is necessary for optimal levels of neuronal rat spiral ganglion cells survival.

Animals ; Cells, Cultured ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; Gene Silencing ; Hydrogen Peroxide ; Oxidation-Reduction ; Oxidative Stress ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley ; Spiral Ganglion ; cytology ; metabolism

Objective To analyze the influencing factors of affected renal function after laparoscopic nephron-sparing nephrectomy with segmental renal artery blocking (SRPN) for low R.E.N.A.L.score small localized renal tumor with follow-up data.Methods The data of 33 patients was collected,who underwent SRPN for small renal tumor from January 2010 to April 2012,and were followed-up for 5 years successfully.In 33 integrated data of 5-year follow-up,there were 14 males and 19 females,aged(56.5 ± 11.8)years.The preoperative affected renal glomerular filtration rate (aGFRpre),postoperative affected renal glomerular filtration rate (aGFRpost),the residual ratio of aGFR (aGFRpost/aGFRpre),preoperative affected renal volume (aVolpre),postoperative affected renal volume (aVolpost) and the residual ratio of aVol (aVolpost/aVolpre) were collected at the postoperative 3rd,6th,12th,24th and 60th month in follow-up period.Preoperative and postoperative data were analyzed by paired-sample T test.Multivariate linear regression analysis determined the influence of body mass index,operation time,localized warm ischemia time and distance from tumor to renal collecting system.Compare the correlation between aGFRpost/aGFRpre and aVolpost/aVolpre.Results All 33 cases were performed successfully.The mean operation time was (108.4 ± 9.1) min,and the mean time of segmental renal artery blocking was (23.3 ± 3.0) min.Postoperative pathologic 17 (51.5％) of clear cell carcinoma,5 (15.2％) of papillary carcinoma,3 (9.1％) of chromophobe cell carcinoma.All of above margins were negative.8 (24.2％) of angiomyolipoma (9.1％).All 33 cases were followed up for more than 5 years.The 5-year overoll survival (OS) and cancer specific survival (CSS) were 100％.In the 3rd,6th,12th,24th and 60th month,aGFRpost were (34.6 ± 4.6) ml/min,(34.7 ± 4.8) ml/min,(34.9 ± 4.4) ml/min,(35.1 ± 4.4) ml/min,(35.2 ± 4.2) ml/min,decreased of (10.4 ± 2.4) ml/min,(10.4 ± 2.6) ml/min,(10.1 ± 2.4) ml/min,(9.9 ± 2.4) ml/min,(9.8 ± 2.5) ml/min compared with aGFRpre (P ＜ 0.05).Multivariate linear regression analysis showed that body mass index,operation time,localized warm ischemia time in SRPN and distance from tumor to renal collecting system were unable to influence aGFRpost (P ＞ 0.05).The correlation coefficients between aGFRpost/aGFRpre and aVolpost/aVolpre were 0.659,0.667,0.663,0.629,0.604 respectively,and the difference was statistically significant (P ＜ 0.05).Conclusions For low R.E.N.A.L.score small localized renal tumor with SRPN,the decrease of postoperative affected renal function is relevant to postoperative affected renal volume,not as to operation time,localized warm ischemia time in SRPN and distance from tumor to renal collecting system.

JS001 (toripalimab) is a humanized IgG monoclonal antibody which strongly inhibits programmed cell death protein 1 (PD1). In this study, we used a different iodine isotype (I) to label JS001 probes to target the human PD1 (hPD1) antigen. , the half maximal effective concentration (EC) value of I-JS001 did not significantly differ from that of JS001. The uptake of I-JS001 by activated T cells was 5.63 times higher than that by nonactivated T cells after 2 h of incubation. The binding affinity of I-JS001 to T cells of different lineages after phytohemagglutinin (PHA) stimulation reached 4.26 nmol/L. Humanized C57BL/6 mice bearing mouse sarcoma S180 cell tumors were validated for immuno-positron emission tomography (immuno-PET) imaging. Pathological staining was used to assess the expression of PD1 in tumor tissues. The homologous Ihuman IgG (IhIgG) group or blocking group was used as a control group. Immuno-PET imaging showed that the uptake in the tumor area of the I-JS001 group at different time points was significantly higher than that of the blocking group or the I-hIgG group in the humanized mouse model. Taken together, these results suggest that this radiotracer has potential for noninvasive monitoring and directing tumor-specific personalized immunotherapy in PD1-positive tumors.