Objective:In order to determine the nature of P 200 autoantigen Methods:12 cases of anti-P 200 pemphigoid sera were collected The skin sections from 6 cases of recessive dystrophic epidermolysis bullosa were studied with these sera by indirect immunofluorescence Results:All the 12 anti-P 200 pemphigoid sera could react with basement membrane zone (BMZ)of 5 cases of recessive dystrophic epidermolysis bullosa, while epidermolysis bullosa acquisita sera were negative in these skins In addition, in a case of recessive dystrophic epidermolysis bullosa, epidermolysis bullosa acquisita sera react with both BMZ and intracytoplasmic deposition of type Ⅶ collagen, while no anti-P 200 pemphigoid sera showed this reactivity Conclusion:These results suggested that the 200 kD antigen is not a component of type Ⅶ collagen, but a specific autoantigen

ObjectiveTo explore the recurrence inhibition of colorectal cancer by adenoviral transducted endostatin gene. Methods The recombinant adenovirus expressing endostatin was constructed. Its biological activities were observed. The levels of endostatin in mice peripheral blood, tumor local recurrence and tumor cell apotosis were analyzed after the endostatin gene transduction by adenovirus. Results The infecting supernatant of recombinant adenovirus significantly inhibited HUVEC proliferation. After the injection of the recombinant adenovirus, persistent high serum levels of endostatin in peripheral blood was observed, local recurrence rate decreased, and apotosis of recurrence tumor cells increased.Conclusions The intraveneously injection of recombinant adenovirus mediated endostatin gene produces high concentration and stable expression of endostatin,which effects prevention of local recurrence after surgical resection of colorectal cancer.

Objective To evaluate the effect of SAHA or/and PTX on survival and apoptosis of human paclitaxel-resist-ant ovarian cancer OC3/P cells, and explore whether the combination of two drugs has a synergistic effect .Methods The morphology of OC3/P cells in different drug-groups was observed by inverted microscope .Cell viability was evaluated by MTT assay.The apoptosis rate was analyzed by Annexin V-FITC/PI assay.Results The morphology change of OC 3/P cells treated with different drug was observed by inverted microscope , and the change in combination group was more signif-icant than one drug alone group .The result of cell survival measured by MTT assay showed that inhibition rate of combina -tion group was more higher than one drug alone group (P<0.05).The analysis of factorial design and gold formula method all proved that the two drugs had synergy .Further the result of flow cytometry showed that apoptosis rate in combination group was significantly higher than SAHA or PTX alone group (P<0.05).Conclusion SAHA and PTX can inhibit the survival and induce apoptosis of OC 3/P cells, and two drugs have synergistic antitumor effects .

Objective To evaluate the effect of suberoylanilide hydroxamic acid(SAHA) or/and paclitaxel(PTX) on lethality and autophagy of human ovarian cancer OC3 cells,and to explore whether the combination of the two drugs has a synergistic function.Methods The morphology of OC3 cells was treated with SAHA and/or PTX, and then the morphology of treated OC3 cells was observed under an inverted microscope, cell proliferation was detected by MTT assay and autoph-agy was analyzed by AO/EB double staining assay.The synergistic effect of SAHA and/or PTX was analyzed by factorial design and gold formula method.Results After treatment with SAHA and/or PTX, the morphology of OC3 cells in the combination group ( SAHA+PTX) displayed significant morphological changes.OC3 cells became less adherent and refrac-tive than in other groups.Cell proliferation by MTT assay demonstrated that the growth inhibition rate of the combination groups was higher than in groups treated with SAHA or PTX respectively( P<0.05) .Furthermore, the synergistic effect af-ter treatment with a combination of SAHA with PTX was proved by the factorial design and gold formula method.The auto-phagy rate of the combined groups was significantly higher than in single treatment groups (P<0.05) by AO/EB double staining.Conclusion SAHA and PTX can inhibit the survival of OC3 cells and induce its autophagy.The two drugs have synergistic antitumor effects.

OBJECTIVE To study the molecular mechanism of cisplatin(DDP)by which HeLa cell growth and proliferation are inhibited. METHODS Cultured HeLa cells were treated with DDP 0.02-75 μmol · L-1 for 24 or 48 h. CCK-8 assay was used to determine the cell proliferation. The wound scratch assay was used to detect the cell migration and invasion. Flow cytometry was used to detect the cell cycle arresting. q-PCR was used to test the expression of metastasis suppressor gene 1 (MTSS1)mRNA. Western blot was used to determine protein levels of MTSS1,phosphorylated-extra?cellular signal-regulated kinase(p-ERK) and phosphorylated-serine-threonine kinase(p-AKT). RESULTS Following the treatment with DDP for 24 or 48 h,the proliferation of HeLa cells was inhibited significantly (P<0.05),the value of the half inhibitory concentration (IC50) of cells was 4.14 and 11.82 μmol · L-1. Migration and invasion activity of HeLa cells were reduced according to the wound scratch assay(P<0.05). Flow cytometry results showed that the cell cycle was arrested at S phase. q-PCR results showed that MTSS1 mRNA expression changed with DDP in a concentration-dependent manner (r24 h=-0.965,P<0.01;r48 h=-0.953,P<0.01). Western blot showed that the protein levels of MTSS1,p-ERK and p-AKT expression declined significantly with the increase in DDP concentrations(p-ERK:r24 h=-0.875,P<0.01;r48 h=-0.966,P<0.01. p-AKT:r24 h=-0.831,P<0.01;r48 h=-0.863,P<0.01. MTSS1:r24 h=-0.969,P<0.01;r48 h=-0.988,P<0.01). CONCLUSION DDP treatment inhibits HeLa growth and proliferation by interfering with the MTSS1 expression and disturbing the activation of ERK and AKT signaling pathways.

Aim To detect the expression of hippocam-pus NGF gene in the mouse model of Parkinson 's dis-ease. Methods By intraperitoneal injection of 1-meth-yl-4-phenyl-1,2, 3, 6-tetrahydropyridine ( MPTP), a mice model of Parkinson ’ s disease was established. The success of PD model was identified by detecting the expression of mesencephalic tyrosine hydroxylase ( TH) with Western blot and immunofluorescence. The movement and cognitive ability of mice were evaluated by foot print analysis and step-through passive avoid-ance test. The expression of NGF gene in hippocampus of mice was detected with RT-PCR and in situ hybrid-ization. Results Compared with the control group, the levels of TH and NGF were reduced in the experi-mental group and the behavioral indexes were deflected significantly. Conclusion NGF may play an impor-tant role in the occurrence and development of the PD cognitive disorder.

Objective To investigate the clinical outcomes derived from Ningmitai combined with tamsulosin to prevent double-J stent syndrome after laser lithotripsy with ureteroscope. Methods 117 patients underwent laser lithotripsy with ureteroscope and then placed a double-J stent for draining were collected from January 2010 to January 2013. Patients with double-J stent placement were divided into four groups determined by dosage regimen. Tamsulosin group (30 cases) was treated with tamsulosin (0.4 mg once daily) lonely, Ningmitai group (29 cases) was treated with Ningmitai (1.52 g, trice time a day) lonely, tamsulosin combined Ningmitai group (30 cases) was treated with tamsulosin and Ningmitai at the same time, operation control group (28 cases) was neither tamsulosin nor Ningmitai. The catheter was removed on the 3rd day post-lithotripsy and then remained double-J stent for 1 month. The scores of urinary tract, pain and the incidence of gross hematuria were assessed. Results The significant differences in the improvement of symptom score (χ2=22.038, P=0.000), pain score (χ2=9.876, P=0.020) and hematuria (χ2=8.000, P=0.046) were found among tamsulosin group, Ningmitai group, and tamsulosin combined Ningmitai group. The number of patients with symptomless, slight symptom in tamsulosin combined Ningmitai group were higher than those of tamsulosin group, Ningmitai group, operation control group (symptomeless:14 vs. 6, 3 and 2 cases;slight symptom:13 vs. 9, 5, 4 cases). The number of patients with>Ⅱpain score (7 vs. 9, 14, 17 cases) and incidence of hematuriag [26.6%(8/30) vs. 56.7%(17/30), 58.6% (17/29), 53.6% (15/28)] were lower in tamsulosin combined Ningmitai group than those of tamsulosin group, Ningmitai group, operation control group. The drug combination of Ningmitai with tamsulosin had the synergism to relived symptom and pain, and showed the more obviousthan lonely use. Conclusion The drug combination of Ningmitai with tamsulosin can be used in clinic for prophylactic purpose to prevent double-J syndrome.

Objective To observe the effects of sodium‐selenite on human hypertrophic scar fibroblasts proliferation in vitro . Methods Human hypertrophic scar fibroblast culture was conducted in vitro ,the status of fibroblast proliferation of the 4th gener‐ation cells was tested by CCK‐8 ,which was divided into experimental group and control group ,the experimental group was divided into six groups (A ,B ,C ,D ,E ,F) ,and were added an equal volume‐containing 2 .5 ,5 .0 ,10 .0 ,20 .0 ,40 .0 ,80 .0 μmol/L concentra‐tions of sodium selenite in 10% FBS culture medium ;the control group added an equal volume of 10% FBS culture medium ,testing cell proliferation by CCK‐8 at 24 ,48 ,72 ,96 h respectively ;testing different concentrations of sodium‐selenite cell survival situation after 24 h by Live/dead reagent ;immunohistochemical was used to test intracellularⅠ ,Ⅲ type collagen expression after 24 h .Re‐sults (1)With the increased of concentration ,the inhibition rate of fibroblasts gradually increased as the concentration of sodium‐selenite ranged in 2 .5-80 .0 μmol/L(P<0 .05);(2) the inhibition rate of sodium‐selenite on fibroblasts gradually increased at the same concentration with time(P<0 .05);(3)Live/dead reagent test results showed that apoptosis cell number increased with the concentration increasing ;(4 ) With concentrations of sodium‐selenite increasing ,typeⅠ ,Ⅲ collagen expression of fibroblast de‐creased gradually .Conclusion Sodium‐selenite can inhibit human hypertrophic scar fibroblast proliferate in vitro and reduce Ⅰ ,Ⅲcollagen expression of fibroblast type.

Objective To evaluate the safety and efficacy of retroperitoneal laparoscopic resec-tion and reconstructive surgeries in urology. Methods Retroperitoneal laparoseopic resection and re-constructive surgeries were performed on 245 patients including 17 cases of adrenalectomies, 32 cases of radical nephrectomies, 12 cases of partial nephrectomies, 53 cases of nephrectomies, 5 cases of nephroureterectomies, 6 cases of unroofing of peripelvie renal cysts, 46 cases of unroofing of renal cysts, 4 cases of unroofing of polyeystic kidneys, 12 cases of pyeloplasties, 58 cases of ureterolithoto-roles. Results All 245 surgeries were successfully completed. The mean operation time was 59 (20-250) min and the estimated blood loss was 5-300 ml with no transfusion. There was no serious complication during perioperative period. Conclusion Retroperitoneal laparoscopic resection and re-conatruetive surgery in urology is safe and effective with the advantages of minimal invasion, quick re-covery and few complications.

Objective To explore the curative effects of mesenchymal stem cells(MSC)that overexpress in murine type 1 diabetes nephropathy (DN).Methods Mice were randomly divided into normal control(NC) group,DN group,C3-treated group,C3-MIGR1-treated group and C3-MIGR1-ICAM-1-treated group.Mice were given streptozotocin until the DN model was set up.The murine DN model was treated with murine MSC(C3H10T1/2),transfection empty vector of murine MSCs(C3H10T1/2-MIGR1/MSC) and murine MSCs (C3H10T1/2-ICAM-1/MSC)that overexpressed ICAM-1.After transplantation, the pathological features of kidneys were observed by Masson staining and the number of homing MSC cells to the kidney was calculated on days 1,3,7 by frozen section, while qPCR was used to analyze the expression of signaling molecules for collagen1, TGF-β1 and SMAD2 after treatment with various MSCs.Results Compared with DN group, the renal fibrosis treated with MSCs overexpressing ICAM-1 was significantly decreased by Masson staining.Three and seven days after transplant, the homing cells of MSC in different groups displayed no difference using tissue freezing section method.Furthermore, TGF-β1/SMAD signaling was lowly activated after the treatment with MSCs that overexpressed ICAM-1 compared with model mice(P<0.01).Conclusion MSCs that overexpress ICAM-1 can protect kidneys in the DN model.