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 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 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 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.

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 effects of cyclopamine (CYP) on endometrial carcinoma (HEC-1A) cell survival and on induction of cell apoptosis .Methods HEC-1A cells were treated with various doses of CYP (0, 5,10, 20 and 40 μmol/L) for 24 h respectively .Then,the inverted microscope was used to observe cell morphology .Cell proliferation and apoptosis were tested by CCK-8 assay and AO/EB bi-labelling assay.The apoptosis rate of HEC-1A was analyzed using flow cytometric analysis , and the key gene expression of Bax and Bcl-2 was detected by quantitative PCR .Results The HEC-1A cells exhibited dramatic morphological changes after treatment with CYP and in a dose-dependent manner .CYP significantly inhibited HEC-1A cell proliferation using CCK8 assays(P<0.05), and induced cell death by AO/EB bi-labelling assay.Moreover,flow cytometry analysis showed that CYP treatment resulted in HEC-1A cell apoptosis, and that a higher concentration of CYP induced severer cell apoptosis (P<0.05).Meanwhile, CYP treated HEC-1A cells exhibited up-regulated expression of Bax and down-regulated expression of Bcl-2 according to Q-PCR.Conclusion Our findings indicatee that CYP can inhibit HEC-1A cell proliferation and induce cell apoptosis .

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.

Objective To detect the expression of metastasis sappressor 1(MTSS1) gene in cervical cancer tissue and to clarify its association with cervical cancer.Methods Totally 103 cases of cervical tissue were collected between Dec 2011 and Dec 2014 and classified according to biopsy and stage .Q-PCR and Western blotting were used to detect the expression of MTSS1 in normal cervical tissue and in different clinical stages of cervical cancer tissue .Results The expression of MTSS1 inⅡB-Ⅳstages of cervical cancer tissue was significantly higher than that of normal tissue or Ⅰ-ⅡA stages through q-PCR (P=0.000).Western blotting results showed that MTSS1 was positively expressed in normal cervical tissue at a rate of 23.3% or 53.3% in cervical cancer tissue.Moreover, the expression of MTSS1 was poorly correlated with age, tumor differentiation and lymphnode metastasis in cervical cancer tissue (P>0.05).The protein level of MTSS1 expressed in ⅡB-Ⅳ stages was significantly higher than that ofⅠ-ⅡA stages(P=0.005).Conclusion The expression of MTSS1 indicates the clinical stage of cervical cancer , suggesting that MTSS1 may play an important role in the development of cervical cancer .

OBJECTIVE To investigate the effect of overexpression of vascular cell adhesion molecule-1(VCAM-1)on the migration in vitro of the murine mesenchymal stem cells(MSCs)and its possible mechanism. METHODS The migration ability of normal mouse MSC (C3) ,empty vector-transfected MSC(C3+N) and VCAM-1 transfected MSC(C3+VCAM-1)was assessed by Transwell culture system in vitro after incubation for 8 and 12 h,respectively. The fetal bovine serum (FBS) was used as the chemotactic agent to induce MSC migration. The transmigrated cells were detected with methylosaniliam chloride(crystal violet)as well as DAPI staining.Furthermore,the specific chemical inhibitors of mitogen-activation protein kinase (MAPK) pathway ( SB203580,PD98059 and JNK inhibitorⅡ)were added to the Transwell system for 12 h and the alteration of the MSC migration ability was evaluated. RESULTS After incubation with FBS for 8 and 12 h,the absolute migrated cell number(7467 ± 485 and 8795 ± 255)and migration rate〔(14.9 ± 1.0)% and(17.6 ± 0.5)%〕of MSC in C3+VCAM-1 group were significantly increased compared with C3 group〔2731±562 and 4779±224, (5.5 ± 1.1)%and(9.6 ± 0.4)%〕and C3+N group〔2539 ± 321 and 5645 ± 1080,(5.1 ± 0.6)%and(11.3 ± 1.1)%〕(P<0.05,P<0.01),but there was no significant difference between C3 and C3+N groups. Moreover,the MSC migration ability of C3+VCAM-1 group was partially suppressed by addition of JNK inhibitorⅡ. The transmigrated cell number(4843 ± 167)and migration rate〔(9.7 ± 0.3)%〕were decreased compared with those of C3+VCAM-1 group without JNK inhibitorⅡ(P<0.01). SB203580 and PD98059,as specific chemical inhibitors of MAPK pathway,had no effect on MSC migration. CONCLUSION VCAM-1 can enhance mouse MSC migration in vitro and th4e mechanism may be related to JNK/MAPK pathway activation.