Objeetive To investigate the expression of claudin-4 in the eutopic and ectopic endometfium of women with endometriosis and evaluate the role of clandin-4 in the pathogenesis of endometriosis.Methods Thiny-five women with endometriosis and 35 controls were studied.Expression of elaudin-4 was investigated using immunohistochemistry,western blot and RT.PCR,respectively.Morphologic change of tight junction was also observed in different kinds of endometria.Results (1)G1andular epithelial cells of control endometrium and eutopic endometrium showed intact tight iunctions in electron micrographs,whereas the morphology of tight junctions in ovarian endometriotic tissue was disrupted and collagen bundles could be easily detected.(2)The immunohistochemical staining of claudin-4 was localized to the glandular epithelial cell membrane.Deftcient or weak staining Wag found in ovarian endometriotic tissues. In control endometrium.eutopic and ectopic endometrium of women with endometriosis,the expression of clandin-4 protein Was 89±24.84±22 and 27±14.respectively.Relative expression of claudin-4 mRNA Was 14.5±6.8,13.8±9.5 and 2.6±2.5.respectively.Expression of claudin-4 Was significantly lower in the ectopic endometriotic tissue than in the eutopic cndometrium and the control at both mRNA and protein levels(P<0.05). N0 significant difference Was foand between eutopic endometrium from women with endometriosis and control endometrium from women without endometriosis (P>0.05).Conclusion Down-regulated expression of claudin-4 might play a pathogenic role in the formation of endometriosis.

OBJECTIVETo explore the impacts on EC50 in the remifentanil inhibition of tracheal intubation response by the transcutaneous electrical acupoint stimulation (TEAS) at Hegu (LI 4) and Neiguan (PC 6).

METHODSForty patients with selective surgery undergoing endotracheal intubation with intravenous general anesthesia were divided into I to II degree by the American Society of Anesthesiologists (ASA) and were randomized into an observation group and a control group, 20 cases in each one. Before general anesthesia induction, in the observation group, the transcutaneous electric stimulation was applied to bilateral Hegu (LI 4) and Neiguan (PC 6) for 30 min, with dense-disperse wave, 2 Hz/100 Hz in frequency; in the control group, the sham-stimulation was applied to the acupoints, with the lamp on, but without electric current output. Afterwards, the general anesthesia induction started. When the target concentration of propofol and remifentanil was stabilized at the preset value, the endotracheal intubation was conducted. Dixon sequential method was applied for the determination of ECs in remifentanil inhibition of tracheal intubation response.

RESULTSThe level of EC50 in remifentanil inhibition of tracheal intubation response was 3. 46 ng/mL, 95% confidence interval was 2. 80 ng/ml to 4. 27ng/mL in the observation group; those were 4. 18 ng/mL and 3. 30 ng/mL to 5. 29 ng/mL in the control group separately. The differences were significant in comparison of the two groups (P<0. 01).

CONCLUSIONTEAS apparently reduces EChe in the remifentanil inhibition of tracheal intubation response by around 17%as.

Acupuncture Points ; Adult ; Aged ; Anesthetics, Intravenous ; administration & dosage ; Female ; Humans ; Intubation, Intratracheal ; adverse effects ; Male ; Middle Aged ; Pain ; etiology ; Pain Management ; Piperidines ; administration & dosage ; Transcutaneous Electric Nerve Stimulation

Objective To construct short hairpin RNA (shRNA) recombinant expression vector for herpes simplex virus typeⅡ(HSV-2) UL29 gene and observe its inhibitory effect on HSV-2. Methods Four interference target sites of HSV-2UL29 gene were selected to construct 4 groups of small hairpin RNA respectively,named shRNA recombinant expression vector. The expression vectors were transfected into HEK293 cells with liposome. HEK293 cells were infected with HSV-2 after expression vector being transfected. The viral titer was estimated by end-point titration assay. The level of transcription was estimated by Real-Time PCR method. The expressing effect of protein was detected by Western-blot. Results Recombinant expression vector pGPU6/GFP/Neo-shRNA was constructed successfully. The result of end-point titration assay showed that the viral titer was reduced comparing with blank control (P<0.05). The result of RT-PCR showed that inhibition rates were respectively 28.80%, 59.95%, 66.08%and 36.27% comparing with blank control, and there were significant differences (P < 0.05). The effect of UL29shRNA1461 group was the best one. The result of Western-blot showed that the expressing quantity of ICP8 was reduced. Conclusion Recombinant expression vector pGPU6/GFP/Neo-shRNA can interfere HSV-2 UL29 gene expression from different cell level in vitro, which can inhibit the replication of HSV-2 genome in HEK293 cells. Thus, RNA interference (RNAi) is conducive to the further exploration of viral therapy.

Objective To observe how ox-LDL impacts the mRNA expressions of MMP-9 and LOX-1 of human umbilical endothelial cells (HUEVC) and how LOX-1 acts as in inflamation course.Methods HUVEC were incubated in vitro.mRNA Expressions of MMP-9 and LOX-1 were determined by reverse transcription polymerase chain reaction (RT-PCR).Results Compared to the control group(0.252±0.032;0.279 ±0.041 ),ox-LDL significantly increased the mRNA expressions of MMP-9 and LOX-1 (25 ng/group 0.486 ± 0.012,0.586 ± 0.02;50 ng/L group 0.668 ± 0.011,0.739 ± 0.014; 100 ng/group 0.817 ±0.030,0.872 ±0.003,P ＜0.01 ).Those expressions were increased by ox-LDL( 1.020 ±0.039)in a concentration-and time-dependent manner.MMP-9 mRNA(0.872 ±0.046) was reduced when LOX-1 was inhibited by polyinosinic acid ( P ＜ 0.01 ).Conclusions The mRNA expressions of MMP-9 and LOX-1 were induced by ox-LDL in HUVEC.Inhibition of LOX-1 may decrease the expression of MMP-9.Those data demonstrate that LOX-1 is involved in the process of ox-LDL-induced MMP-9 expression.

Objective To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma.Methods The expression of claudin-4 in 62 endonetrioid endometrial carcinoma (EEC),30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and realtime PCR.Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells,which were divided into three groups with different intraperitoneal treatments with cisplatin,paclitaxel and saline solution.After the observation period,the tumors were extracted and stained with monoclonal antibody against claudin-4.The messenger RNA expression of claudin-4 was also detected using real-time PCR.Results Among the EEC samples,34％ (21/62) showed medium staining for claudin-4 and 66％ (41/62) showed intense staining.In atypical hyperplasia group,27％ (8/30) showed weak staining,53％ (16/30) showed medium staining and 20％ (6/30) showed intense staining for claudin-4.Of the normal endometrial tissue,47％ (28/60) showed weak staining and 53％ (32/60) showed no staining for claudin-4.According to real-time PCR,the relative quantity of claudin-4 was 170 ± 12 in EEC group,89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium.Compared with those in atypical hyperplasia group and normal endometrium group,the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P ＜ 0.05).In the study of Ishikawa xenografts,no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group.Nevertheless,a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group.After cisplatin treatment,the tumor volume was significantly decreased [(0.51 ±0.21) versus (0.73 ±0.12) cm3],and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27).Conclusion These results demonstrate that claudin-4 is strongly expressed in EEC,which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.

Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019＜0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163～17.890,P=0.000～0.038＜0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ～ 6.308,P=0.000 ～0.016＜ 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.

Objective To induce skin cancer in BALB/c mice using DMBA as initiator and TPA as tumor promot-er. Through optimizing the doses and frequencies of DMBA administration to establish a stable skin cancer model with less time and causing less skin damage. Methods Shaving the back of mice to expose a piece of skin around 2 cm × 2 cm. The mice were divided into a blank control group and four treatment groups randomly. These four groups were given 1, 2, 4, 7 times 100μg/100μL DMBA/acetone, respectively, in the first week, and twice 4μg/100μL TPA/acetone per week in the next 11 weeks. The body weight changes, time of tumor formation and number of tumors formed were recorded during the experiment. The mice were sacrificed at 12th week and samples of tumor tissue and adjacent normal skin tissue were taken for pathological examination using HE staining. Results Tumors were observed at the 7th week in the group with once DMBA administration in the first week and at the 4th week in the group with twice DMBA administration in the first week. Skin cancers were formed also in the group with 4?time DMBA administration in the first week, however, with signif?icantly more severe skin damages. The mice receiving DMBA everyday in the first week died at the 3th week. Conclusions The best induction protocol for skin cancer in BALB/c mice should be twice DMBA in the first week followed by twice TPA

AIM: To investigate the relationship between up-regulation of plasminogen activator inhibitor-1 (PAI-1) expression and activation of p38 mitogen-activated protein kinase (p38 MAPK) and extracellular signal-regulated kinase ( ERK) pathways by TGF-β1 in human ovarian cancer cells .METHODS: PAI-1 expression in human ovarian cancer cells treated with TGF-β1 (10 μg/L)was assayed by real-time PCR and Western blotting.The activation of p38 MAPK and ERK was determined by Western blotting using phosphorylated p 38 MAPK and phosphorylated ERK antibodies . Specific p38 MAPK inhibitor (SB203580) or ERK inhibitor (PD98059) was used to inhibit their activation .RESULTS:TGF-β1 up-regulated the expression of PAI-1, and activated p38 MAPK and ERK pathways in the ovarian cancer cells .In-hibition of p38 MAPK activation by SB203580 resulted in significant inhibition of the mRNA expression of PAI-1 induced by TGF-β1.However, inhibition of ERK activation did not significantly alter TGF-β1-induced increase in PAI-1 mRNA level.CONCLUSION: TGF-β1-activated p38 MAPK pathway contributes to the up-regulation of PAI-1 expression by TGF-β1 in ovarian cancer cells .

AIM:To investigate the expressions of methylthioadenosine phosphorylase(MTAP)and ornithine decarboxylase(ODC)in human ovarian cancer.METHODS:60 fresh samples of ovarian cancer were collected.The expressions of MTAP mRNA and protein were analyzed by using RT-PCR and Western blotting,respectively.ODC activity was measured by high performance liquid chromatography.RESULTS:The expression levels of MTAP mRNA and protein in ovarian cancer were lower than those of control.In 9 of the 60 samples(15%)there were absence of detectable MTAP mRNA and protein.No significant relevance was found between the expression of MTAP and clinical pathologic features.ODC activity in ovarian cancer was(3.82?1.03)U,which was higher than that of normal ovarian tissues(1.38?0.59)U.ODC activity was related with tumor grade.In MTAP-deficiency ovarian cancer tissues ODC activity was significantly increased when compared with that of MTAP-expressing ovarian cancer samples.CONCLUSION:Down-regulated MTAP expression and up-regulated ODC activity really exist in ovarian cancer.Activation of ODC resulting from MTAP deletion may be one of the pathogenetic factors of ovarian cancer.

Objective To study the effect of postoperative adjuvant transarterial chemoembolization (TACE) on prognosis of patients after radical resection of primary hepatocellular carcinoma.Methods A retrospective analysis was conducted on 311 patients with primary hepatocellular carcinoma treated from 2002 to 2008 in Quanzhou First Hospital affiliated to the Fujian Medical University.Utilizing the COX regression model and the Kaplan-Meier analysis,the effects of adjuvant TACE on prognosis of both the high risk group (n =76) and the low risk group (n =91) with tumor ≤5 cm,as well as both the high risk group (n =65) and the low risk group (n =78) with tumor ＞ 5 cm were determined.The low risk group was defined as patients with a single tumor and without vascular invasion,while the high risk group was defined as patients with multiple tumors or with vascular invasion.Results The postoperative overall survival rate of the high risk group who underwent postoperative adjuvant TACE with tumor ＞ 5 cm was higher than that of the high risk group who did not undergo postoperative adjuvant TACE (P ＜ 0.05).However,whether or not postoperative adjuvant TACE was given had no effect on prognosis in the other subgroups of patients (P ＞ 0.05).Conclusion Postoperative adjuvant TACE was beneficial to the high risk group of patients with tumor ＞ 5 cm after radical resection of primary hepatocellular carcinoma.