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Accidents caused by the bites of brown spiders (Loxosceles) generate a clinical condition that often includes a threatening necrotic skin lesion near the bite site along with a remarkable inflammatory response. Systemic disorders such as hemolysis, thrombocytopenia, and acute renal failure may occur, but are much less frequent than the local damage. It is already known that phospholipases D, highly expressed toxins in Loxosceles venom, can induce most of these injuries. However, this spider venom has a great range of toxins that probably act synergistically to enhance toxicity. The other protein classes remain poorly explored due to the difficulty in obtaining sufficient amounts of them for a thorough investigation. They include astacins (metalloproteases), serine proteases, knottins, translationally controlled tumor proteins (TCTP), hyaluronidases, allergens and serpins. It has already been shown that some of them, according to their characteristics, may participate to some extent in the development of loxoscelism. In addition, all of these toxins present potential application in several areas. The present review article summarizes information regarding some functional aspects of the protein classes listed above, discusses the directions that could be taken to materialize a comprehensive investigation on each of these toxins as well as highlights the importance of exploring the full venom repertoire.(AU)
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Animales , Venenos de Araña/toxicidad , Arañas , Serpinas , Serina Proteasas , Mordeduras y PicadurasRESUMEN
Objective To investigate the effect of pigment epithelium-derived factor (PEDF) on the pathogenesis of preeclampsia disease,by detecting the expression of PEDF in the placentas,as well as the relationship between PEDF and the production of placental vessels.Methods A study was performed in 60 cases of pregnant women with preeclampsia in the obstetrical department of Nanfang Hospital affiliated to southern medical university from October 2011 to January 2013,in which 30 cases were patients with mild preeclampsia(mPE) and other 30 cases were those with severe preeclampsia (sPE).40 normal pregnant women who also been hospitalized and delivered were selected as control group.The expression of PEDF and micro-vessel density (MVD) in placentas were assayed by using western blot and SP immunohistochemical method,then the relationship between PEDF and MVD was analyzed.Results (1) The pathological changes of placentas:the placental weight were lightened obviously in the mild preeclampsia and severe preeclampsia groups,the reduced blood vessels and luminal stegnosis were found in chorionic villus,basement membrane of trophocytes were thickening.The hyperplasia syneytiotrophoblast were like nodosity,with focus infarction,fibrinoid necrosis,or thrombogenesis.While there was no the above mentioned pathological alteration in normal control group.(2)The levels of PEDF expression in mild and severe preeclampsia group were 0.63 ± 0.09,0.93 ± 0.07,while 0.47 ± 0.04 in control group,which in mild and sever preeclampsia groups were significantly higher than that in normal group (P ＜ 0.05).Compared to mild preeclampsia group,the expression of PEDF was significantly increased in severe preeclampsia group,there was statistical significance between the difference (P ＜ 0.05).(3) The amount of microvessel density (MVD) in mild and severe preeclampsia group were 106 ±9,93 ±8,while 136 ±9 in control group,which were significantly reduced in mild and severe preeclampsia group,compared to that in normal control group (P ＜ 0.05).And it was significantly lower in severe preeclampsia group than that in mild preeclampsia group (P ＜ 0.05).(4) The expression of PEDF was negatively correlated with the amount of MVD in mild and severe preeclampsia group (r =-0.426,P ＜ 0.05 ; and r =-0.646,P ＜ 0.05 respectively),which was also negative in control group (r =-0.589,P ＜ 0.05).Conclusion Increased PEDF expression in placentas of women with preeclampsia induce the dysfunction of the placental vascular reconstruction and the pathological alteration like ischemic and hypoxia in placentas,which may be involved in pathogenesis and pathogenic progress of preeclampsia.
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Objective To construct eukaryotic expression plasmid pIRESneo3-pigment epithelium-derived factor (PEDF) and detect its transient expression in SP2/0 cells. Methods Specific primers were designed based on the mature peptide sequence of human PEDF cDNA in the GenBank. Human PEDF gene was cloned into the eukaryotic expression vector pIRESneo3. The PEDF DNA was transfected into SP2/0 with LipofectamineTM 2000. The recombinant human PEDF protein expressed in SP2/0 cell culture supernatant was identified by Western blot and enzyme-linked immunosorbent assay. The biological activity of the recombinant human PEDF was measured by 3-(4,5-dimethylthiazol-z-y1)-2,5-diphenytetrazolium bromide(MTT) method. Results PCR amplification, restriction enzyme digestion and DNA sequencing confirmed that the mature peptide sequence of human PEDF cDNA was successfully cloned into the eukaryotic expression vector pIRESneo3. And the plasmid was transfected into SP2/0 cells, which could secret PEDF. Western blot analysis showed that there was only one obvious band at the position of relative molecular weight of 50 000, and it is equivalent to the expected value. Enzyme-linked immunosorbent assay suggested that the content of PEDF began to rise after transfection, and peaked at 36 h [(0.92±0.04) μg/ml]. The proliferation of human umbilical vein endothelial cell line was significantly inhibited by supernatant after transfection of 36 h (P<0.05). Conclusions The eukaryotic expression plasmid pIRESneo3-PEDF had been successfully constructed and active human PEDF was transiently secreted, which made a foundation for further study of stable expression and purification of PEDF. This protein could be a potential medication for preventing and managing retinopathy of prematurity.
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Objective To investigate the impact of squamous cell carcinoma antigen(SCCAg)in patients with recurrent squamous cell carcinoma of the uterine cervix.Methods Totally 72 patients with recurrent squamous cell carcinoma of the uterine cervix treated at the Cancer Hospital,Peking Union Medical College,Chinese Academy of Medical Sciences,between 1999 and 2005 were retrospectively analyzed to investigate the impact of SCCAg on diagnosis and prognosis by univariate and multivariate analysis.Results This study included 30 patients with recurrent disease after primary radical surgery and 42 patients with recurrent cervical cancer after radio-chemotherapy.Sixty one patients(85%)had serum SCCAg elevated (≥1.5 pg/L),and 20 of these(28%)had an increase of SCCAg before clinical manifestation of relapse.The median leading time was 3 months(range:1-13 months).Forty five patients had no symptoms with only SCCAg elevation,and 15 patients experienced leg edema and(or)sciatic pain,7 patients suffered from irregular bleeding and 5 patients had symptoms resulting from distant metastasis.Thirty three patients were diagnosed by histology biopsy and (or) cytology,39 patients were diagnosed with SCCAg elevation and clinical and radiological examinations,29 of these patients were diagnosed only by SCCAg elevation and CT or MRI.Fourteen patients recurred limited to the cervix or to the cervix and adjacent tissues(central recurrence),31 cases recurred at pelvis,and 20 patients with distant metastasis and 7 patients suffered from Pelvic recurrence and distant metastasis.Twenty three cases received salvage therapy including surgery for patients recurring after definitive radiotherapy and radiotherapy and or conform radiotherapy for patients after primary radical surgery,46 patients were given palliative chemotherapy and or radiotherapy,and 3 patients refused any treatment.The median and mean survival time were 11 months and 23 months respectively(2-62 months).The 3-year,5-year overall survival rate were 25%and 19%respectively.Univariate analysis showed SCCAg elevation before primary treatment,grade,recurrent site,treatment method,SCCAg≥10pg/L,SCCAg elevation during treatment,and SCCAg not within normal after treatment were correlated with 3-year survival rate.Twenty patients had an increase of SCCAg before clinical manifestation of relapse compared with other patients who did not,and the 3-year survival rate was not significantly different (22% vs 27%). Multivariate analysis revealed that only grade and treatment methods were independent risk factors. Conclusion The impact of the SCCAg in recurrent squamous cell carcinoma of the uterine cervix needs further study.
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Objective To assess the impact of pigment epithelium-derived factor(PEDF)on the proliferation and apoptosis of the glioma cells by detecting expression of apoptosis related proteins.Methods U87 cells were treated with PEDF(1000μg/ml,U87PEDF),or without PEDF(U87com0),cell proliferation assays were performed by MTT assay to test the effect of PEDF on proliferation of glioma cells;Apoptosis assays were performed by flow-cytometric analysis;Western-blot Was used for evaluating the expression of p16 protein.Results The induced inhibitony rates of glioma cells by PEDF were(54.29±0.62)% Compaxed with the control(t=2.63,P<0.05).The apoptosis assay showed that(21.84±0.36)% of PI- negative/annexin V-positive Was present in the U87 PEDF cells.The appoptosis was associated with the incteases of p16 protein(0.82±0.09)compared with tlle control(0.43±0.03,P<0.05).Conciusion PEDF may play a significant role in apoptosis regulation and proliferation of glioma cell accompanied with the increase of the p16 protein.
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Maspin is a unique serine proteinase inhibitor that has tumor suppressor activity. It has been reported that maspin is expressed in normal human mammary epithelial cells and it is down-regulated during the progression of cancer. However, to date, there is very limited data on the clinical significance of maspin expression in human breast cancer. In this study, maspin expression was assessed immunohistochemically from 80 invasive ductal carcinoma (IDC) specimens of the breast. Also, maspin expression was compared with the clinicopathological factors (age, grade, tumor size and lymph node status), the expression of estrogen receptor (ER), progesterone receptor (PR) and p53, DNA ploidy and the overall survival in an attempt to assess its prognostic value. The maspin expression was positive in 25 IDC cases (31.3%). The maspin expression in IDC was significantly correlated with a higher histologic grade, a larger tumor size, a positive p53 status and shorter survival. There was an inverse association with maspin expression and the PR status. These findings suggest that maspin expression is not down-regulated with the progression of cancer and maspin expression may be associated with a poor prognosis. The immunohistochemical detection of maspin in breast cancers may be helpful for predicting an aggressive phenotype.