With reverse transcriptase PCR, the transcripton of copper homeostasis relative gene Afe0329 in Acidithiobacillus ferrooxians standard strain ATCC23270 was investigated. The further analysis of genes in this transcripton was analyzed employed by Vector NTI, Blast, TMHMM Server, PSORTb software and so on. From the DNA of different strains, the transcripton of Afe0329 was amplified using special primer pairs to identify the universality of it in the genome of A.ferrooxidans strains. The results showed that gene Afe0330 and Afe0331 were cotranscribed with Afe0329, and they were in a single transcripton. Gene Afe0329 was supported to express a P1b3-type ATPase which is a heavy metal ion pumping transmembrane protein, protein AFE0330 which expressed by gene Afe0330 was a cytoplasmic protein, no significant ho- mologous sequences of Afe0330 or Afe0331 had been obtained by Blast analysis. And the transcripton of Afe0329 was universal in genome of A. ferrooxidans strains.

OBJECTIVETo establish an appropriate animal model of uterine leiomyoma and to understand the pathogenesis of this disease.

METHODSMature female rats were intramuscularly injected with estradiol benzoate at 200 μg or 300 μg twice a week. After injection for 8 or 10 weeks, the rats were sacrificed. We measured the serum levels of estrogen (E(2)) and progesterone (P), evaluated ER and PR expression, and calculated the leiomyoma forming rate and mortality of the rats. Histological changes were compared between rat uterine leiomyoma and human uterine leiomyoma with HE staining. The optimal dose and duration of E(2) for induction of uterine leiomyoma in rat were determined.

RESULTSIn the rats treated with estradiol benzoate 200 μg for 8 weeks ìn the serum E(2) level increased significantly (P<0.01). Uterine nodules were visible in some of the tested rats. Based on the pathohistological Results , the uterine leiomyoma developed in the treated rats demonstrated similar features as in human uterine leiomyoma. The expressions of ER and PR were increased in the leiomyoma tissues.

CONCLUSIONThe rat model of uterine leiomyoma can be established by intramuscular injection of estradiol benzoate at 200 μg twice per week for 8 weeks, with similar features as those of human uterine leiomyoma. The high concentrations of ER and PR in uterine tissue might be related with the development of uterine leiomyoma in animal.

Animals ; Disease Models, Animal ; Estrogens ; administration & dosage ; adverse effects ; Female ; Leiomyoma ; chemically induced ; Rats ; Uterine Neoplasms ; chemically induced

BACKGROUNDStathmin was identified as an endometriosis-related protein by comparative proteomics in our previous study. As a microtubule-destabilizing factor, stathmin was shown to participate in the relay and integration of diverse intracellular signaling pathways involved in cell proliferation, differentiation, and many other cellular activities. To investigate whether stathmin is involved in the pathogenesis of endometriosis, we examined the expression of stathmin in eutopic endometrium of women with or without endometriosis.

METHODSEutopic endometrium samples were collected from thirty-six patients who were diagnosed as endometriosis and the nineteen age-matched patients who were confirmed to be free of endometriosis surgically and histologically. The expression of stathmin mRNA was detected by real-time PCR, and its protein was detected by Western blotting and immunohistochemistry.

RESULTSStathmin was overexpressed in eutopic endometrium of women with endometriosis detected by real-time PCR in mRNA levels and by Western blotting in protein levels, without significant difference between proliferative and secretory phase. Immunohistochemistry showed that stathmin protein was localized in both endometrial glandular and stromal cells throughout the menstrual cycle.

CONCLUSIONSStathmin is overexpressed in endometrium of patients with endometriosis and may play a role in the pathogenesis of endometriosis.

Adult ; Blotting, Western ; Endometriosis ; metabolism ; Endometrium ; metabolism ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Polymerase Chain Reaction ; Stathmin ; genetics ; metabolism

OBJECTIVETo explore the effects of adenovirus delivered tissue inhibitor of metalloproteinases-3 (Ad-TIMP-3) on the biological behaviors of cervical cancer cell lines and to evaluate its potential application in cervical cancer gene therapy.

METHODSWe transferred Ad-TIMP-3 into cervical cancer cells. The TIMP-3 mRNA expression was assessed by RT-PCR, and the TIMP-3 and p53 protein expressions were assessed with Western blot. The apoptotic changes of cells were illustrated with morphology and DAPI staining. The viability of cells was determined with MTT assay. The abilities of in vitro invasion and adhesion were evaluated by the invasion and adhesion assays respectively.

RESULTSAfter infection, the TIMP-3 mRNA and protein were significantly upregulated in a time-dependent manner. Overexpression of TIMP-3 markedly increased p53 protein level in spite of the backgrounds of p53 gene in cells. Ad-TIMP-3 infection induced massive apoptosis of cervical cancer cells with a marked bystander effect. The abilities of in vitro invasion and adhesion were inhibited significantly (P < 0.01). The cytotoxicity of Ad-TIMP-3 was significantly stronger than that of Ad-p53 (P < 0.05, P < 0.01).

CONCLUSIONSAd-TIMP-3 infection has cytotoxic effects on cervical cancer cells and can inhibit the expressions of these malignant phenotypes. Ad-TIMP-3 may be a potentially useful agent for cervical cancer gene therapy.

Adenoviridae ; genetics ; Apoptosis ; Cell Adhesion ; Cell Line, Tumor ; Cell Survival ; Female ; Gene Transfer Techniques ; Humans ; Neoplasm Invasiveness ; Tissue Inhibitor of Metalloproteinase-3 ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; Uterine Cervical Neoplasms

OBJECTIVETo investigate the clinical features and management of ureteral endometriosis.

METHODSPatients surgically and histologically diagnosed as ureteral endometriosis from January 2001 to January 2007 in Peking Union Medical College Hospital were retrospectively reviewed.

RESULTSTen patients were diagnosed as ureteral endometriosis among 7561 cases with surgically and histologically proved diagnosis of endometriosis, with an incidence of 0.132%. Nine out of 10 patients were extrinsic ureteral endometriosis and concomitant with severe pelvic endometriosis, and the other was intrinsic ureteral endometriosis. Hormone therapy failed in 2 patients with urinary tract obstruction. Ureterolysis was performed in 6 patients and ureterectomy was performed in 4 patients. One case of ureteral recurrence was observed in a postmenopausal woman without hormonal replacement therapy who received laparoscopic ureterolysis and hysterectomy with bilateral adnexectomy. No relapse was observed in the other 9 patients.

CONCLUSIONSUreteral endometriosis is a rare entity. The upper urinary tract should be evaluated in patients with severe endometriosis, even in postmenopausal women. The treatment of ureteral endometriosis usually requires surgery, while ureterolysis should not be performed in patients with extensive disease. As a form of adjuvant therapy of surgery, hormonal therapy is an appropriate option.

Adult ; Diagnosis, Differential ; Endometriosis ; complications ; pathology ; therapy ; Female ; Humans ; Middle Aged ; Retrospective Studies ; Ureter ; pathology ; Ureteral Obstruction ; etiology

OBJECTIVETo gain insight into the role of Toll-like receptor 2 (TLR2) in graft rejection following penetrating keratoplasty, and investigate the expression of TLR2 mRNA in the corneal graft.

METHODSPenetrating keratoplasty was performed in 3 groups of rats for orthotopic autologous corneal transplantation (group A), allograft corneal transplantation (group B), or allograft corneal transplantation with hormone treatment (C). The transparency and neovascularization of the cornea were observed using a slit-lamp microscope and scored according to the rejection index, with normal cornea serving as the control. The corneal tissues were sampled at 5, 7, and 9 days after the transplantation for histopathological examination and detection of TLR2 mRNA expression using RT-PCR.

RESULTSWith the passage of time, edema, opacities and neovascularization of the corneal graft occurred after the operation in all the groups. Seven days after the operation, the rejection index of group B, but not that of groups A and C, met the diagnostic criteria for graft rejection with also support by histopathological evidence. The expression of TLR2 mRNA was detected in normal corneas and augmented in the corneal grafts in the 3 transplantation groups. TLR2 mRNA expression in group B was significantly higher than that of group A, and the expression in group C decreased significantly in comparison with that in group B (P<0.05).

CONCLUSIONAs the recognition receptors of native immune system, TLR2 in the rejected corneal grafts may recognize the allograft antigen and play a role in acute graft rejection after penetrating keratoplasty.

Animals ; Cornea ; metabolism ; Female ; Graft Rejection ; immunology ; Keratoplasty, Penetrating ; Postoperative Complications ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism

BACKGROUNDOccult stress urinary incontinence may lead to de novo stress urinary incontinence after pelvic floor repair surgery. A measurement of pudendal nerve terminal motor latency can reflect the integrity of the nerves. We aimed to explore the value of pudendal nerve terminal motor latency in the diagnosis of occult stress urinary incontinence in pelvic organ prolapse patients.

METHODSTen patients with stress urinary incontinence (SUI group), 10 with SUI and uterine or vaginal prolapse (POP + SUI group) and 10 with uncomplicated uterine or vaginal prolapse (POP group) were evaluated for their pudendal nerve terminal motor latency using a keypoint electromyogram.

RESULTSThe amplitude of positive waves was between 0.1 and 0.2 mV. The nerve terminal motor latency was between 1.44 and 2.38 ms. There was no significant difference in the wave amplitudes of pudendal nerve evoked action potential among the three different groups (P > 0.05). The pudendal nerve latency of the SUI group, POP + SUI group and POP group were (2.9 ± 0.7) seconds, (2.8 ± 0.7) seconds and (1.9 ± 0.5) seconds respectively. The difference between the SUI group and POP + SUI group was not statistically significant (P > 0.05), whereas the difference between the SUI and POP groups and between the POP + SUI and POP groups were statistically significant (P < 0.05). There was a positive correlation between pudendal nerve latency and the severity of SUI; the correlation coefficient was 0.720 (P < 0.01).

CONCLUSIONSPatients with SUI may have some nerve demyelination injuries in the pudendal nerve but the damage might not involve the nerve axons. The measurement of pudendal nerve latency may be useful for the diagnosis of SUI in POP patients.

Evoked Potentials ; physiology ; Female ; Humans ; Middle Aged ; Pelvic Organ Prolapse ; physiopathology ; Pudendal Nerve ; physiopathology ; Urinary Incontinence, Stress ; diagnosis ; physiopathology ; Uterine Prolapse ; physiopathology

BACKGROUNDAnnexin-1 was identified as an endometriosis-related protein by comparative proteomics in previous study. As an endogenous anti-inflammatory mediator, Annexin-1 has been shown to regulate the immune response, cell proliferation and apoptosis. To investigate whether Annexin-1 is involved in the pathogenesis of endometriosis, we examined the expression of Annexin-1 in eutopic endometrium of women with or without endometriosis, and detected its expression in peritoneal fluids of those with endometriosis.

METHODSEutopic endometrium samples from twenty-five women with endometriosis and those from sixteen age-matched women without endometriosis were collected. Peritoneal fluids were obtained from ten patients with endometriosis. The expression of Annexin-1 protein in eutopic endometrium was detected by immunohistochemistry and Western blotting, and mRNA detected by real-time PCR. Annexin-1 protein in the peritoneal fluids was detected by Western blotting.

RESULTSAnnexin-1 mRNA and protein were overexpressed in eutopic endometrium of endometriosis without significant differences between the proliferative and secretory phase. Immunohistochemistry showed that Annexin-1 protein was expressed mainly in endometrial glandular cells throughout the menstrual cycle. Annexin-1 protein was detected in the peritoneal fluids of all the ten patients with endometriosis.

CONCLUSIONSAnnexin-1 is overexpressed in eutopic endometrium and presents in the peritoneal fluids of patients with endometriosis, and may play a role in the pathogenesis of endometriosis.

Adult ; Annexin A1 ; genetics ; metabolism ; Ascitic Fluid ; metabolism ; Blotting, Western ; Endometriosis ; genetics ; metabolism ; pathology ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the effects of adenovirus-delivered tissue inhibitor of metalloproteinases-3 ( Ad-TIMP-3) on sensitivity of cervical cancer cells to cisplatin and evaluate the potential application of this combined scheme in cervical cancer treatment.

METHODSCells of cervical cancer CaSKi cell line were infected with Ad-TIMP-3 in vitro. Apoptotic effect, cell cycle changes and p53 protein expression were detected. After combined treatment of those cells with cisplatin, colony formation test was performed and cytotoxicity was detected by MTT. The growth curve and tumor growth inhibition in vivo were evaluated.

RESULTSThe expressions of TIMP-3 mRNA and protein were significantly upregulated after transfection. As a result, massive apoptosis was induced and the cells were arrested at G2/M phase. Exogenous overexpression of TIMP-3 increased p53 protein level markedly in spite of the backgrounds of p53 gene in cells. Combined with cisplatin treatment, the cloning efficiency was decreased. A synergism was observed by isobolic method ( D < 1 ) in vitro and tumor growth was significantly inhibited in vivo.

CONCLUSIONAd-TIMP-3 is a powerful proapoptotic agent. It increases sensitivity of the cells to cisplatin and the Ad-TIMP-3 gene therapy in combination with cisplatin could be a promising alternative in cervical cancer treatment.

Adenoviridae ; genetics ; Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; genetics ; Blotting, Western ; Carcinoma, Squamous Cell ; genetics ; pathology ; therapy ; Cell Cycle ; drug effects ; genetics ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Combined Modality Therapy ; Female ; Genetic Therapy ; methods ; HeLa Cells ; Humans ; Mice ; Mice, Nude ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-3 ; genetics ; metabolism ; Transfection ; Tumor Suppressor Protein p53 ; metabolism ; Uterine Cervical Neoplasms ; genetics ; pathology ; therapy ; Xenograft Model Antitumor Assays ; methods

Objective To systematically review the efficacy of angiotensin receptor blocker (ARB) combined with glucocorticoid for patients with idiopathic pulmonary fibrosis (IPF).Methods Electronically searched PubMed,EmBase,the Cochrane Library,CNKI,CBM,VIP,Wanfang Database from the date of establishment to March 2017 for all randomized controlled trials (RCTs) and quasi-RCTs based on ARB combined with glucocorticoid therapy for patients with IPF.Manual search in relevant journals and search of relevant literature on other websites were also performed.The data extraction and quality assessment of included RCTs and quasi-RCTs were conducted by two reviewers independently.Then,Meta-analysis was conducted by using RevMan 5.3 software.Results A total of 12 RCTs involving 1239 patients were included,621 cases in treatment group and 618 cases in control group.The results of Meta-analysis indicated that the treatment group was superior to the control group in clinical effectiveness (OR =2.78,95％ CI:2.12-3.66,P ＜ O.001),partial pressure of oxygen (PaO2,MD =8.76,95％ CI:7.26-10.26,P ＜0.001),partial pressure of carbon dioxide (PaCO2,MD =-14.04,95％ CI:-15.03--13.05,P ＜0.001),forced vital capacity (FVC,MD =0.74,95％ CI:O.55-0.94,P ＜0.001),forced expiratory volume in one second (FEV1,MD =0.74,95％ CI:0.63-0.86,P ＜0.001),diffusion capacity for carbon monoxide (DLCO) value (MD =3.70,95％ CI:1.88-5.51,P ＜ 0.001) and vital capacity (VC,MD=5.23,95％ CI:3.38-7.08,P＜O.001).Conclusion The current evidence suggested that ARB combined with glucocorticoid could increase clinical effectiveness,improve PaO2,FVC,FEV1,DLCO and VC,and decrease PaCO2 in serum of IPF patients.