Whole-exome sequencing and functional validation reveal a rare missense variant in MMP7 that confers ovarian endometriosis risk.

Prior studies have shown that genetic factors play important roles in ovarian endometriosis. Herein, we first analyzed the whole-exome sequencing data from 158 patients with ovarian endometriosis and 385 local control women without endometriosis. Among which, a rare missense variant in the MMP7 (p.I79T, rs150338402) gene exhibited a significant frequency difference. This rare variant was screened in an additional 1176 patients and 600 control women via direct DNA sequencing. Meanwhile, a total of 38 available clinical characteristics were collected. Our results showed 45 out of 1334 (3.37%) patients, while 15 out of 985 control women (1.52%) (P = 0.0076) harbored this rare variant, respectively. This rare variant was associated with clinical features such as follicle-stimulating hormone (Padj = 0.0342), luteinizing hormone (Padj = 0.0038), progesterone (Padj = 1.4e-7), testosterone (Padj = 0.0923), total bilirubin (Padj = 0.0699), carcinoembryonic antigen (Padj = 0.0665) and squamous cell carcinoma antigen (Padj = 0.0817), respectively. Functional assays showed that this rare variant could promote cell migration, invasion, epithelial-mesenchymal transition (EMT) and increase the proteolytic protein activity of MMP7, implicating that the increased capacities of cell invasion, migration and EMT might be mediated by enhanced proteolytic activity of MMP7 mutant. These results showed that the MMP7 rare missense variant (p.I79T) played important roles in the pathogenesis of ovarian endometriosis. In conclusion, we identified, for the first time, a significantly enriched MMP7 rare variant in ovarian endometriosis; this rare variant was closely associated with certain clinical features in ovarian endometriosis; thus, it could be a promising early diagnostic biomarker for this disease.

Endometrial stromal cell proteomic analysis reveals LIM and SH3 protein 1 (LASP1) plays important roles in the progression of adenomyosis.

Adenomyosis is one of the most common gynecological disorders that the molecular events underlying its pathogenesis remain not fully understood. Prior studies have shown that endometrial stromal cells (ESCs) played crucial roles in the pathogenesis of adenomyosis. In this study, we utilized two-dimensional gel electrophoresis combined with protein identification by mass spectrometry (2D/MS) proteomics analysis to compare the differential protein expression profile between the paired eutopic and ectopic ESCs (EuESCs and EcESCs) in adenomyosis, and a total of 32 significantly altered protein spots were identified. Among which, the expression of LIM and SH3 protein 1 (LASP1) was increased significantly in EcESCs compared to EuESCs. Immunohistochemical assay showed that LASP1 was overexpressed in the stromal cells of ectopic endometriums compared to eutopic endometriums; further functional analyses revealed that LASP1 overexpression could enhance cell proliferation, migration and invasion of EcESCs. Furthermore, we also showed that the dysregulated expression of LASP1 in EcESCs was associated with DNA hypermethylation in the promoter region of the LASP1 gene. However, the detailed molecular mechanisms of enhancing cell proliferation, invasion and migration caused by upregulated LASP1 in adenomyosis needs further study. For the first time, our data suggested that LASP1 plays important roles in the pathogenesis of adenomyosis, and could serve as a prognostic biomarker of adenomyosis.

Efficacy of intravitreal Lucentis injection on major and macular branch retinal vein occlusion.

BACKGROUND: The objective of our study was to assess the efficacy of intravitreal Lucentis injection on major and macular branch retinal vein occlusion (BRVO). METHODS: In this retrospective analysis, 43 patients (major BRVO n = 24; macular BRVO, n = 19) were treated with intravitreal injection of Lucentis with a 1 + PRN regimen, which is diagnosed by fluorescein fundus angiography (FFA). "1 + PRN", namely, one intravitreal injection of Lucentis at the baseline, and then continue or stop according to the condition of the patient. The following observation indexes were measured at baseline and follow-up (1-6 months): best corrected visual acuity (BCVA), foveal thickness (CFT), total retinal volume with macular diameter of 6 mm. During the follow-up, repeated injections were given according to patients' demand, and the number of injections was recorded. RESULT: The observation indexes of patients with BRVO were significantly improved after 6 months of Lucentis treatment in both major and macular groups, including BCVA, CFT and the retinal volume of the 6 mm-diameter macula. Interestingly, there were significant differences in the therapeutic effect between the two groups, and the macular group had better therapeutic effect than the major group with the less number of repeated injections. CONCLUSIONS: To sum up, intravitreal injection of Lucentis was effective for both major and macular BRVO, and the efficacy in macular subtype group was better than that in major subtype group with the more obviously improvement and the less number of injections.

Intravitreal injection of conbercept combined with retinal photocoagulation on macular edema secondary to branch retinal vein occlusion.

Effects of Conbercept combined with retinal photocoagulation on macular edema secondary to branch retinal vein occlusion (BRVO) were investigated. A total of 98 patients (98 eyes) with macular edema secondary to BRVO were collected. The central macular thickness (CMT), incidence rate of complications after treatment and best corrected visual acuity (BCVA) were recorded. Also the factors affecting visual recovery of patients were analyzed. At 1 week, 1 month, 3 months and 6 months after treatment, the BCVA in both groups was significantly superior to that before treatment (P<0.05). In the combination group and laser group, the logarithm of the minimum angle of resolution (logMAR) of BCVA increased from 0.84±0.47 to 0.34±0.10 and from 0.89±0.49 to 0.45±0.14, and CMT declined from 559.5±152.7 to 267.8±19.8 µm and from 570.3±172.6 to 314.7±18.4 µm. It was observed that at 1 week, 1 month, 3 months and 6 months after treatment, the BCVA in combination group was obviously better than that in laser group (P=0.008, P<0.001, P=0.004, P<0.001, respectively), while CMT in combination group was obviously smaller than that in laser group (P=0.009, P=0.002, P<0.001, P<0.001). Conbercept with retinal photocoagulation can effectively improve the visual acuity and reduce the CMT. The visual recovery of patients after treatment is related to the BCVA before treatment, decreased value of CMT at 1 month after treatment and integrity of external limiting membrane (ELM).

Efficacy of ranibizumab combined with photodynamic therapy on wet age-related macular degeneration.

Efficacy and safety of intravitreal ranibizumab (IVR) combined with photodynamic therapy (PDT) in treating wet age-related macular degeneration (wAMD) were studied. A total of 130 eyes were collected from 130 wAMD patients treated in Affiliated to Qingdao University Yuhuangding Hospital of Yantai, of which 65 were given IVR combined with PDT (combination therapy group) and the remaining 65 were treated with simple IVR (ranibizumab group). The differences in best corrected visual acuity (BCVA), central macular thickness (CMT), intraocular pressure, choroidal neovascularization (CNV) leakage, levels of serum vascular endothelial growth factor (VEGF) and transforming growth factor-ß1 (TGF-ß1) as well as complication rate were compared before and after treatment between the two groups. At 1, 3, 6 and 12 months after treatment, combination therapy group had remarkably better BCVA and notably smaller CMT than ranibizumab group. Fundus fluorescein angiography (FFA) showed that the area of macular degeneration was reduced markedly after treatment in both groups, and the area in combination therapy group was evidently smaller than that in ranibizumab group at 1, 3 and 6 months after treatment. At 3 months after treatment, the levels of serum VEGF and TGF-ß1 declined obviously in the two groups compared with those before treatment. The IVR combined with PDT can effectively improve the visual acuity, decrease CMT and prominently reduce the area of macular degeneration of wAMD patients, and its therapeutic effects are long-standing and tolerable for the patients, so it is worthy of clinical popularization.

lncRNA SBF2-AS1 promotes epithelial-mesenchymal transition of cervical cancer cells via regulating miR-140-5p/VEGFAaxis / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of lncRNA SBF2-AS1 on epithelial-mesenchymal transition (EMT) of cervical cancer HeLa cell via regulating miR-140-5p/VEGFA (vascular endothelial growth factor A) axis. Methods: After cell culture and transfection, the cells were divided into 5 groups: NC group, miR-140-5p mimic group, miR-140-5p mimic+pcDNA-VEGFA group, si-lncRNA SBF2-AS1+pcDNA-VEGFA group and si-lncRNA SBF2-AS1+miR-140-5p mimic group. The expression level of lncRNA SBF2-AS1 in cervical cancer tissues and cell lines was detected by qPCR. The targeted relationship between lncRNA SBF2-AS1, miR-140-5p and VEGFA was confirmed by Dual luciferase reporter gene assay. The expression levels of VEGFA and EMT-related proteins N-cadherin, Vimentin and E-cadherin in HeLa cells were detected by WB. The invasion and migration of HeLa cells were detected by Transwell. Results: lncRNA SBF2-AS1 was highly expressed in cervical cancer tissues and cell lines (P<0.05 or P<0.01). Dual luciferase reporter gene assay confirmed that lncRNASBF2-AS1 targetedly combined with miR-140-5p and VEGFAwas a target gene of miR-140-5p (P< 0.05). Knockdown of lncRNA SBF2-AS1 inhibited invasion and migration as well as EMT of HeLa cells. Further experiment confirmed that lncRNA SBF2-AS1 up-regulated the expression level of VEGFA via miR-140-5p, thereby promoting invasion, migration and EMT of HeLa cells. Conclusion: lncRNASBF2-AS1 promotes EMT of HeLa cells via miR-140-5p/VEGFAaxis.

Mutational analysis of the RAS/RAF/MEK/ERK signaling pathway in 260 Han Chinese patients with cervical carcinoma.

Prevalent mutations in the mitogen-activated protein kinase 1 (MAPK1)/extracellular signal-regulated kinase 2 (ERK2) pathway have been identified in cervical squamous cell carcinoma in a large-scale genome sequencing effort. Furthermore, mutations in the rat sarcoma viral oncogene homolog (RAS)/Raf/Mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway have also been revealed to have important roles in the pathogenesis of human cancer. However, whether the potential hotspot mutations in ERK2 and other components of the RAS/RAF/MEK/ERK signaling pathway also exist in Chinese patients with cervical carcinoma remains to be elucidated. In the present study, a total of 260 patients with cervical carcinoma of distinct subtypes were analyzed for the presence of potential hotspot mutations in the RAS/RAF/MEK/ERK signaling pathway. No ERK2 mutations were detected in these samples; however, Kirsten RAS (KRAS) p.G12D (c.35G>A) mutation was identified in 2/26 (7.7%) cervical adenocarcinoma cases, including 1/20 cervical mucinous adenocarcinoma and 1/6 cervical endometrioid carcinoma cases. In addition, no mutations in the ERK1, neuroblastoma RAS, Harvey RAS or B-Raf proto-oncogene serine/threonine kinase genes were detected in the present study. These results indicated that ethnic differences may be a primary reason for the discrepancy in ERK2 mutation frequencies between the current study and previous studies. Furthermore, mutation in the KRAS gene, but not other genes in the RAS/RAF/MEK/ERK signaling pathway, may have an active role in the pathogenesis of cervical carcinoma.

Quantification of monohydroxylated polycyclic aromatic hydrocarbons in human urine samples using solid-phase microextraction coupled with glass-capillary nanoelectrospray ionization mass spectrometry.

Polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants with carcinogenic effect raising worldwide concerns. Hydroxylated PAHs (OH-PAHs) could be formed in the body as metabolites of PAHs in human urine samples and thus considered as biomarkers of PAH exposure. In this study, five OH-PAHs including 3-phenanthrol, 1-naphthol, 2-hydroxy fluorene, 1-hydroxprene and 6-hydroxy chrysene in human urine samples were selectively enriched by C18 solid-phase microextraction (SPME), then SPME fiber was connected high voltage and then was inserted into a glass-capillary to elute and ionize the analytes for mass spectrometric (MS) detection. The coupling of SPME-MS showed excellent analytical performance for detection of urinary OH-PAHs under optimal conditions, providing an easy operation for rapid detection of a single sample within minutes. By use of internal standard (i.e., 2-hydroxy fluorene-d9), the limit of detection (LOD) and limit of quantitation (LOQ) of OH-PAHs were found to be less than 0.05 ng L-1 level (S/N > 3) and less than 0.1 ng L-1 level (S/N > 10), respectively. The dynamic ranges of five OH-PAHs were found to be a range at 0.1-5.0 ng L-1 with excellent coefficient (R2 > 0.99). This method also showed good precisions (intra-day: 3.4-5.5%, inter-day: 7.0-9.8%, n = 5) and good accuracy (85.3-95.3%, n = 5). Moreover, ion suppression and matrix effect in detection of OH-PAHs in urine were also investigated. Human urine samples collected from 12 volunteers including 6 non-smokers and 6 smokers have been successfully analyzed, it was found that individual OH-PAHs in smokers were higher than in non-smokers. This study demonstrated that SPME coupled with glass-capillary nanoESI-MS is a sensitive method for rapid detection of urinary OH-PAHs for health risk assessment of PAHs exposure.