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In recent years, long noncoding RNAs (lncRNAs) have been demonstrated to be important tumor-associated regulatory factors. LncRNA growth arrest-specific transcript 5 (Gas5) acts as an anti-oncogene in most cancers. Whether Gas5 acts as an oncogene or anti-oncogene in hepatocellular carcinoma (HCC) remains unclear. In the present study, the expression and role of Gas5 in HCC were investigated in vitro and in vivo. Lower expression levels of Gas5 were determined in HCC tissues and cells by quantitative reverse transcription-polymerase chain reaction. Overexpressed Gas 5 lentiviral vectors were constructed to analyze their influence on cell viability, migration, invasion, and apoptosis. Fluorescence in situ hybridization was used to identify the subcellular localization of Gas5. Protein complexes that bound to Gas5 were isolated from HepG2 cells through pull-down experiments and analyzed by mass spectrometry. A series of novel Gas5-interacting proteins were identified and bioinformatics analysis was carried out. These included ribosomal proteins, proteins involved in protein folding, sorting, and transportation in the ER, some nucleases and protein enzymes involved in gene transcription, translation, and other proteins with various functions.78 kDa glucose-regulated protein (GRP78) was identified as a direct target of Gas5 by Rip-qPCR and Western blot analysis assay. Gas5 inhibited HepG2 cell growth and induced cell apoptosis via upregulating CHOP to activate the ER stress signaling pathway. Further studies indicated that the knockdown of CHOP by shRNA partially reversed Gas5-mediated apoptosis in HepG2 cells. Magnetic resonance imaging showed that the ectopic expression of Gas5 inhibited the growth of HCC in nude mice. These findings suggest that Gas5 functions as a tumor suppressor and induces apoptosis through activation of ER stress by targeting the CHOP signal pathway in HCC.

Esophageal squamous cell carcinoma (ESCC) is the main subtype of esophageal cancer in China, and the prognosis of patients remains poor mainly due to the occurrence of lymph node and distant metastasis. The long non‑coding RNA (lncRNA) maternally expressed gene 3 (MEG3) has been shown to have tumor‑suppressive properties and to play an important role in epithelial‑to‑mesenchymal transition (EMT) in some solid tumors. However, whether MEG3 is involved in EMT in ESCC remains unclear. In the present study, the MEG3 expression level and its association with tumorigenesis were determined in 43 tumor tissues of patients with ESCC and in ESCC cells using reverse transcription‑quantitative PCR analysis. Gene microarray analysis was performed to detect differentially expressed genes (DEGs). Based on the functional annotation results, the effects of ectopic expression of MEG3 on cell growth, migration, invasion and EMT were assessed. MEG3 expression level was found to be markedly lower in tumor tissues and cells. Statistical analysis revealed that MEG3 expression was significantly negatively associated with lymph node metastasis and TNM stage in ESCC. Fluorescence in situ hybridization assay demonstrated that MEG3 was expressed mainly in the nucleus. Ectopic expression of MEG3 inhibited cell proliferation, migration, invasion and cell cycle progression in EC109 cells. Gene microarray results demonstrated that 177 genes were differentially expressed ≥2.0 fold in MEG3‑overexpressing cells, including 23 upregulated and 154 downregulated genes. Functional annotation revealed that the DEGs were mainly involved in amino acid biosynthetic process, mitogen‑activated protein kinase signaling, and serine and glycine metabolism. Further experiments indicated that the ectopic expression of MEG3 significantly suppressed cell proliferation, migration, invasion and EMT by downregulating phosphoserine aminotransferase 1 (PSAT1). In pathological tissues, PSAT1 and MEG3 were significantly negatively correlated, and high expression of PSAT1 predicted poor survival. Taken together, these results suggest that MEG3 may be a useful prognostic biomarker and may suppress EMT by inhibiting the PSAT1‑dependent glycogen synthase kinase‑3β/Snail signaling pathway in ESCC.

Cervical cancer remains a serious health problem till now, with nearly 500,000 women cases diagnosed each year around the world. Long noncoding RNA (lncRNA) is a novel class of RNA transcripts (>200 nucleotides in length) participating in gene transcription, cell proliferation, differentiation, and drug resistance. This study aimed to explore the regulatory relationship among lncRNA NCK1-AS1, miR-134-5p, and MutS protein homolog 2 (MSH2), so that the resistance against cisplatin in cervical cancer treatment could be better understood. Comprehensive lncRNA profiling analysis was performed to screen lncRNAs differentially expressed in cervical cancer. The expression patterns of miR-134-5p, NCK1-AS1, and MSH2 were evaluated in cancerous tissues and adjacent normal tissues obtained from 75 cervical cancer patients. Subsequently, anti-NCK1-AS1 small interfering RNA, miR-134-5p mimics, and miR-134-5p inhibitors were transfected into cervical cancer cells, and the effects of these transcripts on cisplatin resistance and cell apoptosis were investigated. The regulatory relationship among NCK1-AS1, miR-134-5p, and MSH2 was identified using a dual-luciferase reporter gene assay, and the results were further validated by RNA pull-down and RNA immunoprecipitation assays. Based on the microarray data of GSE63514 and GSE27678, NCK1-AS1 was upregulated in cervical cancer. Increased expression of NCK1-AS1, MSH2, and decreased expression of miR-134-5p were observed in cervical cancer tissues. In addition, NCK1-AS1 competitively bound to miR-134-5p to regulate MSH2. Therefore, si-NCK1-AS1 and miR-134-5p mimic both reduced MSH2 activity and increased cisplatin-induced apoptosis in cervical cancer cells. Taken together, NCK1-AS1 may become a novel target in improving the chemotherapeutic response and survival of cervical cancer patients.

The purpose of this study was to measure the interradicular spaces between the maxillary second premolar and first molar in adolescents with skeletal Class Ⅱ malocclusion, so as to obtain scientific knowledge for placing mini-implants. METHODS： Forty four cone-beam computed tomography(CBCT) images were selected, which consisted of 23 males and 21 females (average age, 14.07 years) with skeletal Class II malocclusion. CBCT was used to measure the closest interradicular distance and the shortest distance from cortical bone surface to the narrowest interradicular space apically at 1-mm intervals in 9 axial images from the cementoenamel junction(CEJ). SPSS 20.0 software package was used for statistical analysis.There was no significant difference in the closest interradicular distance and the shortest distance from cortical bone surface to the narrowest interradicular space between the left and right side (P0.05), and no significant difference was found between males and females (P0.05); The shortest distance from cortical bone surface to the narrowest interradicular space of the left apical area had positive correlation with SNA angle; The closest interradicular distance became wider toward the apical area，and the average distance of the shortest distance from cortical bone surface to the narrowest interradicular space was over 5mm in all sections.The distribution of interradicular spaces between the maxillary second premolar and first molar of adolescents with skeletal Class II malocclusion is learned using CBCT. These data might provide a reference for placing mini-implants.

Objectives Cervical cancer (CC) is a common malignant tumor in the female reproductive system that is characterized by a high metastatic potential. LncRNA ANRIL has been found to be a cancer oncogene in multiple tumors. In our study, we altered the expression of ANRIL in CC cells and evaluated its ability on influencing proliferation, migration and invasion of CC cells and associated mechanism. Methods Differentially expressed lncRNAs in CC were identified by microarray and TCGA analyses. CC tissues and adjacent tissues were collected in order to extract CC cells. The expression of ANRIL was determined by RT-qPCR. The CC cells were transfected with siRNA or si-NC against ANRIL to find out whether ANRIL can influence the expression of Cyclin D1, CDK4, CDK6, E-cadherin, vimentin and N-cadherin, as well as affect cell proliferation, cell apoptosis, cell migration and cell invasion of CC cells. Results Based on TCGA and microarray analyses, ANRIL was predicted to be highly expressed in CC and CC with migration. Then further verification was obtained by means of RT-qPCR that ANRIL was highly expressed in CC tissues. In addition, high expression of ANRIL was related to increased E-cadherin expression, high migration of CC as well as decreased cell apoptosis rate. On the other hand, inhibition of ANRIL expression led to decreased expressions of Cyclin D1, CDK4, CDK6, N-cadherin and Vimentin, along with attenuated cell proliferation, migration and invasion of CC cells. Conclusion The key findings of our study demonstrated that the inhibition of lncRNA ANRIL reduces the proliferation, migration and invasion capabilities of CC cells. Down-regulation of ANRIL may serve as a potential therapeutic target in the treatment of CC.

We assessed the prevalence characteristics of single and multiple high-risk human papillomavirus (HR-HPV) infections. A total of 1783 women who underwent colposcopy and cervical biopsy for abnormal ThinPrep Cytology Test and/or HR-HPV subtype genotyping results were enrolled in the study. Among the participants, 770 were diagnosed with cervicitis, 395 with cervical intraepithelial neoplasia grade 1 (CIN1), 542 with CIN2-3, and 76 with squamous cell carcinoma (SCC), with HR-HPV infection rates of 75.8%, 85.8%, 95.9%, and 88.4%, respectively. The prevalence of total and multiple HR-HPV infections exhibited a bimodal age distribution with a peak at ≤25 years, a decline with age and a second peak at ≥55 years, whereas single HR-HPV infections exhibited one peak from 35 to 44 years. The four most dominant HPV genotypes were HPV 16 (29.5%), 52 (15.0%), 58 (14.2%), and 18 (10.4%). In total, 67.0%, 70.4%, and 82.1% of patients with CIN1, CIN2-3, and SCC, respectively, had a single HR-HPV infection, which increased significantly with the aggravation of the cervical lesion grade (P = 0.045). Patients with a single HPV 16 infection had higher incidences of CIN2+ (62.2%) than those with multiple HPV 16 infections (52.4%) (P = 0.021). Patients coinfected with HPV 16 had higher CIN2+ incidence than those with single HPV 52, 31, 33, 35, 39, 45, 51, 56, or 59 infections (P < 0.001). This study provided baseline data on the prevalence characteristics of single and multiple HR-HPV infections in women attending a gynecological outpatient clinic in Beijing.

Objectives: To explore the expression of aquaporin 1 (AQP 1 ) in bladder uroepithelium cell carcinoma (BUCC) and its relevance to recurrence. Materials and Methods: Tissue samples from 45 BUCC patients who underwent total cystectomy or transurethral resection of bladder tumor (TURBT) and from 40 patients with non-bladder cancers who underwent special detection or treatments were collected. The level of expression of AQP 1 in BUCC tissues and normal bladder tissues was assessed by immunohistochemistry so as to analyze the relevance to pathological patterns and time of recurrence in BUCC patients. Results: The expression levels of AQP 1 normal bladder tissues and BUCC tissues were 2.175±0.693 and 3.689±0.701, respectively, and the difference was significant (t=9.99, P

Objective To investigate the effects of oxygen on the proliferation and angiogenesis of endometriosis in vivo. Design Animal studies. Setting Animal research facility. Animal(s) Thirty-six female severe combined immunodeficiency (SCID) mice, implanted with eutopic endometrium from seven endometriosis patients. Intervention(s) Human eutopic endometrial tissues were randomized to normoxia, hyperoxia, or hypoxia pretreatment and were subcutaneously implanted into estrogen-treated ovariectomized SCID mice. Main Outcome Measure(s) The growth and quality of the implants were measured, and the expression of proliferation- and angiogenesis-associated markers (i.e., Ki67, CD31, vascular endothelial growth factor, and hypoxia-inducible factor-1α) were assessed using immunohistochemistry and Western blot analyses. Result(s) The growth curves of the implants were distinct with different oxygen pretreatments. The growth of the implants of the hypoxia group was significantly increased compared with the normoxia group, but the growth of the implants of the hyperoxia group was significantly decreased compared with the normoxia group. Microscopic examination indicated that lesions with hyperplastic cylindrical glandular epithelium were surrounded by the endometrial stroma in the hypoxia group, but the glandular epithelium was partially depauperate in the hyperoxia group. The expression of Ki67, CD31, vascular endothelial growth factor, and hypoxia-inducible factor-1α in the hypoxia-pretreated implants was significantly higher compared with the hyperoxia or normoxia groups. Conclusion(s) Oxygen can alter the growth patterns of endometriosis implants in a SCID mouse model. Hypoxia pretreatment promoted the proliferation and angiogenesis of endometriosis, whereas hyperoxia pretreatment exhibited the opposite effect.

To evaluate the effects of lesion location on adhesion and angiogenesis of transplanted endometriotic lesions in SCID mice.Three groups of female SCID mice included intraperitoneal (i.p.) (n = 12), subcutaneous (s.c.) (n = 12), and mock surgery (control) (n = 12). At 2 weeks after ovariectomy, the mice were transplanted with eutopic endometrium from endometriosis patients either subcutaneously or sutured within the peritoneal, or underwent mock surgery. After 4 weeks, the mice were sacrificed to evaluate the adhesion and volume changes of the implanted lesions. Furthermore, semiquantitative immunohistochemical staining was performed to analyze expression of MMP-2 and TIMP-2 as adhesion makers, and vWF, VEGF, and HIF-1α as angiogenesis markers.Adhesion occurred in 9 of 12 mice in the i.p. group, 3 of 12 mice in the s.c. group, and 3 of 12 mice in the control group. Fisher's exact test showed that the difference of adhesion occurrence between i.p. and s.c. groups was statistically significant (p 0.05). Graft volume changes were higher in the s.c. group than those in the i.p. group. MMP-2 expression was higher in the s.c. group than that in the i.p. group (p 0.01). There was no significant difference of TIMP-2 expression between s.c. and i.p. groups. vWF, VEGF, and HIF-1α expression was significantly higher in the s.c. group than that in the i.p. group (p 0.01).Lesion location might be involved in the pathological changes of endometriosis. The intraperitoneal location is related to endometriotic adhesion, whereas the subcutaneous location is related to the infiltration of endometriotic lesions.

Background Neuromyelitis optica (NMO) is a serious idiopathic inflammatory demyelinating disorder characterized by acute transverse myelitis and optic neuritis. A significant proportion of NMO patients are seropositive for NMO-IgG, an autoantibody targeting aquaporin-4 (AQP4) water channel. Paraneoplastic NMO associated various tumors were recently reported. Aim We studied the expression of AQP4 by thymoma from patients with and without myasthenia gravis (MG). Methods Thymoma obtained from thymomectomy in patients with and without MG were studied by immunohistochemistry and western blot. Results Ten thymoma patients (9 with MG) and two control patients without thymoma or MG were studied. Immunohistochemistry revealed AQP4 immunoreactivity in cell membrane of thymoma cells from all ten thymoma specimens whereas thymic tissues from patients without thymoma or MG were negative for AQP4 immunoreactivity. Western blot revealed that lysates of nine of the ten thymoma specimens reacted with anti-human AQP4 antibody with a band of ~ 30 kDa compatible with the molecular weight of AQP4. Interestingly, immunofluorescence revealed that IgG isolated from 2 NMO patients seropositive for NMO-IgG bound to cell membrane of thymoma cells from all ten thymoma specimens while IgG from healthy control subject did not. Conclusion Thymoma cells of patients with and without MG express AQP4. AQP4 autoantibodies from serum of NMO patients bound to AQP4 expressed on thymoma cell membrane.

AIM: To analyze the composition of the Chinese herbal medicine Sanjie Zhentong Capsule (SJZTC) and test the therapeutic efficacy of each component in a rat model of endometriosis. METHODS: A rapid resolution liquid chromatography (RRLC) method coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS) has been developed for the analysis of SJZTC. Two main ingredients, Drac(h)onis sanguis and saponin, were tested in the endometriosis model. Sixty Lewis female rats were in the estrous cycle stage when endometriosis was experimentally initiated by peritoneal implantation of endometrial tissue. Four weeks later, a second laparotomy was performed and implant volumes were measured. After that, the implanted rats were randomized into five study groups: control group (treatment with saline), anastrozole group (treatment with anastrole, 18 μg per day), loureirin A group (treated with loureirin A, 97.2 mg), ginsenoside Re group (treated with ginsenoside Re, 64.8 mg), and SJZTC groups (treated with SJZTC, 86.4 mg) administered once a day for 4 weeks via gastric gavage. After four weeks of treatment, a third laparotomy was performed, implant volumes were re-measured, and the levels of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) were tested. RESULTS: A total of 38 compounds including, both the target and unknown compounds, were rapidly predicted in the capsule extract by the developed method. Compared with the control group, the anastrozole group, loureirin A group, ginsenoside Re group, and SJZTC treated group showed smaller implant volumes, as well as lower levels of VEGF and TNF-α in the peritoneal focus (P < 0.01 for all comparisons). Furthermore, parameters in the groups treated with SJZTC, loureirin A and ginsenoside Re were significantly better than the control group and the anastrozole group. These results indicate that SJZTC and its two main components are effective in reducing the development of endometriosis.

To compare the effects of sanjie zhentong capsule (SZC) and danazol on rats with endometriosis (EMT).Totally 48 adult female Lewis rats were selected, 12 as the blank control group, and the rest 36 rats in the estrus cycle were used to establish the EMT model. After modeling they were randomly divided into 3 groups, i.e., the model control group, the SZC treatment group, and the danazol treatment group, 12 in each group. Four weeks later the focus was measured by a second laparotomy. The normal saline at 1 mL/day was administered to rats in the model control group, SZC at 86.4 mg/day to those in the SZC treatment group, and danazol at 7.2 mg/day to those in the danazol treatment group. All the treatment lasted for 4 weeks. At the end of the treatment, a third laparotomy was performed to measure the size of focus. The expression of proliferating cell nuclear antigen (PCNA) was detected using immunohistochemical assay. The cell apoptosis rate was detected using terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick-end labeling (TUNEL). The concentration of prostaglandin E2 (PGE2) in the peritoneal fluid and the serum were detected using ELISA.There was statistical difference in the change of the focus volume between the SZC treatment group (-23.27 +/- 18.18) and the danazol treatment group (-12.28 +/- 10.04) and the model control group (13.97 +/- 7.54, P0.01). The expression of ectopic PCNA significantly decreased and the positive expression rate of TUNEL obviously increased in the two treatment groups when compared with the model control group (P0.05, P0.01). The expression of ectopic PCNA decreased and the positive expression rate of TUNEL increased more obviously in the SZC treatment group than in the danazol treatment group (P0.01). The concentration of PGE2 in the peritoneal fluid and the serum was significantly lower in the two treatment group when compared with the model control group (P0.01). The concentration of PGE2 in the peritoneal fluid and the serum was significantly lower in the SZC treatment group than in the danazol treatment group (P0.01).SZC and danazol both could inhibit the focus growth in EMT rats. SZC showed better effects. It was an effective drug for treating EMT.

To observe the effect and mechanism of acupuncture on infertility of rats with polycystic ovarian syndrome (PCOS).PCOS rat model was induced by subcutaneous injection of oil solution of dehydroepiandrosterone (DHEA) in immature (24-day-old) female rats for continuous 20 days. Rats for control were given with same dose of oil for instead. PCOS rats were randomly divided into the model group untreated and the acupuncture group treated by needling acupoints of Guanyuan (CV4), Zhongji (CV3), Sanyin-jiao (SP6) and Zigong (CX-CA1), 15 min once a day for 5 continuous days, starting from the 80th day after birth. All rats were sacrificed at terminal of the treatment, their uterus and bilateral ovaries were dissected for observation and blood levels of sex hormones were measured.compared with the model group, the number of implanted blastocyte and blastocyte implantation rate were higher and the blood levels of testosterone (T) and estradiol (E2) were lower in the acupuncture group (P0.05); but the difference between groups in serum levels of follicular stimulating hormone, luteinizing hormone and progesterone were of statistical insignificance (P0.05). Moreover, the wet weight of ovary was lower and the equipotent diameter and area of glandular organ and cavity, area ratio of gland and the stroma, and mean thickness of endometria were higher in the acupuncture group than those in the control group (P0.05).Acupuncture can conspicuously downregulate the expressions of serum levels of T and E2, improve the development of ovaries and uterus, promote ovulation, enhance endometrial receptivity, and advance blastocyte implantation.