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Supplemental Digital Content is available in the text, Background: Fusion genes may play an important role in tumorigenesis, prognosis, and drug resistance; however, studies on fusion genes in endometrial cancer (EC) are rare. This study aimed to identify new fusion genes and to explore their clinical significance in EC. Methods: A total of 28 patients diagnosed with EC were enrolled in this study. RNA sequencing was used to obtain entire genomes and transcriptomes. STAR-comparison and STAR-fusion prediction were applied to predict the fusion genes. Chi-square tests and Student t tests were used to verify the clinical significance with SPSS 13.0 software. Results: New fusion genes were found, and the number of fusion genes varied from 3 to 110 among all patients with EC. The type of fusion genes varied and included messenger RNA (mRNA)-mRNA, long non-coding RNA (lncRNA)-lncRNA, and lncRNA-mRNA. There were six fusion genes with high fusion rates, namely, RP11–123O10.4–GRIP1, RP11–444D3.1–SOX5, RP11–680G10.1–GSE1, NRIP1–AF127936.7, RP11–96H19.1–RP11–446N19.1, and DPH7–PTP4A3. Further studies showed that these fusion genes are related to stage, grade, and recurrence, in which NRIP1–AF127936.7 and DPH7–PTP4A3 were found only in stage III patients with EC. DPH7–PTP4A3 was found in grades 2 and 3, and recurrent patients with EC. Conclusion: Fusion genes play an essential role in EC. Six genes that are overexpressed with high fusion rates are identified. NRIP1–AF127936.7 and DPH7–PTP4A3 might be related to stage, and DPH7–PTP4A3 be related to grade and recurrence.

To evaluate real dynamic assessment of tear film optical quality for monitoring and prevention of dry eye. Right eyes of 62 normal and 39 dry eye subjects were included. Dynamic measurement of objective scatter index (OSI) was performed by using the Optical Quality Analysis System II (OQAS II), correlation coefficient between OSI and time (CCOT) was calculated. According to whether the CCOT was significantly ascending, normal and dry eye groups were further subdivided for comparison. By using Scheimpflug-Placido topographer, non-invasive tear break-up time (NITBUT) was recorded, and a 2-dimensional precorneal tear film map was reconstructed and divided into central, middle, and peripheral corneal zones, distribution of tear break-up spots in the 3 corneal zones were analyzed. The numbers of tear break-up spots were higher in all the 3 corneal zones of the dry eye subjects (P

The monitoring of cancer biomarkers is crucial to the early detection of cancer. However, a limiting factor in biomarker analysis is the ability to obtain the multilayered information of various biomarker molecules located at different parts of cells from the plasma membrane to the cytoplasm. A two-stage dissociation nanoparticle system based on multifunctionalized polydopamine-coated gold nanoparticles (Au@PDA NPs) is reported, which allows for the two-stage imaging of cancer biomarkers in single cells. We demonstrate the feasibility of this strategy on sialic acids (SAs), p53 protein, and microRNA-21 (miRNA-21) in MCF-7 breast cancer cells by two custom-built probes. Furthermore, the multicolor fluorescence information extracted is used for the monitoring of biomarker expression changes under different drug combinations, which allows us to investigate the complex interactions between various cancer biomarkers and to describe the cancer biomarker-synergic networks in single cells.

CMS-1, mainly composed of imperatorin as its active compound, is a partially purified fraction of a Chinese herbal medicine, Semen Cnidium monnieri. CMS-1 has the potential to be further developed as a new treatment for hypertension. Thus, we studied its toxicity in both Sprague-Dawley rats and beagle dogs. Rats (0-900mg/kg/day) and dogs (0-450mg/kg/day) received CMS-1 orally for 30 consecutive days, followed by a 15-day recovery period. The major target organs of CMS-1 toxicity are the GI (inappetence), liver (hepatocellular necrosis, enzyme elevation), thymus (atrophy), cardiovascular (hypotension), changes in ECG T and P waveforms, elevation of nitrous oxide levels and hematological (RBC parameters disturbances) systems. Most treatment-induced adverse effects were reversible or showed a progressive recovery upon discontinuation of the treatment. The No Observed Adverse Effect Level (NOAEL) was 100mg/kg/day for rats and 50mg/kg/day for dogs. This non-clinical study suggests that clinical monitoring of CMS-1 in patients should focus on the gastrointestinal system, blood tests for liver functions, electrolytes, and blood homeostasis, cardiovascular functions, and immune functions.

Epidemiologic studies have demonstrated that long-term exposure to relatively low levels of particulate air pollution is associated with adverse cardiovascular outcomes in Europe and North America. However, few studies have assessed the association with high level air pollutants. We aimed to assess the cardiovascular effects of long-term exposure to high level concentrations of inhalable particulate and to identify the characteristics of the Chinese population that are susceptible to the health effects. A retrospective cohort, containing 39,054 subjects from four cities in northern China, was followed for mortality of all cause and specific cardiovascular diseases from 1998 to 2009. Information on concentrations of PM10 (particulate matter

Objectives: To establish a taxol-resistant cell line of human ovarian carcinoma (A2780/Taxol) and investigate its biological features. Methods: The drug-resistant cell line (A2780/Taxol) was established by continuous stepwise selection with increasing concentrations of Taxol. Cell morphology was assessed by microscopy and growth curves were generated with in vitro and in vivo tumor xenograft models. With rhodamine123 (Rh123) assays, cell cycle distribution and the apoptotic rate were analyzed by flow cytometry (FCM). Drug resistance-related and signal associated proteins, including P-gp, MRPs, caveolin-1, PKC-α, Akt, ERK1/2, were detected by Western blotting. Results: A2780/Taxol cells were established with stable resistance to taxol. The drug resistance index (RI) was 430.7. Cross-resistance to other drugs was also shown, but there was no significant change to radioresistance. Compared with parental cells, A2780/Taxol cells were significantly heteromorphous, with a significant delay in population doubling time and reduced uptake of Rh123 (p < 0.01). In vivo, tumor take by A2780 cells was 80%, and tumor volume increased gradually. In contrast, with A2780/Taxol cells in xenograft models there was no tumor development. FCM analysis revealed that A2780/Taxol cells had a higher percentage of G0/G1 and lower S phase, but no changes of G2 phase and the apoptosis rate. Expression of P-gp, MRP1, MRP2, BCRP, LRP, caveolin-1, PKC-α, Phospho-ERK1/2 and Phospho-JNK protein was significantly up-regulated, while Akt and p38 MARK protein expression was not changed in A2780/Taxol cells. Conclusion: The A2780/Taxol cell line is an ideal model to investigate the mechanism of muti-drug resistance related to overexpression of drug-resistance associated proteins and activation of the PKC-α/ERK (JNK) signaling pathway.

Tracheal glomus tumor is an extremely rare neoplasm resected mostly by open surgery or through rigid bronchoscopy. We report two cases presenting with polypoid masses arising from the tracheal membrane in the posterior wall of the lower tracheal segment. The tumor was removed by high-frequency electrocautery and flexible bronchoscopic argon-plasma coagulation, and follow-up bronchoscopy and chest CT did not reveal tumor recurrence 12 months after the operation. In patients with tracheal glomus tumor who have poor surgical tolerance or are not willing to receive an open surgery, flexible bronchoscopic tumor removal can be a good alternative to relieve the airway obstruction symptoms.

Gliding adaptations in thoracopterid flying fishes represent a remarkable case of convergent evolution of overwater gliding strategy with modern exocoetid flying fishes, but the evolutionary origin of this strategy was poorly known in the thoracopterids because of lack of transitional forms. Until recently, all thoracopterids, from the Late Triassic of Austria and Italy and the Middle Triassic of South China, were highly specialized ‘four-winged’ gliders in having wing-like paired fins and an asymmetrical caudal fin with the lower caudal lobe notably larger than the upper lobe. Here, we show that the new genus Wushaichthys and the previously alleged ‘peltopleurid’ Peripeltopleurus , from the Middle Triassic (Ladinian, 235–242 Ma) of South China and near the Ladinian/Anisian boundary of southern Switzerland and northern Italy, respectively, represent the most primitive and oldest known thoracopterids. Wushaichthys , the most basal thoracopterid, shows certain derived features of this group in the skull. Peripeltopleurus shows a condition intermediate between Wushaichthys and Thoracopterus in having a slightly asymmetrical caudal fin but still lacking wing-like paired fins. Phylogenetic studies suggest that the evolution of overwater gliding of thoracopterids was gradual in nature; a four-stage adaption following the ‘cranial specialization–asymmetrical caudal fin–enlarged paired fins–scale reduction’ sequence has been recognized in thoracopterid evolution. Moreover, Wushaichthys and Peripeltopleurus bear hooklets on the anal fin of supposed males, resembling those of modern viviparious teleosts. Early thoracopterids probably had evolved a live-bearing reproductive strategy.

To explore the mechanism resistance of medroxyprogesterone 17-acetate(MPA) on the endometrial cancer side-population(SP) cells.(1) Ishikawa-SP cells from endometrial cancer cell lines Ishikawa were be separated by Hoechst 33342 dyeing method and flow cytometry analysis. The clone formation efficiency between Ishikawa-SP cells and Ishikawa-non-SP cells were performed by clone formation assay. Breast cancer resistance protein (BCRP) was examined by immunocytochemistry method. (2) Ishikawa, Ishikawa-SP, Ishikawa-non-SP cells were treated with various concentrations of MPA at 5, 10, 15, 20 µmol/L. After cultured for 24, 48, and 72 hours, cells growth were measured by methanethiosulfomate (MTS) assay. (3) The groups of Ishikawa, Ishikawa-SP, Ishikawa-non-SP cells incubated with MPA at the half maximal inhibitory concentration (IC50) were selected for cell apoptosis assay by using flow cytometry. After MPA treatment, the expression of caspase-3 was examined by immunocytochemistry method.(1) There were few proportion of Ishikawa-SP cells in Ishikawa endometrial carcinoma, which were 2.7%. There were stronger clone formation efficiency for Ishikawa-SP cells than that for Ishikawa-non-SP cells in Ishikawa [(6.02 ± 1.17)% vs.(0.53 ± 0.20)%, P = 0.001]. And there were higher level expression of BCRP (P = 0.001) and also more resistant Taxol and radiation between Ishikawa-SP cells and Ishikawa-non-SP cells. (2) The inhibitory effect of MPA was concentration-dependent and time-dependent. (3)After MPA treatment, the apoptosis rates of Ishikawa-SP, Ishikawa-non-SP,Ishikawa were (4.01 ± 0.43) %, (9.30 ± 0.67) %, and (4.64 ± 0.18) %, respectively(P0.05). The level expression of caspase-3 in Ishikawa group after MPA treated were higher than that in Ishikawa-SP group.MPA may be inhibit the growth of endometrial cancer, Ishikawa-SP and Ishikawa-non-SP cells, while Ishikawa-SP may be more resistant to MPA than Ishikawa-non-SP, which mechanism of resistance on MPA may be related to the properties of cancer stem-like cells and cell apoptosis.

Tumors with different gene expression develop and progress in different ways. To deepen our understanding of the progression in endometrial cancer, and provide a useful tool for accurate diagnosis and prognosis assessment, we identified the new molecular prognostic markers in endometrial carcinoma and analyzed the relationship of them with clinical and pathological features of endometrial carcinoma.Ninety-four cases of endometrial endometrioid adenocarcinoma with complete data from the Peking University People's Hospital from 2000 to 2008 and 40 cases of normal endometrium were enrolled. Among these, 30 endometrial endometrioid adenocarcinoma samples of different International Federation of Gynecology and Obstetrics (FIGO) stage were selected for further Agilent genome-wide microarray analysis. Significance analysis of microarrays (SAM) was used to identify genes that are significantly associated with tumor progress. Immunohistochemistry was utilized to identify the genes of interest in endometrial carcinoma and normal endometrium. The relationship between the genes and the age, clinical stage, histological grade, myometrium invaded depth, lymph node metastasis status, and the expression of ER, PR, P53, and PTEN were analyzed by χ(2) test.Analysis between FIGO 1988 stage I and stage III identified a 362-gene "progress signature"; 171 down-regulated and 191 up-regulated genes. Among the alterative genes, TARP (T cell receptor gamma alternate reading frame protein) and KRT5 (keratin 5) decreased 3.57 fold and 5.8 fold in FIGO stage III patients. The expression of TARP in endometrial carcinoma increased compared to normal endometrium, while that of KRT5 decreased (P0.05). The expression of TARP and KRT5 decreased when stage, histological grading, myometrium invaded depth increased (P0.05). In the cases with lymph node metastasis, the expression of TARP decreased, while the expression of KRT5 did not differ (both P0.05) both. The expression of P53 had a negative relationship with the expression of KRT5 (P0.05), but not with the expression of TARP (P0.05). There was no correlation between the expression of TARP and KRT5 and the expression of ER, PR, PTEN (all P0.05). There was no significant difference in TARP and KRT5 expression in patients aged 50 or younger and patients older than 50 (P0.05).The expression of TARP and KRT5 was correlated with the progress of endometrial cancer and their role needs further study.

We performed a case-control study to investigate whether SNPs of CHIP might affect the development of IA in Chinese Han nationality. We believe we are the first to have screened IA patients for mutations in the CHIP gene to determine the association with these variants. The study group comprised 224 Chinese Han nationality patients with at least one intracranial aneurysm and 238 unrelated healthy Han nationality controls. Genomic DNA was isolated from blood leukocytes. The entire coding regions of CHIP were genotyped by PCR amplification and DNA sequencing. Differences in genotype and allele frequencies between patients and controls were tested by the chi-square method. Genotype and allele frequencies of the SNP rs116166850 was demonstrated to be in Hardy-Weinberg equilibrium. No significant difference in genotype or allele frequencies between case and control groups was detected at the SNP. Our data do not support the hypothesis of a major role for the CHIP gene in IA development in the Chinese Han population.

Hyperinsulinemia, insulin-like growth factor (IGF)-I and -II (IGF-II) are associated with increased risk of endometrial carcinoma. Insulin receptor isoform A (IR-A) is more frequently expressed in endometrial carcinoma than in normal endometrial tissues. To better understand their roles in endometrial carcinoma, we investigated the effects of insulin, IGF-I, and IGF-II in endometrial carcinomas cells with different IR-A expression levels.To explore the role of IR-A in mediating the activity of IGF-I, IGF-II, and insulin, we investigate the cellular proliferation of endometrial carcinoma cell lines RL95-2 and RL95-2-IR-A by MTS assays. Then we examined the protein kinase Akt phosphorylation and extracellular signal-regulated kinase (ERK) 1/2 phosphorylation in both cell lines by Western blotting. The effect of IGF-II and AG1024 on cell cycle progression and apoptosis was assessed by flowcytometry. To examine whether the effects of IGFs were mediated by IR-A, we blocked IGF-I receptor (IGF-IR) in both cell lines using AG1024, an IGF-IR-specific inhibitor.IGF-I and IGF-II significantly enhanced proliferation of both cell lines (P0.05). By contrast, insulin significantly increased proliferation of RL95-2-IR-A cells only (P0.05). IGF-I and IGF-II significantly increased pAkt levels in RL95-2 cells and pERK1/2 levels in RL95-2-IR-A cells (all, P0.05). Insulin increased pERK1/2 levels in RL95-2-IR-A cells only (P0.05). LY294002 and PD98059 inhibited the specific signaling activities and cellular proliferation. After AG1024 pretreatment, neither IGF-I nor IGF-II affected pAkt levels in RL95-2 cells. IGF-II, but not IGF-I, increased pERK1/2 levels in RL95-2-IR-A cells. After AG1024 pretreatment, the proliferation rate and DNA content corresponding to the S phase increased and apoptosis decreased significantly in IGF-II-treated RL95-2-IR-A cells only (P0.05).The proliferation effect of insulin is mediated by IR-A. When IR-A dominates in a cell line, IGF-II activated cell proliferation mainly through the ERK1/2 pathway. On the other hand, IGF-II activated cell proliferation mainly through the Akt pathway. IR-A can at least partly mediate the proliferative and anti-apoptotic effects of IGF-II through the ERK1/2 pathway.

To explore the clinical significance of KRAS mutation detection in colorectal adenocarcinoma.Paraffin-embedded tissue specimens were obtained from 440 patients with colorectal adenocarcinoma. The genomic DNA was extracted. Mutations of exon 2 of KRAS gene were examined by PCR and direct sequencing.Somatic mutations of KRAS gene were identified in 146 cases, with the mutation rate of 33.2% (146/440). Among these 146 patients, KRAS mutation involved codon 12 in 118 patients, including 35GA (Gly12Asp, 62 cases), 35GT (Gly12Val, 35 cases), 34GT (Gly12Cys, 9 cases), 34GA (Gly12Ser, 6 cases), 35GC (Gly12Ala, 5 cases), and 34GC (Gly12Arg, 1 case); in 27 patients the mutation involved codon 13, including 38GA (Gly13Asp, 25 cases), 38GC (Gly13 Val, 1 case) and 37GT (Gly13 Cys, 1 case); and in one patient, the mutation involved codon 14 with 40GA (Val14Ile). The status of KRAS or codon 12 mutations in colorectal adenocarcinoma was related to patients' gender (P = 0.021 and P = 0.030, respectively), and this significant correlation to females was conserved in clinical stage III (P = 0.007 and P = 0.003, respectively), but not in stages I, II, and IV. The status of KRAS or codon 12 mutations was also related to tumor stage. Between stage II and stage IV, the mutation rate of KRAS and codon 12 showed significant difference (P = 0.028 and 0.034, respectively). Between stage III and stage IV, only the codon 12 mutation rate showed significant difference (P = 0.011). Codon 13 mutation was not related to tumor stage.About one third of patients with colorectal adenocarcinoma have KRAS gene mutation, which might be related to patients' gender; and could be consistently detected by PCR and direct sequencing.

Objective: To determine whether silence of PKC-α expression by small interference RNA (siRNA) might regulate MDR1 expression and reverse chemoresistance of ovarian cancer. Methods: We measured gene and protein expression of MDR1 and PKC-α in ovarian cancer cells and assessed their correlation with cell drug resistance. We also examined whether blocking PKC-α by RNA interference (RNAi) affected MDR1 expression and reversed drug resistance in drug sensitivity tests. Results: The drug resistance cell lines, OV1228/DDP and OV1228/Taxol, had higher gene and protein expression of MDR1 and PKC-α than their counterpart sensitive cell line, OV1228. SiRNA depressed PKC-α gene protein expression, as well as MDR1 and protein expression and improved the drug sensitivity in OV1228/DDP and OV1228/Taxol cells. Conclusion: These results indicated that decreasing PKC-α expression with siRNA might be an effective method to improve drug sensitivity in drug resistant cells with elevated levels of PKC-α and MDR1. A new siRNA-based therapeutic strategy targeting PKC-α gene could be designed to overcome the chemoresistance of ovarian cancer.

Histone deacetylase (HDAC) inhibitors are a group of small chemical molecules that inhibit histone deacetylase. At cell level, HDAC inhibitors have multiple biological effects such as cell cycle arrest, apoptosis, cell differentiation and auotophagy. At molecular level, HDAC inhibitors cause histone and nonhistone acetylation and induce gene expression. HDAC inhibitors are widely used in cancer therapy because of its function of inducing apoptosis. However, the mechanisms of apoptosis effect are not fully understood. TSA is a classical HDAC inhibitor and widely used in epigenetic and anti-cancer research. In this study, we selected Trichostatin A (TSA) to investigate the mechanisms of HDAC inhibitors apoptotic effect on cancer cells.Cervical cancer cell lines such as Hela, Caski and normal human keratinocyte line HaCaT were treated with various concentrations of TSA. Crystal violent assay and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay were performed to determine cell number. PARP cleavage and FITC-AnexinV were performed to determine apoptosis. DNA-methyltransferase (DNMT)1, DNMT3A and DNMT3B were determined by regular PCR, qPCR and Western Blotting. Small interfering RNA (SiRNAi) was used to knock down DNMT3B.HDAC inhibitors only induce cervical cancer cell apoptosis. At 1 µmol/L of TSA, 86% of Hela cell and 76% of Caski went apoptosis. For normal cells, HDAC inhibitors have no cytotoxic effect at therapeutic dosage, (90.0 ± 8.4)% of normal cell survive after treated with 1 µmol/L of TSA. We compared 1 µmol/L group with untreated control with t-test. There was no significance between 1 µmol/L group and untreated control for normal cell (P0.05). HDAC inhibitors decreased DNMT3B in cancer cell but not in normal cell. Manually knock-down of DNMT3B induced Hela and Caski cell apoptosis. More than 99% of Hela and Caski cell went apoptosis after deprived of DNMT3B.DNMT3B was essential to cervical cancer cell survival. Down-regulated DNMT3B by HDAC inhibitors may play an important role in the toxicity of HDAC inhibitors on cervical cancer cells.

To investigate if a-keto/amino acid supplemented low protein diet can slow down the progression of diabetic nephrophathy in comparison with non-supplemented diabetes diet.A prospective, randomized, controlled clinical study was conducted. Twenty three cases of type 2 diabetic nephropathy in IV stage were randomly divided into alpha-keto/amino acid supplemented diet group (trial group) and conventional diabetes diet group (control group), The treatment duration was 52 weeks. 24 h urine protein was measured at 0, 12, 20, 36 and 52 weeks. Before and after the 52 weeks treatment, all the patients received the measurement of glomerular filtration rate (GFR), blood glucose, blood lipids, inflammatory markers, as well as nutritional status.After the treatment for 20, 36, 52 weeks, mean 24 h urine protein decreased significantly in trial groups (P0.05), and 24 h urine protein in trial group were significantly decreased (P0.05) compared with control group in 20 weeks after treatment. Either in trial group or in control group, GFR remained relatively stable during the observation period. Nutrition status, inflammatory markers, and serum calcium, phosphorus levels between the two groups were no significantly difference. The adverse events experienced by the patients in trial group were similar and consistent with the patients underlying renal diseases.Alpha-keto/amino acid can reduce proteinuria more effectively, while improve renal function and nutritional status in diabetic nephropathy patients with well-toleration.

Sporadic multiple gastrointestinal stromal tumors (GISTs) are rare events especially those developed metachronously. This study aimed to investigate the clinico-pathologic and genetic features defining multiple GISTs.624 cases of GISTs were retrieved for retrospective review. 15 cases were identified as multiple GISTs including 13 synchronous and 2 metachronous ones. 32 tumors and 15 normal tissues were obtained from these cases each containing 2-3 tumor nodules and the genomic DNA was extracted for mutational analysis of KIT and PDGFRA genes. The associated patients were recruited for clinical follow-up studies, including 5 males and 10 females at 49 to 84 years of age.Multiple GISTs comprised of 2.4% of GIST cases in our consecutive series. Twenty-six tumors showed mutations at KIT gene in exon 11 and one at PDGFRA gene in exon 18. In seven synchronous cases, different tumors from the same patients displayed different genotypes of KIT or PDGFRA, suggesting their polyclonal origin. In the two multiple GISTs occurring metachronously, the tumors from each patient showed different KIT mutations, suggesting that the second tumors were not the relapse or metastasis of the primary GISTs.Based on types of KIT or PDGFRA mutations and other pathological features, multiple primary GISTs can be differentiated from multiple GISTs resulting from recurrence or metastasis of a single primary tumor. Unlike recurrence or metastasis of GISTs that are malignant, most multiple GISTs are mostly benign and do not require aggressive adjuvant therapy. Therefore, correct diagnosis is critical for proper treatment.

The objective is to study the safety and effectiveness of implantation of posterior chamber phakic intraocular contact lens (ICL) by observing the changes in anterior segment using ultrasound biomicroscopy (UBM).It was a perspective study. The study sampled 30 high myopia patients (30 eyes) who were treated with posterior chamber phakic ICL implant. Central anterior chamber depth (ACD), trabecular-iris angle (TIA), angle opening distance (AOD500), trabecular-ciliary processes distance (TCPD) and iris-ciliary processes distance (ICPD) were measured using UBM preoperatively, 3 months and 1 year postoperatively. The distance from ICL to the central surface of lens and peripheral lens and intra-ocular pressure were measured postoperatively and examined using slit-lamp biomicroscope. One-way ANOVA was used to analyze the distance between peripheral surface of ICL and the lens. One-way repeated measures ANOVA and Bonferroni were conducted.Preoperatively, 3 months and 1 year postoperatively, ACD were (3.16±0.08) mm, (2.76±0.13) mm, (2.74±0.14) mm; AOD500 were (0.45±0.04) mm, (0.41±0.04) mm, (0.41±0.03) mm; TIA were (35.00±3.24)°, (32.47±3.56)°, (32.40±3.23)°, respectively. There were significant difference in TIA, ACD and AOD (P0.05) between preoperative and postoperative data. There was no significant difference between the two postoperative periods tested. TCPD and ICPD showed no significant difference between various time points (F=0.49, F=0.57; P0.05).The decrease in ACD depth and correction in TIA and AOD were the noticeable changes observed in morphological structure of the ocular anterior segment after the ICL treatment. The incidence of complication did not increase as the result of the minor changes in morph structure during the course of the study. However, the long-term effects would require further long-term observation.

To establish the cisplatin (DDP)-resistant cell line from human endometrial cancer cell line Ishikawa and to investigate its resistant mechanism to DDP.A resistant endometrial cancer cell line ISH/DDP was established by gradually increasing dose of cisplatin and high-dose stimulation. The resistant index was estimated by 3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Cell growth curve, doubling time and cell cycle phase distribution were measured; drug-resistant protein of breast cancer resistance protein (BCRP), P-glycoprotein (P-gp) and glutathione-S-transferase-π (GST-π) were examined by immuocytochemistry. Results The DDP-resistant cell line ISH/DDP was established with the resistant index of 3.77. The proliferation of ISH/DDP got slow, doubling time prolonged, which were 40.1 hours, while it was 34.1 hours in Ishikawa (P0.05); and the cell number of G0/G1 phase [(44.3±5.7)% and (39.0±10.7)%, P0.05] and G2/M phase [(11.9±0.7)% and (5.7±2.4)%, P0.05] decreased, while S-phase [(44.2±6.1)% and (55.3±8.4)%, P0.05] increased compared with parent cells. The comprehensive score of the expression of BCRP in ISH/DDP was 16.3±2.0, while it was 13.4±1.5 in Ishikawa (P0.05). The score of the expression of P-gp in ISH/DDP and Ishikawa were 15.5±1.2 and 16.1±1.0 (P0.05), respectively. The score of the expression of GST-π in ISH/DDP and Ishikawa were 15.2±1.9 and 14.9±1.1 (P0.05).ISH/DDP cell line showed a typical resistant phenotype and biological characteristics, which may be accounted for high BCRP expression.

Ethnopharmacological relevance Angelicae Dahurica(Hoffm.)Benth.&Hook.f.ex Franch.&Sav combined with Scutellaria baicalensis Georgi. has been widely used as herb-pairs in traditional Chinese medicine (TCM) to treat migraine headache and cataract, but the underlying compatibility mechanism of the two herbs remains unknown. Aim of study In the present work, we investigated the additive or synergistic effects of absorption behavior of Radix Angelicae dahuricae extracts on baicalin, and the absorption-enhancing mechanism of Radix Angelicae dahuricae extracts on baicalin. Materials and methods Total coumarins (Cou) and volatile oil (VO), as the two main components of Radix Angelicae dahuricae , were extracted by supercritical fluid extraction (SFE) further treated with liquid–liquid separation method. The absorption behavior was investigated by applying the everted gut sac technique and in situ single-pass intestinal perfusion method. Results and conclusions The results showed that both the Cou and the VO could improve the intestinal absorption of baicalin in vivo, and had synergistic action the enhanced absorption of baicalin. Since verapamil did not affect the P app and K a of baicalin significantly, we concluded that the absorption of Baicalin could not be an active transportation in dependent of P-glycoprotein-Mediated efflux systems. Based on intestinal absorption of drug studying was one of the efficacious methods to clarify the compatibility of principles of herb-pairs. The everted gut sac technique and in situ single-pass intestinal perfusion technique model were the effective methods to study the absorption of drug, the application of the animal model to investigating the absorption of herb–drug interactions or other relevant research purposes is envisioned.

To observe the therapeutic efficacy of bronchoscopic interventional therapy on severe tuberculous main bronchial stenosis or atresia complicated with unilateral atelectasis.Ninety patients with severe tuberculous main bronchial stenosis or atresia complicated with unilateral atelectasis, who had received bronchoscopic interventional therapy, were divided into group A and B according to whether stents had been implanted or not. Patients in group A had been treated with electrocautery, balloon dilatation and cryotherapy. Group B had been treated with metallic stent implantation on the basis of the above interventional management. In order to observe the effectiveness, the time needed for taking effect and restenosis rate were noted. The efficacy between patients with different disease courses, radiology, bronchoscopy and dyspnea index were evaluated before treatment and after the patients' conditions were stable.Three months after treatment, the good response rate and the total effective rate of group B were higher than those of group A, 97% vs 42% (χ(2) = 29.595, P0.05), 100% vs 81% (χ(2) = 6.060, P0.05), respectively. The time needed for taking effect in group B was significantly shorter than that in group A, 0.25 month vs 1.6 month. The dyspnea indexes of both groups were significantly improved after treatment, but the improvement of group B was more significant than that of group A (u = -2.478, P0.05). The disease course of patients with different therapeutic efficacy was evaluated, and the median disease course was 2 months in good response efficacy patients, 3.5 months in improved patients, and 5 months in ineffective patients; the difference being significant between ineffective and good response efficacy patients (u = -3.079, P0.01). The restenosis rate of group B was significantly higher than that of group A, 72% vs 32% (χ(2) = 9.090, P0.01). The median restenosis time was 4 months in group A, and 6 months in group B. Bronchoscopy follow-up 12 months after the initial effective treatment showed that the good response rate and the total effective rate of group B were better than those of group A, 60% vs 29% (χ(2) = 10.559, P0.01), 88% vs 60% (χ(2) = 10.261, P0.01, respectively), and the total effective rate of main bronchial atresia patients in group B was significantly higher than that in group A, 90% vs 50% (Fisher's exact test, P0.05). There was no significant difference in effectiveness between severe stenosis and atresia patients in group A and B.Electrocautery, balloon dilatation, cryotherapy and stent implantation were effective methods to treat severe tuberculous main bronchial stenosis or atresia complicated with unilateral atelectasis. Among them, the therapeutic efficacy was better and the symptoms improved more quickly in patients with stent implantation. The efficacy of stent implantation was better than that of conventional interventional therapy, but the incidence of restenosis was also higher. Following-up should be emphasized in this group of patients. Disease courses were associated with the therapeutic efficacy; longer disease course was related to worse therapeutic efficacy, and restenosis occurred earlier. So interventional therapy should be initiated earlier for bronchial tuberculosis with dyspnea, especially for that complicated by atelectasis.

Type I gonadotropin-releasing hormone (GnRH-I) agonists have been applied for the treatment of steroid-dependent tumors such as breast carcinoma, ovarian cancer and prostatic carcinoma. But the mechanism has not been clarified yet. There are few reports about the treatment of endometrial carcinoma using GnRH-I agonists. Type II GnRH (GnRH-II) is a new subtype of GnRH. Our aim was to investigate the effects of GnRH-I agonists and GnRH-II on estrogen receptor-negative human endometrial carcinoma cells and the effect from phosphatase and tensin homolog gene (PTEN) to them.A lentiviral vector-mediated RNAi method was used to establish a PTEN-negative HEC-1A cell clone (HEC-1A-ND). MTT and flow cytometry were used to detect the cell proliferation, cell cycle and apoptosis of HEC-1A, HEC-1A-NC and HEC-1A-ND cells after treatment with GnRH-I agonist Triptorelin (10(-11) mol/L to 10(-5) mol/L) or GnRH-II (10(-11) mol/L to 10(-5) mol/L). Western blotting was used to detect AKT and ERK1/2 activation after treatment with different concentrations of Triptorelin or GnRH-II for 30 minutes in the above mentioned three kinds of cells.Triptorelin and GnRH-II induced apoptosis and inhibited proliferation of HEC-1A, HEC-1A-ND and HEC-1A-NC in a dose-dependent manner. This effect was augmented in HEC-1A-ND cells in which PTEN gene was knocked-down. Furthermore, Triptorelin and GnRH-II inhibited the AKT and ERK activity in HEC-1A-ND cells.Triptorelin and GnRH-II can promote apoptosis rate and inhibit cell proliferation of estrogen receptor-negative endometrial carcinoma cells in a dose-dependent manner. PTEN gene can inhibit the effects of Triptorelin or GnRH-II on human endometrial carcinoma cells.

To analysis the activity of transcriptional factors in endometrial cancer cell lines with different estrogen receptor subtypes.The mRNA levels of estrogen receptor (ER) was detected by quantitative RT-PCR, and the activity of transcriptional factors was also analysed by 345-channel protein/DNA array in RL-952 (the expression status of ERalpha and ERbeta both positive), HEC-1A [ERalpha (+/-), while ERbeta negative] and HEC-1B (ERalpha and ERbeta both negative). The transcription factors of NFkappaBp65 and p38MAPK with different activity were tested by enzyme-linked immunosorbent assay (ELISA) to confirm the results of protein/DNA array.The mRNA levels of ERalpha in RL-952, HEC-1A and HEC-1B were (6780+/-282), (684+/-84) and (168+/-38) copy/ng, respectively. Among 345 candidate transcriptional factors, there were 28 factors associated with ER status. Compared with RL-952 cells, 13 transcriptional activity factors were concomitantly up-regulation, while 15 concomitantly down-regulation in HEC-1A and HEC-1B cells. Transcriptional activities of TTF (1)-1, NRF-1, TCE were significantly correlated with the high-expression status of ERalpha mRNA (r=0.523, P=0.037), while RFX123 and Ikaros were significantly correlated with the low-expression status of ERalpha mRNA (r=-0.312, P=0.041).Transcriptional factors of TTF (1)-1, NRF-1, TCE may be associated with ER-mediated signal pathway, while RFX123 and Ikaros may be associated with non ER-mediated signal pathway in endometrial cancer.

To study the effect of gonadotropin-releasing hormone-I (GnRH-I) agonist triptorelin and gonadotropin-releasing hormone-II (GnRH-II) on human endometrial carcinoma with different states of PTEN.The endometrial carcinoma cells (Ishikawa, Ishikawa-PTEN, and Ishikawa-neo) were treated with different concentrations of triptorelin (10(-11) to 10(-5) mol/L) or GnRH-II (10(-11) to 10(-5) mol/L). Thirty min later, serine/threonine protein kinase (Akt) and extracellular signal-regulated kinase (ERK) 1/2 activation were detected using western blot method. 48 h later, the cell proliferation, cell cycle and apoptosis were detected using methyl thiazolyl tetrazolium (MTT) and flow cytometry. 17beta-estradiol(17beta-E2, 10(-8) mol/L) or the specific estrogen receptor (ER) antagonist, ICI182780I (10(-6) mol/L) was added. After using the two drugs: triptorelin or GnRH-II, the above parameters were detected again.After treated with different concentrations (10(-11), 10(-9), 10(-7), 10(-5) mol/L) of triptorelin and GnRH-II, the cell growth was slowed, the percentage of G0/G1 phase cells increased, the percentage of G2/M and S phase cells decreased and the apoptosis rate increased in a dose-dependent manner (P0.01, P0.05). These changes were more obvious in Ishikawa. The apoptosis rate induced by GnRH-II was higher than that by the same concentration of triptorelin in the three cell lines. Triptorelin and GnRH-II inhibited the Akt and ERK1/2 activity in the endometrial carcinoma cells. 17beta-E2 counteracted the effect of triptorelin and GnRH-II on the endometrial carcinoma cells (P0.01, P0.05).Triptorelin and GnRH-II can promote apoptosis rate of endometrial carcinoma cells and inhibit cell proliferation in a dose-dependent manner which may be caused by ERK1/2 and Akt activity inhibition, and is related to the status of PTEN and could be offset by 17beta-E2.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitously distributed endocrine-disrupting chemical and reproductive toxicant. In order to elucidate low-dose TCDD-mediated effects on reproductive or endocrine functions, female Sprague-Dawley rats were orally administered various concentrations (20, 50, or 125 ng/kg once weekly) TCDD for 29 wk. A proteomic analysis of the ovaries by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization (MALDI) tandem mass spectrometry showed distinct changes in the levels of several proteins that are relevant markers of TCDD toxicity. Serum estradiol (E2) levels of TCDD-treated animals were markedly lower than control. There were no significant differences in bone mineral density (BMD) of femurs. The body weight of the 125-ng/kg TCDD group was significantly decreased relative to control and there was also a significant reduction in absolute and relative ovarian weights. Expressions of selenium binding protein 2, glutathione S-transferase mu type 3, Lrpap1 protein, NADPH, and peptidylprolyl isomerase D were upregulated, while prohibitin and N-ethylmaleimide-sensitive factor expression levels were downregulated. Data provide further insight into the mechanisms by which TCDD disrupts ovarian function by indicating which differential protein expressions following low-dose TCDD exposure.

To investigate the genomic amplification of the human telomerase RNA component (hTERC) gene in cervical cytology and evaluate its role in screening of cervical lesions.A total of 301 cases were recruited, with liquid-based cytology diagnoses as normal (n = 203), atypical squamous cells (ASC, n = 66), low-grade squamous intraepithelial lesions (LSIL, n = 18), and high-grade squamous intraepithelial lesions (HSIL, n = 14). Following cytological examination, the slides were analyzed using a two-color fluorescence in situ hybridization (FISH) probe targeted to chromosome 3q26 containing hTERC. The hTERC findings were compared to the cytologic and histologic results, as well as high-risk human papilloma viruses (HPV) results.Genomic amplification of hTERC was found in 3.0% (6/203) of normal specimens, 21.2% (14/66) of ASC, 44.4% (8/18) of LSIL and 92.9% (13/14) of HSIL, with a significant difference in each pair wise (all P0.05). Significantly more cells with 3q26 gain were found in cervical intraepithelial lesion (CIN)II than in CINI (75.0% vs. 20.0%), as well as in CINIII (86.7% vs. 20.0%) and squamous cervical cancer (SCC) than in CINI (100.0% vs. 20.0%)(all P0.01). The sensitivity of hTERC amplification was significantly higher than cytological screening (82.6% vs. 17.4%, P0.01), and its specificity was higher than high-risk HPV test (67.8% - 73.5% vs. 25.6% - 27.7%, P0.01) in the diagnosis of HSIL (CINII - III). The abnormal hTERC signal type mostly was 2:3 in CINI (84.9%); whereas in CINII - III, 2:3, 2:4 and 4:4 accounted for 44.6%, 24.8% and 17.8%, respectively.Testing the gain of chromosome 3q26 in cytological specimens using specific probe for hTERC is powerful in screening of HSIL, and the amplification patterns of 2:4 and 4:4 may serve as potential prognosis markers.

To analyze the causes of benign central airway stenoses and to evaluate the efficacy of interventional treatments through flexible bronchoscopy.Three hundred and eighty-six outpatients and inpatients with benign central airway stenoses in our hospital from January 1999 to December 2006 were retrospectively analyzed. Interventional treatments through flexible bronchoscopy were used to treat the benign central airway stenoses. The endoscopic interventional treatments included laser, electrocautery, argon plasma coagulation, cryotherapy, balloon dilation and stent insertion. Airway diameters, FEV1 and dyspnea index of patients were evaluated before and immediately after the last treatment procedure.The main causes of benign central airway stenoses were as follows: tuberculosis in 64.25% (248/386), secondary to prolonged orotracheal intubation or tracheotomy in 15.03% (58/386), injury in 3.63% (14/ 386) and inhalation burns in 3.11% (12/386), others were 54 cases. All the 386 patients received endoscopic interventional treatments. 89.89% (347/386) of the patients experienced improvement in dyspnea and cough. The average airway diameter increased from (2.49 +/- 1.57) mm to (6.41 +/- 1.70) mm (t = 47.427, P0.01). Dyspnea index decreased from 2.40 +/- 0.79 to 0.64 +/- 0.50 (t = 44.226, P0.01). The average value of FEV1 evaluated in 115 inpatients increased from (2.11 +/- 0.60) L to (3.46 +/- 0.75) L (t = 20.128, P0.01). Most patients needed multiple interventional treatments except 26 patients who received a single endoscopic treatment. Stable control of the diseases was achieved in 65.54% (253/ 386) patients 3 months after the last operation.Tuberculosis is the most common cause of benign central airway stenoses in this series. Utilization of interventional methods through flexible bronchoscopy is effective in treating benign central airway stenoses.

Background The NLRP3 inflammasome (NOD-like receptor family, pyrin domain containing 3) is an intracellular protein complex that plays an important role in innate immune sensing. Its activation leads to the maturation of caspase-1 and regulates the cleavage of interleukin (IL)-1β and IL-18. Various studies have shown that activation of the immune system plays a pivotal role in the development of fatigue. However, the mechanisms underlying the association between immune activation and fatigue remained elusive, and few reports have described the involvement of NLRP3 inflammasome activation in fatigue. Methods We established a mouse fatigue model with lipopolysaccharide (LPS, 3 mg/kg) challenge combined with swim stress. Both behavioural and biochemical parameters were measured to illustrate the characteristics of this model. We also assessed NLRP3 inflammasome activation in the mouse diencephalon, which is the brain region that has been suggested to be responsible for fatigue sensation. To further identify the role of NLRP3 inflammasome activation in the pathogenesis of chronic fatigue syndrome (CFS), NLRP3 KO mice were also subjected to LPS treatment and swim stress, and the same parameters were evaluated. Results Mice challenged with LPS and subjected to the swim stress test showed decreased locomotor activity, decreased fall-off time in a rota-rod test and increased serum levels of IL-1β and IL-6 compared with untreated mice. Serum levels of lactic acid and malondialdehyde (MDA) were not significantly altered in the treated mice. We demonstrated increased NLRP3 expression, IL-1β production and caspase-1 activation in the diencephalons of the treated mice. In NLRP3 KO mice, we found remarkably increased locomotor activity with longer fall-off times and decreased serum IL-1β levels compared with those of wild-type (WT) mice after LPS challenge and the swim stress test. IL-1β levels in the diencephalon were also significantly decreased in the NLRP3 KO mice. By contrast, IL-6 levels were not significantly altered. Conclusions These findings suggest that LPS-induced fatigue is an IL-1β-dependent process and that the NLRP3/caspase-1 pathway is involved in the mechanisms of LPS-induced fatigue behaviours. NLRP3/caspase-1 inhibition may be a promising therapy for fatigue treatment. Electronic supplementary material The online version of this article (doi:10.1186/s12974-016-0539-1) contains supplementary material, which is available to authorized users.