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Objective: To find out a better criterion to identify septic shock patients after flexible ureteroscopic lithotripsy (FUL). Materials and Methods: In total, 2364 patients who underwent the FUL proc...

Knotted suture bridge repair (KSBR) has been widely proven to be an effective method for rotator cuff repairs. However, the occurrence of type 2 failure after suture bridge repair remains a frequent problem because of the stress concentration and disturbance of tendon perfusion in the medial row. The authors have developed the H-loop knotless double-row repair (HLDR) to counteract these problems.To compare the biomechanical and histological outcomes of HLDR and KSBR for rotator cuff tear in the rabbit model.Controlled laboratory study.Acute bilateral supraspinatus tears were created on the shoulders of 46 New Zealand White rabbits. HLDR and KSBR were randomly performed on the left side or right side. Thirteen animals each were sacrificed at 2, 4, and 8 weeks after surgery (n = 39), with 6 rabbits used for histological evaluation and the other 7 rabbits for biomechanical testing. The remaining 7 animals from the original 46 were only used for initial biomechanical evaluation at week 0.Macroscopically, all repaired tendons were connected to their footprint on the greater tuberosity without postoperative complications at 8 weeks after surgery. The HLDR group had significantly better histological bone-to-tendon integration compared with the KSBR group in terms of fibrocartilage regeneration, collagen composition, and fiber organization. The biomechanical outcomes in the HLDR group were demonstrated to be better than those of the KSBR group at time 0 and 8 weeks after surgery.Both repair techniques were effective for rotator cuff tears in a rabbit rotator cuff tear model; however, HLDR demonstrated more advantages in improving biomechanical properties and histological tendon-to-bone healing compared with KSBR.This animal study suggested that HLDR might be an alternative choice for rotator cuff tears in humans to increase tendon-to-bone healing and reduce the rate of failure to heal.

Background: Mothers are the primary source of bacteria for newborns, but it is unclear whether mother-to-newborn transmission occurs prior to, during or after birth. Similarly, the effect of the delivery mode on neonatal microorganisms has been the focus of controversy. Methods: Healthy maternal and neonatal pairs that underwent vaginal birth and caesarean section were enrolled in this study. Meconium, placenta, membrane and amniotic fluid samples for newborns and vaginal, rectal and oral samples for mothers were collected. All samples were amplified and sequenced by a 16S rRNA gene primer set targeting bacteria and archaea. Findings: A total of 550 samples from 36 mother-neonate pairs with vaginal births and 42 mother-neonate pairs with caesarean sections were included in this study. The negative controls showed that the data analysis in this study was not affected by contamination. There was a high diversity of microbial communities in the pregnancy environment of the foetus. Meconium samples could be divided into three distinct types that were not influenced by the delivery method. Interpretation: The distribution patterns of bacterial communities in the meconium, placenta, and foetal membranes were highly similar and had nothing to do with the mode of delivery. For approximately half of the placental microorganisms, the same sequence could be found in the vaginal, rectal, and oral samples of the mother. Keywords: Meconium, Microbiota, Delivery mode, Archaea

The tetrapeptide Arg-Leu-Tyr-Glu (RLYE), a vascular endothelial growth factor (VEGF) receptor-2 antagonist, has been used previously either alone or in combination with chemotherapeutic drugs for treating colorectal cancer in a mouse model. We analyzed the half-life of the peptide and found that because of degradation by aminopeptidases B and N, it had a short half-life of 1.2 hours in the serum. Therefore, to increase the stability and potency of the peptide, we designed the modified peptide, N-terminally acetylated RLYE (Ac-RLYE), which had a strongly stabilized half-life of 8.8 hours in serum compared with the original parent peptide. The IC50 value of Ac-RLYE for VEGF-A-induced endothelial cell migration decreased to approximately 37.1 pM from 89.1 pM for the parent peptide. Using a mouse xenograft tumor model, we demonstrated that Ac-RLYE was more potent than RLYE in inhibiting tumor angiogenesis and growth, improving vascular integrity and normalization through enhanced endothelial cell junctions and pericyte coverage of the tumor vasculature, and impeding the infiltration of macrophages into tumor and their polarization to the M2 phenotype. Furthermore, combined treatment of Ac-RLYE and irinotecan exhibited synergistic effects on M1-like macrophage activation and apoptosis and growth inhibition of tumor cells. These findings provide evidence that the N-terminal acetylation augments the therapeutic effect of RLYE in solid tumors via inhibition of tumor angiogenesis, improvement of tumor vessel integrity and normalization, and enhancement of the livery and efficacy of the coadministered chemotherapeutic drugs. SIGNIFICANCE STATEMENT: The results of this study demonstrate that the N-terminal acetylation of the tetrapeptide RLYE (Ac-RLYE), a novel vascular endothelial growth factor receptor-2 (VEGFR-2) inhibitor, significantly improves its serum stability, antiangiogenic activity, and vascular normalizing potency, resulting in enhanced therapeutic effect on solid tumors. Furthermore, the combined treatment of Ac-RLYE with the chemotherapeutic drug, irinotecan, synergistically enhanced its antitumor efficacy by improving the perfusion and delivery of the drug into the tumors and stimulating the conversion of the tumor-associated macrophages to an immunostimulatory M1-like antitumor phenotype.

Microwave assisted extraction (MAE) technology was used to extract polysaccharides from Porphyra Haitanensis (PHP) with water. Polysaccharide yield was used as index to evaluate the extraction process. Effects of ratio of water to raw material (X1), microwave power (X2) and extraction time (X3) on polysaccharide yield were investigated. Based on single factor experiment, MAE process of PHP was optimized using response surface methodology. Chemical characterization of PHP was investigated based on analysis of chemical compositions, monosaccharide and thermal gravimetric. The antitumor activities of PHP in vivo and in vitro were evaluated. The results indicated that the optimal conditions for MAE of PHP were ratio of water to raw material 28.98(mL/g), microwave power 77.84 W and extraction time 14.14 min. MAE was a suitable and efficient technique for PHP extraction compared with traditional hot water extraction. Analysis of chemical characterization showed that PHP contained 75.36 ± 1.48% of total carbohydrates, 24.63 ± 1.69% of uronic acid residue and no proteins with monosaccharides such as rhamnose, arabinose, xylose, mannose, glucose, and galactose in a molar ratio of 10.25:9.38:1:12.45:9.9:11.55. Thermal gravimetric analysis suggested that PHP was relatively stable below 170℃. PHP had obvious effect on inhibiting proliferation, inducing apoptosis of SGC-7901 tumor cells in vitro and antitumor effect on SGC-7901 tumor-bearing mice in vivo. The results suggested that PHP had the potential for clinical use in cancer prevention and treatment.

To compare the therapeutic effect between long-snake moxibustion combined with western medication and simple medication on diarrhea type irritable bowel syndrome (IBS-D) of spleen and kidneyA total of 60 patients with IBS-D of spleen and kidneyCompared before treatment, the symptom scores of abdominal pain, defecation frequency, mucous stool and appetite reduction were decreased (Long-snake moxibustion combined with western medication can effectively treat the IBS-D of spleen and kidney

It has been shown previously that a novel tetrapeptide, Arg-Leu-Tyr-Glu (RLYE), derived from human plasminogen inhibits vascular endothelial growth factor (VEGF)-induced angiogenesis, suppresses choroidal neovascularization in mice by an inhibition of VEGF receptor-2 (VEGFR-2) specific signaling pathway. In this study, we report that a modified tetrapeptide (Ac-RLYE) showed improved anti-choroidal neovascularization (CNV) efficacy in a number of animal models of neovascular age-related macular degeneration (AMD) which include rat, rabbit, and minipig. The preventive and therapeutic in vivo efficacy of Ac-RLYE via following intravitreal administration was determined to be either similar or superior to that of ranibizumab and aflibercept. Assessment of the intraocular pharmacokinetic and toxicokinetic properties of Ac-RLYE in rabbits demonstrated that it rapidly reached the retina with minimal systemic exposure after a single intravitreal dose, and it did not accumulate in plasma during repetitive dosing (bi-weekly for 14 weeks). Our results suggested that Ac-RLYE has a great potential for an alternative therapeutics for neovascular (wet) AMD. Since the amino acids in human VEGFR-2 targeted by Ac-RLYE are conserved among the animals employed in this study, the therapeutic efficacies of Ac-RLYE evaluated in those animals are predicted to be observed in human patients suffering from retinal degenerative diseases.

To explore the method and effect of small incision TightRope fixation in the treatment of fresh acromioclavicular joint dislocation.From January 2016 to May 2018, 28 cases of fresh acromioclavicular dislocation were treated, including 20 males and 8 females, aged 26 to 87 years with an average age of 51.3 years. The modified Rockwood classification included 1 case of typeⅡ, 22 cases of typeⅢand 4 cases of type V. The average time from injury to operation was 2.4 days. The operative time, shoulder function recovery time and postoperative complications were recorded, and the immediate reduction effect and Karlsson function of shoulder joint were evaluated.In 28 patients, only one Rockwood typeⅡ was used to reconstruct the pyramidal ligament, and the other 27 were used to reconstruct the pyramidal ligament and the trapezoid ligament. The average operation time was (66.50±12.62) min (including intraoperative fluoroscopy time). Twenty-eight cases were followed up for 11 to 20 (16.7±4.6) months. The recovery time of shoulder function was 2 to 7 months with an average of 4 months. During the follow-up period, 1 case had osteolysis and loss of reduction at the clavicular plate site, and the rest had no complications such as re-dislocation and button plate prolapse. Immediate reduction effect after operation:6 cases with reduction insufficiency, 17 cases with complete reduction and 5 cases with excessive reduction;Karlsson function evaluation of shoulder joint in the last follow-up:excellent in 21 cases, good in 6 cases and poor in 1 case;Pearson analysisTightRope fixation through a small incision in the base of coracoid process is a simple and effective method for the treatment of dislocation of acromioclavicular joint. There was no significant correlation between the slight difference of immediate reduction effect within 5 mm and Karlsson function evaluation of shoulder joint in the last follow-up. It is suggested to pay attention to the loss of reduction and osteolysis of clavicular plate in clinical follow-up.

An association between animals and volatile anaesthetic requirements has been shown; however, evidence related to the postoperative outcome of human patients is lacking. Our aim was to investigate whether there is a difference in the requirement for sevoflurane among people undergoing gastrointestinal surgery. We observed 390 adult patients who underwent gastrointestinal surgery with an American Society of Anesthesiologists physical status of I or II with an expected surgery duration of > 2 hours. We used the bispectral index (BIS) to guide the regulation of end-tidal sevoflurane concentration (ETsevo). The mean ETsevo from 20 minutes after endotracheal intubation to 2 hours after the start of surgery was calculated for all patients. Differential sevoflurane requirements were identified according to ETsevo. The BIS, ETsevo, heart rate, mean arterial pressure, dose of sufentanil and cisatracurium, tracheal extubation time, incidence of intraoperative awareness, and incidence of postoperative nausea and vomiting were compared between patients with a low requirement for sevoflurane (group L) and patients with a high requirement for sevoflurane (group H). The mean ETsevo of the 390 patients was 1.55% ± 0.26%. Based on our definition, patients with an ETsevo of < 1.29% were allocated to the low requirement group (group L; n = 69), while patients with an ETsevo of > 1.81% were allocated to the high requirement group (group H; n = 78). The ETsevo of group L was significantly lower than the ETsevo of group H (1.29% ± 0.014% vs 1.82% ± 0.017%, P

Ethnopharmacological relevance Laminaria japonica, a brown seaweed, has been used in Traditional Chinese Medicine (TCM) to treat a variety of diseases including lung cancer. Aim of the study To demonstrate the effects of Fucoxanthin (FX), a major active component extracted from Laminaria japonica on metastasis and Gefitinib (Gef) sensitivity in human lung cancer cells both in vitro and in vivo. Materials and methods Invasion and migration of lung cancer cells were detected using the wound healing assay and transwell assay. Epithelial-to-mesenchymal transition (EMT) factors and PI3K/AKT/NF-κB pathways were analyzed by western blotting. RNA interference (RNAi) technology was used to silence TIMP-2 gene expression in A549 cells. The anti-metastatic effect of FX was evaluated in vivo in an experimental lung metastatic tumor model. On the other hand, cell counting kit-8 assay was used to study the cell viability of human lung cancer PC9 cells and Gef resistant PC9 cells (PC9/G) after Gef, FX or FX combined with Gef treatment. PC9 xenograft model was established to explore the anti-tumor effect of FX or combined with Gef. Immunohistochemistry staining assay and immunofluorescence staining assay were used to reveal the effects of FX on lung cancer cell proliferation and apoptosis. Results FX was able to significantly inhibit lung cancer cells migration and invasion in vitro. FX suppressed the expressions of Snail, Twist, Fibronectin, N-cadherin, MMP-2, PI3K, p-AKT and NF-κB, and increased the expression of TIMP-2. Furthermore, knockdown of TIMP-2 attenuated FX-mediated invasion inhibition. Additionally, we demonstrated that FX inhibited lung cancer cells metastasis in vivo. The anti-metastatic effects of FX on lung cancer cells might be attributed to inhibition of EMT and PI3K/AKT/NF-κB pathway. We further demonstrated that the anti-tumor activity of FX was not only limited to the drug sensitive cell lines, but also prominent on lung cancer cells with Gef resistant phenotype. Furthermore, in vivo xenograft assay confirmed that FX inhibited tumor growth and enhanced the sensitivity of lung cancer cells to Gef and this effect may be due to inhibition of tumor cell proliferation and activation of apoptosis. Conclusion Collectively, our findings suggested that FX suppresses metastasis of lung cancer cells and overcomes EGFR TKIs resistance. Thus, FX is worthy of further investigation as a drug candidate for the treatment of lung cancer.

This study validated the effectiveness and safety of the treatment for residual stones using flexible ureteroscopy (fURS) and holmium laser (0.6–1.2 J, 20–30 Hz) lithotripsy via a fiber with a 200-μm core diameter and 0.22 numerical aperture (NA) after the management of complex calculi with single-tract percutaneous nephrolithotomy (PCNL). Between January 2014 and June 2016, 27 consecutive patients with complex calculi underwent fURS and holmium laser lithotripsy after a planned single-tract PCNL. Among the 27 patients with complex calculi, 9 had full staghorn calculi, 7 had partial staghorn calculi, and 11 had multiple calculi. After the first single-tract PCNL session, the mean stone size and mean stone surface area were 18.0 ± 10.7 mm and 181.9 ± 172.2 mm2, respectively. Treatment for residual stones with fURS and holmium laser lithotripsy was successfully completed and was performed without intraoperative complications. The mean operative time of the fURS procedure was 69.1 ± 23.6 min, and the mean hospital stay was 5.3 ± 2.4 days. The mean decrease in the hemoglobin level was 7.3 ± 6.5 g/l. After the fURS procedure, the overall stone-free rate was 88.9%. The overall postoperative complication rate was 14.8% (Clavien grade I 11.1%; Clavien grade II 3.7%). The current approach tested here combines the advantages of both PCNL and fURS and effectively manages complex calculi with a high stone-free rate (SFR) (88.9%). This approach also reduced the number of treatment sessions, the number of percutaneous access tracts, and the blood loss and potential morbidity associated with multiple tracts.

Ethnopharmacological relevance Larix laricina , a native tree of North America, is a highly respected medicinal plant used for generations by Indigenous Peoples across its range, including the Cree of northern Quebec who use the bark to treat symptoms of diabetes. This study investigates the antioxidant capacity and bioavailability of active constituents identified in L. laricina bark extracts. Materials and methods (1) Oxygen radical absorbance capacity (ORAC) assay was employed to test antioxidant capacity of organic extracts (80% ethanol) from bark of L. laricina as well as fractions, isolated compounds, and media samples collected during permeability assays. (2) Caco-2 cell monolayer cultures were used to determine the permeability of identified antioxidants, which were quantified in basolateral media samples using liquid chromatography – tandem mass spectrometry (HPLC-ESI-MS/MS). Results Crude ethanolic extract possessed strong antioxidant potential in vitro (7.1±0.3 Trolox equivalents (TE) μM/mg). Among the 16 L. laricina fractions obtained by chromatographic separation, fraction 10 (F10) showed the highest antioxidant capacity (21.8±1.7 µm TE/mg). Among other identified antioxidants, the stilbene rhaponticin (isolated from F10) was the most potent (24.6±1.1 µm TE/mg). Caco-2 transport studies revealed that none of the identified compounds were detectable in basolateral samples after 2-h treatment with crude extract. In monolayers treated with F10 (60% rhaponticin), small quantities of rhaponticin were increasingly detected over time in basolateral samples with an apparent permeability coefficient (P app ) of 1.86×10 −8 cm/s (0–60 min). To model potential effects on blood redox status, we evaluated the antioxidant capacity of collected basolateral samples and observed enhanced activity over time after exposure to both extract and F10 (75 μg/mL) relative to control. Conclusions By profiling the antioxidant constituents of L. laricina bark, we identified rhaponticin as the most potent oxygen radical scavenger and observed low permeability in Caco-2 cell monolayers but an increase in basolateral antioxidant capacity.

Staphylococci are Gram-positive bacteria that are ubiquitous in the environment and able to form biofilms on a range of surfaces. They have been associated with a range of human health issues such as medical device-related infection, localized skin infection, or direct infection caused by toxin production. The extracellular material produced by these bacteria resists antibiotics and host defence mechanism which complicates the treatment process. The commonly reported

The overuse and misuse of antibiotics has resulted in the emergence of drug-resistant pathogenic bacteria, including meticillin-resistant Staphylococcus aureus (MRSA), the primary pathogen responsible for human skin and soft-tissue infections. Antibacterial peptides are known to kill bacteria by rapidly disrupting their membranes and are deemed plausible alternatives to conventional antibiotics. One advantage of their membrane-targeting mode of action is that bacteria are unlikely to develop resistance as changing their cell membrane structure and morphology would likely involve extensive genetic mutations. However, major concerns in using peptides as antibacterial drugs include their instability towards plasma proteases, toxicity towards human cells due to their membrane-targeting mode of action and high manufacturing cost. These concerns can be mitigated by developing peptides as topical agents, by the judicial selection of amino acids and developing very short peptides respectively. In this preliminary report, we reveal a linear, non-hemolytic tetrapeptide with rapid bactericidal activity against MRSA developed from a structure-activity relationship study based on the antimicrobial hexapeptide WRWRWR-NH2. Our finding opens promising avenues for the development of ultra-short antibacterials to treat multidrug-resistant MRSA skin and soft tissue infections.

The Chinese Dietary Guidelines (CDGs) were first released to the public in 1989 by the Chinese Nutrition Society (CNS). In 2016, the Ministry of Health commissioned the CNS to revise and publish new CDGs.The CNS convened an expert committee of leaders in the fields of nutrition, epidemiology, public health, preventive medicine, and food science. The CDGs were revised according to the World Health Organization Handbook for Guideline Development procedures. The expert committee recommended key inclusions for the CDGs on the basis of the current status of public health and priority health challenges affecting the Chinese population, as well as the quality of scientific evidence.The CDGs (2016) provide six key dietary recommendations for the general Chinese population aged 2 years and above. In addition to a newly revised Chinese Food Guide Pagoda, both the Chinese Food Guide Plate and Chinese Food Guide Abacus are newly created pictorial models in the CDGs (2016); the Chinese Food Guide Abacus is specifically designed for children. Seven additional dietary guidelines targeting specific populations are included; these guidelines emphasize the different physiological and nutritional requirements of particular populations, including dietary guidelines for vegetarians for the first time.The CDGs (2016) is an official document published by the National Health Commission of China. Its content highlights the current status of public health and offers advice to address health concerns faced by the Chinese population. The CDGs (2016) have been widely disseminated and accepted in the Chinese population; the National Nutrition Week is a major national event in which the CDG are used as its core to increase public awareness towards a healthy diet and lifestyle.

Kringle 5, derived from plasminogen, is highly capable of inhibiting angiogenesis. Here, we have designed and synthesized 10 tetrapeptides, based on the amino acid properties of the core tetrapeptide Lys-Leu-Tyr-Asp (KLYD) originating from anti-angiogenic kringle 5 of human plasminogen. Of these, Arg-Leu-Tyr-Glu (RLYE) effectively inhibited vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation, migration and tube formation, with an IC{sub 50} of 0.06–0.08 nM, which was about ten-fold lower than that of the control peptide KLYD (0.79 nM), as well as suppressed developmental angiogenesis in a zebrafish model. Furthermore, this peptide effectively inhibited the cellular events that precede angiogenesis, such as ERK and eNOS phosphorylation and nitric oxide production, in endothelial cells stimulated with VEGF. Collectively, these data demonstrate that RLYE is a potent anti-angiogenic peptide that targets the VEGF signaling pathway. - Highlights: • The tetrapeptide RLYE inhibited VEGF-induced angiogenesis in vitro. • RLYE also suppressed neovascularization in a zebrafish model. • Its effect was correlated with inhibition of VEGF-induced ERK and eNOS activation. • RLYE may be used as a therapeutic drug for angiogenesis-related diseases.

Background/purpose A common trait among cancers is the increased level of glycolysis despite adequate oxygen levels to support aerobic respiration. This has been shown repeatedly in different human malignancies. Glycolysis inhibitors, especially 3-bromopyruvate, have been shown to be effective chemotherapeutic agents. The effect of glycolysis inhibition upon chemotherapy resistance is relatively unknown. Methods Wild-type and doxorubicin-resistant lines of neuroblastoma (SK-N-SH and SK-N-Be(2)C) were used in this study. Using an MTT assay, the IC 50 of 3-BrPA was determined. Subsequently, doxorubicin-resistant cell lines were treated with 3-bromopyruvate, doxorubicin, and 3-bromopyruvate with doxorubicin. Additionally, a luminescence ATP detection assay was used to measure intracellular ATP levels, and a lactate assay was used to determine intracellular lactate levels. All experiments were repeated in hypoxic conditions. Results Treatment with 3-bromopyruvate and doxorubicin significantly decreased the mean cell viabilities at 24, 48, and 72hours in normoxic conditions. A similar response was replicated in hypoxic conditions. Treatment with 3-bromopyruvate significantly decreased intracellular ATP and lactate levels. Conclusion Glycolysis inhibitors such as 3-bromopyruvate could prove to become an effective means by which chemotherapy resistance can be overcome in human neuroblastoma.

Vulvovaginal candidiasis/candidosis is a common fungal infection afflicting approximately 75% of women globally caused primarily by the yeast Candida albicans. Fluconazole is widely regarded as the antifungal drug of choice since its introduction in 1990 due to its high oral bioavailability, convenient dosing regimen and favourable safety profile. However, its widespread use has led to the emergence of fluconazole-resistant C. albicans, posing a universal clinical concern. Coupled to the dearth of new antifungal drugs entering the market, it is imperative to introduce new drug classes to counter this threat. Antimicrobial peptides (AMPs) are potential candidates due to their membrane-disrupting mechanism of action. By specifically targeting fungal membranes and being rapidly fungicidal, they can reduce the chances of resistance development and treatment duration. Towards this goal, we conducted a head-to-head comparison of 61 short linear AMPs from the literature to identify the peptide with the most potent activity against fluconazole-resistant C. albicans. The 11-residue peptide, P11-6, was identified and assayed against a panel of clinical C. albicans isolates followed by fungicidal/static determination and a time-kill assay to gauge its potential for further drug development. Copyright © 2017 European Peptide Society and John WileySons, Ltd.

Myeloperoxidase (MPO) is an endogenous oxidant enzyme and can generate reactive oxygen species. The MPO G463A polymorphism influences MPO transcription levels and has been proposed to be associated with risk of lung cancer. To assess the effect of MPO G463A polymorphism on lung cancer risk in Asians, a pooled analysis of published case-control studies was performed. PubMed, Embase, China Biomedical Literature, and Wanfang Medicine databases were searched for eligible studies. The strength of the association between MPO G463A polymorphism and lung cancer risk was measured by odds ratio (OR) with 95 % confidence interval (95% CI). Finally, eight studies with a total of 1,679 lung cancer cases and 1,876 non-cancer controls were included. Overall, MPO G463A polymorphism was associated with decreased risk of lung cancer risk in Asians under two genetic models (OR AA vs. GG = 0.58, 95% CI 0.36-0.96, P = 0.033; OR AA vs. GG+AG = 0.60, 95% CI 0.37-0.98, P = 0.040). There was no obvious risk of publication bias in this meta-analysis. In conclusion, the pooled analysis suggests that MPO G463A polymorphism is associated with decreased risk of lung cancer risk in Asians.

Carbon monoxide (CO), derived by the enzymatic reaction of heme oxygenase (HO), is a cellular regulator of energy metabolism and cytoprotection; however, its underlying mechanism has not been clearly elucidated. Astrocytes pre-exposed to the CO-releasing compound CORM-2 increased mitochondrial biogenesis, mitochondrial electron transport components (cytochrome c, Cyt c; cytochrome c oxidase subunit 2, COX2), and ATP synthesis. The increased mitochondrial function was correlated with activation of AMP-activated protein kinase α and upregulation of HO-1, peroxisome proliferators-activated receptor γ-coactivator-1α (PGC-1α), and estrogen-related receptor α (ERRα). These events elicited by CORM-2 were suppressed by Ca

Supplemental Digital Content is available in the text, As chronic inflammation is involved in the pathogenesis and progression of hepatocellular carcinoma (HCC), we investigated the prognostic accuracy of a cluster of inflammatory scores, including the Glasgow Prognostic Score, modified Glasgow Prognostic Score, platelet to lymphocyte ratio, Prognostic Nutritional Index, Prognostic Index, and a novel Inflammation-Based Score (IBS) integrated preoperative and postoperative neutrophil to lymphocyte ratio in 2 independent cohorts. Further, we aimed to formulate an effective prognostic nomogram for HCC after hepatectomy. Prognostic value of inflammatory scores and Barcelona Clinic Liver Cancer (BCLC) stage were studied in a training cohort of 772 patients with HCC underwent hepatectomy. Independent predictors of survival identified in multivariate analysis were validated in an independent set of 349 patients with an overall similar clinical feature. In both training and validation cohorts, IBS, microscopic vascular invasion, and BCLC stage emerged as independent factors of overall survival (OS) and recurrence-free survival (RFS). The predictive capacity of the IBS in both OS and RFS appeared superior to that of the other inflammatory scores in terms of C-index. Additionally, the formulated nomogram comprised IBS resulted in more accurate prognostic prediction compared with BCLC stage alone. IBS is a novel and validated prognostic indicator of HCC after curative resection, and a robust HCC nomogram including IBS was developed to predict survival for patients after hepatectomy.

Keap1 is a cytoplasmic repressor of the transcription factor Nrf2, and its degradation induces Nrf2 activation, leading to upregulation of antioxidant phase II genes. We investigated the roles of phase II genes in vascular inflammation and septic injury using Keap1 siRNA and elucidated its underlying mechanism. Selective knockdown of Keap1 with siRNA promoted Nrf2-dependent expression of phase II genes in endothelial cells, such as heme oxygenase-1 (HO-1), glutamate-cysteine ligase (GCL), and peroxiredoxin-1 (Prx1), resulting in the elevation of cellular glutathione levels and suppression of tumor necrosis factor (TNF)-α-induced intracellular H(2)O(2) accumulation. Keap1 knockdown inhibited TNF-α-induced expression of intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by suppressing NF-κB activation via inhibition of its upstream modulators, Akt, NIK, and IKK, resulting in the elevation of monocyte adhesion to endothelial cells. Importantly, these events were reversed by HO-1 and GCL inhibitors and Prx1-specific siRNA. Keap1 knockdown also inhibited endotoxin-induced expression of inducible nitric oxide synthase (iNOS) and TNF-α by upregulating HO-1, GCL, and Prx1 expression in macrophages. Moreover, in vivo Keap1 knockdown increased the expression of phase II genes and suppressed the expression of ICAM-1, VCAM-1, iNOS, and TNF-α in an endotoxemic mouse model, resulting in significant protection against liver and lung injuries and lethality. Our results indicate that Keap1 knockdown prevents NF-κB-mediated vascular inflammation and endotoxic shock by suppressing NF-κB-mediated inflammatory gene expression via upregulation of Nrf2-mediated antioxidant genes. Thus, siRNA targeting Keap1 may provide a new therapeutic approach for inflammation-associated vascular diseases and sepsis.

article i nfo Taurine, a non essential sulfur-containing amino acid, plays a critical role in cardiovascular functions. We here examined the effect of taurine on angiogenesis and its underlying signal pathway. Taurine treatment in- creased angiogenesis in vitro and in vivo, which was followed by activation of the phosphatidylinositol 3- kinase (PI3K)/Akt, MEK/ERK, and Src/FAK signaling pathways. Further, taurine promoted endothelial cell cycle progression to the S and G2/M phases by up-regulating the positive cell cycle proteins, particularly cyclins D1 and B, as well as down-regulating the negative cell cycle proteins, p53 and p21 WAF1/CIP1 , resulting in Rb phosphorylation. This angiogenic event was inhibited by inhibitors of PI3K and MEK. In addition, a PI3K inhibitor blocked the activation of Akt and ERK, while Akt knockdown did not affect taurine-induced ERK ac- tivation, indicating that PI3K is an upstream mediator of both MEK and Akt. Taurine-induced endothelial cell migration was suppressed by Src inhibitor, but not by other inhibitors, suggesting that the increase in cell mi- gration is regulated by Src-dependent pathway. Moreover, inhibition of cellular taurine uptake by β-alanine and taurine transporter knockdown promoted taurine-induced cell proliferation, ERK and Akt activation, and in vivo angiogenesis, suggesting that extracellular taurine induces angiogenesis. However, taurine did not in- duce vascular inflammation and permeability in vitro and in vivo. These data demonstrate that extracellular taurine promotes angiogenesis by Akt- and ERK-dependent cell cycle progression and Src/FAK-mediated cell migration without inducing vascular inflammation, indicating that it is potential use for the treatment of vascular dysfunction-associated human diseases.

To study the demographic characteristics of ocular trauma in Central Sarawak and identify the determinant factors of poor visual outcome.A retrospective study of ocular trauma cases presenting at the referral hospital in 2013.Patients were identified and recruited from hospital records. Those presenting for follow-up review were excluded. Case records were retrieved and reviewed after recruitment.We studied 168 patients with 179 ocular injuries, of which 44% were work related. Compared with non-work-related cases, work-related cases were more likely to be male [odds ratio (OR), 19.7; 95% confidence interval (CI), 2.6-150.9] and foreign (OR, 18.0; 95% CI, 2.3-142.0). Open globe injuries constituted a higher percentage of impaired visual acuity (VA) during the first visit: 84.6% compared with 18.1% for closed globe injuries (OR, 25.0; 95% CI, 5.3-118.4; P0.001). Of the open globe injuries, 61.5% worsened or showed no improvement in VA after 3 months compared with closed globe injuries (28.9%) (OR, 3.9; 95% CI, 1.2-12.7; P = 0.015). Of cases presenting 7 or more days after trauma, 76.9% worsened or showed no improvement in VA after 3 months compared with those presenting in less than 7 days (27.7%) (OR, 8.7; 95% CI, 2.3-33.0; P0.001). Among those with work-related injuries, 23.1% had used eye protective devices (EPDs).Ocular injuries in Central Sarawak were predominantly work related, occurring at industrial premises, and involving males and foreigners. Both open globe injuries and a delay in seeking treatment resulted in significantly poorer visual outcomes.

The objective of the study was to evaluate fluid-electrolyte and renal pelvic pressure (RPP) changes during ureteroscopic lithotripsy. Fifteen patients were detected with residual ureteral calculi after minimally invasive percutaneous nephrolithotomy (MPCNL), distal ureter calculi in three, midureter calculi in four, proximal calculi in eight. RPP was measured via the percutaneous nephrostomy tube by urodynamic study at irrigation pressures of 50, 100 and 200 mmHg. Haemoglobin (Hb), haematocrit (Hct), blood urea mitrogen (BUN), creatinine (Cre), serum sodium (Na(+)), potassium (K(+)), chlorine (Cl(-)) were recorded before and after ureteroscopic lithotripsy. There were no significant differences between Hb, Hct, BUN, Cre, Na(+), K(+), Cl(-) values. Baseline RPP was (16.37 ± 3.14) cmH(2)O, RPPs were 46.06 ± 6.85 cmH(2)O, 99.07 ± 14.62 cmH(2)O and 166.27 ± 33.08 cmH(2)O at irrigation pressures of 50, 100 and 200 mmHg, they were much higher than baseline RPP (p0.0001). RPP in the proximal ureter was much higher than in the distal and middle ureter, 50.98 ± 4.52 cmH(2)O versus 40.44 ± 4.07 cmH(2)O (p = 0.0004), 110.26 ± 2.39 cmH(2)O versus 86.29 ± 11.60 cmH(2)O (p = 0.0014), 193.21 ± 5.88 cmH(2)O versus 135.47 ± 20.95 cmH(2)O (p = 0.0002) at irrigation pressures of 50, 100 and 200 mmHg. There were no significant changes in fluid-electrolyte. RPP was significantly increased during ureteroscopic lithotripsy, it was correlated with the irrigation pressure and the position in the ureter.

Purpose The purpose of the study was to determine if nerve growth factor (NGF) stimulation induces apoptosis in the BE(2)C neuroblastoma cell line in vitro. Methods The LPCX retroviral vector was used to achieve stable transduction of NGF complementary DNA into BE(2)C neuroblastoma cells. Wild-type and NGF-transduced cells were then incubated with varying concentrations of NGF for varying periods. A laddering assay was performed to determine the presence of DNA fragments characteristic of apoptosis. The expression of various cleaved and total caspases was determined by Western immunoblotting. Results p75 receptor expression in the NGF-transduced cell line was equivalent to that in the wild-type cell line, but Trk A receptor expression was significantly decreased in BE(2)C-NGF cells. DNA laddering assay demonstrated that only BE(2)C-NGF cells underwent apoptosis after stimulation with exogenous NGF. BE(2)C-NGF cells have increased expression of cleaved caspase-3 when compared with wild-type cells. Cleaved caspase-3 expression is further increased with exogenous NGF stimulation in the transduced cells. Conclusion This study confirms that NGF stimulation of BE(2)C neuroblastoma cells can induce apoptosis through activation of the caspase cascade in vitro. The differential expression of the receptors Trk A and p75 between the wild-type and NGF-transduced cell lines may explain the differing effects observed.

To evaluate the therapeutic effect of heated (60 degrees C) lipiodol via hepatic artery administration in a rabbit model of VX2 liver cancer.Thirty male New Zealand white rabbits were randomly divided into three groups with 10 rabbits assigned to each group. VX2 carcinoma cells were surgically implanted into the left hepatic lobe. The tumors were allowed to grow for 2 weeks, and studies were performed until the diameter of the tumors detected by ultrasonograph reached 2-3cm. Under anesthesia, trans-catheter hepatic arterial embolization was performed and doxorubicin-lipiodol (37 degrees C) (1mL), lipiodol (60 degrees C) (1mL) or control (physiological saline (37 degrees C) (1mL)) solution was injected into the hepatic arteries of animals in the three groups. One week later, the volume of the tumor was measured by ultrasonograph again. The serum of all rabbits was collected before injection and at 4 and 7 days after injection, and the level of aspartate aminotransferase (AST) was checked. The survival period of the three groups of rabbits after treatment was also recorded. During the last course of their disease, the rabbits were given analgesics to relieve suffering.The tumor growth rate in the lipiodol (60 degrees C) group (0.92+/-0.21, tumor volume from 1811+/-435 to 1670+/-564mm(3)) was significantly lower than that in the control group (3.48+/-1.17, tumor volume from 1808+/-756 to 5747+/-1341mm(3)) (P0.05) and in the doxorubicin-lipiodol (37 degrees C) group (1.69+/-0.26, tumor volume from 1881+/-641 to 2428+/-752mm(3)) (P0.05). Consequently, the survival period of the animals in the lipiodol (60 degrees C) group (41.0+/-3.0 days) was significantly greater than that in the doxorubicin-lipiodol (37 degrees C) group (38.0+/-2.5 days) (P0.05). On the other hand, there was no statistically significant difference in serum AST levels between the lipiodol (60 degrees C) group (148.2+/-11.3UL(-1)) and the doxorubicin-lipiodol (37 degrees C) group (139.7+/-12.3UL(-1)) (P0.05). However, the serum AST level in the lipiodol (60 degrees C) group was significantly higher at 4 days after injection (P0.05) than in the control group (68.6+/-6.6UL(-1)).Treatment with lipiodol (60 degrees C) resulted in an effect on serum AST levels similar to that caused by treatment with doxorubicin-lipiodol (37 degrees C). Thus, lipiodol (60 degrees C) treatment could greatly prolong the survival period of rabbits with VX2 cancer by inhibiting tumor growth.

Strontium ranelate is a new effective anti-osteoporotic treatment having a unique mode of action, reducing bone resorption while promoting continued bone formation, with a broad range of anti-fracture efficacy at vertebral as well as peripheral sites. In Phase III studies, it has proven its early and sustained efficacy against vertebral fractures in Caucasians along with a significant increase in lumbar bone mineral density (BMD). The aim of this randomized double-blind study was to demonstrate the efficacy of strontium ranelate (2 g/day) on lumbar spine bone mineral density and the clinical and biological safety in Asian postmenopausal osteoporotic patients compared to placebo over 1 year. Three hundred and twenty-nine eligible women from mainland China, Hong Kong and Malaysia were randomized into the study. The baseline characteristics were similar in the treatment and placebo groups: mean age of 66.2+/-6.5 years, time since menopause 17.6+/-7.2 years. In the Full Analysis Set (FAS, N=302), the mean baseline lumbar L2-L4 BMD was 0.715+/-0.106 g/cm(2) in the strontium ranelate group and 0.708 +/- 0.109 g/cm2 in the placebo group. The mean baseline femoral neck BMD was 0.575+/-0.074 g/cm2 and 0.566+/-0.069 g/cm2 respectively and mean total hip BMD was 0.642+/-0.080 g/cm2 and 0.631 +/-0.088 g/cm2 respectively. The overall compliance was 91.4% in the study drug group, and 97.4% in the placebo group. After 1 year of treatment, the lumbar spine, femoral neck and total hip BMD in the treated group was significantly increased by 3-5% as compared to placebo. Strontium ranelate was well tolerated. The most frequently reported emergent adverse events were comparable in both groups (60.4% versus 60.0%), with majority of them being mild gastrointestinal disorders. There were no clinically relevant changes in laboratory tests, such as blood routine, hepatic and renal function. It is thus concluded that the effects of 2 g/day strontium ranelate on BMD and its safety profile in this cohort of postmenopausal osteoporotic Asian women were consistent with results obtained from Caucasian women in which the efficacy on the reduction in risk of fracture has been proven.

Objective To investigate the clinical characteristics, treatment options and causes of misdiagnosis of ipsilateral femoral neck and shaft fractures. Methods Among 20 patients with ipsilateral femoral neck and shaft fractures, 19 were treated operatively and 1 was treated conservatively. Sixteen cases of femoral shaft fractures were treated by open reduction and internal fixation with compressive plate, and 2 cases were treated with interlocking intramedullary nailing. Eighteen femoral neck fractures were treated with cannulated screws. Another patient was treated with proximal femoral nail to fix both the neck and shaft. Delayed diagnosis for femoral neck fractures occurred in 2 cases preoperatively. Results A total of 19 patients were followed up. The follow up period ranged from 5 to 48 months with an average of 15 months. All the fractures were healed. Conclusion For case of femoral shaft fracture caused by high energy injury, an AP pelvic film should be routinely taken. Once the femoral neck fracture is recognized, operative reduction and fixation should be performed in time. Femoral neck and shaft fractures should be fixed separately.

Prolonged exposure to high level of estrogen is a known risk factor for breast carcinogenesis. In cells, estrogens, in particular estrone (E1) and 17 beta-estradiol (E2), can be converted to catecholestrogens (CEs) which may be oxidized to form CE-semiquinones and CE-quinones that are capable of binding to DNA to induce mutations, followed by carcinogenesis. Whether the body is equipped with protective mechanisms against potentially harmful CEs, therefore, is an important issue. The present study was designed to examine the role of sulfation in the metabolism of CEs. MCF-7 breast cancer cells and MCF 10A human mammary epithelial cells were metabolically labeled with [35S]sulfate in the presence of individual CEs. Analysis of the labeling media showed the generation and release of exclusively [35S]sulfated 2-methoxy-E1 or [35S]sulfated 2- or 4-methoxy-E2 by cells labeled in the presence of 2-OH-E1 or 2- or 4-OH-E2. Whereas both [35S]sulfated 4-methoxy-E1 and [35S]sulfated 4-OH-E1 were detected in the labeling media of cells labeled in the presence of 4-OH-E1. These results indicated a concerted action of catechol-O-methyltransferase (COMT) and the cytosolic sulfotransferase (SULT) enzyme(s) in the metabolism of CEs. Enzymatic assays revealed that, five (SULT1A1, SULT1A2, SULT1A3, SULT1C4, and SULT1E1) of eleven known human SULTs tested could use CEs and methoxyestrogens (MEs) as substrates, with SULT1E1 displaying the strongest sulfating activity.

BACKGROUND: The advent of wavefront technology in the past five years has provided some insight into the optical outcomes of cataract surgery. The Tecnis Z9001 intraocular lens (Tecnis IOL, AMO) with a modified prolate anterior surface was claimed to reduce or even eliminate ocular spherical aberration to improve the visual quality. The purpose of this study was to determine whether Tecnis IOLs can improve the quality of vision as evaluated by measuring visual acuity, wavefront aberration, and contrast sensitivity. METHODS: In an intraindividual prospective study, 20 patients with bilateral cataract were randomly assigned to receive a modified prolate anterior surface IOL (Tecnis Z9001, AMO) in one eye and a biconvex spherical surface IOL (CeeOn 911A, AMO) in the other. After 3 months, the following were investigated: best corrected visual acuity, pupil diameter, photopic and mesopic contrast sensitivity, and wavefront aberration of the whole eye (ocular), cornea, and internal plane. RESULTS: The differences in the best corrected visual acuity and pupil diameter between the two groups were not statistically significant. Negative 4th-order spherical aberration (Z(4)(0)) was found in the Tecnis group, whereas positive Z(4)(0) found in the CeeOn group for the internal and ocular plane. Statistically significant differences were found at the ocular higher-aberrations between the two IOLs. Contrast sensitivity testing showed significantly better results in the Tecnis group at visual angles higher than 1.0 degree under photopic conditions and at visual angles higher than 1.6 degree under mesopic conditions both without glare and with glare in comparison with the CeeOn group. CONCLUSIONS: The Tecnis Z9001 IOL with a modified prolate anterior surface produces negative spherical aberration and consequently reduces the higher-order aberrations in pseudophakic eyes. This leads to enhanced contrast sensitivity and improved functional vision compared to conventional spherical IOLs.

To investigate the expression and clinical significance of gelatinases (MMP-2 and MMP-9) in patients with laryngeal squamous cell carcinoma (LSCC).In a retrospective study of 72 consecutive patients with LSCC hospitalized in a single cancer center, immunohistochemistry was used to examine the expression of MMP-2 and MMP-9 in surgical samples. The results were compared to clinicopathological features and prognosis.The positive expression of MMP-2 and MMP-9 in patients with LSCC was 50% (36/72) and 73.6% (53/72), respectively. According to the expression scale, there were 36 patients of -, 26 patients of +, 7 patients of ++, and 3 patients of +++ expression of MMP-2; 19 patients of -, 26 patients of +, 16 patients of ++, and 11 patients of +++ expression of MMP-9. There was no significant relationship found between the expression of MMP-2 or MMP-9 and clinicopathological features of LSCC, such as histological grade, primary site, T stage, N stage, and clinical stage. The 5-year overall survival (OS) and disease-free survival (DFS) rate calculated by Kaplan-Meier method in patients with negative and positive expression of MMP-9 and MMP-2 was 73.68%, 50.94%, 73.68%, and 49.06% in MMP-9 and 72.22%, 41.67%, 72.22%, and 38.89% in MMP-2, respectively. Significant 5-year survival difference was found between patients with negative and positive expression of MMP-2 (log rank = 6.74, P = 0.0094). There was significant lower survival rate in patients with higher positive expression of MMP-2 (log rank = 11.77, P = 0.0028). In glottic laryngeal cancer, positive expression of MMP-2 could predict poor survival and was more likely to present primary recurrence.The expression of MMP-2 could be used as a potential predictor for poor prognosis in patients with LSCC.

To investigate the causes and prevention of the complications about treatment of acromioclavicular joint dislocation (Tossy III) and unstable distal clavicular fracture (Neer II) with clavicular hook plate.From January 2001 to December 2011, 246 patients with acromioclavicular joint dislocation (Tossy III) and 222 patients with unstable distal clavicular fracture (Neer II) were treated with acromioclvicular hook plate fixation,including 348 males and 120 females with an average age of 45.4 years old ranging from 21 to 80 years old. The mean time from injury to operation was 30.8 hours (ranged from 1 h to 15 d). All patients had normal shoulder function before injury. According to Karlsson evaluation standard, the cases with excellent and good function of the shoulder joint were regarded as the normal group, and the cases with poor function of shoulder joint as the abnormal group. The comparison of the range of forward flexion,backward stretch, adduction, abduction and elevation of shoulder joints between two groups was performed. The data of impingement, subacromial osteolysis, acromioclavicular arthritis, clavicular stress fracture, downward acromioclavicular joint subluxation, hook cut-out and hook break were summarized.All patients were followed up from 8 to 48 months with an average of 12.5 months. The results were excellent in 308 cases,good in 76,and poor in 84 according to Karlsson evaluation. The excellent and good rate was 82.1%. The difference of the range of forward flexion, backward stretch, adduction, abduction and elevation of shoulder joints between two groups had a statistically significant difference (P0.01). Among 84 poor cases, there were 41 (8.76%) in acromial impingement or inadequate place of plate hook, 12 (2.56%) with subacromial osteolysis or/and bursitis, 10 (2.14%) with acromioclavicular arthritis or painful shoulder caused by delayed dirigation,7 (1.50%) with clavicular stress fracture or interal plate upward, 6 (1.28%) with downward acromioclavicular joint subluxation, 5 (1.07%) with hook cut -out and 3 (0.64%) in hook break.The clavicular hook plate is useful for the treatment of acromioclavicular joint dislocation (Tossy III) and unstable distal clavicular fracture (Neer II). The correct place and suitable preflex of plate hook,the restoration of fiber structure around the acromioclavicular joint and the advisable dirigation contribute to the modified rate of complications.

To investigate the characteristics of menopause of Chinese women with the age of 40-60 years concerning gynecologic clinics in China.From Mar.2008 to Sept.2008, a face-to-face questionnaire was conducted in gynecological clinic in perimenopausal and postmenopausal women in 14 hospitals in China, which included general demographic data, menstrual change process, climacteric symptoms and knowledge about menopause. Modified Kupperman index were used to evaluate climacteric symptoms during the recent week and awareness of hormonal replacement therapy were studied.A total of 1641 women were investigated. The ages of onset of menopause transition, climacteric symptoms and natural menopause were (47 ± 4), (46 ± 4), (49 ± 3) years old respectively. Climacteric symptoms could be found in 78.43% (1287/1641) women during menopausal transition, which were mainly mild to moderate symptoms. The top 5 symptoms were fatigue and weakness (71.48%, 1173/1641), irritability (68.68%, 1127/1641), insomnia (67.65%, 1110/1641), muscle and joint pain (64.11%, 1052/1641) and hot flush (57.90%, 950/1641). The climacteric symptoms were not constant during menopausal transition, usually more severe in late transition and postmenopausal periods, during which the moderate and severe symptoms were 59.1% (189/320) and 51.1% (291/570) respectively. Although most symptoms primarily appeared along with menstruation change, there are about 17.5% (172/981) patients experienced climacteric symptoms before menstruation change occurrence. There were 56.39% (733/1300) women had ever heard (mostly from gynecologist) about hormone replacement therapy from Obstetrician and Gynecologist.Most of the women during menopausal transition had climacteric symptoms, usually mild and moderate ones. Although most symptoms primarily appeared along with menstruation change, there are other patients' experienced climacteric symptoms before menstruation change occurrence.

In diabetic retinopathy (DR), visual deterioration is related with retinal neovascularization and vascular hyperpermeability. Anti-vascular endothelial growth factor (VEGF) agents are currently utilized to suppress retinal neovascularization and macular edema (ME); however, there are still concerns on the widespread use of them because VEGF is a trophic factor for neuronal and endothelial cells in the retina. As an alternative treatment strategy for DR, it is logical to address hypoxia-related molecules to treat DR because the retina is in relative hypoxia as DR progresses. In this study, we demonstrate that destabilization of hypoxia-inducible factor-1α (HIF-1α) by SH-1242 and SH-1280, novel heat shock protein 90 (hsp90) inhibitors, leads to suppression of hypoxia-mediated retinal neovascularization and vascular leakage in diabetic retina. In vitro experiments showed that these inhibitors inhibited hypoxia-induced upregulation of target genes of HIF-1α and further secretion of VEGF. Furthermore, these inhibitors effectively suppressed expression of target genes of HIF-1α including vegfa in the retina of oxygen-induced retinopathy (OIR) mice. Interestingly, despite hsp90 inhibition, these inhibitors do not induce definite toxicity at the level of gene expression, cellular viability, and histologic integrity. We suggest that SH-1242 and SH-1280 can be utilized in the treatment of DR, as an alternative treatment of direct VEGF inhibition. Key message: SH-1242 and SH-1280 are novel hsp90 inhibitors similar to deguelin. HIF-1α destabilization by hsp90 inhibition leads to anti-angiogenic effects. Despite hsp90 inhibition, both inhibitors do not induce definite toxicity. HIF-1α modulation can be a safer therapeutic option than direct VEGF inhibition.

DNA G-quadruplexes are four-stranded DNA structures and are widely distributed in functional regions of the human genome, such as telomeres and gene promoter regions. G-quadruplex structures can play important roles, including in immunoglobulin gene rearrangements, DNA replication, gene transcription, and are viewed as valid therapeutic targets in human cancer diseases. Design of G-quadruplex binders that target these structures and regulate related gene functions through stabilization of these structures are emerging as an exciting new class of potential anticancer agents. Besides as drug candidate, DNA G-quadruplex binders can also serve as excellent probes, helping the further exploration of biological functions of G-quadruplex and early diagnosis of G-quadruplex-related disease.This review provides an overview of current knowledge on patents of DNA G-quadruplex binders from 2008 to 2013. Information is collected from an extensive search, covering Derwent Innovations Index, Espacenet, SciFinder, and Google patent search.With the accumulating evidence of G-quadruplex as an effective drug target, an increasing number of DNA G-quadruplex binders with diverse structural features were developed. These binders are either used as drug candidates targeting G-quadruplex, or as probes for diagnostic purpose in genomic study. This review would mainly focus on the patents published after 2008 (including 2008). In order to cover all the diverse structural types of DNA G-quadruplex binders, some patents published before 2008 would be mentioned as well.

To observe the effect of rapamycin on the MG-63 osteosarcoma cells (OC), osteosarcoma stem cells (OSC) and on mTOR signaling pathway, and explore the feasibility of rapamycin as a novel therapeutic measure in osteosarcoma chemotherapy regimens.OC and OSC were cultured in vitro. Immunofluorescence assay was used to detect the expression of Nanog and Oct4 in OC and OSC. OC and OSC were treated with rapamycin in concentrations of 0, 20, 50 and 100 nmol/L. Semi-quantitative PCR and RT-PCR were used to detect the mTOR mRNA and CCK-8 assay was used to detect cell proliferation, and the cell morphology was observed under an inverted microscope.The cores of MG-63 cellular spheres exhibited embryonic stem cell characteristics such as Nanog and Oct4 expession. The mTOR pathway was activated in the OSC and the expression of mTOR mRNA was higher in OSC (0.761 ± 0.080) than that in OS (0.406 ± 0.090, P0.05) by semi-quantitative PCR. RT-PCR showed that the expression of mTOR mRNA was lower in OSCs treated with 100 nmol/L rapamycin (0.961 ± 0.060) than that with 0 nmol/L rapamycin (1.654 ± 0.246, P0.05). Cell counting kit-8 (CCK-8) assay showed that the proliferation of OC treated with 20, 50 and 100 nmol/L rapamycin was significantly inhibited, compared with that with 0 nmol/L rapamycin (P0.05). Compared with 0 nmol/L rapamycin, the proliferation of OSC treated with 20 and 50 nmol/L rapamycin was not significantly inhibited (P0.05), but that with 100 nmol/L rapamycin was significantly inhibited (P0.05). The invert microscopic observation revealed that rapamycin inhibited the formation of OSC spheres.Rapamycin can effectively inhibit cell proliferation and the ability of sphere formation of OSCs. It will provide a basis for a novel therapeutic approach in osteosarcoma chemotherapy regimens.

To observe the correlation between the decline of cognitive function and the level of plasma homocysteine in patients with Alzheimer's disease (AD).Thirty six AD patients were selected from hospitals in Tianjin. The enrolled patients were in accord with the diagnosis criteria. Thirty two control subjects were corresponding patients without AD in the period. Blood samples were extracted from each subject to determine the levels of homocysteine (Hcy) and folate. Cognitive status was evaluated by the mini- mental state examination (MMSE) and clinical dementia rating scale (CDR).The mean value of serum Hcy concentration [(17.51 +/- 5.62) micromol/L] of AD group was higher than that of control group [(12.38 +/- 4.25)micromol/L]. The serum [(5.17 +/- 1.76) microg/L] and diet folate [(206.94 +/- 44.51) microg/d] concentration of AD group were lower than those of control group [(7.92 +/- 2.22) microg/L, (259.74 +/- 41.92) microg/ d]. The incidence of hyperhomocysteinemia in AD group (64%) was higher than that in control group (22%). A significant relation between Hcy concentrations and the CDR was observed. With the increase of Hcy concentrations the CDR raised, and with the increase of Hcy concentrations the MMSE decreased.Hyperhomocysteinemia is one of the risk factors inducing the onset of AD. There is a significant negative correlation between Hcy levels and cognitive levels in AD group. Folate deficiency is an important reason to cause elevated Hcy levels in AD.

The study of blood flow improvement of local estrogen on the low genitourinary tract and its safety.The color Doppler flow imaging technique was employed to observe the flow spectrum of genitourinary tract. During the period of February 2007 to December 2010, there were 78 cases of postmenopausal females on local estrogen. The Doppler parameters included vaginal wall, urethra resistance index (RI) and systolic/diastolic ratio and others. And the lower genital urinary tract symptoms, such as sexual satisfaction, vulvovaginitis, urinary frequency, nocturia and other improvements, as well as adverse drug reactions before treatment and after 1 month, 3 months were recorded and analyzed.The results before treatment and after 1 month, 3 months, vaginal wall artery RI were 0.9 ± 0.2, 0.6 ± 0.2 and 0.7 ± 0.3; S/D 8.6 ± 3.7, 6.0 ± 2.9 and 6.4 ± 2.1; urethral artery RI 0.7 ± 0.2, 0.6 ± 0.2 and 0.6 ± 0.2, S/D 6.6 ± 1.9, 3.5 ± 0.7 and 3.4 ± 0.5 respectively. When the pre- and post-treatment indices were compared, the differences were statistically significant (P0.01). After using local estrogen in postmenopausal women, the resistance of lower genital urinary tract vasodilation decreased. Sexual satisfaction increased while the morbidities of vulvovaginitis, urinary frequency and nocturia decreased. The difference was statistically significant (P0.01).Postmenopausal females on topical estrogen may promote the blood circulation of lower genitourinary tract, red.

Epithelial-mesenchymal transition (EMT) is an essential step for cancer metastasis. MicroRNAs (miRNAs) are small non-coding RNAs that regulate target-mRNAs post-transcriptionally. The expression and function of miRNAs in EMT of HT-29 colonic cells remain elusive. This study looks at expression of miRNAs in EMT and explores the effects of miRNAs on EMT in HT-29 cell line. HT-29 was treated with TGF β to establish an EMT model, in which a collection of miRNAs was dynamically regulated by real-time PCR (qPCR) analysis. Among them, miR-21 and miR-27 were significantly upregulated, while miR-22, miR-26, miR-30, miR-181, miR-200b, miR-200c and miR-214 were markedly downregulated. MiRNA-inhibitors were used to knockdown miRNAs in HT-29 and EMT markers were determined by qPCR to monitor the effects of miRNAs on EMT process. Results showed that miR-22 could not alter the expression of EMT markers, while knockdown of miR-200b could significantly increase that of epithelial markers, N-cadherin, Vimentin, α-Sma and Twist1 and decrease that of mesenchymal marker, E-cadherin. Bioinformatic analysis and Western blot showed that ZEB1 was directly suppressed by miR-200b. In conclusion, miRNAs are dynamically regulated in TGF β-induced EMT of HT-29 and miR-200b was essential for EMT by suppressing the expression of ZEB1 in HT-29.

The C-5-methylation of cytosine in the CpG islands is an important pattern for epigenetic modification of gene, which plays a key role in regulating gene transcription. G-quadruplex is an unusual DNA secondary structure formed in G-rich regions and is identified as a transcription repressor in some oncogenes, such as c-myc and bcl-2. In the present study, the results from CD spectrum and FRET assay showed that the methylation of cytosine in the CpG islands could induce a conformational change of the G-quadruplex in the P1 promoter of bcl-2, and greatly increase the thermal-stability of this DNA oligomer. Moreover, the methylation of cytosine in the G-quadruplex could protect the structure from the disruption by the complementary strand, showing with the increasing ability to arrest the polymerase in PCR stop assay. This data indicated that the stabilization of the G-quadruplex structure in the CpG islands might be involved in the epigenetical transcriptional regulation for specific genes through the C-5-methylation modification pattern.

Radiofrequency thermocoagulation (RFT) is widely used to treat trigeminal neuralgia (TN); however, the optimal temperature at which RFT is most efficacious remains under much debate. Thus, the aim of the present study was to determine the lowest temperature at which morbidity could be minimized and patient outcomes maximized. A multivariate analysis was used to study 1354 patients who underwent computed tomography (CT)-guided RFT for V2/V3 idiopathic trigeminal neuralgia (ITN) during from June 2006 to May 2015. RFT was carried out at 62, 65, and 68°C, while keeping all other RF parameters the same. This was a prospective cohort study, in which we assessed intra- and postoperative complications, pain relief, and long-term health-related quality of life (HRQoL). The intraoperative and in-hospital complications of patients were mainly facial hematoma, mouth and external auditory meatus penetration, nausea, vomiting, dizziness, and headache, which were all treated symptomatically. In long-term follow-up, patients with pain relief (defined as no pain and no required drug intervention) at 62, 65, and 68°C accounted for 94.2%, 98.3%, and 98.8% (at discharge); 83.8%, 90.1%, and 91.4% (at 1 year); 66.7%, 80.5%, and 88.2% (at 3 years); 59.0%, 64.3%, and 77.2% (at 5 years); 48.7%, 57.8%, and 72.3% (at 7 years); 40.6%, 53.7%, and 60.3% (at 9 years), respectively. The number of patients with facial numbness, masticatory atonia, or corneal hypoesthesia was increased with the elevation of temperature, but these complications were all mild. No blindness, deafness, intracranial hemorrhage, or death as a result of the surgical intervention occurred in any patients. SF-36 scores showed highest HRQoL in the group treated at 68°C, followed by the 65 and 62°C groups, respectively. Our results demonstrate that 68°C is a good choice for RFT of V2/V3 ITN. The alternative option is 65 or 62°C for RFT to minimize the occurrence of complications including facial numbness, yet which often yields a higher recurrence rate.

To construct pCEP4/hIL-17B recombinant expression vector and express it stably in eukaryotic cells and investigate the biological activity in vitro.The CDS region of human IL-17B gene was cloned by RT-PCR. After identification by sequencing, the hIL-17B gene was inserted into expression vector of pCEP4 to construct the recombinant vector pCEP4/hIL-17B, then transfected into 293T cells. The transgenic 293T cell line stably expressing rhIL-17B protein was selected in the presence of Hygromycin B. After FCS-free cultivation and sub-cloning, The IL-17B/mFc gene and protein expression was confirmed by RT-PCR, ELISA and Western blot analysis. To investigate the ability of combination with IL-17B receptor on human leukemic monocytic cell line, THP-1, by Flow cytometrical analysis (FACS) and of stimulation to secret cytokines in vitro.The recombinant pCEP4/hIL-17B and its transgenic 293T cells stably expressing rhIL-17B protein were obtained successfully. FACS analysis showed its high affinity with its receptor and it can stimulated THP-1 cell line to excrete IL-1β and TNF-α in vitro and consistently caused a dose-dependent influx of neutrophil into the peritoneal cavity by intraperitoneal injection in vivo.The obtainment of transgenic 293T cell line stably expressing rhIL-17B protein paved the way for further study on biological functions of hIL-17B.

Histone deacetylase (HDAC) inhibitors, especially vorinostat, are currently under investigation as potential adjuncts in the treatment of neuroblastoma. The effect of vorinostat co-treatment on the development of resistance to other chemotherapeutic agents is unknown. In the present study, we treated two human neuroblastoma cell lines [SK-N-SH and SK-N-Be(2)C] with progressively increasing doses of doxorubicin under two conditions: with and without vorinsotat co-therapy. The resultant doxorubicin-resistant (DoxR) and vorinostat-treated doxorubicin resistant (DoxR-v) cells were equally resistant to doxorubicin despite significantly lower P-glycoprotein expression in the DoxR-v cells. Whole genome analysis was performed using the Ilumina Human HT-12 v4 Expression Beadchip to identify genes with differential expression unique to the DoxR-v cells. We uncovered a number of genes whose differential expression in the DoxR-v cells might contribute to their resistant phenotype, including hypoxia inducible factor-2. Finally, we used Gene Ontology to categorize the biological functions of the differentially expressed genes unique to the DoxR-v cells and found that genes involved in cellular metabolism were especially affected.

To study the different proportions of intermediate epithelial cells in human prostate cancer tissue and their clinical significance.We performed immunohistochemical staining for Cytokeratin 5 (CK5) and Cytokeratin 8 (CK8) on 60 samples of human prostate cancer, determined the proportions of intermediate epithelial cells in the cancer tissue, and classified the samples into 2 types, one with a majority of intermediate epithelial cells (CaP-INT, n = 32), and the other composed mostly of luminal epithelial cells (CaP-LUM, n = 28). Then we compared the 2 types of prostate cancer in the expression of the androgen receptor (AR), age of the patient, serum t-PSA, prostate volume, Gleason score, clinical stage, androgen resistance, and incidence of distant metastasis.CaP-INT showed a significantly lower expression of AR ([24.42 +/- 11.41] %) and a higher incidence of distant metastasis (n = 14) than CaP-LUM ([77.21 +/- 10.22] % and n = 4) (P0.05). In the CaP-INT group, 6 of the 26 endocrinologically treated cases developed into androgen-independent prostate cancer (AIPC), while in the CaP-LUM group, only 1 out of 23 (P0.05). The former also showed remarkably higher clinical stages than the latter (P0.05), but no significant differences were found in age, serum t-PSA, prostate volume and Gleason score between the two groups (P0.05).A higher proportion of intermediate epithelial cells may lead to increased invasiveness and metastasis of human prostate cancer.

Mg(2+) is one of the most important cations in cells, affecting the structures and functions of the proteins and nucleic acids. It should be noted that Mg(2+) is indispensable in DNA transcription, where G-quadruplex is believed to be actively involved. Therefore, it is important to investigate the influence of Mg(2+) on G-quadruplex. Here we studied the effect of Mg(2+) on G-quadruplex DNA with CD, FRET, EMSA, and PCR-stop assay. We found that various G-quadruplexes could be differentiated through simultaneous addition of both K(+) and Mg(2+), which could be used for selective identification of G-quadruplexes in promoter oncogene but not in telomere. Mg(2+) at physiological relevant concentration not only greatly enhanced the thermostability of oncogene G-quadruplexes but also efficiently protected them from unfolding by their complementary strands, which revealed the great impact of Mg(2+) on the equilibrium between promoter G-quadruplex and duplex DNA. The PCR-stop assay further confirmed that Mg(2+) could affect gene transcription by stabilizing promoter G-quadruplex. The above studies were carried out for various G-quadruplexes of varying sequences in promoter oncogenes and telomeric region. Our results suggest that Mg(2+) may be a key regulator for G-quadruplexes of oncogene promoter, which can subsequently affect the expression of related genes.