Quorum sensing mediated response mechanism of anammox consortia to anionic surfactant: Molecular simulation and molecular evidence.

The widespread use of surfactants raise challenges to biological wastewater treatment. Anaerobic ammonium oxidation (anammox) process has the potential to treat wastewater containing anionic surfactants, but the response of anammox consortia at the molecular level under long-term exposure is unclear. Using high-throughput sequencing and gene quantification, combined with molecular docking, the effect of sodium dodecyl sulfonate (SDS) on anammox consortia were investigated. Levels of reactive oxygen species (ROS) might be lower than the threshold of oxidative damage, while the increase of lactate dehydrogenase (LDH) represented the cell membrane damage. Decreased abundance of functional genes (hdh, hzsA and nirS) indicated the decrease of the anammox bacterial abundance. Trace amounts of N-acyl homoserine lactone (AHL, C6-HSL, C8-HSL and C12-HSL) contained in influent could induce endogenous quorum sensing (QS), which could regulate the correlation between functional bacteria to optimize the microbial community and strengthen the resistance of anammox consortia to SDS. In addition, the proliferation of disinfectant resistance genes might increase the environmental pathogenicity of sewage discharge. This work highlights the potential response mechanism of anammox consortium to surfactants and provides a universal microbial-friendly bioenhancement strategy based on QS.

[Regulatory effect of five-element music therapy of traditional Chinese medicine for suboptimal health status of liver stagnation and spleen deficiency based on the temperature changes of acupoints].

OBJECTIVE: To observe the effect of five-element music therapy of traditional Chinese medicine (TCM) on the clinical symptoms and the quality of life in the patients with suboptimal health status (SHS) of liver stagnation and spleen deficiency and explore the corresponding specificity changes in the temperature of acupoints when zangfu functions are of dysfunction and recovered to be balanced, separately. METHODS: Sixty patients with SHS of liver stagnation and spleen deficiency were randomized into an observation group and a control group, 30 cases in each one. In the control group, the conventional health education was provided. In the observation group, on the base of the therapeutic regimen as the control group, the patients received the five-element music therapy to pacify the liver qi and strengthen the spleen functions, once every two days, 30 min each time, 3 treatments a week. The course of treatment consisted of 4 weeks. Before and after treatment, the TCM syndrome score and the MOS 36-item short form healthy survey (SF-36) score were compared between the two groups and the clinical therapeutic effect was evaluated. Using infrared thermal imaging, the temperature at the acupoints of the affected organs (liver, spleen), the related organs (gallbladder, stomach) and the other non-related zangfu organs (pericardium, lung) was detected before and after treatment in the two groups. RESULTS: After treatment, the TCM syndrome scores were reduced when compared with those before treatment in the two groups (P<0.01, P<0.05); the reduction in the observation group was larger than that of the control group (P<0.01). The score of each domain for the SF-36 in the observation group and the score of role-emotional domain in the control group were all increased when compared with the scores before treatment (P<0.01, P<0.05); and the score of each domain for the SF-36 in the observation group was higher than that in the control group (P<0.01). The total effective rate was 66.7% (20/30) in the observation group, which was higher than 10.0% in the control group (3/30, P<0.05). In the observation group, the temperature of the yuan-primary point, the back-shu point and the front-mu point related to the liver, as well as those related to the gallbladder after treatment was reduced when compared with the temperature before treatment; and the changes were larger than those of the control group (P<0.01). The temperature of the yuan-primary point, the back-shu point and the front-mu point related to the spleen, as well as the back-shu point and the front-mu point related to the stomach in the observation group was increased when compared with the temperature before treatment (P<0.01); and the changes were larger than those of the control group (P<0.01). For the temperature of the non-specific points related to the liver and spleen, as well as the yuan-primary point, the back-shu point and the front-mu point related to the pericardium and the lung, there was no significant differences when compared with the temperature at the above-mentioned acupoints before and after treatment (P>0.05). CONCLUSION: TCM five-element music therapy associated with the conventional health education may effectively relieve the clinical symptoms and improve the quality of life in the patients with suboptimal health status of liver stagnation and sleep deficiency; and the therapeutic effect is better than the simple health education. The changes in the temperature of acupoints may reflect the functional regulation of the related zangfu organs in the body.

Doxorubicin and rhein loaded nanomicelles attenuates multidrug resistance in human ovarian cancer.

Tumor targeting delivery system has been suggested as an attractive strategy against tumor progression. Combination chemotherapy is essential and effective in preventing ovarian cancer. Rhein (4, 5-dihydroxyanthraquinone-2-carboxylic acid) is a lipophilic anthraquinone. Emerging evidence indicates that rhein has many pharmacological effects, such as nephroprotective, hepatoprotective, anti-inflammatory, antioxidant, and anticancer activities. In our study, doxorubicin (DOX) and rhein (RHE) co-loaded polymeric micelle (nano-DOX/RHE) were prepared to attenuate drug resistance in ovarian cancer cells while promoting the therapeutic efficiency of DOX. The morphology, particle size (about 25 nm), zeta potential, release profile in vitro, cell proliferation and cytotoxicity effects were calculated. The results suggested that DOX and RHE could be efficiently loaded into micelle nanoparticles, and in vitro study indicated that they could be released from the nanoparticles in an extended period into DOX-resistant SKOV3 cells (SKOV3/DOX). Nano-DOX/RHE exerted an enhanced cytotoxicity and high apoptosis-inducing activities in SKOV3/DOX cells. Importantly, nano-DOX/RHE exhibited better cancer targeting ability, enhancing the anti-tumor efficacy with little toxicity. In conclusion, nano-DOX/RHE promoted the drug target on tumor site with preferable anti-tumor effects, which could be a promising therapeutic strategy against human ovarian cancer.

MicroRNA-497 accelerates apoptosis while inhibiting proliferation, migration, and invasion through negative regulation of the MAPK/ERK signaling pathway via RAF-1.

The aim of this study is to explore the various modes of action miR-497 has on human cervical cancer (CC) cell behavior. We also speculate that miR-497 achieves its anti-tumor role by governing RAF-1 via MAPK/ERK signaling pathway. CC tissues with corresponding adjacent normal tissues were collected from 168 CC patients. RAF-1-positive cells were identified by means of immunohistochemistry in tissues. A series of inhibitors, mimics and siRNA against RAF-1 were introduced to validate regulatory mechanisms for miR-497 and RAF-1. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assay were employed for evaluating alternations of miR-497, RAF-1, and MAPK/ERK signaling pathway. HeLa cell proliferation, invasion, migration, cycle progression, and apoptosis were assessed by means of CCK-8, wound-healing, transwell invasion assays, and flow cytometry, respectively. The target prediction program and luciferase activity determination were used to identify miR-497 targeting RAF-1. We determined reduced miR-497 expression and elevated expression of RAF-1 in CC tissues as opposed to adjacent tissues. Transfection of miR-497 mimics and siRNA-RAF-1 both decreased levels of MEK1, ERK1, and p38 phosphorylation in HeLa cells, inhibited cell proliferation, migration and invasion, induced more cells arrested in the G0/G1 phase, and promoted cell apoptosis; while miR-497 inhibitors led to opposite results. These findings indicate miR-497 as a tumor suppressor results from negative regulation of the MAPK/ERK signaling pathway via RAF-1 in CC.

Carnosic acid cooperates with tamoxifen to induce apoptosis associated with Caspase-3 activation in breast cancer cells in vitro and in vivo.

Tamoxifen is known as a standard therapeutic treatment for estrogen receptor-positive breast cancer, which down-regulates breast cancer mortality by 31% approximately. Carnosic acid is a phenolic diterpene, which has anti-cancer, anti-inflammation, anti-diabetic and anti-bacterial properties, generated by various species coming from Lamiaceae family. The breast cancer is reported as one of the most common tumors among women worldwide. In our study, the possible benefits of carnosic acid cooperation with tamoxifen for breast cancer treatment in vitro and in vivo were investigated. Carnosic acid and tamoxifen cooperation led to apoptosis in breast cancer cells. Caspase-3 signaling pathway was promoted for carnosic acid and tamoxifen co-treatment. Consistently, anti-apoptotic molecules Bcl-2 and Bcl-xl were down-regulated, while pro-apoptotic signals Bax and Bad were up-regulated. The elevation of decoy receptor 1 and 2 (DcR1 and DcR2) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) were enhanced for carnosic acid and tamoxifen cooperation. Furthermore, the mouse xenograft model in vivo suggested that carnosic acid and tamoxifen combined therapy inhibited breast cancer growth in comparison to the carnosic acid or tamoxifen monotherapy. Our study supplies a novel therapeutic strategy to induce apoptosis for suppressing breast cancer, which was relied on Caspase-3/TRAIL activation.

High expression of G-protein signaling modulator 2 in hepatocellular carcinoma facilitates tumor growth and metastasis by activating the PI3K/AKT signaling pathway.

The aim of this study was to investigate the role of G-protein signaling modulator 2 in the carcinogenesis and progression of hepatocellular carcinoma. We previously showed that G-protein signaling modulator 2 was upregulated in hepatitis B virus-related hepatocellular carcinoma tissues through a hierarchical clustering analysis. With this study, we first assessed the expression pattern of G-protein signaling modulator 2 in hepatocellular carcinoma specimens and adjacent noncancerous tissues; clinical data were analyzed, along survival times, utilizing the Kaplan-Meier method. Moreover, the functions of G-protein signaling modulator 2 were examined using small-interfering RNAs in vitro. The results showed that G-protein signaling modulator 2 was clearly overexpressed in hepatocellular carcinoma tissues and cell lines and that the G-protein signaling modulator 2 expression level was related to tumor size and hepatitis B virus infection. Furthermore, G-protein signaling modulator 2 knockdown studies suggested that G-protein signaling modulator 2 accelerates cell growth, cell cycle, migration, and invasion and inhibits apoptosis, acting as an oncogene in hepatocellular carcinoma. Western blotting indicated that silencing of G-protein signaling modulator 2 in HepG2 and SMMC-7721 cells increased the expression levels of Bax, caspase-3, and E-cadherin, while notably suppressing the cyclin-dependent kinase 4, cyclin-dependent kinase 6, CyclinD1, Snail1, Vimentin, and matrix metallopeptidase 9 expression levels, compared with that in the control groups. In addition, we found that G-protein signaling modulator 2 can affect the expression of key proteins involved in protein kinase B activation. In conclusion, high expression of G-protein signaling modulator 2 was involved in the pathological processes of hepatocellular carcinoma through activation of the phosphatidylinositol 3-kinase/protein kinase B signaling pathway, which may provide an attractive potential diagnostic biomarker and therapeutic target for treatment of hepatocellular carcinoma.

Effect of silencing NEK2 on biological behaviors of HepG2 in human hepatoma cells and MAPK signal pathway.

To investigate the expression level of NEK2 in 40 tissue specimens of primary liver cancer and to search for clues whether the effect of NEK2 depletion plays a role on biological behaviors of HepG2 cells and the relevant molecular mechanism are the objectives of this study. Real-time PCR and immunohistochemistry assessed expression level of NEK2 in specimens of cancerous tissues and carcinoma-adjacent tissues. The NEK2 expression level in HepG2, Huh7, SMMC, and 7402 cells was detected by real-time PCR and western blot to screen experimental cell line. To assess the expression levels of NEK2 mRNA and protein, an effective siRNA transfected into the HepG2 cells was designed. CCK8 and colony-forming assays were performed to verify short-term and long-term proliferative activities, respectively. Capacity of apoptosis and cell cycle changes were assessed by flow cytometry. Ability of transference and invasion was measured by Transwell Chambers. Western blot approach was used to determine the protein expression levels. There was significantly high expression level of NEK2 in cancerous tissues compared to adjacent tissues. The expression of NEK2 was higher in HepG2 cells than other cell lines. Real-time PCR and western blot shown there were obviously down-regulated NEK2 expression in the NEK2-siRNA group compared to control groups. The capacity of amplification and invasion was inhibited distinctly, and FCM revealed the apoptosis rate was increased and G1 phase was arrested in NEK2-siRNA group. Western blot indicated that low expression of NEK2 in HepG2 cells could increase the expression levels of Bax, caspase-3, P21, and TIMP-1, but significantly suppressed the c-myc, c-jun, Bcl-2, cyclinD1, CDK4, MMP2, and MMP9 expression levels and the phosphorylation levels of ERK, JNK, and P38 compared with the control groups. Our findings demonstrated that NEK2 could be a valuable carcinogenic factor and a promising therapeutic target for primary liver cancer; NEK2 may regulate proliferation, apoptosis, and other biological behaviors of HepG2 cells via MAPK signal pathway.

Long noncoding RNAs in hepatitis B virus-related hepatocellular carcinoma.

AIM: To study the expression of long noncoding RNAs (lncRNAs) in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). METHODS: The lncRNA profiles between HBV-related HCC tissues and corresponding normal liver tissues were generated using microarray analysis. Datasets were analyzed using multiple algorithms to depict alterations in gene expression on the basis of gene ontology (GO), pathway analysis, and lncRNA levels. RESULTS: The microarray revealed that 1772 lncRNAs and 2508 mRNAs were differently expressed. The pathway analysis demonstrated that the cell cycle, cytokine-cytokine receptor interaction, chemokine signaling pathway, and phosphoinositide 3-kinase-protein kinase B signaling pathway may play important roles in HCC. Several GO terms, such as cell cycle, DNA replication, immune response, and signal transduction, were enriched in gene lists, suggesting a potential correlation with HBV-related HCC. The upregulated large intergenic noncoding RNA ULK4P2 was physically combined with enhancer of zeste homolog 2. Therefore, the lncRNAs may participate in regulating HBV-related HCC. CONCLUSION: lncRNAs play important roles in HCC, future studies should verify whether large intergenic noncoding ULK4P2 functions by combining with enhancer of zeste homolog 2 in HCC.

Pretreatment with low dose etomidate prevents etomidate-induced rat adrenal insufficiency by regulating oxidative stress-related MAPKs and apoptosis.

Etomidate is frequently used as an anesthetic and sedation agent in the clinic setting. This study determined that a low-dose pre-infusion followed by a continuous dose infusion of etomidate could reduce etomidate-induced adrenal gland insufficiency. Sixty adult male Wistar rats were used, with six rats per group. Based on preliminary experiments, 0.6mg/kg etomidate was selected as the low dose for this study. Oxidative stress and apoptosis-related proteins in the adrenal glands were assayed using Western blot, and serum levels of CORT and 11ß-hydroxylase were detected using ELISA. Pretreatment with a single bolus of low dose etomidate significantly increased the levels of CORT and 11ß-hydroxylase as well as the activities of superoxide dismutase (SOD), catalase (CAT) and glutathioneperoxidase (GPx) in the adrenal glands, but reduced nitric oxide (NO) production when compared to the positive group. Furthermore, Western blot data showed that pretreatment with low dose etomidate increased extracellular signal-regulated kinase1/2 (ERK1/2), CREB and bcl-2 activation, but suppressed the p-p38, c-JunN-terminal kinase (JNK), inducible NO synthase (iNOS), cleaved-caspase3, cleaved-poly-ADP-ribose polymerase (PARP), bax, and AKT activation. The ERK inhibitor PD98059 and the p38MAPK inhibitor SB203580 abolished the protective effect of low dose etomidate pretreatment. These data demonstrated that pretreatment with low dose etomidate attenuated etomidate-induced adrenal insufficiency to rat adrenal glands. Oxidative stress-related MAPKs and apoptosis proteins might be responsible for mediating the etomidate preconditioning effect in rats.

Inhibition of autophagy alleviates the senescent state of rat mesenchymal stem cells during long-term culture.

Following a limited number of cell divisions, mesenchymal stem cells (MSCs) undergo senescence, and these senescent cells maintain metabolic modification and remain viable for long periods. Autophagy, an intracellular bulk degradation process, provides a survival effect for cells under stress. In this study, the effect of autophagy on senescent MSCs was analyzed. Following serial passaging, rat MSCs underwent replicative senescence, characterized by positive staining for senescence-associated ß-galactosidase (SA-ß-gal), and increased expression levels of p16 and p21. During MSC senescence, the levels of autophagic activity were increased, a greater number of autophagic vacuoles were observed in senescent MSCs by transmission electron microscopy, acridine orange staining was elevated and the expression levels of autophagy­related proteins (microtubule­associated protein 1A/1B­light chain 3-II, Atg7 and Atg12) were increased. The role of autophagy in MSC senescence was further investigated through pharmacological inhibition of autophagy with bafilomycin A1 and 3-methyladenine. Inhibition of autophagy by pharmacological means reduced the rate of positive staining for SA-ß-gal and the expression levels of senescence­related proteins. In conclusion, these findings suggest that autophagy is activated during senescence and the autophagic activity may be a requirement for maintaining the senescent state of MSCs.

[Application of carbon nanoparticles in the laparoscopic sentinel lymph node detection in patients with cervical cancer].

OBJECTIVE: To explore the role of laparoscopic sentinel lymph node(SLN) detection with carbon nanoparticles tracer in cervical carcinoma. METHODS: Totally 21 patients with confirmed early cervical cancer were enrolled in this study.Before laparoscopic extended hysterectomy and pelvic lymphadenoetomy(and para-aortic lymphadenoectomy) , they were injected with carbon nanoparticles suspension injection tracer from cervical neck before surgery. The black-staining lymph nodes were cut as SLN under the laparoscope for routine pathological examination. RESULTS: Of these 21 patients, at least one SLN was successfully detected in 20 patients(95.24%) , and a total of 158 SLNs were detected.The conventional pathology results suggested that 5 patients(23.81%) had positive lymph nodes(n=16, including 14 in 4 patients) . The new approach showed a sensitivity of 80.0%(4/5) , accuracy of 100.0%(20/20) , and negative predictive value of 100.0%(16/16) for SLN detection. CONCLUSION: Laparoscopic SLN detection with carbon nanoparticles tracer is a relative safe and sensitive method for in cervical carcinoma.

[Short term clinical outcomes of laparoscopic fertility preserving radical hysterectomy in the management of early stage cervical cancer].

OBJECTIVE: To investigate the feasibility and effectiveness of laparoscopic radical trachelectomy and lymphadenectomy in the treatment of early-stage cervical cancer. METHODS: The clinical data of 6 patients (stage 1a2 to 1b1), who underwent laparoscopic fertility-preserving radical operation for cervical cancer in our department from February 2009 to October 2010, were retrospectively analyzed in terms of operation duration, intra-operative blood loss, postoperative pathology, complications, and pregnancy. RESULTS: Both radical resection of cervical and pelvic lymph node dissection were completed under laparoscopy, and only the cervical and vaginal cuffs were closed from vagina. The operation duration ranged 155-210 min (mean: 185 min) and the intra-operative blood loss was approximately 60-120 ml(mean: 105 ml). The average length of hospital stay was 18 days without complications, postoperative infection, and bleeding. Postoperative pathology showed no lymph node metastasis, and no ligament, blood vessels, vaginal cutting margin, or upper part of cervix was invaded by tumor cells. During the 8-20-month follow-up, 1 patient had become pregnant for 4 months and no case experienced tumor recurrence. CONCLUSION: Laparoscopic fertility-preserving lymphadenectomy and radical trachelectomy is feasible for patients with early-stage cervical cancer who have strong wish to have a child.

[Effectiveness and safety of laparoscopic presacral neurectomy in treating endometriosis-associated pain].

OBJECTIVE: To explore the effectiveness and safety of laparoscopic presacral neurectomy (LPN) in treating endometriosis-associated pain. METHODS: Totally 64 patients with endometriosis were divided into two groups using prospective non-random method. Patients in the control group received only the conventional laparoscopic resection of endometriosis lesions, while patients in the LPN group underwent LPN in addition to the resection of endometriosis lesions. The pre-operative pain scores, intra-operative staging results, surgical duration, intra-surgical blood loss, post-operative pain relief were compared between these two groups. RESULTS: These two groups showed no significant differences in terms of age, body weight, pre-operative pain score, surgery staging, surgical duration, and intra-operative blood loss (all P > 0.05). All patients were followed up for 6 to 18 months (median: 12.8 months). The post-operative pain relief rate was 89.28% (25/ 28) in LPN group and 61.29% (19/31) in the control group (P = 0.030). CONCLUSION: LPN can effectively and safely in treating endometriosis and its associated pain.