In vivo Bidirectional Modulation of Cannabinoid on the Activity of Globus Pallidus in Rats.

Endocannabinoids are bioactive substances which participate in central motor control. The globus pallidus (GP) is a major nucleus in the basal ganglia circuit, which plays an important function in movement regulation. Both cannabinoid receptor type 1 (CB1R) and cannabinoid receptor type 2 (CB2R) are expressed in the GP suggesting GP as a main action area of endocannabinoids. To investigate the direct electrophysiological and behavioral effects of cannabinoids in GP, in vivo single unit extracellular recordings and behavioral tests were performed in rats. Administration of WIN 55,212-2 exerted three neuronal response patterns from all sampled neurons of GP, including (1) increase of the firing rate; (2) decrease of the firing rate; (3) increase and then decrease of the firing rate. Selectively blocking CB1R by AM 251 decreased the firing rate and increased the firing rate. Selectively blocking CB2R by AM 630 did not change the firing rate significantly, which suggested that endocannabinoids modulated the spontaneous firing activity of pallidal neurons mainly via CB1R. Furthermore, co-application of AM 251, but not AM 630, blocked WIN 55,212-2-induced modulation of firing activity of pallidal neurons. Finally, both haloperidol-induced postural behavioral test and elevated body swing test (EBST) showed that unilateral microinjection of WIN 55,212-2 mainly induced contralateral-biased swing and deflection behaviors. Meanwhile, AM 251 produced opposite effect. The present in vivo study revealed that cannabinoids produced complicated electrophysiological and behavioral effects in the GP, which further demonstrated that the GP is a major functional region of endocannabinoid.

Tumor suppressor miR-192-5p targets TRPM7 and inhibits proliferation and invasion in cervical cancer.

Cervical cancer is the fourth highest mortality cancer among women worldwide. Many researchers have discovered the major anticancer role of miR-192-5p. However, no study has revealed the effect of miR-192-5p on cervical cancer and its molecular mechanism. Therefore, in this study, we aimed to explore the role of miR-192-5p in proliferation, invasion of cervical cancer, and its regulatory mechanism. Firstly, the expression level of miR-192-5p was examined by real-time quantitative polymerase chain reaction. Cell counting kit-8 analysis was applied to detect the proliferation of transfected Caski and SiHa cells. Flow cytometry assay was applied to detect the apoptosis of transfected Caski and SiHa cells. Our result showed that miR-192-5p restrained cervical cancer cell proliferation and induced apoptosis. Then we employed wound healing and transwell assays to analyze the migration and invasion abilities of Caski and SiHa cells in vitro. The results showed that miR-192-5p had an inhibitory effect on cervical cancer migration and invasion. The results of in vivo experiment demonstrated that miR-192-5p also inhibited tumor development in nude mice. We further detected that the binding of transient receptor potential melastatin-subfamily member 7 (TRPM7) to miR-192-5p using bioinformatic methods and dual-luciferase reporter assay. Finally, we found that TRPM7 overexpression reversed the inhibitory effects of miR-192-5p on proliferation, migration, and invasion on cervical cancer cells. In conclusion, the findings of the present study revealed that miR-192-5p performs an inhibitory role in cervical cancer proliferation and invasion by targeting TRPM7.

Consistency of postoperative pain assessments between nurses and patients undergoing enhanced recovery after gynaecological surgery.

AIMS AND OBJECTIVES: To explore the consistency of pain intensity and pain location assessed by nurses and patients in gynaecology undergoing enhanced recovery after surgery pathway. BACKGROUND: Several studies have shown that clinical nurses' assessment of patients' pain is not always accurate. Little is known about the accuracy of nurses' pain assessments for gynaecological patients. Postoperative pain assessment and management is an essential part of enhanced recovery after surgery. DESIGN: Comparative cross-sectional study. METHODS: A total of 160 patients were recruited and only 85 patients and 17 nurses participated. Patients and nurses recorded pain scores (using an 11-point Numeric Rating Scale) and pain location (incision pain, surgical area pain in the abdominal cavity, other pain or no pain) on Pain Assessment Forms at 4 hr after surgery and on the first and second postoperative days. We used the STROBE guidelines to report our study. RESULTS: The patients' pain score was higher than that of nurses from 4 hr to second day after laparoscopic surgery at rest. The pain scores of both nurses and patients decreased over this period of time. All the intraclass correlation coefficients were between 0.214-0.296. At the three time points, surgical area pain in the abdominal cavity and abdominal incision pain were the main pain areas. All the kappa coefficients were between 0.164-0.255. CONCLUSIONS: The consistency of postoperative pain assessment about pain score and pain location between nurses and patients was not high. We should attach importance to systematic pain assessment, and more detailed enhanced recovery after surgery pathways should be developed about pain assessment. RELEVANCE TO CLINICAL PRACTICE: Continuing education for nurses regarding pain assessment is necessary. Nurses should accept the patient's self-reported pain. There should be a step that gives more time for pain assessment in enhanced recovery after surgery pathways.

Protopanaxadiol and metformin synergistically inhibit estrogen-mediated proliferation and anti-autophagy effects in endometrial cancer cells.

Metformin is commonly used for treating type II diabetes and has recently been reported to possess anti-proliferative properties that can be exploited for the prevention and treatment of a variety of cancers. Ginsenosides are the main effective biological components of ginseng. It has been reported that ginsenoside-Rb2 inhibit the invasiveness of endometrial cancer cells (ECC). The aim of this study was to investigate whether protopanaxadiol (PPD, a metabolite of ginsenosides) and metformin could synergistically regulate the biological behavior of ECC and analyze its possible mechanism. We here found that either metformin or PPD treatment led to a decreased viability and increased apoptosis and autophagy levels in ECC lines (Ishikawa and RL95-2 cells), and combination of PPD and metformin could enhance these effects induced by metformin or PPD in vitro. PPD and metformin significantly decreased the expression of estrogen receptor alpha (ERα) in Ishikawa and RL95-2 cells. Estrogen promoted the viability and restricted the apoptosis and autophagy of Ishikawa and RL95-2 cells, and PPD and metformin reversed these effects. In vivo trials showed that combination of PPD and metformin had the strongest activity of anti-tumor growth compared with PPD alone and metformin alone. These data suggest that PPD and metformin can be used together to play a more powerful anti-EC effect. Our study provides a scientific basis for the clinical application of PPD and metformin in the treatment of EC, especially in estrogen-dependent patients.

The cross talk between cervical carcinoma cells and vascular endothelial cells mediated by IL-27 restrains angiogenesis.

PROBLEM: To explore whether cervical carcinoma cell-derived interleukin-27 (IL-27) modulates the angiogenesis of vascular endothelial cells. METHOD OF STUDY: The expression of IL-27 in cervical cancer tissues and cervical cell lines was analyzed by immunohistochemistry, ELISA and flow cytometry. Then, the effects of IL-27 on the proliferation and apoptosis-related molecules and angiogenesis in vitro of human umbilical vein endothelial cells (HUVECs) were investigated. Finally, in vivo experiment was performed to further confirm the effects of IL-27. RESULTS: Compared with cervicitis, the cervical cancer tissues highly expressed IL-27. Both HeLa and CaSki cells secreted IL-27, and HUVECs expressed low levels of IL-27 receptors (IL-27R). However, the co-culture of cervical cell lines and HUVECs led to a significant elevation of IL-27R on HUVECs. Co-culturing with IL-27-overexpressed HeLa cells downregulated Ki-67 and Bcl-2 and upregulated Fas expression in HUVECs. In addition, overexpression of IL-27 in HeLa cells and CasKi cells secreted less IL-8 and could further restrict angiogenesis compared with control cells in vitro. In the subcutaneous tumorous model of C57/BL6 mouse, there were decreased vessel density and tumor volume when inoculation with IL-27-overexpressed TC-1 cells. CONCLUSION: This study indicates that IL-27 secreted by cervical carcinoma cells restricts the angiogenesis in a paracrine manner in the pathogenesis of cervical cancer.

Cisplatin inhibits the growth, migration and invasion of cervical cancer cells by down-regulating IL-17E/IL-17RB.

Interleukin (IL)-17E mainly produced by immune cells, is a distinct member of the IL-17 cytokine family, which has multifarious immunomodulatory activities. As a potent anticancer drug, cisplatin is commonly used against various types of solid tumors. The present study was performed to investigate whether cisplatin regulates the expression of IL-17E and it receptor IL-17RB, and the role of IL17E in cervical cancer cells in vitro. The expression of IL-17E and IL-17RB in cervical cancer cells was detected by flow cytometry and ELISA. The viability, apoptosis, migration and invasion of cervical cancer cells were analyzed by CCK8, Annexin V-7AAD apoptosis, transwell migration, wound healing, and matrigel invasion assays. Here, we found that cervical cancer cells co-expressed IL-17E and IL-17RB, especially HeLa and SiHa cells. Recombinant human IL-17E protein (rhIL-17E) enhanced the viability, migration and invasion of HeLa and SiHa cells, and blocking IL-17E with anti-human IL-17RE neutralizing antibody promoted the apoptosis of HeLa and SiHa cells. Cisplatin significantly down-regulated the expression of IL-17E and IL-17RB, and further reversed the regulatory effects of rhIL-17E on viability, apoptosis, migration and invasion of HeLa and SiHa cells. The results suggest that cisplatin inhibits the viability, migration, invasion, and promotes the apoptosis of cervical cancer cells possibly by down-regulating IL-17E/17RB signaling. Cisplatin may be the first choice for cervical cancer patients with abnormally high IL-17E expression.

TSLP promotes angiogenesis of human umbilical vein endothelial cells by strengthening the crosstalk between cervical cancer cells and eosinophils.

Our previous study demonstrated that thymic stromal lymphopoietin (TSLP) secreted by cervical cancer cells promotes angiogenesis and recruitment, and regulates the function of eosinophils (EOS). However, the function of TSLP in the crosstalk between EOS and vascular endothelial cells in cancer lesions remains unknown. The aim of the present study was to investigate the effect of EOS caused by TSLP in in vitro angiogenesis of human umbilical vein endothelial cells (HUVECs). The results of the present study revealed that recombinant human TSLP protein (rhTSLP) increased the secretion of vascular endothelial growth factor (VEGF), but not fibroblast growth factors, in HL-60-eosinophils (HL-60E). Compared with cervical cancer cells (HeLa or CasKi cells) or HL-60E alone, there were increased levels of interleukin (IL)-8 and VEGF in the co-culture system between cervical cancer cells, and HL-60E cells. This effect was strengthened by rhTSLP, but inhibited by inhibiting the TSLP signal with anti-human TSLP or TSLP receptor neutralizing antibodies. The results of the tube formation assays revealed that treatment with the supernatant from cervical cancer cells and/or HL-60E resulted in an increase in angiogenesis in HUVECs, which could be decreased by TSLP or TSLPR inhibitors. The results of the present study suggested that TSLP derived of cervical cancer cells may indirectly stimulate angiogenesis of HUVECs, by upregulating IL-8 and VEGF production, in a co-culture model between cervical cancer cells and EOS, therefore promoting the development of cervical cancer.

RANKL/RANK interaction promotes the growth of cervical cancer cells by strengthening the dialogue between cervical cancer cells and regulation of IL-8 secretion.

Receptor activator for nuclear factor κB ligand (RANKL) is a member of the tumor necrosis factor (TNF) family. The interaction between RANKL and its receptor RANK plays an important role in the development and function of diverse tissues. However, the expression and role of RANKL in cervical cancer are still unknown. In the present study, we found that RANKL and RANK were highly co-expressed in cervical cancer. HeLa and SiHa cells secreted soluble RANKL (sRANKL), expressed member RANKL (mRANKL) and RANK. Recombinant human RANKL protein had no effect on the viability of HeLa and SiHa cells. Yet, blocking RANKL with an anti-human RANKL neutralizing antibody (α-RANKL) or recombinant human osteoprotegrin (OPG) protein resulted in the downregulation of Ki-67 and B-cell lymphoma 2 (Bcl-2) expression and an increase in Fas and Fas ligand (FasL) expression, as well as a high level of viability and a low level of apoptosis in the HeLa and SiHa cells. In addition, α-RANKL led to a decrease in IL-8 secretion. Recombinant human IL-8 protein reversed the effect of α-RANKL on the expression of proliferation- and apoptosis­related molecules, and proliferation and apoptosis in the HeLa and SiHa cells. The present study suggests that a high level of mRANKL/RANK expression in cervical cancer lesions plays an important role in the rapid growth of cervical cancer cells possibly through strengthening the dialogue between cervical cancer cells and regulation of IL-8 secretion, which may be a possible target for cervical cancer therapy.

Enhancement of anammox performance by Cu(II), Ni(II) and Fe(III) supplementation.

This study explored the influence of metal ion addition on specific anaerobic ammonium oxidation activity (SAA). Batch assays were used to demonstrate the enhancement of the SAA upon the addition of Cu2+, Ni2+ and Fe3+. The SAA was enhanced by 41.0% when the Cu2+ concentration was below 1 mg L−1, while it was improved by 63.5% at Ni2+ concentrations below 1.74 mg L−1. An enhancement of 533.2% was obtained when 3.68 mg L−1 Fe3+ was supplied. The effects of Fe3+, Cu2+ and Ni2+ on the SAA were analyzed and optimized by a response surface methodology, which demonstrated that the interaction between Fe3+ and Cu2+ was significant and that 6.61 mg Fe3+ L−1, 1.18 mg Cu2+ L−1 and 1.11 mg Ni2+ L−1 were the optimal values for metal dosing. Subsequently, an Fe3+­Cu2+­Ni2+ continuous test was carried out under optimal conditions and revealed that the addition of Fe3+, Cu2+ and Ni2+ could stimulate the reactor potential at ambient temperature. The maximum nitrogen removal rate (NRR) of the test reactor was 52.8% higher than that of the control reactor (8.1 versus 5.3 kg N m−3 d−1). Moreover, a continuous test conducted by adding Fe3+ achieved an average nitrogen removal efficiency and maximum NRR of 67.4% and 4.9 kg N m−3 d−1, respectively, while the corresponding values of the control test were 64.7% and 4.1 kg N m−3 d−1, respectively. Altogether, appropriate dosages of Cu2+, Ni2+ and Fe3+ can significantly enhance the SAA and improve the reactor capacity at ambient temperature.

Hypoxia promotes the proliferation of cervical carcinoma cells through stimulating the secretion of IL-8.

To explore whether hypoxia and interleukin 8 (IL-8) regulate the viability and apoptosis of cervical carcinomas cells and the possible mechanism. We evaluated the expression of hypoxia inducible factor-1α (HIF-1α), IL-8 and its receptors (CXCR1 and CXCR2) in cervical cancer and cervicitis tissues by immunohistochemistry. Then the effects of hypoxia and IL-8 on the viability and apoptosis of HeLa and SiHa cells were detected by the SRB and apoptosis assays. Here we observed that the expression of HIF-1α, IL-8 and CXCR1 in cervical cancer tissues was significantly higher than that in cervicitis tissues. Hypoxic condition stimulated the secretion of IL-8 and the expression of CXCR1 and CXCR2 on HeLa and SiHa cells. Recombinant human IL-8 enhanced the viability and reduced the apoptosis in HeLa and SiHa cells. HeLa and SiHa cells cultured in 1% oxygen showed the increased viability and apoptosis, and the former effect could be partly reversed by anti-human IL-8 neutralizing antibody. This data suggested that IL-8 secreted by cervical carcinomas cells induced by hypoxia can stimulate the viability of cervical carcinomas cells in an autocrine dependent manner, and contribute to the pathogenesis of cervical cancer.

Transient and long-term effects of bicarbonate on the ANAMMOX process.

In this study, the effects of both transient and long-term inorganic carbon (IC) addition on the anaerobic ammonium oxidation (ANAMMOX) process under pseudo-steady-state and substrate inhibitions were analyzed using reactor performance and measures of sludge activity. Compared with the nitrogen removal rate (NRR) of 3.42 kg N m(-3) day(-1) in the control bioreactor (ICDR) without IC, the peak NRR reached 21.0 kg N m(-3) day(-1) in the reactor (ICAR) with sufficient IC added. It was revealed that the long-term addition of bicarbonate significantly enhanced the performance of the ANAMMOX reactor. The optimum HCO3 (-)/TN ratio was considered to be 1.20, which is lower than that in normal conditions. The IC concentration affected biomass activity, and the transient addition or removal of IC to differing sludge media caused a significant loss of activity. Sufficient addition of IC alleviated the inhibition of excess substrates, while the inhibition was aggravated by the IC limitation. The half-maximal (50 %) inhibitory concentrations of substrate for the sludge were 295 mg L(-1) NO2 (-)-N and 361 mg L(-1) NH4 (+)-N with 120 mg L(-1) of fixed HCO3 (-) and 346 mg L(-1) NO2 (-)-N and 456 mg L(-1) NH4 (+)-N with unlimited IC, respectively. Changing the HCO3 (-)/TN (in milligrams per milligram) ratio resulted in the variation of ANAMMOX stoichiometric ratios. Sludge characterization parameters in the ICDR, including biomass, extracellular polymeric substances, heme C, and so on, were lower than those in ICAR. Filamentous bacteria and spherical bacteria were also observed in the reactor with limited IC.

Enhancement of ANAMMOX activity by low-intensity ultrasound irradiation at ambient temperature.

This paper aims to investigate the enhancement effect of low intensity intermittent ultrasound irradiation on the efficiency of anaerobic ammonium oxidation (ANAMMOX) process at ambient temperature. With intermittently irradiated (ultrasound intensity of 0.19 w/cm(2), exposure time of 0.2 min), the reactor (RU) had a nitrogen removal rate (NRR) of 5.49 kgTN/m(3)/d at 14.8°C, while the NRR was 1.53 kgTN/m(3)/d in the control reactor (RC). At the end of operation, the contents of polysaccharide, protein, TTC-dehydrogenase and VSS were 6.82 mg/mgVSS, 26.79 mg/mgVSS, 0.58 mgTF/L/H and 10.11 gVSS/L in RU, higher than the levels in the RC. These results demonstrated that it is possible to achieve stable and highly efficient operation in an ANAMMOX reactor at low ambient temperature by implementation of ultrasonication.

The effect of sulfide inhibition on the ANAMMOX process.

The feasibility of anaerobic ammonium oxidation (ANAMMOX) process to treat wastewaters containing sulfide was studied in this work. Serum bottles were used as experimental containers in batch tests to analyze the short-term response of the ANAMMOX process under sulfide stress. The IC(50) of sulfide-S for ANAMMOX biomass was substrates-dependent and was calculated to be 264 mg L(-1) at an initial total nitrogen level of 200 mg L(-1) (molar ratio of ammonium and nitrite was 1:1). The long-term effects and the performance recovery under sulfide stress were continuously monitored and evaluated in an upflow anaerobic sludge blanket reactor. The performance of the ANAMMOX system was halved at an sulfide-S level of 32 mg L(-1) within 13 days; however, the nitrogen removal rate (NRR) decreased by only 17.2% within 18 days at an sulfide-S concentration of 40 mg L(-1) after long-time acclimatization of sludge in the presence of sulfide. The ANAMMOX performance recovered under sulfide-S level of 8 mg L(-1) with a steady NRR increasing speed, linear relationship between the NRR and operation time. The synchronic reduce in the specific ANAMMOX activity and the biomass extended the apparent doubling time of the nitrogen removal capacity and decreased biomass growth rate.

Analysis of diferentially expressed protein from primary and recurrent ovarian cancer serum.

OBJECTIVE: To study the value of the differentially expressed proteins from primary and recurrent ovarian cancer serum for early diagnosis of primary and recurrent ovarian cancer. METHODS: WCX kit (Bruker Daltonics GraBH) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technology were used to detect serum samples from 49 patients with primary ovarian cancer and 21 patients with recurrent disease. RESULTS: In the mass range (Mr) from 1,000 to 12,000 Da, eight differentially expressed protein peaks were screened from primary ovarian cancer serum. Among them, four protein peaks with Mr 1,457, 1,857, 2,202, 7,761 were lowly expressed and the others with Mr 2,946, 5,333, 5,859, 5,901 were highly expressed. Ten diferentially expressed protein peaks were screened from recurrent ovarian cancer serum. Among them, 1,944, 1,980, 2,080, 2,661, 2,993, 4,450, 4,659, 5,359 Da protein expressions were increased significantly, and 1897, 7868 Da protein expressions were decreased significantly. The pattern of primary ovarian cancer was applied to 8 early-stage ovarian cancer serum samples, and 7 serum samples were successfully predicted with the accuracy of 87.5%. The pattern of recurrent ovarian cancer was applied to 9 without pelvic or abdominal mass recurrent ovarian cancer serum samples, and 8 serum samples were successfully predicted with the accuracy of 88.9%. CONCLUSION: Combination of MALDI-TOF-MS and WCX kit technology can directly screen the diferrential expressed protein from primary and recurrent ovarian cancer serum. They have clinical significance for enhancement of sensitivity and specificity of ovarian cancer diagnosis.

The ANAMMOX reactor under transient-state conditions: process stability with fluctuations of the nitrogen concentration, inflow rate, pH and sodium chloride addition.

The process stability of an anaerobic ammonium oxidation (ANAMMOX) was investigated in an upflow anaerobic sludge blanket reactor subjected to overloads of 2.0- to 3.0-fold increases in substrate concentrations, inflow rates lasting 12 or 24h, extreme pH levels of 4 and 10 for 12h and a 12-h 30 g l(-1) NaCl addition. During the overloads, the nitrogen removal rate improved, and the shock period was an important factor affecting the reactor performance. In the high pH condition, the reactor performance significantly degenerated; while in the low pH condition, it did not happen. The NaCl addition caused the most serious deterioration in the reactor, which took 108 h to recover and was accompanied by a stoichiometric ratio divergence. There are well correlations between the total nitrogen and the electrical conductivity which is considered to be a convenient signal for controlling and monitoring the ANAMMOX process under transient-state conditions.

Performance and robustness of an ANAMMOX anaerobic baffled reactor subjected to transient shock loads.

The impacts of transient overloads on the performance of a laboratory-scale anaerobic ammonium oxidation (ANAMMOX) anaerobic baffled reactor was studied by increasing the substrate concentration or inflow rate to 1.5-3.0 times above normal values. These shocks, with the exception of the highest substrate shock, weakened the nitrogen removal efficiency (NRE) but improved the nitrogen removal rate by 0.01-0.18 g l(-1) h(-1). The communities and the location of the sludge may be altered by distinct types of shocks. The substrate vibration data showed that the reactor was unresponsive to hydraulic shocks but sensitive to substrate shocks and the former compartments were more susceptible to the shocks. In the inhibition period, the pH and NRE of the reactor were related to the residual ammonium and free ammonia (FA) and FA was a factor in the reactor fluctuations. The Gaussian model proposed to describe the shocks response fits the experimental data well.

Impacts of transient salinity shock loads on Anammox process performance.

The effect of salinity shock (5-60 g l(-1) NaCl) on anaerobic ammonium oxidation (Anammox) process performance was investigated. The response to the shock loads can be divided into three stages: a sensitive period, an interim stable period and a recovery period which lasted 6-26 d. When exposed to NaCl shocks for 12h, the sludge retention time (SRT) of the reactor decreased with increasing NaCl shock loads, ranging between 2.9 and 22.5d, meanwhile the biomass decreased by 0.8-37.4%. When shock loads were higher than 10 g l(-1) NaCl, the reactor was at risk of losing too much biomass. The granular surface was rough due to rapid growth of filamentous bacteria and extracellular polymeric substances (EPS), also the EPS amount changed during all NaCl shock loads. In the latter of shocks, the microorganisms in the reactor showed a little adaption to the NaCl shock.