[Quality assessment of global colorectal cancer screening guidelines and consensus].

Objective: To systematically evaluate the methodology and reporting quality of colorectal cancer (CRC) screening guidelines/consensus and provide lights for drafting CRC screening guidelines in China. Methods: The literature retrieval for all the Chinese and English guidelines published before September 1st, 2020 was conducted by using Chinese/English databases, such as China National Knowledge Infrastructure, Wanfang Data, VIP, SinoMed, PubMed, Embase, Web of Science, Cochrane Library, Guideline International Network, and supplement with the official website of multiple regions, such as the U.S. Preventive Services Task Force and American Cancer Society. We utilized The Appraisal of Guidelines for Research & Evaluation Ⅱ (AGREE Ⅱ) and Reporting Items for Practice Guidelines in Healthcare (RIGHT) approaches to assess the quality of CRC screening guidelines/consensus comprehensively. Results: After quality control, a total of 19 guidelines/consensus released by the United States, China, Australia, Canada, Britain, South Korea, and International organizations are successfully included, and strikingly, most of those belong to the United State(7). The results of the AGREE Ⅱ quality evaluation show that the average scores of scope and purpose (87.5%) and clarity of presentation (89.6%) are high. In contrast, there are deficient in stakeholder involvement (47.0%), the rigor of development (42.3%), applicability (47.5%), and editorial independence (50.2%). Among all the guidelines, there are 12 with an overall score of 50 or more, 13 with a recommendation level of "A", 2 with a rating of "B" and 4 with a rating of "C". Additionally, the RIGHT evaluation revealed that the average report rate in each field is necessary information (76.3%), background (77.0%), evidence (55.8%), recommendations (59.4%), review and quality assurance (26.3%), funding and declaration and management of interests (43.4%), other information (49.1%). Among all the guidelines, six have good reporting quality, whereas the additional 13 have general or weak evidence. Furthermore, subgroup analysis indicates that the quality of guidelines in developed countries is superior to that of China. Conclusion: The number of CRC screening guidelines/consensus is increasing gradually, and the overall quality of those is high, but the normative nature is warranted to be strengthened.

Long noncoding RNA DDX11-AS1 induced by YY1 accelerates colorectal cancer progression through targeting miR-873/CLDN7 axis.

OBJECTIVE: Increasing studies have confirmed long non-coding RNAs (lncRNAs) as novel regulators in tumorigenesis. LncRNA DDX11 antisense RNA 1 (DDX11-AS1) has been found to be abnormally expressed in several tumors. In this work, we aimed to evaluate its expressions and functions in colorectal cancer (CRC). PATIENTS AND METHODS: The Cancer Genome Atlas (TCGA) datasets were used for the identification of dysregulated lncRNA in CRC. The levels of DDX11-AS1 were determined in tumor tissues and cell lines by Real Time-Polymerase Chain Reaction (RT-PCR). The clinical significance of DDX11-AS1 in CRC patients was analyzed using Chi-square test and Kaplan-Meier analysis. Functional assays for the exploration of DDX11-AS1 and miR-873 were performed using a series of cells experiment. ChIP assay and luciferase reporter assays were used to explore the mechanism of actions of DDX11-AS1 in CRC cells. RESULTS: We identified DDX11-AS1 as a new CRC-related lncRNA whose levels were distinctly up-regulated in CRC specimens and cell lines, partly induced by YY1. Clinical explorations suggested that increased expressions of DDX11-AS1 in CRC were positively associated with lymph nodes metastasis and TNM stage and had a distinct influence on the overall survival. Further multivariate assays indicated that DDX11-AS1 was an independent prognostic parameter implying a poorer clinical outcome for patients with CRC. Functional assays revealed that the knockdown of DDX11-AS1 suppressed the proliferation, migration, and invasion of CRC cells, and stimulate apoptosis. Mechanistic studies showed that the up-regulation of DDX11-AS1 competitively bound to miR-873 prevented CLDN7 from miRNAs-mediated degradations, thus facilitated the CRC progress. Further rescue assays were carried out to achieve confirmation. CONCLUSIONS: Our present findings may enhance our understanding of the pathogenesis of CRC and revealed DDX11-AS11 as a potential therapeutic target for CRC.

Efficacy and safety of LCI699 for hypertension: a meta-analysis of randomized controlled trials and systematic review.

OBJECTIVE: This study reviews the available data from randomized controlled trials on efficacy and safety of LCI699, a novel inhibitor of aldosterone synthase, as treatment of hypertension. MATERIALS AND METHODS: We performed a meta-analysis of phase II randomized, controlled trials comparing the efficacy/safety of LCI699 with placebo in hypertension patients. For this purpose, PubMed, Embase, Cochrane Library database, ISI-Science Citation Index, and the Chinese Biomedicine Literature Database were searched until August 2013. The available data on mean sitting systolic blood pressure (MSSBP), mean sitting diastolic blood pressure (MSDBP), adverse effects, renin-angiotensin-aldosterone system biomarkers (RAASB) and adrenocorticotropic hormone-stimulated cortisol concentration (AHSC) were collected. All data were analyzed using Review Manager, version 5.2. RESULTS: The present study finally included three randomized controlled trials, comprising of 623 patients in total. The daily use of ≥ 1 mg LCI699 was associated with a significant reduction of MSSBP (Weighted mean difference/WMD = -8.80, 95% CI: -11.31 to -5.68, p < 0.00001, I2 = 0%) and MSDBP (WMD = -4.94, 95% CI: -7.49 to -2.40, p = 0.00001, I2 = 9%). Adverse reactions occurred in 73 of the 139 patients (52.51%) treated with LCI699 and in 34 of the 63 patients (53.96%) treated with placebo. Pooled meta-analysis showed that the use of LCI699 was associated with no increased risk of side effects compared with placebo (RR = 0.90; 95% CI: 0.68 to 1.18, p = 0.43, I2 = 0%). Suppression of plasma aldosterone was measured at all doses of LCI699 treatment groups. LCI699 suppressed the ACTH-stimulated cortisol response in a dose- and time-dependent manner. CONCLUSIONS: Current evidence indicates that the novel aldosterone inhibitor LCI699 is an effective and well-tolerated antihypertensive agent that lowers plasma aldosterone concentration and produces a mild ACTH-stimulated cortisol response suppressive effect.

Stimulation of the inferior olivary complex alters the distribution of the type 1 corticotropin releasing factor receptor in the adult rat cerebellar cortex.

In a previous study, it was shown that populations of climbing fibers, derived from the inferior olivary complex (IOC) contain the peptide corticotropin releasing factor (CRF) and that the expression of this peptide in climbing fibers could be modulated by the level of activity in olivary afferents. The intent of this study was to determine if there was comparable plasticity in the distribution of the type 1 CRF receptor (CRF-R1) in the cerebellum of the rat. Our results indicate that CRF-R1 was localized primarily to Purkinje cell somata and their primary dendrites and granule cells. In addition, scattered immunolabeling was present over the somata of Golgi cells, basket cells and stellate cells, as well as Bergmann glial cells and their processes. IOC stimulation for 30 min at 1 Hz increased CRF-R1 expression in molecular layer interneurons and processes of Bergmann glial cells. Little to no effect on CRF receptor distribution was observed in Purkinje cells, granule cells, or Golgi cells. IOC stimulation at 5 Hz however, increased CRF-R1 expression in the processes of Bergmann glial cells while decreasing its expression in basket, stellate and, to some extent, in Purkinje cells. The present results suggest that there is activity-dependent plasticity in CRF-R1 expression that must be considered in defining the mechanism by which the CRF family of peptides modulates activity in cerebellar circuits. The present results also suggest that the primary targets of CRF released from climbing fibers are Bergmann glial cells and interneurons in the molecular layer. Further, interneurons responded with a decrease in receptor expression following more intense levels of stimulation suggesting the possibility of internalization of the receptor. In contrast, Bergmann glial cells showed an increased expression in receptor expression. These data suggest that CRF released from climbing fibers may modulate the physiological properties of basket and stellate cells as well as having a heretofore unidentified and potentially unique effect on Bergmann glia.

Presynaptic localization of a truncated isoform of the type 2 corticotropin releasing factor receptor in the cerebellum.

It is now well established that corticotropin releasing factor is present in two major excitatory afferent systems to the cerebellum, namely climbing fibers and mossy fibers. Two major classes of corticotropin releasing factor receptors, each with unique binding characteristics, have been identified as type 1 and type 2. In this study we used an antibody made to the n-terminus of the type 2 corticotropin releasing factor receptor. Characterization of this antibody showed that it strongly labeled a protein with a molecular weight of 16-32 kDa and only faintly labels a 62-83 kDa protein. The lower molecular weight protein corresponds to the weight of a recently described truncated isoform of this receptor that is designated corticotropin releasing factor-type 2alpha-truncated isoform. We carried out transfection paradigms using corticotropin releasing factor-type 2alpha-truncated isoform constructs and confirmed that the antibody recognized the truncated isoform of the type 2 corticotropin releasing factor receptor. Further, light and electron microscopic studies were carried out in mice and rats to define the distribution of the truncated receptor. Immunoreactivity is evident in the basal region of many, but not all Purkinje cell bodies and their initial axonal segments, as well as the initial axonal segments of isolated Golgi cells, and cerebellar nuclear neurons. In addition, punctate elements in the molecular layer were immunolabeled. The localization of the receptor to the initial segment of Purkinje cells was confirmed with electron microscopy. Further, the punctate labeling in the molecular layer was localized to parallel fibers and their terminals. In conclusion, evidence has been presented to show that distinct isoforms of the corticotropin releasing factor receptor are present in the cerebellum. The complex interactions between corticotropin releasing factor and other members of the corticotropin releasing factor family of peptides with both pre- and postsynaptic receptors support a growing concept that corticotropin releasing factor plays an important role in modulating activity in cerebellar circuits and ultimately in controlling motor behavior.

Evidence for an axonal localization of the type 2 corticotropin-releasing factor receptor during postnatal development of the mouse cerebellum.

Previous studies have described the embryonic and postnatal development of CRF, as well as the type 1 CRF receptor in the mouse cerebellum. The present immunohistochemical study localizes the cellular distribution of the type 2 CRF receptor (CRF-R2) during postnatal development of the mouse cerebellum. Western blot analysis indicates that the antibody used in this analysis recognizes both a full-length and a truncated isoform of the type 2 receptor. We propose that each isoform has a unique cellular distribution. In the present study, the postnatal (P) development (P0-P14) and cellular localization of CRF-R2 in different cell types was analyzed using PAP and double-label fluorescent immunohistochemistry; cell-specific antibodies were used to identify cells expressing CRF-R2 at different stages of postnatal development. At P0, CRF-R2 immunoreactivity was localized within the somata of Purkinje cells and migrating GABAergic interneurons. CRF-R2 was first observed in the initial axonal segments of some Purkinje cells at P5, and was evident in many Purkinje cell axon hillocks at P8. Punctate immunoreactivity is present in the molecular layer by P5 and is interpreted to be immunolabeled parallel fibers. Between P8 and P14, CRF-R2 immunostaining is present in the initial axonal segments of Golgi cells, within the internal granule cell layer. Finally, CRF-R2 is present in both radial glia in the molecular layer as well as in astrocytes in the white matter and internal granule cell layer from P5 to P14. The present results suggest that CRF-R2, both the truncated and the full-length isoforms, are present in the developing cerebellum, each with a unique cellular distribution. The immunohistochemical evidence indicates that the truncated isoform of the type 2 CRF receptor is in the axons of several different types of cerebellar cortical neurons, and suggests that CRF could play a role in cerebellar development by modulating the release of transmitters from excitatory and/or inhibitory interneurons, which in turn could directly alter the maturation of cerebellar circuits. In contrast, the binding of a ligand to the full-length isoform of CRF-R2 or to CRF-R1, both in a postsynaptic location, may have a more direct effect on regulating the responsiveness of these cells to growth factors or neurotransmitters released from afferent axons by regulating permeability of ion channels or altering second messenger systems.

Frequency-dependent expression of corticotropin releasing factor in the rat's cerebellum.

Corticotropin releasing factor (CRF), localized in extrinsic afferents in the mammalian cerebellum, is defined as a neuromodulator within cerebellar circuits, and appears to be an essential element in the generation of long term depression, a proposed mechanism for motor learning. These physiological studies are based on exogenous application of CRF and do not address potential mechanisms that may influence endogenous release of the peptide. In the present study, immunohistochemistry was used to analyze changes in the lobular distribution of CRF-like immunoreactivity (LIR). In addition radioimmunoassay (RIA) was used to quantify changes in levels of the peptide in the cerebellum following stimulation of the inferior cerebellar peduncle (ICP) at 10 or 40 Hz or the inferior olivary nucleus (ION) at 1, 5, 10, or 20 Hz. Results indicate that there is a greater distribution of CRF-like-immunolabeled climbing fibers, mossy fibers, and astrocytes in all lobules of the cerebellum that is directly related to stimulation frequency. Maximal effects were elicited with 40 Hz ICP and 5-10 Hz ION stimulation. Quantitatively, the RIA studies indicate that there is a significant increase in CRF levels in the vermis, hemispheres and flocculus that correlates closely with stimulation frequency. In conclusion, stimulation of cerebellar afferents induces a significant change in the distribution and levels of CRF-LIR in climbing fibers, mossy fibers and glial cells. This suggests that the modulatory effects ascribed to CRF may influence a greater number of target neurons when levels of activity in afferent systems is increased.

Suppression of morphine withdrawal by electroacupuncture in rats: dynorphin and kappa-opioid receptor implicated.

Our previous work has demonstrated that 100-Hz electroacupuncture (EA) or 100-Hz transcutaneous electrical nerve stimulation (TENS) was very effective in ameliorating the morphine withdrawal syndrome in rats and humans. The mechanism was obscure. (1) Rats were made dependent on morphine by repeated morphine injections (5-140 mg/kg, s.c., twice a day) for eight days. They were then given 100-Hz EA for 30 min 24 h after the last injection of morphine. A marked increase in tail flick latency (TFL) was observed. This effect of 100-Hz EA could be blocked by naloxone (NX) at 20 mg/kg, but not at 1 mg/kg, suggesting that 100-Hz EA-induced analgesia observed in morphine-dependent rats is mediated by kappa-opioid receptors. (2) A significant decrease of the concentration of dynorphin A (1-17) immunoreactivity (-ir) was observed in the spinal perfusate in morphine-dependent rats, that could be brought back to normal level by 100-Hz EA. (3) 100-Hz EA was very effective in suppressing NX-precipitated morphine withdrawal syndrome. This effect of EA could be prevented by intrathecal administration of nor-BNI (2.5 micrograms/20 microliters), a kappa-opioid receptor antagonist, or dynorphin A (1-13) antibodies (25 micrograms/20 microliters) administered 10 min prior to EA. In conclusion, while the steady-state spinal dynorphin release is low in morphine-dependent rats, it can be activated by 100-Hz EA stimulation, which may be responsible for eliciting an analgesic effect and ameliorating morphine withdrawal syndrome, most probably via interacting with kappa-opioid receptor at spinal level.

[Frequency dependence of somatostatin and calcitonin gene related peptide release induced by electroacupuncture in rat spinal cord].

Radioimmunoassay (RIA) was used to determine the changes of the immunoreactivity (ir) of somatostatin (SOM) and calcitonin gene-related peptide (CGRP) in the spinal perfusate of rat induced by electroacupuncture (EA) of different frequencies. The frequency of EA was chosen to be 2, 15 and 100 Hz and the spinal perfusate was collected in three periods of 30 min before, during and after EA. The results indicate: (1) low frequency EA (2 Hz) increased the spinal SOM-ir level by 39% (P < 0.05) but decreased that of CGRP-ir by 47% (P < 0.05); (2) conversely, 15 Hz EA decreased spinal fluid SOM-ir level by 37% (P < 0.05) but increased that of CGRP-ir by 92% (P < 0.05); (3) 100 Hz EA behaved like 15 Hz in inhibiting SOM-ir level (P < 0.01), but without effect on CGRP-ir. The above results show that specific frequency is required for peripheral electrical stimulation to activate or suppress the release of spinal neuropeptides SOM and CGRP, a fact that may have clinical implications.

[Frequency dependence of substance P release by electroacupuncture in rat spinal cord].

Previous studies in our laboratory have shown that electroacupuncture (EA) using different frequencies produced differential opioid peptides release in the spinal cord of rats and human beings. In the present study we observed the frequency dependence of substance P (SP) release from rat spinal cord, with the frequencies of EA set at 2, 4, 8, 15, 30 and 100 Hz. The spinal perfusate was collected in 3 periods of 30 min before, during and after EA, and the immunoreactive SP (SP-ir) was measured by radioimmunoassay (RIA). The effectiveness of EA-induced analgesia was assessed by tail flick latency (TFL). Rats showing an increase of TFL over 40% was considered as EA responder. The results showed that in the responders, SP-ir in spinal perfusate showed a moderate decrease during 2 Hz EA, (P < 0.01 compared with baseline level), no change in the 4 Hz EA group, and a marked increase during 8, 15, 30 and 100 Hz EA (P < 0.01), with maximal increase occurring at 15 Hz (P < 0.001). The above results suggest that EA may induce upward or downward modulation in SP-ir release depending on the frequency of EA. However in the non-responder rats no change in spinal fluid SP-ir content was observed. This suggests that changes in SP-ir release have same causal relation with the analgesia induced by EA stimulation.