A Neural Integrity Monitor Electromyography Endotracheal Tube Causes More Severe Postoperative Sore Throat Than a Standard Endotracheal Tube in Adults: A Prospective Cohort Study.

BACKGROUND: This study aimed at figuring out the different effects of a neural integrity monitor electromyography endotracheal tube (NIM-EMG-ETT) and a standard endotracheal tube (ETT) on postoperative sore throat (POST). METHODS: This prospective cohort study enrolled 143 patients scheduled to undergo general anesthesia with endotracheal intubation. Patients were allocated into three groups: Group A, non-thyroid surgery with a standard ETT; Group B, thyroid surgery with a standard ETT; Group C, thyroid surgery with a NIM-EMG-ETT. The incidence, the severity and visual analog scale (VAS) of POST were recorded. The incidence and the severity of POST were tested by χ2 test or Fisher's exact test. And VAS of POST was tested by Kruskal-Wallis test. RESULTS: The incidences of POST in Group B and Group C were significantly higher than that of Group A at all the time points after extubation (P < 0.001). The incidences of POST in Group C was significantly higher than that in Group B at 8 h, 24 h and 48 h after extubation (89.4% vs. 68.8%, P = 0.014, relative risk (RR) 1.30, 95% confidence interval (CI) 1.05-1.61; 89.4% vs. 58.3%, P = 0.001, RR 1.53, 95% CI 1.18-1.98; 76.6% vs. 45.8%, P = 0.002, RR 1.67, 95% CI 1.18-2.36). Moreover, there was a significant higher VAS of POST and more serious POST with Group C than with Group B. CONCLUSIONS: A NIM-EMG-ETT may induce higher incidence of POST and more serious POST than a standard ETT. TRAIL REGISTRATION: Chinese Clinical Trail Registry ( http://www.chictr.org.cn/index.aspx , ChiCTR2200058896, 2022-4-18).

[Retracted] MicroRNA­136 inhibits prostate cancer cell proliferation and invasion by directly targeting mitogen­activated protein kinase kinase 4.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that certain of the cell migration and invasion assay data shown in Fig. 5C were strikingly similar to data appearing in different form in other articles by different authors at different research institutes, some of which have been retracted. Owing to the fact that the contentious data in the above article were already under consideration for publication, or had already been published, prior to its submission to Molecular Medicine Reports, the Editor has decided that this paper should be retracted from the Journal. After having been in contact with the authors, they agreed with the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 17: 4803­4810, 2018; DOI: 10.3892/mmr.2018.8417].

LncRNA PEG11as silencing sponges miR-874-3p to alleviate cerebral ischemia stroke via regulating autophagy in vivo and in vitro.

Long non-coding RNAs (lncRNAs) are reportedly involved in the regulation of physiological and pathophysiological processes. However, the potential role of lncRNAs in stroke remains largely undefined. Here, RNA-Seq analysis of lncRNAs found that the lncRNA PEG11as (PEG11as) levels were significantly increased in ischemic brain tissue in a transient middle cerebral artery occlusion/reperfusion (tMCAO/R) mouse model of stroke. To explore the role of PEG11as in stroke, the lentivirus containing PEG11as silencing construct(siRNA-PEG11as) was microinjected intracerebroventricularly into male or transfected to N2a cells and then exposed to tMCAO/R or oxygen-glucose deprivation/reoxygenation (OGD/R). Knockdown of PEG11as expression significantly reduced infarct volume, alleviated neuronal deficits and inhibited neuronal apoptosis in tMCAO/R mice. Mechanistically, as an endogenous microRNA-874-3p (miR-874-3p) sponge, PEG11as silencing inhibited miR-874-3p activity, resulting in downregulation of ATG16L1 expression and subsequent inhibition of neuronal apoptosis by regulating autophagy. Overall, the results of this current study indicate that PEG11as is involved in the pathophysiology of cerebral ischemia, thus providing translational evidence that PEG11as can be envisioned as a novel biomarker or/and therapeutic target for stroke.

The FOXP4-AS1/miR-3130-3p/SP4 feedback loop is associated with prostate cancer.

The vital function of mounting long noncoding RNAs (lncRNAs) in prostate cancer (PCa) has been illustrated in increasing reports. However, the roles of many lncRNAs in PCa have not been deciphered. A total of 62 pairs of PCa and adjacent normal tissue samples were provided by PCa patients undergoing surgery. Extensive assays were conducted in this study to investigate the role of FOXP4 antisense RNA 1 (FOXP4-AS1) in PCa tumorigenesis. This study elucidated that FOXP4-AS1 expression was elevated in PCa tissue samples and cell lines. Loss-of-function experiments revealed that depleted FOXP4-AS1 inhibited PCa cell proliferation in vitro and retarded tumor growth in vivo. Mechanically, FOXP4-AS1 functioned as a competing endogenous RNA (ceRNA) of miR-3130-3p, releasing SP4 from the inhibitory effect of miR-3130-3p. Rescue assays validated that FOXP4-AS1 modulated PCa progression via SP4. Interestingly, SP4 is known as a transcription factor and was predicted to bind with the promoter region of FOXP4-AS1. This current research confirmed that SP4 activated the transcription activity of FOXP4-AS1 and thus positively regulated its expression. To conclude, we discovered that FOXP4-AS1, miR-3130-3p, and SP4 constitute a feedback loop and contribute to PCa tumorigenesis, providing a new valuable diagnosis and therapeutic strategy for PCa.

Bacterial networks mediate pentachlorophenol dechlorination across land-use types with citrate addition.

Soil microorganisms play a crucial role in the bioremediation of pentachlorophenol (PCP)-contaminated soils. However, whether and how soil bacterial networks with keystone taxa affect PCP dechlorination is not well understood. The present study investigated the effects of citrate on soil bacterial networks mediating PCP dechlorination by direct and indirect transformation in iron-rich upland and paddy soils. The rates of PCP dechlorination and Fe(II) generation were accelerated by citrate addition, particularly in the paddy soils. Network analysis revealed that the topological properties of bacterial networks were changed by citrate addition; more modules and keystone taxa were significantly correlated with PCP dechlorination and Fe(II) generation in the networks. Random forest modeling indicated that Clostridiales was the most important bacterial order; it was significantly involved in both the direct and indirect pathways of PCP dechlorination. Citrate addition had less influence on the balance between the direct and indirect pathways of PCP dechlorination in the upland soils, whereas it enhanced biological PCP dechlorination more directly and efficiently in the paddy soils. Our results suggested that land-use type and citrate addition play a critical role in controlling the biogeochemical mechanisms of PCP dechlorination.

Inhibition of miR-1247 on cell proliferation and invasion in bladder cancer through its downstream target of RAB36.

Recently, microRNA-1247 (miR-1247) has been reported to function as tumour suppressor in several cancer types, including pancreatic cancer, hepatocellular cancer and lung cancer. However, the biological function of miR-1247 in bladder cancer and the underlying mechanisms have remained largely uncovered. In this study, the expression of miR-1247 was significantly downregulated, while RAB36 protein was remarkably upregulated in bladder cancer tissues and cell lines compared with that in paired adjacent normal tissues or normal cell line (SU-HUC-1). The function of miR-1247 and RAB36 in the cell viability, proliferation and invasion of bladder cancer cells (T24 and J82) was assessed by CCK-8, colony formation and Transwell assay, respectively. Gain of function studies showed that upregulation of miR-1247 significantly inhibited cell proliferation and invasion capacity of bladder cancer cells. Consistently, downregulation of RAB36 mimicked the suppressive effects of miR-1247 overexpression in bladder cancer cells. Importantly, miR-1247 was confirmed to target the 30untranslated region (UTR) of RAB36 and downregulated its expression using luciferase reporter assay and Western blot assays. In conclusion, these results provide the first clues regarding the role of miR-1247 might be a potential therapeutic agent and diagnostic marker of bladder cancer by inhibiting RAB36 expression.

MicroRNA­136 inhibits prostate cancer cell proliferation and invasion by directly targeting mitogen­activated protein kinase kinase 4.

Prostate cancer (PCa) is the second most common type of cancer and the 6th leading cause of cancer­associated mortality worldwide. Accumulated evidence suggests that PCa initiation and progression are controlled by microRNAs (miRNAs). Therefore, investigating PCa­associated miRNAs may provide novel biomarkers for the diagnosis and treatment of patients with PCa. In the present study it was demonstrated that miRNA­136 (miR­136) expression was significantly downregulated in PCa tissues and cell lines. The resumption of miR­136 expression suppressed cell proliferation and invasion in PCa cells. Bioinformatics analysis predicted that mitogen­activated protein kinase kinase 4 (MAP2K4) was a direct target of miR­136. This prediction was experimentally confirmed by a luciferase reporter assay, RT­qPCR and western blot analysis. MAP2K4 was highly expressed in PCa tissues and inversely correlated with the miR­136 expression level. Additionally, the restoration of MAP2K4 expression significantly blocked the inhibitory effects of miR­136 on cell proliferation and invasion in PCa cells. Therefore, miR­136 may suppress the proliferation and invasion of PCa cells by targeting MAP2K4 and may be a novel candidate target for cancer therapy against PCa.

Human mesenchymal stromal cell-derived extracellular vesicles alleviate renal ischemic reperfusion injury and enhance angiogenesis in rats.

BACKGROUND: Mesenchymal stromal cells (MSCs) derived extracellular vesicles (EVs) were regarded as a potent medium for kidney injury repair and angiogenesis were regarded as an important step in tissue regeneration. However, the pro-angiogenesis effect of MSC-EVs in ischemia-reperfusion induced kidney injury and its potential mechanisms have yet to be determined. METHODS: EVs were isolated from the medium of human umbilical cord-derived MSCs (huMSCs) were injected in rats intravenously after unilateral kidney ischemia. Animals were sacrificed at 24 h and 2 weeks after injury. The renal functions and histology staining were examined to assess the therapeutic effect of the EVs. Moreover, we investigated the pro-angiogenesis effects of EVs in injured kidneys and tested the angiogenesis-related factors to further illuminate the probable mechanisms. RESULTS: It was observed that EVs could reduce cell apoptosis and enhances proliferation 24 h after kidney injury, meanwhile renal function was improved and the histological lesion was mitigated. Moreover, renal VEGF was up-regulated by EVs and HIF-1α was down-regulated. Further, the increase of capillary vessel density and reduce of renal fibrosis was observed after 2 weeks. In vitro, EVs could deliver human VEGF directly to renal tubular epithelial cells (TECs) and increase VEGF levels. Most important, all the beneficial effects of EVs were abrogated by RNase treated except for the delivery of human VEGF. CONCLUSIONS: Human MSC-EVs could protect against ischemic/reperfusion injury induced kidney injury through pro-angiogenesis effects in HIF-1α independent manner, and both the delivery of pro-angiogenesis related VEGF and RNAs were involved in this process.

NK Cell Regulatory Property is Involved in the Protective Role of MSC-Derived Extracellular Vesicles in Renal Ischemic Reperfusion Injury.

Immunomodulation has been regarded as an important therapeutic aspect of mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) in renal ischemic reperfusion injury (IRI), and the specific mechanism still unclear. Here, we attempt to test the function of human MSC-EVs on renal IRI by targeting the natural killer (NK) cells and to investigate the possible mechanism. Data indicated that EVs decreased NK cells in spleen and ischemic kidney. Both the EVs and antibody-dependent depletion of NK cells displayed a protective role in IRI rats. Moreover, the splenectomy model was established to evaluate the role of spleen in this process. It showed that the NK cell regulatory ability and renal protective effects by EVs still exist without spleen, which is unlike MSC properties published previously. Further, the down-regulation of chemokines in injured kidney and the delivery of RNAs through EVs in vitro were also observed. Through the microRNA array test, various inflammation-related microRNAs highly expressed in MSC-EVs compared with fibroblast EVs were tested. Thus, these results indicated that MSC-EVs could ameliorate renal ischemic reperfusion injury by decreasing NK cells and the spleen is not necessary in this process. The regulation of chemokines in injured kidney was the other factor, and the transfer of various microRNAs in the MSC-EVs may be involved. This provides direction for future clinical applications.

Microvesicles derived from human Wharton's jelly mesenchymal stem cells promote human renal cancer cell growth and aggressiveness through induction of hepatocyte growth factor.

In our previous study, microvesicles (MVs) released from human Wharton's jelly mesenchymal stem cells (hWJ-MSCs) retard the growth of bladder cancer cells. We would like to know if MVs have a similar effect on human renal cell carcinoma (RCC). By use of cell culture and the BALB/c nu/nu mice xeno-graft model, the influence of MVs upon the growth and aggressiveness of RCC (786-0) was assessed. Cell counting kit-8 (CCK-8) assay, incidence of tumor, tumor size, Ki-67 or TUNEL staining was used to evaluate tumor cell growth in vitro or in vivo. Flow cytometry assay (in vitro) or examination of cyclin D1 expression (in vivo) was carried out to determine the alteration of cell cycle. The aggressiveness was analyzed by Wound Healing Assay (in vitro) or MMP-2 and MMP-9 expression (in vivo). AKT/p-AKT, ERK1/2/p-ERK1/2 or HGF/c-MET expression was detected by real-time PCR or western blot. Our data demonstrated that MVs promote the growth and aggressiveness of RCC both in vitro and in vivo. In addition, MVs facilitated the progression of cell cycle from G0/1 to S. HGF expression in RCC was greatly induced by MVs, associated with activation of AKT and ERK1/2 signaling pathways. RNase pre-treatment abrogated all effects of MVs. In summary, induction of HGF synthesis via RNA transferred by MVs activating AKT and ERK1/2 signaling is one of crucial contributors to the pro-tumor effect.

Microvesicles derived from human Wharton's Jelly mesenchymal stromal cells ameliorate renal ischemia-reperfusion injury in rats by suppressing CX3CL1.

INTRODUCTION: Studies have demonstrated that mesenchymal stromal cells (MSCs) could reverse acute and chronic kidney injury by a paracrine or endocrine mechanism, and microvesicles (MVs) have been regarded as a crucial means of intercellular communication. In the current study, we focused on the therapeutic effects of human Wharton-Jelly MSCs derived microvesicles (hWJMSC-MVs) in renal ischemia/reperfusion injury and its potential mechanisms. METHODS: MVs isolated from conditioned medium were injected intravenously in rats immediately after ischemia of the left kidney for 60 minutes. The animals were sacrificed at 24 hours, 48 hours and 2 weeks after reperfusion. The infiltration of inflammatory cells was identified by the immunostaining of CD68+ cells. ELISA was employed to determine the inflammatory factors in the kidney and serum von Willebrand Factor (VWF). Tubular cell proliferation and apoptosis were identified by immunostaining. Renal fibrosis was assessed by Masson's tri-chrome straining and alpha-smooth muscle actin (α-SMA) staining. The CX3CL1 expression in the kidney was measured by immunostaining and Western blot, respectively. In vitro, human umbilical vein endothelial cells were treated with or without MVs for 24 or 48 hours under hypoxia injury to test the CX3CL1 by immunostaining and Western blot. RESULTS: After administration of hWJMSC-MVs in acute kidney injury (AKI) rats, renal cell apoptosis was mitigated and proliferation was enhanced, inflammation was also alleviated in the first 48 hours. MVs also could suppress the expression of CX3CL1 and decrease the number of CD68+ macrophages in the kidney. In the late period, improvement of renal function and abrogation of renal fibrosis were observed. In vitro, MVs could down-regulate the expression of CX3CL1 in human umbilical vein endothelial cells under hypoxia injury at 24 or 48 hours. CONCLUSIONS: A single administration of MVs immediately after ischemic AKI could ameliorate renal injury in both the acute and chronic stage, and the anti-inflammatory property of MVs through suppression of CX3CL1 may be a potential mechanism. This establishes a substantial foundation for future research and treatment.

Urothelial differentiation of human umbilical cord-derived mesenchymal stromal cells in vitro.

Human umbilical cord-derived mesenchymal stromal cells (hUCMSCs) are the most primitive of those isolated from other post-natal tissue source. The hUCMSCs possess the capability of differentiating along multi-lineage. This study aimed to investigate whether hUCMSCs can differentiate into urothelium-like cells. The hUCMSCs were isolated from fresh human umbilical cord postpartum and expanded at least to passage 3 in vitro. Subsequently, they were cultured with conditioned medium from urothelial cells (UC-CM) supplemented with 20 ng/ml exogenous epidermal growth factor (EGF). Urothelial cell specific marker uroplakin II (UPII) and cytokeratins were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunofluorescence technology. During culture, hUCMSCs started to express UPII and cytokeratins weakly at 7 days and were significantly up-regulated at 2 weeks post-induction. Additionally, morphology of hUCMSCs changed from spindle-shape to a polygonal epithelial-shape similar to that of urothelial cells after 7 days. The study results indicated that hUCMSCs can differentiate into urothelium-like cells in a defined micro-environment in vitro constituted by UC-CM and exogenous EGF.

Effects of transfection of ICAP-1α and its mutants on adhesion and migration of 2H-11 cells.

This study examined the effect of integrin cytoplasmic domain-associated protein 1α (ICAP-1α) and its mutatants T38A and I138A on the adhesion, migration and tube formation of 2H-11 cells. rAAV-ICAP-1α, rAAV-T38A and rAAV-I138A were constructed. After infection, the expression of ICAP-1α and p-ERK1/2, p-c-Jun protein was measured by Western blotting. Adhesion ability was evaluated by using MTT. Cell migration was determined by using Boyden chamber method. Tube formation test was conducted on Matrigel. The results showed that in ICAP-1α, T38A and I138A groups, ICAP-1α protein expression was increased. In T38A and I138A groups, phospho-ERK1/2, phospho-c-Jun protein expressions were significantly increased as compared with the control group and the GFP group. ICAP-1α group protein expression was obviously decreased when compared with the control group and the GFP group. Cell adhesion ratio was 0.1429±0.0080 in control group, 0.1434±0.0077 in GFP group and the ratio in T38A and I138A groups increased to 0.3210±0.0082 and 0.3250±0.0079, respectively. In ICAP-1α group, the ratio was decreased to 0.1005±0.0073. In T38A and I138A groups, the number of migrating 2H-11 cells was increased to 31.45±3.20 and 33.10±5.40 against 18.51±2.80 in control group and 20.47±3.12 in GFP group. In ICAP-1α group, the number was decreased to 12.06±1.72. The number of tube-like structures was increased to 20.41±2.54 in T38A and to 22.26±3.07 in I138A groups as compared to those of control group 12.45±1.84 and GFP group 13.63±2.71. In ICAP-1α group, the number of tube-like structures was decreased to 8.32±1.24. It was suggested that rAAV-T38A and rAAV-I138A transfection can substantially increase 2H-11 cell adhesion, migration and angiogenisis, while rAAV-ICAP-1α can greatly inhibit the effect. These effects might be correlated with ERK1/2 and c-Jun protein phosphorylation.

Effects of Transfection of ICAP-1α and Its Mutants on Adhesion and Migration of 2H-11 Cells / 华中科技大学学报（医学）（英德文版）

This study examined the effect of integrin cytoplasmic domain-associated protein la (ICAP-la) and its mutatants T38A and I138A on the adhesion, migration and tube formation of 2H- 11 cells. rAAV-ICAP- 1 α, rAAV-T38A and rAAV-I 138A were constructed. After infection, the expression of ICAP-la and p-ERK1/2, p-c-Jun protein was measured by Westerrr blotting. Adhesion ability was evaluated by using MTT. Cell migration was determined by using Boyden chamber method. Tube formation test was conducted on Matrigel. The results showed that in ICAP-lα, T38A and I138A groups, ICAP-la protein expression was increased. In T38A and I138A groups, phospho-ERK1/2, phospho-c-Jun protein expressions were significantly increased as compared with the control group and the GFP group. ICAP-la group protein expression was obviously decreased when compared with the control group and the GFP group. Cell adhesion ratio was 0.1429±0.0080 in control group, 0.1434±0.0077 in GFP group and the ratio in T38A and I138A groups increased to 0.3210±0.0082 and 0.3250±0.0079, respectively. In ICAP-la group, the ratio was decreased to 0.1005±0.0073. In T38A and I 138A groups, the number of migrating 2H-11 cells was increased to 31.45±3.20 and 33.10±5.40 against 18.51±2.80 in control group and 20.47±3.12 in GFP group. In ICAP-la group, the number was decreased to 12.06±1.72. The number of tube-like structures was increased to 20.41±2.54 in T38A and to 22.26±3.07 in I138A groups as compared to those of control group 12.45±1.84 and GFP group 13.63±2.71. In ICAP-la group, the number of tube-like structures was decreased to 8.32±1.24. It was suggested that rAAV-T38A and rAAV-I138A transfection can substantially increase 2H-11 cell adhesion, migration and angiogenisis, while rAAV-ICAP-1 α can greatly inhibit the effect. These effects might be correlated with ERK1/2 and c-Jun protein phosphorylation.