YAP silencing by RB1 mutation is essential for small-cell lung cancer metastasis.

Small cell lung cancer (SCLC) is highly lethal due to its prevalent metastasis. Most SCLCs have inactivating mutations in TP53 and RB1. We find that loss of YAP expression is key for SCLC cells to acquire rapid ameboid migration and high metastatic potential. YAP functions through its target genes CCN1/CCN2 to inhibit SCLC ameboid migration. RB1 mutation contributes to YAP transcriptional silencing via E2F7, which recruits the RCOR co-repressor complex to YAP promoter. We discover that benzamide family HDAC inhibitors stimulate YAP expression by inhibiting the RCOR-HDAC complex, thereby suppressing SCLC metastasis and improving survival in a mouse model. Our study unveils the molecular and cellular basis underlying SCLC's high metastatic potential, the previously unrecognized role of YAP in suppressing ameboid migration and tumor metastasis, and the mechanism of YAP transcription regulation involving E2F7, RCOR, and Sin3 HDAC. This study reveals a therapeutic potential of benzamides for SCLC treatment.

The Arf family GTPases: Regulation of vesicle biogenesis and beyond.

The Arf family proteins are best known for their roles in the vesicle biogenesis. However, they also play fundamental roles in a wide range of cellular regulation besides vesicular trafficking, such as modulation of lipid metabolic enzymes, cytoskeleton remodeling, ciliogenesis, lysosomal, and mitochondrial morphology and functions. Growing studies continue to expand the downstream effector landscape of Arf proteins, especially for the less-studied members, revealing new biological functions, such as amino acid sensing. Experiments with cutting-edge technologies and in vivo functional studies in the last decade help to provide a more comprehensive view of Arf family functions. In this review, we summarize the cellular functions that are regulated by at least two different Arf members with an emphasis on those beyond vesicle biogenesis.

Hippo pathway regulation by phosphatidylinositol transfer protein and phosphoinositides.

The Hippo pathway plays a key role in development, organ size control and tissue homeostasis, and its dysregulation contributes to cancer. The LATS tumor suppressor kinases phosphorylate and inhibit the YAP/TAZ transcriptional co-activators to suppress gene expression and cell growth. Through a screen of marine natural products, we identified microcolin B (MCB) as a Hippo activator that preferentially kills YAP-dependent cancer cells. Structure-activity optimization yielded more potent MCB analogs, which led to the identification of phosphatidylinositol transfer proteins α and ß (PITPα/ß) as the direct molecular targets. We established a critical role of PITPα/ß in regulating LATS and YAP. Moreover, we showed that PITPα/ß influence the Hippo pathway via plasma membrane phosphatidylinositol-4-phosphate. This study uncovers a previously unrecognized role of PITPα/ß in Hippo pathway regulation and as potential cancer therapeutic targets.

Defining the proximal interaction networks of Arf GTPases reveals a mechanism for the regulation of PLD1 and PI4KB.

The Arf GTPase family is involved in a wide range of cellular regulation including membrane trafficking and organelle-structure assembly. Here, we have generated a proximity interaction network for the Arf family using the miniTurboID approach combined with TMT-based quantitative mass spectrometry. Our interactome confirmed known interactions and identified many novel interactors that provide leads for defining Arf pathway cell biological functions. We explored the unexpected finding that phospholipase D1 (PLD1) preferentially interacts with two closely related but poorly studied Arf family GTPases, ARL11 and ARL14, showing that PLD1 is activated by ARL11/14 and may recruit these GTPases to membrane vesicles, and that PLD1 and ARL11 collaborate to promote macrophage phagocytosis. Moreover, ARL5A and ARL5B were found to interact with and recruit phosphatidylinositol 4-kinase beta (PI4KB) at trans-Golgi, thus promoting PI4KB's function in PI4P synthesis and protein secretion.

Protective Effect of Targeted Fluid Therapy on Patients with One-Lung Ventilation.

Objective: To evaluate the protective effect of target-directed fluid therapy on the lungs and postoperative rehabilitation in elderly patients with single-lung ventilation undergoing total endoscopic radical resection of esophageal cancer. Methods: Seventy elderly patients who underwent total endoscopic radical resection of esophageal cancer from January 2017 to December 2019 in our hospital were selected and divided into two groups by the random number table method: the goal-directed fluid treatment group (group G, n = 35) and the control group (group C, n = 35). Venous blood was extracted before surgery (T1), at the end of free esophagus (T2) by thoracoscopy, at the end of abdominal surgery (T3), and at the end of surgery (T4). IL-6 and IL-10 levels were detected by ELISA. The clinical pulmonary infection score (CIPS) was used to evaluate the pulmonary inflammation on the second day after surgery and the occurrence of complications. Duration of antibiotic use and length of hospital stay were recorded. Results: At T1, there were no significant differences in IL-6 and IL-10 levels between the two groups (P > 0.05). At T2, the IL-6 level in group G increased to 26.65 ± 1.80 pg/ml but was significantly lower than that in group C (32.28 ± 3.22 pg/ml) (P < 0.01). At T3 and T4, IL-6 and IL-10 levels in group G were significantly lower than those in group C (P < 0.01). The CIPS score of group G was lower than that of group C (1.5 ± 1.0 vs 2.7 ± 1.4), and the duration of antibiotic use in group G was shorter than that in group C (211.2 ± 15.4 vs 232.6 ± 18.7 h), with statistical significance (P < 0.01). The incidence of complications in group G was lower than that in group C (28.6% vs 40.0%), and the length of hospital stay in group G was shorter than that in group C (10.5 ± 1.7 vs 11.2 ± 1.9 days), but there was no significant difference between the two groups (P > 0.05). Conclusion: Target-directed fluid therapy inhibited inflammatory cytokine levels and had better lung protection, but no significant benefit in the complications or the length of hospital stay was observed.

The Hippo pathway mediates Semaphorin signaling.

Semaphorins were originally identified as axonal guidance molecules, but they also control processes such as vascular development and tumorigenesis. The downstream signaling cascades of Semaphorins in these biological processes remain unclear. Here, we show that the class 3 Semaphorins (SEMA3s) activate the Hippo pathway to attenuate tissue growth, angiogenesis, and tumorigenesis. SEMA3B restoration in lung cancer cells with SEMA3B loss of heterozygosity suppresses cancer cell growth via activating the core Hippo kinases LATS1/2 (large tumor suppressor kinase 1/2). Furthermore, SEMA3 also acts through LATS1/2 to inhibit angiogenesis. We identified p190RhoGAPs as essential partners of the SEMA3A receptor PlexinA in Hippo regulation. Upon SEMA3 treatment, PlexinA interacts with the pseudo-guanosine triphosphatase (GTPase) domain of p190RhoGAP and simultaneously recruits RND GTPases to activate p190RhoGAP, which then stimulates LATS1/2. Disease-associated etiological factors, such as genetic lesions and oscillatory shear, diminish Hippo pathway regulation by SEMA3. Our study thus discovers a critical role of Hippo signaling in mediating SEMA3 physiological function.

The two sides of Hippo pathway in cancer.

The Hippo signaling pathway was originally characterized by genetic studies in Drosophila to regulate tissue growth and organ size, and the core components of this pathway are highly conserved in mammals. Studies over the past two decades have revealed critical physiological and pathological functions of the Hippo tumor-suppressor pathway, which is tightly regulated by a broad range of intracellular and extracellular signals. These properties enable the Hippo pathway to serve as an important controller in organismal development and adult tissue homeostasis. Dysregulation of the Hippo signaling has been observed in many cancer types, suggesting the possibility of cancer treatment by targeting the Hippo pathway. The general consensus is that Hippo has tumor suppressor function. However, growing evidence also suggests that the function of the Hippo pathway in malignancy is cancer context dependent as recent studies indicating tumor promoting function of LATS. This article surveys the Hippo pathway signaling mechanisms and then reviews both the tumor suppressing and promoting function of this pathway. A comprehensive understanding of the dual roles of the Hippo pathway in cancer will benefit future therapeutic targeting of the Hippo pathway for cancer treatment.

Comparison of Autologous Platelet-Rich Plasma and Chitosan in the Treatment of Temporomandibular Joint Osteoarthritis: A Retrospective Cohort Study.

PURPOSE: The purpose of this article is to study the effect of autologous platelet-rich plasma (PRP) injected into the upper cavity of the temporomandibular joint (TMJ) on the treatment of TMJ osteoarthritis. PATIENTS AND METHODS: We retrospectively analyzed the data of 27 patients with TMJ osteoarthritis treated at the China Medical University Hospital of Stomatology from September 2018 to September 2019. Maximal interincisal opening, pain intensity, and TMJ sounds were recorded and compared before treatment and at the 3rd and 6th months after the treatment. SPSS 24.0 software was used to analyze the data of each group, and the imaging changes in the condylar bone were compared before and 6 months after the treatment. The P-value was set at .05. RESULTS: Better results were observed in the group treated with PRP on maximal interincisal opening and pain intensity than in the group receiving chitosan treatment. Regarding TMJ sounds, relief was observed in both groups, with no significant difference. CONCLUSIONS: The effect of PRP on the improvement of the maximal interincisal opening and pain intensity of patients with TMJ osteoarthritis is better than that of chitosan. However, it should be noted that the incidence of complications associated with the injection of PRP may be higher than that with injection of chitosan.

Sialendoscopy-Assisted Treatment of Stensen's Duct Injury: A Case Series.

PURPOSE: To evaluate the clinical value of sialendoscopy in the treatment of Stensen's duct injury. PATIENTS AND METHODS: A total of 5 patients with Stensen's duct injuries who had been treated from December 2017 to April 2019 were included in the present study. The operations were performed with the help of a sialendoscope. All patients were followed for 6 months. RESULTS: The distal end of the ductal system was found precisely with the use of the sialendoscope, and the proximal end was identified by the location of the distal end. The end-to-end anastomosis was performed successfully. None of the patients complained of salivary gland fistula at the 6-month follow-up examination. CONCLUSIONS: The stumps of the ductal system could be precisely and effectively located with the help of a sialendoscope.

Intravenous lidocaine infusion for pain control after laparoscopic cholecystectomy: A meta-analysis of randomized controlled trials.

BACKGROUND: This meta-analysis aimed to assess the efficiency and safety of intravenous infusion of lidocaine for pain management after laparoscopic cholecystectomy (LC). METHODS: A systematic search was performed in PubMed (August 1966-2017), Medline (August 1966-2017), Embase (August 1980-2017), ScienceDirect (August 1985-2017), and the Cochrane Library. Only randomized controlled trials (RCTs) were included. Fixed/random effect model was used according to the heterogeneity tested by I2 statistic. Meta-analysis was performed using Stata.11.0 software. RESULTS: A total of 5 RCTs were retrieved involving 274 patients. The present meta-analysis indicated that there were significant differences between groups in terms of visual analog scale scores at 12hours (weighted mean difference [WMD]=-0.743, 95% CI: -1.246 to -0.240, P = .004), 24hours (WMD=-0.712, 95% CI: -1.239 to -0.184, P = .008), and 48hours (WMD=-0.600, 95% CI: -0.972 to -0.229, P = .002) after LC. Significant differences were found regarding opioid consumption at 12hours (WMD=-3.136, 95% CI: -5.591 to -0.680, P = .012), 24hours (WMD=-4.739, 95% CI: -8.291 to -1.188, P = .009), and 48hours (WMD=-3.408, 95% CI: -5.489 to -1.326, P = .001) after LC. CONCLUSION: Intravenous lidocaine infusion significantly reduced postoperative pain scores and opioid consumption after LC. In addition, there were fewer adverse effects in the lidocaine groups. Higher quality RCTs are still required for further research.

Acetylation accumulates PFKFB3 in cytoplasm to promote glycolysis and protects cells from cisplatin-induced apoptosis.

Enhanced glycolysis in cancer cells has been linked to cell protection from DNA damaging signals, although the mechanism is largely unknown. The 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) catalyzes the generation of fructose-2,6-bisphosphate, a potent allosteric stimulator of glycolysis. Intriguingly, among the four members of PFKFB family, PFKFB3 is uniquely localized in the nucleus, although the reason remains unclear. Here we show that chemotherapeutic agent cisplatin promotes glycolysis, which is suppressed by PFKFB3 deletion. Mechanistically, cisplatin induces PFKFB3 acetylation at lysine 472 (K472), which impairs activity of the nuclear localization signal (NLS) and accumulates PFKFB3 in the cytoplasm. Cytoplasmic accumulation of PFKFB3 facilitates its phosphorylation by AMPK, leading to PFKFB3 activation and enhanced glycolysis. Inhibition of PFKFB3 sensitizes tumor to cisplatin treatment in a xenograft model. Our findings reveal a mechanism for cells to stimulate glycolysis to protect from DNA damage and potentially suggest a therapeutic strategy to sensitize tumor cells to genotoxic agents by targeting PFKFB3.

Thromboxane A2 Activates YAP/TAZ Protein to Induce Vascular Smooth Muscle Cell Proliferation and Migration.

The thromboxane A2 receptor (TP) has been implicated in restenosis after vascular injury, which induces vascular smooth muscle cell (VSMC) migration and proliferation. However, the mechanism for this process is largely unknown. In this study, we report that TP signaling induces VSMC migration and proliferation through activating YAP/TAZ, two major downstream effectors of the Hippo signaling pathway. The TP-specific agonists [1S-[1α,2α(Z),3ß(1E,3S*),4 α]]-7-[3-[3-hydroxy-4-(4-iodophenoxy)-1-butenyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid (I-BOP) and 9,11-dideoxy-9α,11α-methanoepoxy-prosta-5Z,13E-dien-1-oic acid (U-46619) induce YAP/TAZ activation in multiple cell lines, including VSMCs. YAP/TAZ activation induced by I-BOP is blocked by knockout of the receptor TP or knockdown of the downstream G proteins Gα12/13 Moreover, Rho inhibition or actin cytoskeleton disruption prevents I-BOP-induced YAP/TAZ activation. Importantly, TP activation promotes DNA synthesis and cell migration in VSMCs in a manner dependent on YAP/TAZ. Taken together, thromboxane A2 signaling activates YAP/TAZ to promote VSMC migration and proliferation, indicating YAP/TAZ as potential therapeutic targets for cardiovascular diseases.

Impact of acetylation on tumor metabolism.

Acetylation of protein lysine residues is a reversible and dynamic process that is controlled by histone acetyltransferases (HATs) and deacetylases (HDACs and SIRTs). Recent studies have revealed that acetylation modulates not only nuclear proteins but also cytoplasmic or mitochondrial proteins, including many metabolic enzymes. In tumors, cellular metabolism is reprogrammed to provide intermediates for biosynthesis such as nucleotides, fatty acids, and amino acids, and thereby favor the rapid proliferation of cancer cells and tumor development. An increasing number of investigations have indicated that acetylation plays an important role in tumor metabolism. Here, we summarize the substrates that are modified by acetylation, especially oncogenes, tumor suppressor genes, and enzymes that are implicated in tumor metabolism.

Mitogenic and oncogenic stimulation of K433 acetylation promotes PKM2 protein kinase activity and nuclear localization.

Alternative splicing of the PKM2 gene produces two isoforms, M1 and M2, which are preferentially expressed in adult and embryonic tissues, respectively. The M2 isoform is reexpressed in human cancer and has nonmetabolic functions in the nucleus as a protein kinase. Here, we report that PKM2 is acetylated by p300 acetyltransferase at K433, which is unique to PKM2 and directly contacts its allosteric activator, fructose 1,6-bisphosphate (FBP). Acetylation prevents PKM2 activation by interfering with FBP binding and promotes the nuclear accumulation and protein kinase activity of PKM2. Acetylation-mimetic PKM2(K433) mutant promotes cell proliferation and tumorigenesis. K433 acetylation is decreased by serum starvation and cell-cell contact, increased by cell cycle stimulation, epidermal growth factor (EGF), and oncoprotein E7, and enriched in breast cancers. Hence, K433 acetylation links cell proliferation and transformation to the switch of PKM2 from a cytoplasmic metabolite kinase to a nuclear protein kinase.

[Changes in lymph circulation in rats with experimental disseminated intravascular coagulation].

OBJECTIVE: To explore changes in lymph circulation in rats with experimental disseminated intravascular coagulation (DIC). METHODS: Thirty-two male Wistar rats were randomly divided into two groups: DIC group and sham-operation group, with 16 rats in each group.In each group, 8 rats were used to observe the lymphatic circulation of intestinal mesentery, and the others were used to observe lymphatic dynamics. Experimental DIC model was replicated by injecting dextran 500 via jugular vein in Wistar rats. The changes in lymph circulation were observed in these rats with lymphology methods. RESULTS: During the phase of DIC, the contractility of mesenteric micro-lymphatic (ML), the intestinal lymph flux, lymphocytes output were obviously decreased, and there was a few monocytes in the lymph fluid, but the lymph fluid viscosity was increased. After treatment with normal saline (NS), the contractility of ML, the intestinal lymph flux, lymphocytes output were obviously increased, and there was a large amount of monocytes in the lymph fluid, with significant difference compared with sham-operation group (all P<0.05). Moreover, the lymph fluid viscosity was decreased compared with sham-operation group (P<0.05). CONCLUSION: The disturbance in lymph circulation mainly includes depression of contractility of lymphatics, the transport function of lymph circulation, and increased lymph fluid viscosity.