A YAP/TAZ-CD54 axis is required for CXCR2-CD44- tumor-specific neutrophils to suppress gastric cancer

As an important part of tumor microenvironment, neutrophils are poorly understood due to their spatiotemporal heterogeneity in tumorigenesis. Here we defined, at single-cell resolution, CD44-CXCR2- neutrophils as tumor-specific neutrophils (tsNeus) in both mouse and human gastric cancer (GC). We uncovered a Hippo regulon in neutrophils with unique YAP signature genes (e.g., ICAM1, CD14, EGR1) distinct from those identified in epithelial and/or cancer cells. Importantly, knockout of YAP/TAZ in neutrophils impaired their differentiation into CD54+ tsNeus and reduced their antitumor activity, leading to accelerated GC progression. Moreover, the relative amounts of CD54+ tsNeus were found to be negatively associated with GC progression and positively associated with patient survival. Interestingly, GC patients receiving neoadjuvant chemotherapy had increased numbers of CD54+ tsNeus. Furthermore, pharmacologically enhancing YAP activity selectively activated neutrophils to suppress refractory GC, with no significant inflammation-related side effects. Thus, our work characterized tumor-specific neutrophils in GC and revealed an essential role of YAP/TAZ-CD54 axis in tsNeus, opening a new possibility to develop neutrophil-based antitumor therapeutics.

Systematic Evaluation of Clinical Efficacy and Safety of Traditional Chinese Medicine for Post Stroke Cognitive Impairment / 中国实验方剂学杂志

Objective:To evaluate the efficacy and safety of traditional Chinese medicine (TCM) in the treatment of post stroke cognitive impairment (PSCI). Method:Seven databases, including CNKI, WanFang, VIP, CBM, PubMed, The Cochrane library and ClinicalTrials.gov, were electronically searched for relevant randomized controlled trials (RCTs) of TCM in the treatment of PSCI. The Cochrane risk of bias assessment tool was used to evaluate the methodological quality of the included studies, descriptive analysis was carried out on the included studies, and the Meta quantitative analysis was carried out with RevMan 5.3 software. Result:A total of 16 RCTs were included with 1 296 participants, and they were assigned to the intervention group (n=649) and the control group (n=647). The results showed that TCM combined with western medicine group and TCM group were better than western medicine group in improving the scores of Montreal Cognitive Assessment (MoCA), Mini-mental State Examination (MMSE), Barthel Index (BI), Activity of Daily Living (ADL), Chinese stroke scale (CSS) and National Institutes of Health stroke scale (NIHSS) of PSCI patients, and no serious adverse events were observed. Conclusion:TCM has potential advantages in improving the cognitive function of patients with PSCI, and it also has certain efficacy in improving the daily living ability and neurological impairment symptoms, and no serious adverse events have been observed. Due to the low quality of methodology included in the studies, in order to provide reliable basis for clinical decision-making, high-quality of RCTs are still needed to study the efficacy and safety of TCM for PSCI.

Electroacupuncture ameliorate learning and memory by improving N -acetylaspartate and glutamate metabolism in APP/PS1 mice

OBJECTIVE: To explore the precise mechanism of electroacupuncture (EA) to delay cognitive impairment in Alzheimer disease. Methods N -Acetylaspartate (NAA), glutamate (Glu) and myoinositol (mI) metabolism were measured by magnetic resonance spectroscopy, learning and memory of APP/PS1 mouse was evaluated by the Morris water maze test and the step-down avoidance test, neuron survival number and neuronal structure in the hippocampus were observed by Nissl staining, and BDNF and phosphorylated TrkB detected by Western blot. RESULTS: EA at DU20 acupuncture significantly improve learning and memory in behavioral tests, up-regulate NAA, Glu and mI metabolism, increase the surviving neurons in hippocampus, and promote the expression of BDNF and TrkB in the APP/PS1 transgenic mice. CONCLUSION: These findings suggested that EA is a potential therapeutic for ameliorate cognitive dysfunction, and it might be due to EA could improve NAA and Glu metabolism by upregulation of BDNF in APP/PS1 mice.

Evaluation of teaching-in-English reform in five-year clinical tropical medicine program – Case analysis of curriculum reform of clinical medicine in Hainan Medical University

Objective To investigate the model, quality as well as effect of teaching-in-English in five-year clinical medicine program of Hainan Medical University. Methods The questionnaire was carried out among clinical medicine undergraduates of 2012–2015 grades in Hainan Medical University, to investigate studying time, studying habits and the impact of teaching in English. Additionally results of CET-4, CET-6 and overseas internship from undergraduates of 2012–2015 grade, as well as the result of phased medical licensing examination and post-graduate entrance examination from undergraduates of 2012 were accordingly collected from the Teaching Management Department. Results For the Chinese students in international classes, the average time of self-study was 161.49 min, 58.3% had preview before classes, and 90.7% had habit of review after classes. Thus the first time pass rate, total pass rate, first time excellent rate and total excellent rate of CET-4 and CET-6 of international classes were significantly higher than those of regular classes. The result of post-graduate entrance examination in 2016 showed that the score, pass rate and acceptance rate of international classes of 2012 grade were significantly higher those of regular classes (P < 0.01). Conclusions Teaching-in-English reform in Hainan Medical University has achieved initial success. Chinese students from international classes are superior to those from regular classes in many aspects. However, there are still many problems, and effective measures should be implemented to promote teaching quality continuously.

Clinical Observation of Enhanced Immunosuppressive Therapy in the Treatment of Refractory Nephrotic Syndrome / 中国药房

OBJECTIVE:To observe the clinical efficacy and safety of enhanced immunosuppressive therapy in the treatment of refractory nephrotic syndrome (RNS). METHODS:Totally 76 RNS patients were selected from 2 hospitals during Jan. 2012-Mar. 2015,and then divided into control group and observation group according to random number table,with 38 cases in each group. Two groups were given Prednisone acetate tablet 50 mg,qd;decreasing to 15 mg,qd,after urine protein returned to normal. Based on it,control group was given Leflunomide tablets 50 mg,qd;decreasing to 20 mg,qd,3 days later. Based on con-trol group,observation group was additionally given Mycophenolate mofetil dispersible tablet 750 mg,bid;decreasing to 500 mg, qd,3 months later. Both groups were treated for 6 months. Clinical efficacies,follow-up recurrence rate as well as renal function in-dexes and inflammatory cell factors before and after treatment,and the occurrence of ADR were compared between 2 groups. RE-SULTS:The total response rate of observation group(92.11%)was significantly higher than control group(73.68%),and fol-low-up recurrence rate(5.26%)was significantly lower than control group(23.68%),with statistical significance(P0.05). After treatment,24 h urinary protein quantification,urinary IL-6 and IL-8 levels of 2 groups decreased significantly,while the content of serum protein increased significantly;the observation group was significantly better than the control group,with sta-tistical significance(P0.05). The incidence of ADR in the control group and the observation group was 34.21% and 44.74% respec-tively,without statistical significance between 2 groups(P>0.05). CONCLUSIONS:Enhanced immunosuppressive therapy in the treatment of RNS can improve renal function,reduce inflammatory reaction and long-term recurrence risk,and have good therapeu-tic efficacy and safety.

Evaluation of teaching-in-English reform in five-year clinical tropical medicine program-Case analysis of curriculum reform of clinical medicine in Hainan Medical University

Objective:To investigate the model,quality as well as effect of teaching-in-English in five-year clinical medicine program of Hainan Medical University.Methods:The questionnaire was carried out among clinical medicine undergraduates of 2012-2015 grades in Hainan Medical University,to investigate studying time,studying habits and the impact of teaching in English.Additionally results of CET-4,CET-6 and overseas internship from undergraduates of 2012-2015 grade,as well as the result of phased medical licensing examination and post-graduate entrance examination from undergraduates of 2012 were accordingly collected from the Teaching Management Department.Results:For the Chinese students in international classes,the average time of self-study was 161.49 min,58.3％ had preview before classes,and 90.7％ had habit of review after classes.Thus the first time pass rate,total pass rate,first time excellent rate and total excellent rate of CET-4 and CET-6 of international classes were significantly higher than those of regular classes.The result of post-graduate entrance examination in 2016 showed that the score,pass rate and acceptance rate of international classes of 2012 grade were significantly higher those of regular classes (P ＜ 0.01).Conclusions:Teaching-in-English reform in Hainan Medical University has achieved initial success.Chinese students from international classes are superior to those from regular classes in many aspects.However,there are still many problems,and effective measures should be implemented to promote teaching quality continuously.

Structural and biochemical studies of RIG-I antiviral signaling

Retinoic acid-inducible gene I (RIG-I) is an important pattern recognition receptor that detects viral RNA and triggers the production of type-I interferons through the downstream adaptor MAVS (also called IPS-1, CARDIF, or VISA). A series of structural studies have elaborated some of the mechanisms of dsRNA recognition and activation of RIG-I. Recent studies have proposed that K63-linked ubiquitination of, or unanchored K63-linked polyubiquitin binding to RIG-I positively regulates MAVS-mediated antiviral signaling. Conversely phosphorylation of RIG-I appears to play an inhibitory role in controlling RIG-I antiviral signal transduction. Here we performed a combined structural and biochemical study to further define the regulatory features of RIG-I signaling. ATP and dsRNA binding triggered dimerization of RIG-I with conformational rearrangements of the tandem CARD domains. Full length RIG-I appeared to form a complex with dsRNA in a 2:2 molar ratio. Compared with the previously reported crystal structures of RIG-I in inactive state, our electron microscopic structure of full length RIG-I in complex with blunt-ended dsRNA, for the first time, revealed an exposed active conformation of the CARD domains. Moreover, we found that purified recombinant RIG-I proteins could bind to the CARD domain of MAVS independently of dsRNA, while S8E and T170E phosphorylation-mimicking mutants of RIG-I were defective in binding E3 ligase TRIM25, unanchored K63-linked polyubiquitin, and MAVS regardless of dsRNA. These findings suggested that phosphorylation of RIG inhibited downstream signaling by impairing RIG-I binding with polyubiquitin and its interaction with MAVS.

ERK, JNK/AP-1 pathway was involved in silica-induced cell cycle changes / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the alteration of activator protein-1 (AP-1) luciferase activity in human embryo lung fibroblasts (HELF) after exposed to silica, and the role of mitogen activated protein kinase (MAPK)/AP-1 pathway on silica-induced cell cycle changes.</p><p><b>METHODS</b>After HELF cells were treated with 200 microg/ml silica, immunofluorescence assays were employed to detect the translocation and the phosphorylation level of extracellular signal-regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK), flow cytometry was used to detect the distributions of cell cycle, the dominant negative mutant of ERK, JNK and p38 were applied to detect the upstream or downstream relationship of signaling pathways.</p><p><b>RESULTS</b>After HELF-AP-1 cells were exposed to 200 microg/ml silica 6, 12, 24 h respectively, silica exposure lead to AP-1 activation in a time-dependent manner, inducing significant AP-1 activation at 6 h, reaching a maximum activation at 12 h, and having a little decrease at 24 h. After silica exposure 1 h, phosphorylation level of ERK and JNK increased mainly in cytoplasm, however, after exposure 2 h, they translocated to nucleus. The proportion of cells in G1 phases was decreased from (63.80 +/- 9.57)% to (32.23 +/- 7.22)%, and the proportion of cells in S phases was increased from (35.17 +/- 10.33)% to (66.00 +/- 8.07)% after exposed to silica 24 h. Curcumin, a chemical inhibitor of AP-1, impaired the decrease of cells in G1 phases. Furthermore we found expression of dominant-negative mutant of ERK and JNK impaired silica-induced AP-1 activation, whereas, dominant-negative mutant of p38 did not show the effect.</p><p><b>CONCLUSION</b>These result suggested that 200 microg/ml silica exposure can induce AP-1 activation, induce cell cycle changes through ERK, JNK/AP-1-dependent pathway.</p>

Activated protein 1-cyclin D1/E2F 1 pathways involved in cell cycle changes induced by benzo (a) pyrene / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the roles of activated protein 1 (AP-1) in cell cycle changes on human embryo lung fibroblasts (HELF) induced by benzo (a) pyrene [B (a) P], and relationships between AP-1 and cyclin D1/CDK4-E2F-1/4.</p><p><b>METHODS</b>Cells transfected with AP-1 luciferase reporter plasmid (AP-H) were cultured with serum-free RPMI1640 for 48 h, and treated with 2 micromol/L B (a) P for 24 h. AP-1 relative activity was detected by luciferase assay. Changes of cell cycle and the expression of cyclin D1, CDK4 and E2F-1/4 were checked using the flow cytometer and Western blot assay.</p><p><b>RESULTS</b>After B (a) P was treated for 24 h, the ratio of G1 phase cells (71 +/- 2)% was decreased to (48 +/- 3)% (P < 0.05), and an increase was observed in the ratio of S phase. AP-1 activity and cyclin D1/E2F-1 expression were increased significantly, but CDK4/E2F-4 expression did not change after B (a) P treatment. When AP-1 activity was inhibited by curcumin, decreases of G1 phase in response to B (a) P treatment were blocked, and overexpression of cyclin D1/E2F-1 was attenuated, but CDK4/E2F-4 expression was not changed significantly.</p><p><b>CONCLUSION</b>AP-1 is involved in B (a) P induced cell cycle changes, and is the upstream signals of cyclin D1/E2F-1, but not CDK4/E2F-4.</p>

Effects of p53 in benzo (a) pyrene induced p21 and E2F-1 expression and cell cycle changes / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the roles of p53 in cell cycle changes on human embryo lung fibroblasts (HELF) induced by benzo(a) pyrene[ B(a) P], and relationships between p53 and p21, E2F-1.</p><p><b>METHODS</b>Cells transfected with p53 siRNA plasmid (p53-H) and CMV vector (HELF/CMV) were cultured with serum-free R/MINI-1640 for 48 hours, then treated with 2 micromol/L B(a)P for 24 hours. Flow cytometry assay was used for detecting the cell cycle alteration after being exposed to B(a)P. Changes of p53 and p21 expressions were checked using Western blot assay, and the cytoplasmic and nuclear extraction was used to observe the subcellular localizations of p53 and p21. The immunofluorescence assay was used to check changes of E2F-1 expression and the distribution of E2F-1 in nuclear and cytoplasm after exposed to B (a)P. p53siRNA plasmid and the chemical inhibitor of p53 [pifithrin-alpha (PFT)] were used to observe effects of p53 in B(a)P induced cell cycle changes and the relationships of p53 and p21, E2F-1.</p><p><b>RESULTS</b>After 2 micromol/L B(a)P exposure, the ratio of G1 phase cells (71 +/- 5)% was decreased to (39 +/- 4)% (P < 0.05). p53, p21 and E2F-1 expressions were increased significantly, and over expressed proteins were mostly located in nuclear after B(a)P treatment. When p53 expression was inhibited by p53 siRNA or PFT, the decreases of G1 phase in response to B(a)P treatment still existed, and over expression of p21 induced by B(a)P was attenuated, especially in nuclear, but E2F-1 over expression was not changed significantly.</p><p><b>CONCLUSION</b>B(a)P could induce cell cycle changes through p53 independent pathways. And p53 could regulate p21 expression positively, but not E2F-1.</p>

c-Jun NH2-terminal kinase and extracellular signal-regulated protein kinase signaling pathways in regulation of benzo(a)pyrene-induced c-Jun activation in human embryo lung fibroblasts / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the role of mitogen activated protein kinases (MAPKs) signaling pathways in the regulation of benzo(a)pyrene (B(a)P)-induced c-Jun activation in human embryo lung fibroblasts (HELFs).</p><p><b>METHODS</b>HELFs were cultured with 2.0 micromol/L B(a)P for various time (0, 3, 6, 12, 24 h) or with various concentration of B(a)P (0.0, 0.5, 1.0, 2.0 micromol/L) for 12 h. Western blot was performed to examine the effect of B(a)P on c-Jun activation. The dominant negative mutants of p38, c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated protein kinase (ERK) were applied to establish stable transfectant, and to detect the relationship of MAPK signal molecules and c-Jun activation in B (a) P-treated cells.</p><p><b>RESULTS</b>B(a)P treatment resulted in a marked activation of c-Jun in time-dependent manner with a peak at 12 h (the densitometric ratios of phosphorylated c-Jun Ser63, Ser73 to actin were 20.1, 15.2 times for control respectively) and in dose-dependent manner. However, there was no evident change on total c-Jun expression in B(a)P-treated HELFs. Moreover, B(a)P-induced activation of c-Jun was inhibited by stable expression of dominant negative mutants of JNK or ERK, but not by dominant negative mutant of p38.</p><p><b>CONCLUSION</b>JNK and ERK signaling pathways, but not p38 pathway regulate B(a)P-induced c-Jun activation in HELFs.</p>