AIM: To explore the anticancer function of Shp2 in lung adenocarcinoma A549 cells and the related molecular mechanisms.METHODS: The viability and proliferation of A549 cells treated with Shp2 specific inhibi-tor Phps-1 or cisplatin (DDP) were measured by CCK-8 assay and EdU assay.Annexin V-FITC/PI double staining was ap-plied to detect apoptotic rate of A549 cells with different interventions.The protein levels of caspase-3-17p, Bcl-2, Bax, p-STAT3 /STAT3 and p-ERK/ERK were determined by Western blot.RESULTS: Compared with control group, Phps-1 at the concentration of 20 μmol/L significantly increased the viability of A549 cells after 24 h of treatment ( P <0.05). Meanwhile, the proliferation rate of A549 cells in Phps-1 20 μmol/L group was significant increased compared with control group (P <0.05).The apoptotic rate of A549 cells in DDP treatment group decreased from 13.01% ±2.62% to 3.67%±0.93% after adding Phps-1 (P <0.05).Phps-1 down-regulated the protein levels of caspase-3-17p, Bax and p-ERK, but up-regulated p-STAT3.CONCLUSION: Shp2 is a tumor suppressor in A549 cells, which may be associated with the activation of STAT3 signal pathway.

We analyszed 108 examination papers that were taken when the students of 8 groups had finished theexercitation. The attainment was 73.6?4.4 points. Among the 400 selective questions, the difficult questions whichwere concentrated only on a few diseases accounted for 35.3%. There was mush difference between the proportion of theselective questions and that of the demands of the teaching program in different system of diseases. There was few or noselective question in the important diseases of the teaching program. It is suggested that the proposition of difficultquestions must include the important diseases in the teaching program. The diseases that are not commonly encountereddiseases may be deleted from the teaching program, but the commonly encountered must be put in the teaching program.It is necessary to reinforce the ability of students to analyse and resolve problems[

Objective To investigate the relation between the characteristics of CD8+T lymphocyte infiltration and the prognosis of triple-negative breast cancer patients. Methods We retrospectively analyzed the clinicopathological data of 126 patients with triple-negative breast cancer undergoing preoperative neoadjuvant chemotherapy. Immunohistochemical staining was used to analyze the relation between CD8+T lymphocyte infiltration and clinicopathological characteristics. Kaplan-Meier method was used to draw the survival curve, and Cox risk ratio regression model was used to analyze the prognostic factors affecting disease-free survival time (DFS). Results High-density CD8+Tils was associated with age < 60 years old, high pathological grade and high clinical stage (P < 0.05). The pCR rate of high-density CD8+Tils group was higher than that of the low-density group (66.7% vs. 19.8%, P=0.000). The median DFS of the high-density group was significantly longer than that of the low-density group (49 vs. 25 months, P < 0.05). Multivariate analysis showed that high pathological grade, tumor diameter > 2 cm, lymph node metastasis, vascular invasion and CD8+Tils low-density infiltration were factors for poor prognosis (P < 0.05), and CD8+Tils was an independent prognostic factor. Conclusion CD8+Tils may be an independent prognostic indicator for triple-negative breast cancer. The patients with high-density infiltration have high postoperative pCR rate, long DFS and better long-term efficacy.

BACKGROUND:Previous studies have found that qi deficiency and blood stasis syndrome is the main syndrome among various TCM syndromes of cervical spondylotic myelopathy.However,there is no report on proteomic markers as early diagnosis indicators for the transformation of developmental cervical spinal stenosis with qi deficiency and blood stasis syndrome to cervical spondylotic myelopathy. OBJECTIVE:To explore serum proteomics difference between developmental cervical spinal stenosis and cervical spondylotic myelopathy and to find and identify the potential serum biomarkers between them. METHODS:Serum samples of nine patients with cervical spondylotic myelopathy of qi deficiency and blood stasis syndrome(experimental group)and nine patients with developmental cervical spinal stenosis of qi deficiency and blood stasis syndrome(control group)were collected.The proteomic analysis was carried out by Tandem Mass Tag combined with liquid chromatography tandem mass spectrometry,so as to find and identify differentially expressed proteins. RESULTS AND CONCLUSION:A total of 1027 significantly differential proteins were initially screened by TMT technology and 89 significantly differential proteins were finally identified(P<0.05).Compared with the control group,there were 45 up-regulated proteins in the experimental group,such as α-actinin-4,α-actinin-1,cell division control protein 42 homolog,integrin-linked protein kinase and B-actin.Conversely,there were 44 down-regulated proteins in the experimental group compared with the control group,such as fibronectin,fibrinogen γ chain,fibrinogen α chain,fibrinogen β chain.Gene ontology enrichment analysis indicated that these differential proteins were involved in signal receptor binding,kinase binding,protein kinase activity,integrin binding,actin filament binding and other molecular functions.Based on the Kyoto Encyclopedia of Genes and Genomes pathway analysis,20 common differential signal/metabolic pathways were identified,including Rap1 signaling pathway,adherens junction,tight junction,platelet activation,and regulation of actin cytoskeleton.Protein-protein interaction analysis showed that ILK,FGA,FGB,FGG,FN1,Cdc42,ACTN1,ACTN4 and ACTB were located at the nodes of protein-protein interaction network and were closely related to bone formation and destruction system,nervous system,coagulation system,cellular inflammation and other systems.To conclude,the serum differentially expressed proteins between developmental cervical spinal stenosis and cervical spondylotic myelopathy can be successfully screened by Tandem Mass Tag combined with liquid chromatography tandem mass spectrometry.ILK,FN1,CDC42 and ACTN 4 are identified as specific markers for the transformation of developmental cervical spinal stenosis with qi deficiency and blood stasis syndrome into cervical spondylotic myelopathy.These findings provide a basis for further clarifying the transformation mechanism.

BACKGROUND:Serum-specific biomarkers between normal healthy individuals and populations with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome)have not been fully defined. OBJECTIVE:To screen and identify the potential biomarkers of developmental cervical canal stenosis with Qi deficiency and blood stasis. METHODS:Serum samples were collected from nine patients with developmental cervical canal stenosis with Qi deficiency and blood stasis and eight healthy people.Differentially expressed proteins in serum were screened and identified using isotope relative labeling and absolute quantification combined with liquid chromatography tandem mass spectrometry.Western blot was used to verify some significant differentially expressed proteins. RESULTS AND CONCLUSION:A total of 61 differentially expressed proteins(P<0.05)were identified using tandem mass spectrometry techniques.Compared with the healthy normal population group,14 differentially expressed proteins such as complement component C1q receptor,apolipoprotein A4,and C-C motif chemokine ligand 18 were significantly upregulated,while 47 differentially expressed proteins such as myosin light chain 3,mitochondrial translation elongation factor,and nucleolar phosphoprotein 1 were significantly downregulated.The results of gene ontology enrichment analysis indicated that these differentially expressed proteins might participate in molecular functions such as regulation of chromosomal tissue,mitochondrial membrane tissue,and muscle system processes.Protein-protein interaction network analysis showed that 38 common differential proteins,including complement component C1q receptor,apolipoprotein A4,C-C motif chemokine ligand 18,myosin light chain 3,mitochondrial translation elongation factor,and nucleolar phosphoprotein 1,were located at functional network nodes between healthy normal individuals and those with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome),and were closely related to the local energy metabolism of the cervical spine,the production of cervical vertebral osteocytes,and the formation of osteoclasts.The main differentially expressed protein myosin light chain 3 was validated using western blot assay,and the validation results were consistent with the proteomic results.To conclude,the preliminary discovery of differentially expressed proteins in serum between healthy normal individuals and those with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome)through absolute quantitative technology combined with liquid chromatography tandem mass spectrometry technology suggests that myosin light chain 3 may be a specific serum marker for developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome).

To investigate the effects of airway epithelial cells on macrophages chemotaxis and inflammatory cytokine expression under hypoxic conditions.
 Methods: Human bronchial epithelial cells (HBE) treated with different concentrations (0, 100, 200, 400, 800 μmol/L) of CoCl2 or transfected with HIF-1α siRNA were co-cultured with THP-1-derived M1 macrophages or M2 macrophages. The chemotactic effects on macrophages were analyzed by Transwell assay. The levels of TNF-α, IFN-γ, IL-4, IL-13 and IL-10 in the supernatants of macrophages were detected by ELISA, and HIF-1α or Cav-1 mRNA expression in HBE or macrophages was detected by RT-qPCR.
 Results: HBE cells promoted macrophages chemotaxis in a time- and concentration-dependent manner. Compared to un-transfected group, the chemotactic ability of HBE transfected with HIF-1α siRNA was significantly weakened (P<0.01). Under the same culture conditions, the chemotaxis of M2 macrophages was greater than that in THP1-derived M1 macrophages. The concentrations of TNF-α, IFN-γ, IL-4, IL-13 and IL-10 in the supernatants of macrophages were increased in a time-and concentration-dependent manner. The concentrations of TNF-α and IFN-γ were increased further after co-culturing for 8 and 12 h; while IL-4, IL-13 and IL-10 concentrations were increased further during 24 h of co-culture. The levels of cytokines in the supernatants of macrophages co-cultured with HBE and transfected with HIF-1α siRNA were significantly lower than those in un-transfected cells (P<0.05 or P<0.01). The reduction of TNF-α or IFN-γ was more obvious. The expression of HIF-1α or Cav-1 mRNA in HBE or macrophages was increased in a concentration-dependent manner after 8 or 12 h co-culture, which was significantly reduced when HBE was transfected with HIF-1α siRNA.
 Conclusion: Airway epithelial cells can enhance macrophages chemotaxis and pro-inflammatory cytokines expressions under hypoxic condition. HIF-1α and Cav-1 may be the important mediators in these processes.
Cell Hypoxia ; Chemotaxis ; Cytokines ; Epithelial Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; Macrophages

OBJECTIVETo explore the spatial and temporal distributions of animal plague in Junggar Basin natural plague focus.

METHODSData regarding plague antibody (F1) in serum of Great Gerbil (Rhombomys opimus, R. opimus) which were collected from 2005 to 2012 in Junggar Basin and analyzed. The changing rates on the positivity of F1 that appeared spatially and temporally were also analyzed.

RESULTSA total of 4 825 R. opimus serum samples were collected in 13 administrative regions in Junggar Basin.

RESULTSshowed that plague R. opimus existed in two areas-Gurbantonggut desert in the eastern-center and the clay desert of western Junggar Basin. However, in these two areas, the intensity of animal plague prevalence was different. In the former region where Yesinia pestis positive serum was detected from R. opimus, the detected rate of R. opimus was 8.39%. However, in the latter areas, the average positive rate was 1.56%. The changing trends of R. opimus plague prevalence were also varied annually. In the western Junggar Basin, the trend showed a slowly downward profile. The serum positive rate of R. opimus for Yesinia pestis decreased, from 7.59% in 2005 to 0.61% in 2008, and appeared as a resting state that none of the positive sample could be found since then. However, in the eastern-center Junggar Basin area-also named as Gurbantonggut desert which had been divided into 3 segments(western, central and eastern, according to related geographical characteristics), the changing trends of animal plague seemed quite complex. In the western segment, the animal plague had two epidemic peaks-in 2006 and 2010, with the interval of 4 years, with the higher peak of all the three geographic segments as 45.65% in 2010 and the positive serum of R. opimus for plague could be detected each year from 2006 to 2012. However, there were 3 epidemic peaks in the same period in the central and eastern segments. In the central segment, the peaks appeared in 2006, 2009 and 2011, with the intervals as 2.5 years and the average positive rate 8.92% was seen the lowest in Gurbantonggut desert. In the eastern segment, the first 2 peaks appeared the same season as in the central segment, but the third peak appeared in 2012, with the peak interval as 3 years. The positive rate of R. opimus for plague was also different in seasons, with the positive rate higher in autumn than in spring. These findings showed that the animal plague could be continuously prevalent from spring to autumn in the natural foci of plague in the Junggar Basin.

CONCLUSIONBoth geographical and temporal fluctuations of animal plague existed in the natural foci of Junggar Basin which was also named as geographical heterogeneity. Consequently, animal plague could be divided into two areas-the clay plains desert in the western and the Gurbantonggut desert in the eastern-center Junggar Basin.

As confusion mounts over RNA isoforms involved in phenotypic plasticity, aberrant CpG methylation-mediated disruption of alternative splicing is increasingly recognized as a driver of intratumor heterogeneity (ITH). Protease serine 3 (PRSS3), possessing four splice variants (PRSS3-SVs; PRSS3-V1-V4), is an indispensable trypsin that shows paradoxical effects on cancer development. Here, we found that PRSS3 transcripts and their isoforms were divergently expressed in lung cancer, exhibiting opposing functions and clinical outcomes, namely, oncogenic PRSS3-V1 and PRSS3-V2 versus tumor-suppressive PRSS3-V3, by targeting different downstream genes. We identified an intragenic CpG island (iCpGI) in PRSS3. Hypermethylation of iCpGI was mediated by UHRF1/DNMT1 complex interference with the binding of myeloid zinc finger 1 (MZF1) to regulate PRSS3 transcription. The garlic-derived compound diallyl trisulfide cooperated with 5-aza-2'-deoxycytidine to exert antitumor effects in lung adenocarcinoma cells through site-specific iCpGI demethylation specifically allowing MZF1 to upregulate PRSS3-V3 expression. Epigenetic silencing of PRSS3-V3 via iCpGI methylation (iCpGIm) in BALF and tumor tissues was associated with early clinical progression in patients with lung cancer but not in those with squamous cell carcinoma or inflammatory disease. Thus, UHRF1/DNMT1-MZF1 axis-modulated site-specific iCpGIm regulates divergent expression of PRSS3-SVs, conferring nongenetic functional ITH, with implications for early detection of lung cancer and targeted therapies.