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Objective To investigate the capillarization of liver sinusoidal endothelial cells (LSECs) and its association with hepatic fibrosis during the development of alveolar echinococcosis, so as to provide the basis for unraveling the mechanisms underlying the role of LSEC in the development and prognosis of hepatic injuries and hepatic fibrosis caused by alveolar echinococcosis. Methods Forty C57BL/6 mice at ages of 6 to 8 weeks were randomly divided into a control group and 1-, 2- and 4-week infection groups, of 10 mice in each group. Each mouse in the infection groups was intraperitoneally injected with 2 000 Echinococcus multilocularis protoscoleces, while each mouse in the control group was given an equal volume of phosphate-buffered saline using the same method. All mice were sacrificed 1, 2 and 4 weeks post-infection and mouse livers were collected. The pathological changes of livers were observed using hematoxylin-eosin (HE) staining, and hepatic fibrosis was evaluated through semi-quantitative analysis of Masson’s trichrome staining-positive areas. The activation of hepatic stellate cells (HSCs) and extracellular matrix (ECM) deposition were examined using immunohistochemical staining of α-smooth muscle actin (α-SMA) and collagen type I alpha 1 (COL1A1), and the fenestrations on the surface of LSECs were observed using scanning electron microscopy. Primary LSECs were isolated from mouse livers, and the mRNA expression of LSEC marker genes Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf was quantified using real-time fluorescence quantitative PCR (qPCR) assay. Results Destruction of local liver lobular structure was observed in mice 2 weeks post-infection with E. multilocularis protoscoleces, and hydatid cysts, which were surrounded by granulomatous tissues, were found in mouse livers 4 weeks post-infection. Semi-quantitative analysis of Masson’s trichrome staining showed a significant difference in the proportion of collagen fiber contents in mouse livers among the four groups (F = 26.060, P < 0.001), and a higher proportion of collagen fiber contents was detected in mouse livers in the 4-week infection group [(11.29 ± 2.58)%] than in the control group (P < 0.001). Immunohistochemical staining revealed activation of a few HSCs and ECM deposition in mouse livers 1 and 2 weeks post-infection, and abundant brown-yellow stained α-SMA and COL1A1 were deposited in the lesion areas in mouse livers 4 weeks post-infection, which spread to surrounding tissues. Semi-quantitative analysis revealed significant differences in α-SMA (F = 7.667, P < 0.05) and COL1A1 expression (F = 6.530, P < 0.05) in mouse levers among the four groups, with higher α-SMA [(7.13 ± 3.68)%] and COL1A1 expression [(13.18 ± 7.20)%] quantified in mouse livers in the 4-week infection group than in the control group (both P values < 0.05). Scanning electron microscopy revealed significant differences in the fenestration frequency (F = 37.730, P < 0.001) and porosity (F = 16.010, P < 0.001) on the surface of mouse LSECs among the four groups, and reduced fenestration frequency and porosity were observed in the 1-[(1.22 ± 0.48)/μm2 and [(3.05 ± 0.91)%] and 2-week infection groups [(3.47 ± 0.10)/μm2 and (7.57 ± 0.23)%] groups than in the control group (all P values < 0.001). There was a significant difference in the average fenestration diameter on the surface of mouse LSECs among the four groups (F = 15.330, P < 0.001), and larger average fenestration diameters were measured in the 1-[(180.80 ± 16.42) nm] and 2-week infection groups [(161.70 ± 3.85) nm] than in the control group (both P values < 0.05). In addition, there were significant differences among the four groups in terms of Stabilin-1 (F = 153.100, P < 0.001), Stabilin-2 (F = 57.010, P < 0.001), Ehd3 (F = 31.700, P < 0.001), CD209b (F = 177.400, P < 0.001), GATA4 (F = 17.740, P < 0.001), and Maf mRNA expression (F = 72.710, P < 0.001), and reduced mRNA expression of Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf genes was quantified in three infection groups than in the control group (all P values < 0.001). Conclusions E. multilocularis infections may induce capillarization of LSECs in mice, and result in a reduction in the expression of functional and phenotypic marker genes of LSECs, and capillarization of LSECs occurs earlier than activation of HSC and development of hepatic fibrosis.
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OBJECTIVES@#This study aims to investigate the effects of tumor-stromal fibroblasts (TSFs) on the proliferation, invasion, and migration of salivary gland pleomorphic adenoma (SPA) cells in vitro.@*METHODS@#Salivary gland pleomorphic adenoma cells (SPACs), TSFs, and peri-tumorous normal fibroblasts (NFs) were obtained by tissue primary culture and identified by immunocytochemical staining. The conditioned medium was obtained from TSF and NF in logarithmic phase. SPACs were cultured by conditioned medium and treated by TSF (group TSF-SPAC) and NF (group NF-SPAC). SPACs were used as the control group. The proliferation, invasion, and migration of the three groups of cells were detected by MTT, transwell, and scratch assays, respectively. The expression of vascular endothelial growth factor (VEGF) in the three groups was tested by enzyme linked immunosorbent assay (ELISA).@*RESULTS@#Immunocytochemical staining showed positive vimentin expression in NF and TSF. Results also indicated the weak positive expression of α-smooth muscle actin (SMA) and fibroblast activation protein (FAP) in TSFs and the negative expression of α-SMA and FAP in NFs. MTT assay showed that cell proliferation in the TSF-SPAC group was significantly different from that in the NF-SPAC and SPAC groups (P<0.05). Cell proliferation was not different between the NF-SPAC and SPAC groups (P>0.05). Transwell and scratch assays showed no difference in cell invasion and migration among the groups (P>0.05). ELISA showed that no significant difference in VEGF expression among the three groups (P>0.05).@*CONCLUSIONS@#TSFs may be involved in SPA biological behavior by promoting the proliferation of SPACs but has no effect on the invasion and migration of SPACs in vitro. Hence, TSF may be a new therapeutic target in SPA treatment.
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Humans , Adenoma, Pleomorphic/metabolism , Vascular Endothelial Growth Factor A , Culture Media, Conditioned/metabolism , Fibroblasts/metabolism , Salivary Glands/metabolismABSTRACT
Objective To evaluate the immunoprotective effect of active immunization with recombinant peptidyl-prolyl cis-trans isomerase from Babesia microti against B. microti infection in mice. Methods Female BALB/c mice at 6 weeks of age, each weighing approximately 20 g, were divided into the recombinant protein immunization group, the infection control group and the normal control group, of 25, 18, 15 mice in each group, respectively. Mice in the recombinant protein immunization group were given active immunization with recombinant BmPPIase protein, and 18 mice with the highest antibody titers were intraperitoneally injected with 100 μL of B. microti-infected whole blood 2 weeks after the last immunization. Mice in the infection control group were intraperitoneally injected with 100 μL of B. microti-infected whole blood, while 15 mice in the normal control group received no treatment. Blood samples were collected from mice in the recombinant protein immunization group and the infection control group on days 0 to 30 post-immunization for detection of B. microti infection, and blood samples were collected on days 0, 7, 14, 21, and 28 post-immunization for routine blood tests with a blood cell analyzer and for detection of serum cytokines using cytometric bead array. Results Anti-BmPPIase antibodies were detected in 25 mice in the recombinant protein immunization group 2 weeks after the last immunization, with titers of 5 × 103 to 8 × 104. B. microti infection rate peaked in mice in both the recombinant protein immunization and the infection control group on day 7 post-immunization, with positive infection rates of 13.3% and 50.0%, and there were significant differences between the two groups in terms of B. microti infection rate on days 3 (χ2= 113.18, P < 0.01), 5 (χ2 = 475.22, P < 0.01), 7 (χ2 = 465.98, P < 0.01) and 9 post-infection (χ2= 18.71, P < 0.01), while the B. microti infection rate tended to be 0 in both groups on day 11 post-immunization. Routine blood tests showed higher red blood cell counts [(5.30 ± 0.50) × 1012 to (9.87 ± 0.24) × 1012 counts/L)] and hemoglobin levels [(89.67 ± 22.80) to (148.60 ± 3.05) g/L)] in the recombinant protein immunization group than in the infection control group on days 0 to 28 post-immunization. Cytometric bead array detected higher serum interferon-γ [(748.59 ± 17.56) to (3 858.28 ± 1 049.10) fg/mL], tumor necrosis factor-α [(6 687.34 ± 1 016.64) to (12 708.13 ± 1 629.79) fg/mL], interleukin (IL)-6 [(611.05 ± 75.60) to (6 852.68 ± 1 554.00) fg/mL] and IL-17a [(167.68 ± 185.00) to (10 849.27 ± 355.40) fg/mL] and lower IL-10 levels [(247.65 ± 138.00) to (18 787.20 ± 2 830.22) fg/mL] in the recombinant protein immunization group than in the infection control group during the study period. Conclusions Recombinant BmPPIase protein induces up-regulation of interferon-γ, tumor necrosis factor-α and presents a high immunoprotective activity against B. microti infection in mice, which is a potential vaccine candidate protein.
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Objective:To explore and analyze the influencing factors of Helicobacter pylori (HP) infection in obese preschool children.Methods:From May 2020 to May 2022, 50 obese preschool children with HP infection and 50 obese preschool children without HP infection were prospectively selected from Tangshan Maternal and Child Health Hospital. We collected and collected basic data on two groups of children, including gender, age, eating habits, and family related information. Single factor analysis was used to screen for possible influencing factors of HP infection in obese preschool children, and logistic regression analysis was performed on the selected influencing factors.Results:Single factor analysis of variance showed that there were statistically significant differences in the prevalence of HP infection among obese preschool children of different ages, body mass index (BMI), sharing tableware, per capita monthly income of families, using public chopsticks, washing hands before meals, partial eating, habit of biting fingers, family population, binge eating, family history of HP infection, etc. (all P<0.05). Multivariate logistic analysis showed that BMI ( OR=1.576, 95% CI: 1.119-2.221), shared tableware ( OR=1.317, 95% CI: 1.018-1.702), per capita monthly income of households ( OR=1.330, 95% CI: 1.021-1.733), no use of public chopsticks ( OR=1.408, 95% CI: 1.019-1.945), washing hands before meals ( OR=1.206, 95% CI: 1.041-1.397), habit of biting fingers ( OR=1.470, 95% CI: 1.064-2.031), binge eating ( OR=1.443, 95% CI: 1.004-2.074) and family history of HP infection ( OR=1.317, 95% CI: 1.051-1.649) were the influencing factors of HP infection in obese preschool children (all P<0.05). Conclusions:BMI, shared tableware, per capita monthly income of families, failure to use public chopsticks, washing hands before meals, habit of biting fingers, binge eating, family history of HP infection, etc. are the influencing factors of HP infection in obese preschool children. If there are children with abnormal related factors, attention should be paid to them. If there are related infection symptoms, targeted interventions can be taken in time to prevent and control children′s HP infection.
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With the widespread application of low-dose CT in lung cancer screening, more and more pulmonary nodules are detected. Early diagnosis and treatment of pulmonary nodules can significantly reduce the risk of death from lung cancer. However, a series of problems still need to be solved in the early diagnosis and treatment of lung cancer, such as how to avoid over-diagnosis or under-diagnosis through necessary and accurate examination, how to determine the timing of surgery and how to adopt the best surgical method. This article will focus on the diagnostic value of imaging examination, tumor markers and invasive examination in the evaluation of benign and malignant pulmonary nodules, as well as the key clinical issues such as timing, method and extent of resection in the surgical treatment of pulmonary nodules, sorting out the clinical diagnosis and treatment ideas, and discussing management strategies, so as to provide references for the standardized clinical diagnosis and treatment of pulmonary nodules.
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Objective:To summarize the clinical characteristics of primary retroperitoneal tumors (PRT).Methods:All PRT cases undergoing surgical resection during recent 10 years at our center were retrospectively analyzed.Results:Tumors in all 92 cases were of malignant in 64 cases, borderline in 10 and benign PRT in 18, among which liposarcoma and leiomyosarcoma were the most common types. The tumor size and Ki-67 was significantly higher in malignant compared to borderline or benign PRT. The multifocal rate was 50%, en-bloc resection rate was 72%, R 0 rate was 61%, and combined organ resection rate was 41% in malignant PRTs. Small intestine and the colon were the most frequently resected organs. During 9.3 years of follow-up period, the 1-, 3- and 5-year cumulative reoperation rate of malignant PRT was 10.6%, 44.7% and 62.9%, respectively, and the median re-operation period was 4.0 years. The 1-, 3- and 5-year cumulative survival rate was 90.1%, 73.0% and 64.2%, respectively, and the median survival period was 6.1 years. None of postoperative borderline or benign PRT recurred or needed re-operation or deceased. Conclusion:Most of PRTs were malignant, presenting themsehies as a challenge to surgery with unfaror prognosis.
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ArcScan Insight 100 very high-frequency(VHF)digital ultrasound scanner is a new ocular ultrasonic measuring instrument, which can detect and measure the anterior segment. It can be used for screening before corneal refractive surgery and follow-up after corneal refractive surgery, measuring anterior segment parameters before implantable collamer lens(ICL)implantation, predicting preoperative vault, measuring postoperative vault, early screening keratoconus, and diagnosing glaucoma, cataract and eye injuries, etc. Taking the advantages of a wide range examination of ultrasound biomicroscope(UBM)and simple operation of optical coherence tomography(OCT), it has a broad prospect for clinical application. In this paper, the measurement principle, application method, parameters and clinical application progress of ArcScan Insight 100 VHF digital ultrasound scanner are reviewed in detail.
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Echinococcosis is a zoonotic parasitic disease caused by Echinococcus infections, and this disorder may cause fibrosis of multiple vital organs, which may further progress into cirrhosis. Early-stage hepatic fibrosis is reversible, and unraveling the mechanisms underlying hepatic fibrosis induced by Echinococcus infections is of great significance for the prevention and treatment of early-stage hepatic fibrosis. Recently, the studies pertaining to hepatic fibrosis associated with Echinococcus infections focus on cytokines and immune cells. This review summarizes the advances in the mechanisms underlying host immune cells- and cytokines-mediated hepatic fibrosis in humans or mice following Echinococcus infections.
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Objective@#To understand the prevalence of low vision among Tujia and Han children and adolescents in Tujia inhabited areas, and to provide reference for the prevention and control of myopia in children and adolescents in minority areas.@*Methods@#A cluster sampling of Tujia and Han primary school students from two primary schools in Lichuan City, Enshi Tujia and Miao Autonomous Prefecture, Hubei Province (2 466 Tujia and 971 Han) were selected for visual acuity assessment. Univariate χ 2 test and multivariate Logistic analysis were used. Low vision and associated factors between Tujia and Han nationality were compared.@*Results@#The overall detection rate of low vision among children and adolescents in Tujia inhabited areas was 44.9%. There were differences in the degree of low vision in the left and right eyes of individuals, and the detection rate of low vision varied significantly by ethnic, gender and grade ( χ 2=22.10, 18.43, 19.06, 17.97 for the left eye, 17.52, 20.44, 21.49, 18.61 for the right eye, P < 0.05). There were many factors affecting low vision among children and adolescents in Tujia inhabited areas, overweight and obesity were negatively associated with low vision ( OR=1.81, 1.70, 95%CI=1.76-1.92, 1.66-1.82, P <0.01).@*Conclusion@#Low vision is highly prevalent in Tujia children and adolescents. Effective intervention measures should be taken to treat and prevent myopia in children and adolescents.
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Human sparganosis is a food-borne parasitic disease caused by the plerocercoids of Spirometra species. Clinical diagnosis of sparganosis is crucial for effective treatment, thus it is important to identify sensitive and specific antigens of plerocercoids. The aim of the current study was to identify and characterize the immunogenic proteins of Spirometra erinaceieuropaei plerocercoids that were recognized by patient sera. Crude soluble extract of the plerocercoids were separated using 2-dimensional gel electrophoresis coupled with immunoblot and mass spectrometry analysis. Based on immunoblotting patterns and mass spectrometry results, 8 antigenic proteins were identified from the plerocercoid. Among the proteins, cysteine protease protein might be developed as an antigen for diagnosis of sparganosis.
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Objective To construct a cDNA library of Sparganum mansoni and immunoscreen antigen candidates for immunodiagnosis of sparganosis mansoni. Methods Total RNA was extracted from S. mansoni, and reversely transcribed into cDNA, which was ligated into the phage vector. These recombinant vectors were packaged in vitro to construct the SMART cDNA library of S. mansoni. Then, the cDNA library was immunoscreened with sera from patients with sparganosis mansoni to yield positive clones. The inserted fragments of positive clones were sequenced and subjected to homology analyses, and the structure and functions of the coding proteins were predicted. Results The SMATR cDNA library of S. mansoni was successfully constructed. The titer of the cDNA library was 6.25 × 106 pfu/mL, with a recombinant efficiency of 100%, and the mean length of the inserted fragments in the library was larger than 1 100 bp. A total of 12 positive clones were obtained by immunoscreening, and were categorized into Sm-I (Sm60-1), Sm-II (Sm58-1), Sm-III (Sm20-1) and Sm-IV (Sm22-3), with 1 134, 1 063, 883 bp and 969 bp long inserted fragments. Their coding proteins were highly homologous with the Spirometra erinaceieuropaei antigenic polypeptide, cytoplasmic antigen, ribosomal protein S4-like protein and unnamed protein product, respectively. Conclusions A SMART cDNA library of S. mansoni has been successfully constructed and 4 categories of positive clones have been identified, which provides a basis for further studies on diagnostic antigens for sparganosis mansoni.
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OBJECTIVE@#To observe the effects of tripterine on adhesion molecules and cell biological characteristics in mice with acute promyelocytic leukemia (APL) tumor.@*METHODS@#Eighteen SCID beige mice were caudal vein injected with NB4 cell lines (5×10@*RESULTS@#The neutrophil decrased and promyelocytes, NB4 cells, B lymphocytes and white blood cells increased in tumor-bearing group as compared with control group (P<0.05), and the expressions of serum P-selectin (P-selectin), soluble vascular adhesion molecule-1 (soluble vascular adhesion molecule-1, sVCAM-1) and soluble intercellular adhesion molecule-1 (soluble intercellular adhesion molecule-1, sICAM-1) all increased (P<0.05). The cell cycle showed that the proportion of G@*CONCLUSION@#Tripterine may not only inhibit the expression of sVCAM-1 and sICAM-1 proteins in APL tumor-bearing mice and reduce the adhesion of tumor cells, but also block tumor cells at G
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Animals , Humans , Mice , Cell Cycle , Cell Division , Intercellular Adhesion Molecule-1 , Leukemia, Promyelocytic, Acute/drug therapy , Mice, SCID , Triterpenes , Vascular Cell Adhesion Molecule-1ABSTRACT
Objective To identify the species of common necrophagous flies in Fujian Province by gene fragment sequences of mitochondrial cytochrome c oxidase subunit Ⅰ （COⅠ） and 16S ribosomal deoxyribonucleic acid （16S rDNA）, and to explore the identification efficacy of these two molecular markers. Methods In total 22 common necrophagous flies were collected from the death scenes in 9 different regions in Fujian Province and DNA was extracted from the flies after morphological identification. The gene fragments of COⅠ and 16S rDNA were amplified and sequenced. All the sequences were uploaded to GeneBank and BLAST and MEGA 10.0 software were used to perform sequence alignment, homology analysis and intraspecific and interspecific genetic distance analysis. The phylogenetic trees of DNA fragment sequences of COⅠ and 16S rDNA of common necrophagous flies in Fujian Province were established by unweighted pair-group method with arithmetic means （UPGMA）, respectively. Results The flies were classified into 6 species, 5 genera and 3 families by morphological identification. The results of gene sequence analysis showed that the average number of interspecific and intraspecific genetic distance of 16S rDNA ranged from 1.8% to 8.9% and 0.0% to 2.4%, respectively. The average number of interspecific and intraspecific genetic distance of COⅠ ranged from 7.2% to 13.6% and 0.0% to 6.3%, respectively. Conclusion The gene sequences of COⅠ and 16S rDNA can accurately identify the species of different necrophagous flies, and 16S rDNA showed higher value in species identification of common calliphoridae necrophagous flies in Fujian Province.
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Animals , Humans , DNA, Ribosomal/genetics , Diptera/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Species SpecificityABSTRACT
Gene mutations can impair the sensitivity of cancer cells to targeted drugs, and lead to individual differences of clinical therapeutic effects. Epidermal growth factor receptor (EGFR) mutation plays an important role in therapeutic decision-making. Furthermore, some co-existing gene mutations, such as TP53 mutation, can also affect the therapeutic effect and prognosis of patients. Whether EGFR mutation combined with TP53 mutation affects the sensitivity of lung cancer cells to tyrosine kinase inhibitor (TKI) and long-term prognosis of non-small cell lung cancer (NSCLC) patients is still unknown and has attracted more attentions. However, in the current clinical practice, TP53 mutation is not a key factor of therapeutic decision-making, so further studies are needed to clarify the impact of TP53 mutation (including each subtype) on the potential benefits of EGFR-targeted therapy of NSCLC.
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Immunological checkpoint inhibitors have been approved for a short period of time in China, and real-world clinical data are still in the collection stage. Reports of domestic programmed death-1 (PD-1) treatment-related adverse reactions are rare. The author reported a case of hypotension in the process of Pembrolizumab infusion and successful infusion after blood pressure recovery, hoping to provide reference for the application of immunological checkpoint inhibitors, to provide patients with the greatest clinical benefit.
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The application of immunological checkpoint inhibitors (ICIs) has modified many treatment strategies of malignant tumors, which has become a milestone in cancer therapy. The principle of action can be explained as "brake theory". After releasing the brakes by ICIs, unprecedented systemic toxicities, even some refractory and fatal immune-related adverse effects (irAEs) may develop. In this article, we summarized the recommended treatments of grade 3-4 severe irAEs in the latest European Society for Medical Oncology (ESMO), National Comprehensive Cancer Network (NCCN)/American Society of Clinical Oncology (ASCO), Society for Immunotherapy of Cancer (SITC) and Chinese Society of Clinical Oncology (CSCO) guidelines and consensus. We also performed a systemic review of case reports and reviews of irAEs up to May 20, 2019 in PubMed and Chinese journals. Successful applications of specific immunosuppressive drugs and stimulating factors beyond the above guidelines and consensus were supplemented and highlighted, including agents blocking interleukin 6 (IL-6), rituximab, anti-tumor necrosis factor-α (TNFα) monoclonal antibody (mAb), anti-integrin 4 mAb, Janus kinase inhibitors, thrombopoietin receptor agonists and antithymocyte globulin (ATG) etc. We put some concerns of using high-dose steroids for long-term, and emphasize the secondary infections, tumor progression, and unavailability of ICI re-challenge during steroid treatment. We propose the "De-escalation Therapy" principle for severe and refractory irAEs, and suggest that immunosuppressive drugs specifically targeting cytokines should be used as early as possible. Many irAEs in the era of immunotherapy are unprecedented compared with traditional chemotherapy and small-molecule targeted therapy, which is a big challenge to oncologists. Therefore, the establishment of multidisciplinary system is very important for the management of cancer patients.
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Small cell lung cancer (SCLC) is a refractory cancer with high degree of malignancy, rapid disease progression, poor prognosis and easy recurrence. In the past 30 years, the traditional treatment of SCLC, mainly chemotherapy and radiotherapy, has not changed significantly, and the effective treatment method for clinical needs is extremely urgent. The rapid development of precision medicine has revealed the molecular biological characteristics of SCLC, so its diagnosis and treatment will into a new era. At present, some studies have shown that anti-angiogenic drugs, immunotherapy and so on have improved the efficacy of SCLC treatment to some extent, and there are more studies on the diagnosis and treatment of SCLC, so a new field of SCLC treatment are coming and bringing more survival benefits to patients. New studies on targeted therapy, anti-angiogenesis drugs and immunotherapy of molecular pathology of SCLC are emerging. This paper reviews the new diagnosis and treatment methods of SCLC to provide new guidance for its clinical treatment. .
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Animals , Humans , Angiogenesis Inhibitors , Immunologic Factors , Immunotherapy , Lung Neoplasms , Diagnosis , Drug Therapy , Small Cell Lung Carcinoma , Diagnosis , Drug TherapyABSTRACT
Objective To identify the expression of ribosomal protein S9(RPS9)in multiple myeloma(MM)and explore its effect on the biological characteristics of myeloma cells and the corresponding mechanisms. Methods Bone marrow mononuclear cells were harvested in 10 healthy volunteers(CON group)and bone marrow CD138 +cells from 30 MM patients(CD138+group).Quantitative polymerase chain reaction(qPCR)was performed to detect RPS9 expression at mRNA level.In three cases from CON group and 11 cases from CD138+group,Western blot was performed to detect RPS9 at protein level.GSE19784 dataset was employed to detect the relationships of RPS9 expression with the overall survival rate,nuclear factor-κB(NF-κB),small ubiquitin-like modifier(SUMO),and ubiquitin pathway.After the RPS9 knock-down vector was constructed,flow cytometry was performed to detect the infection efficiency and qPCR and Western blot to detect the knock-down efficiency.RPMI8226 was divided into CON group and RPS9-short hairpin RNA(shRNA)group,in which annexin V allophycocyanin/propidium iodide(PI)double staining was performed to detect the change of apoptosis,CCK8 to detect the proliferation change,and PI staining to detect cell cycle change.After sentrin-specific protease 1(SENP1)overexpression vector was constructed,Western blot was performed to detect the phosphorylation of P65 and inhibitory subunit-κBα(IκBα)from NF-κB pathway in CON,RPS9-shRNA,and RPS9-shRNA-SENP1 cells;in addition,annexin V/PI double staining was also performed to detect the apoptosis in these three cells. Results The relative expression of RPS9 in CON group and CD138+group was(1.00±0.12)and(5.45±0.71),respectively(t=4.291,P=0.0036).Western blot showed RPS9 expression was high in most myeloma CD138+cells.The high expression of RPS9 was associated with both extramedullary invasion and overall survival in GSE19784 dataset.After RPMI8226 was infected with CON or RPS9-shRNA lentivirus for 48 hours,flow cytometry confirmed that the infection efficiencies were above 90% in both groups.qPCR and Western blot confirmed that RPS9 expression was inhibited at both mRNA and protein levels.After RPMI8226 CON and RPS9-shRNA infected with virus for 48 hours,the proportion of annexin V-positive cells in CON and RPS9-shRNA cells was(3.47±0.37)% and(18.60±64.00)%(t=9.015,P=0.0008).The proliferation index significantly differed between CON group and RPS9-shRNA group at 72 hours(t=6.846,P=0.0024).When CON and RPS9-shRNA were infected with virus for 48 hours,the proportion of G2 phase cells was(29.28±3.42)% and(10.43±1.43)%,respectively(t=9.329,P=0.0007).The RPS9 expression was positively correlated with SENP1 in GSE19784 dataset and negatively correlated with IκBα coding gene NFKBIA.Western blot further confirmed that RPS9 knockdown inhibited the expression of SENP1,inhibited the phosphorylation of NF-κB subunit P65 and inhibitor IκBα,and promoted the expression of IκBα.Overexpression of SENP1 not only impeded this effect but also reduced RPS9-induced apoptosis. Conclusions RPS9 is highly expressed in MM CD138+cells and is associated with overall survival and extramedullary infiltration.Inhibition of RPS9 can promote apoptosis,cell cycle arrest,and proliferation of myeloma cells.RPS9 can affect the activation of NF-κB pathway and cell apoptosis through SENP1,suggesting that SENP1 may be a key factor in the biological effect of RPS9.
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Humans , Apoptosis , Cell Line, Tumor , Cell Proliferation , Cysteine Endopeptidases , Metabolism , Multiple Myeloma , Metabolism , Ribosomal Proteins , Metabolism , Signal TransductionABSTRACT
Objective: To investigate the anticancer effect of isoliquiritigenin (ISL) on human clear cell renal cell carcinoma 786-O cells, and explore its possible molecular mechanism. Method: Thiazolyl blue tetrazolium bromide (MTT) assay was used to detect effect of ISL (0, 10, 25,50, 75, 100 μmol·L-1) on proliferation of 786-O cells. The effect of ISL on migration and invasion of 786-O cells was detected by cell scratch test and Transwell assay. The autophagy was observed under the fluorescence microscope through acridine orange staining and Ad-GFP-LC3 transfection experiment. Western blot was used to detect the expression of autophagy related protein and analyze the changes of phosphatidylinositol-3-kinase (PI3K)/protein kinase B(Akt)/mammalian target of rapamycin (mTOR) signaling pathway to explore the possible mechanism. Result: MTT results showed that ISL could significantly inhibit the proliferation of 786-O cells in a time-dose dependent manner (PPPPPPPPConclusion: ISL can inhibit the proliferation, migration and invasion of clear cell renal carcinoma 786-O cells, and induce autophagy by inhibiting the PI3K/Akt/mTOR signaling pathway.
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BACKGROUND@#Simultaneity multiple primary lung cancer always is a rare disease, but in recent years due to the progress of the diagnosis and treatment means detection rate increased. This study summarized the clinical data of 31 cases of synchronous multiple primary lung cancer (sMPLC) analysis, further to explore the diagnosis, treatment and prognosis of MPLC.@*METHODS@#Sum up the clinical data of 31 cases of simultaneous multiple primary lung cancer, the diagnostic method, surgical methods, pathology, were retrospectively analyzed.@*RESULTS@#All the patients are thoracoscope surgery, no deaths. Thin high resolution computed tomography (CT) in preoperative found multiple lung nodules. Lesions located in the same side of the same period surgical treatment, the operation method is given priority to with under the thoracoscope lung + the lobectomy; Lesions located in bilateral, all staged operation, the time interval for 3 months-4 months.@*CONCLUSIONS@#Thin layer distinguish CT preoperative diagnosis is the best way to simultaneous multiple primary lung cancer. Multiple primary lung cancer incidence of ipsilateral lung at the same time higher than that of bilateral lung (23:8), type, around 94%, the most common histology to adenocarcinoma, 80.6% (25/31). Primary lesions under thoracoscope lobectomy plus the lobectomy of secondary lesions is the most commonly used.