This study investigated the "homing" phenomenon in hepatocellular carcinoma (HCC). The "homing" specificity of endothelial progenitor cells (EPC) by establishing an orthotopic implantation model in nude mice. EPCs harvested from the marrow cells were separated by density gradient centrifugation. Fluorescence microscope, flow cytometry (FCM) and double fluorescence staining with FITC-UEA-I and DiI-ac-LDL, were employed to identify the cells. 4',6-diamidino-2-phenylindole (DAPI) labelling and real-time PCR were used for detecting the expression of CD133 and chemokines to trace and observe the distribution of EPCs. Our results showed that the distribution rate of EPCs was obviously higher than that in other important organs and the negative control group. Detection of CD133 and chemokines yielded similar results in difference tissues. Our experiment confirmed that the chemotaxis of EPCs does exist in HCC. Moreover, HIF-1α, SDF-1 and VEGF might play important roles in the "homing" of EPCs in HCC. EPCs might be a potential candidate for targeting vector of HCC for gene therapy.

Objective:To evaluate the effect of tropisetron on pyroptosis in cardiomyocytes subjected to hypoxia-reoxygenation (H/R) and the relationship with α7 nicotinic acetylcholine receptor (α7nAChR).Methods:Routinely cultured H9C2 cardiomyocytes were divided into 4 groups ( n=6 each) using a random number table method: control group (C group), H/R group, tropisetron plus H/R group (Tro+ H/R group), and α7nAchR antagonist MLA plus tropisetron plus H/R group (MLA+ Tro+ H/R group). H/R was produced by 12 h exposure of cells to hypoxia followed by 6 h reoxygenation in the other three groups except group C. Tropisetron at the final concentration of 10 nmol/L was added at 1 h before hypoxia in group Tro+ H/R.In group MLA+ Tro+ H/R, MLA was added at 2 h before hypoxia, and then 1 h later tropisetron at the final concentration of 10 nmol/L was given.At 6 h of reoxygenation, the pyroptosis rate of cardiomyocytes was determined by fluorescence immunostaining of caspase-1-AlexaFluor 488/DAPI, the concentrations of interleukin-1beta (IL-1β) and IL-18 in the supernatant were determined by enzyme-linked immunosorbent assay, the activity of LDH in the supernatant was measured by 2, 4 dinitrophenylhydra-zine colorimetric method, and the expression of α7nAchR, NLRP3 and caspase-1 in cardiomyocytes was detected by Western blot. Results:Compared with group C, the pyroptosis rate, activity of LDH and concentrations of IL-1β and IL-18 in the supernatant were significantly increased, the expression of NLRP3 and caspase-1 was up-regulated, and the expression of α7nAchR was down-regulated in group H/R ( P<0.05). Compared with group H/R, the pyroptosis rate, activity of LDH and concentrations of IL-1β and IL-18 in the supernatant were significantly decreased, the expression of NLRP3 and caspase-1 was down-regulated, and the expression of α7nAchR was up-regulated in group Tro+ H/R ( P<0.05). Compared with group Tro+ H/R, the pyroptosis rate, activity of LDH and concentrations of IL-1β and IL-18 in the supernatant were significantly increased, the expression of NLRP3 and caspase-1 was up-regulated, and the expression of α7nAchR was down-regulated in group MLA+ Tro+ H/R ( P<0.05). Conclusion:α7nAchR is involved in the process of tropisetron inhibiting pyroptosis in cardiomyocytes subjected to H/R.

Objective To study the relationship between Ephrin-A1 and its receptor with angiogenesis in hepatocellular carcinoma(HCC).Methods Immunohistochemistry staining method(S P methods)and reverse transcription polymerase chain reaction(RT-PCR) were used to determine the protein and mRNA expression of Ephrin-A1 and its receptor EphA1、EphA2 in tumor tissues and their corresponding adjacent liver tissues from 52 HCC patients;then,analyse of the relationship between Ephrin-A1 and clinicopathologyfactor and microvessel density(MVD) in HCC was made.Results The protein expression rate of Ephrin-A1 and EphA1,EphA2 in HCC was 59.6%(31/52),53.8%(28/52)and 17.3%(9/52),respectively,but in the paired liver tissues adjacent to HCC the expression rate was 23.1%(12/52),and respectively.The protein expression rate of Ephrin-A1 and EphA1 was significantly higher than that in the paired liver tissues adjacent to HCC(P0.05).The mRNA express rate of Ephrin-A1 and EphA1 in HCC [67.3%(35/52) and 73.7%(38/52)] were prominently higher than those in the paired liver tissues adjacent to HCC [42.3%(22/52) and 48.1%(25/52)](P0.05).The higher expression of Ephrin-A1 was correlated with the AFP level and thrombus in the portal vein(P

Objective To explore the risk factors for lung cancer-related cerebral infarction . Methods The hospitalized active lung cancer patients on anti-cancer therapy with no traditional stroke risk factors, who experienced an acute cerebral infarct in the First Affiliated Hospital of Guangxi Medical University from January 2005 to December 2015, were consecutively collected as the LCRS ( lung cancer-related stroke) group.The active lung cancer patients without cerebral infarction hospitalized at the same peroid matched with the LCRS group for age and gender were collected as the LC ( lung cancer ) group. Clinical data from the two groups were analyzed .Results A total of 139 LCRS patients and 139 LC patients were enrolled in the study , with 110 male and 29 female in each group , and there were no significant difference for the mean age between the LCRS group (52.1 ±10.4 years old ) and the LC group (52.1 ± 10.1 years old).Two or more acute ischemic lesions of the brain were showed by MRI in most patients in the LCRS group (117 cases, 84.2%).Compared with the LC group, more patients in the LCRS group were found with adenocarcinoma , metastasis, elevated plasma D-dimer, CA125 and CA199 levels [ 88 cases (63.3%) vs 47 cases (33.8%);98 cases (70.5%) vs 56 cases (40.3%);(468.38 ±291.37) μg/L vs (277.59 ±191.22) μg/L;(221.42 ±146.34) U/ml vs (106.84 ±69.97) U/ml;(254.68 ±185.84) U/ml vs (97.15 ±63.64) U/ml;with all P<0.001].By logistic regression analysis of multiple factors , the elevated plasma D-dimer, CA125 and CA199 levels were showed to be independent risk factors for the cerebral infarction (OR=1.003, 95%CI 1.001 -1.004; OR=1.006, 95%CI 1.003 -1.010; OR=1.011, 95%CI 1.007-1.015).Conclusions The elevated plasma D-dimer, CA125 and CA199 levels are the risk factors for the lung cancer related cerebral infarction , which may lead to hypercoagulation and induce cerebral infarction eventually .

This study investigated the "homing" phenomenon in hepatocellular carcinoma (HCC). The "homing" specificity of endothelial progenitor cells (EPC) by establishing an orthotopic implantation model in nude mice. EPCs harvested from the marrow cells were separated by density gradient centrifugation. Fluorescence microscope, flow cytometry (FCM) and double fluorescence staining with FITC-UEA-I and DiI-ac-LDL, were employed to identify the cells. 4',6-diamidino-2-phenylindole (DAPI) labelling and real-time PCR were used for detecting the expression of CD133 and chemokines to trace and observe the distribution of EPCs. Our results showed that the distribution rate of EPCs was obviously higher than that in other important organs and the negative control group. Detection of CD133 and chemokines yielded similar results in difference tissues. Our experiment confirmed that the chemotaxis of EPCs does exist in HCC. Moreover, HIF-1α, SDF-1 and VEGF might play important roles in the "homing" of EPCs in HCC. EPCs might be a potential candidate for targeting vector of HCC for gene therapy.
Animals ; Endothelial Cells ; pathology ; Liver Neoplasms ; pathology ; Mice ; Mice, Inbred C57BL ; Mice, Nude ; Stem Cells ; pathology

OBJECTIVE: To explore the better operational methods by the laser surgery to treat diseases involving the laryngeal anterior commissure. It can excise the diseases as well as avoid anterior commissure adhesion, laryngostenosis and dyspnea after operations. METHOD: Twelve dogs were divided into 4 groups at random. There were three dogs in one group. A: excising experimental dog's anterior commissure by twice operations, the interval time was two weeks; B: excising experimental dog's anterior commissure in one time, at the same time suturing the silica gel sheet on the anterior commissure; C: excising experimental dog's anterior commissure in one time and then applying MMC on the wound of the anterior commissure; D: excising experimental dog's anterior commissure in one time, without any treatment. RESULT: All of the dog's surgery were completed successfully by laser. Four weeks later, we observed the raw surfaces. A: the neonatal membrane covered the wound, inflammatory reaction slight, we could not see obvious adhesion in the anterior commissure. B: the membrane covered the wound, appearing the dark chronic inflammation, we could see the adhesion in the anterior commissure slight. C: the membrane covered the wound, edematization, we could see the moderate adhesion in the anterior commissure. D: edematization, we could see the adhesion in the anterior commissure obviously. Four groups were all appeared hoarsenesses, the most slightly in group A, secondly in B and C, the worst in D. The analysis of vocal cord length of 4 groups, we used matched-pairs t-test, A, B, C groups' P>0.05, the vocal cord length didn't become shorter than before obviously. Group D's P<0.01, that meant the vocal cord length became shorter obviously. Between each group,we used reiterature-measurement analysis of variance (P<0.05), the change of vocal cord length had disparity in different groups. The comparison in two groups suggested that group A is the best. B and C are inferior ,but no disparity between them. The analysis of glottis area of 4 groups, as the same method above, A,B groups' P>0.05, the glottis area didn't shrink than before obviously. C and D groups' P<0.05, that meant having statistical significance and glottis area shrinked obviously. Between each group, we used reiterature-measurement analysis of variance (P>0.05), we could not think that the changes of four groups have disparity. CONCLUSION Excising the experimental dog's anterior commissure by laser, compared the 4 different operation methods, group A is the best method. B and C are inferior. The results are valuable in clinic when we perform operations by laser to treat the disease involving the anterior commissure.
Animals ; Disease Models, Animal ; Dogs ; Laryngeal Diseases ; etiology ; prevention & control ; Larynx ; surgery ; Laser Therapy ; methods ; Male ; Postoperative Complications ; prevention & control ; Tissue Adhesions ; prevention & control

Currently,the ethical review model for organ donation and transplantation in domestic hospitals is generally characterized by suddenness,unpredictability,tight time,difficulty in convening meetings and training committee members,as well as generally low quality and efficiency of ethical review,which cannot meet clinical needs and cause the waste of some scarce resources.The team of the Clinical Application Center of Human Organ Transplantation and the Ethics Committee of the First People's Hospital of Kunming combine more than 10 years of review practice experience,as well as continuously explore and optimize the ethical review process and operating procedures for organ donation and transplantation.The special application has been approved and jointly developed with Soochow University and the Medical Ethics Committee of Fujian Province to build a full-process information software system management platform for organ ethical review of donation and transplantation,giving the full play the advantages of the review information system in improving work efficiency and review quality,facilitating full-process information management,and conducting online training and learning for committee members,with a view to providing a specialized practical model for addressing the difficulties and challenges related to ethical review of human organ donation and transplantation.

OBJECTIVE: To investigate the effect of exosomes derived from Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC) cells on lymphangiogenesis and lymph node metastasis of NPC. METHODS: Exosomes from NP69 cells and EBV-positive HK1 (HK1-EBV) cells were obtained by ultracentrifugation and identified by Western blotting and nanoparticle tracking analysis. Dio dye phagocytosis test was performed to observe exosome uptake by lymphatic endothelial cells. Lymphatic endothelial cells were treated with exosomes from nasopharyngeal epithelium (NP69), HK1-EBV, and C666-1 cells or exosome-free supernatant of HK1-EBV and C666-1 cells, and tube formation and migration of the cells were observed. In a nude mouse model of popliteal lymph node metastasis of NPC, the effects of normal saline, NP69 cell-derived exosomes, HK1-EBV cell-derived exosomes, exosome-free supernatant of HK1-EBV cells, and HK1-EBV exosome-free supernatant protein on lymphangiogenesis and lymph node metastasis of the tumor were observed. RESULTS: The exosomes obtained by ultracentrifugation contained abundant exosome-specific proteins and showed a normal size range. The exosomes from NPC cells and NP69 cells could be taken up by lymphatic endothelial cells. Compared with the blank control and exosomes form NP69 cells, exosomes derived from HK1-EBV and C666-1 cells significantly promoted tube formation and migration of lymphatic endothelial cells ( CONCLUSIONS Exosomes from EBV-positive NPC cells can significantly promote lymphangiogenesis and lymph node metastasis of NPC.
Animals ; Cell Line, Tumor ; Endothelial Cells ; Epstein-Barr Virus Infections ; Exosomes ; Herpesvirus 4, Human ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Mice ; Mice, Nude ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms

ObjectiveMetabolomics was used to reveal the mechanism of Aconiti Lateralis Radix Praeparata（ALRP） in attenuating toxicity by processing from the aspects of amino acid metabolism， oxidative stress and energy metabolism by analyzing multiple metabolic pathways. MethodTwenty-four rats were randomly divided into control group， raw group and processed group， 8 rats in each group. The raw and processed group were given with 0.64 g·kg^-1 of raw ALRP and processed ALRP respectively every day， the control group was given with an equal amount of normal saline once a day. After continuous administration for 7 days， the urine， serum and heart tissue of rats were collected. Pathological examination of the heart was carried out using hematoxylin-eosin（HE） staining， and the activities of lactate dehydrogenase（LDH） and creatine kinase-MB（CK-MB） in serum and cardiac tissues were detected by microplate assay and immunoinhibition assay. The effects of ALRP on rat heart before and after processing were compared and analyzed. Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used to perform urine metabolomics analysis， and multivariate statistical analysis was used to screen for differential metabolites related to ALRP in attenuating toxicity by processing， and pathway enrichment analysis was carried out to explore the processing mechanism. ResultHE staining showed that no obvious pathological changes were observed in the heart tissue of the control group， while obvious infiltration of inflammatory cells such as plasma cells and granulocytes was observed in the heart tissue of the raw group， indicating that the raw ALRP had strong cardiotoxicity. There was no significant difference in HE staining of heart tissue between the processed group and the control group， indicating that the toxicity of ALRP was significantly reduced after processing. Compared with the control group， the activities of LDH and CK-MB were significantly increased in serum and heart tissue of the raw group， and those were significantly decreased in serum and heart tissue of the processed group， suggesting that the myocardial toxicity of processed ALRP was reduced. A total of 108 endogenous differential metabolites associated with the raw ALRP were screened using multivariate statistical analysis in positive and negative modes， of which 51 differential metabolites were back-regulated by the processed ALRP. Biological analysis of the key regulatory pathways and associated network changes showed that the pathways related to toxicity of ALRP mainly included tryptophan metabolism， arginine and proline metabolism， phenylalanine metabolism， aminoacyl-tRNA biosynthesis， alanine， aspartate and glutamate metabolism， etc. The metabolic pathways related to the attenuation of processed ALRP mainly included aminoacyl-tRNA biosynthesis， tryptophan metabolism， phenylalanine， tyrosine and tryptophan biosynthesis， phenylalanine metabolism and caffeine metabolism. ConclusionThe processing technology of ALRP in Guilingji can significantly attenuate the cardiotoxicity of raw products， the mechanism mainly involves amino acid metabolism， oxidative stress and energy metabolism， which can provide experimental bases for the research related to the mechanism of toxicity reduction of ALRP by processing and its clinical safety applications.

The voltage-gated Na channel subtype Nav1.7 is important for pain and itch in rodents and humans. We previously showed that a Nav1.7-targeting monoclonal antibody (SVmab) reduces Na currents and pain and itch responses in mice. Here, we investigated whether recombinant SVmab (rSVmab) binds to and blocks Nav1.7 similar to SVmab. ELISA tests revealed that SVmab was capable of binding to Nav1.7-expressing HEK293 cells, mouse DRG neurons, human nerve tissue, and the voltage-sensor domain II of Nav1.7. In contrast, rSVmab showed no or weak binding to Nav1.7 in these tests. Patch-clamp recordings showed that SVmab, but not rSVmab, markedly inhibited Na currents in Nav1.7-expressing HEK293 cells. Notably, electrical field stimulation increased the blocking activity of SVmab and rSVmab in Nav1.7-expressing HEK293 cells. SVmab was more effective than rSVmab in inhibiting paclitaxel-induced mechanical allodynia. SVmab also bound to human DRG neurons and inhibited their Na currents. Finally, potential reasons for the differential efficacy of SVmab and rSVmab and future directions are discussed.
Animals ; Antibodies, Monoclonal ; therapeutic use ; Biotin ; metabolism ; Cells, Cultured ; Disease Models, Animal ; Female ; Ganglia, Spinal ; cytology ; HEK293 Cells ; Humans ; Hybridomas ; chemistry ; Hyperalgesia ; drug therapy ; Male ; Mice ; Mice, Inbred C57BL ; NAV1.5 Voltage-Gated Sodium Channel ; metabolism ; NAV1.7 Voltage-Gated Sodium Channel ; chemistry ; immunology ; metabolism ; Neuralgia ; drug therapy ; metabolism ; Protein Binding ; drug effects ; Recombinant Proteins ; biosynthesis ; therapeutic use ; Sensory Receptor Cells ; drug effects ; physiology