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OBJECTIVE@#Here, we explored molecular changes that could potentially mediate healing effects of Gua Sha - a method employed by the Chinese traditional medicine with proven track records of safe and efficient applications dating back to ancient times as well as support from randomized controlled trials performed by modern medical studies - yet remaining almost entirely unexplored by the modern-day high-throughput methods of the -omics sciences.@*METHODS@#We investigated transcriptome changes occurring shortly after Gua Sha treatment in the whole blood of healthy volunteers using bulk RNA-seq analysis. We applied various analytical tools to identify genes with consistent expression changes in multiple individuals in response to Gua Sha and their networks.@*RESULTS@#We found that while the changes were very subtle and individual-specific, we could identify consistent upregulation of three histone genes. Further analysis of the potential regulatory networks of these histone genes revealed the enrichment of functions involved in the immune response and inflammation.@*CONCLUSION@#The significance of these results in the context of potential effects of Gua Sha and the next steps in exploring the molecular mechanisms of action of this technique are discussed.
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Humans , Medicine, Chinese Traditional/methods , Histones , Gene ExpressionABSTRACT
Objective To investigate the predictors for the preoperative diagnosis of pancreatic mucinous cystic neoplasm with invasive carcinoma (MCN-IC).Methods Clinical data of 132 patients with pancreatic mucinous cystic neoplasm ( MCN ) who underwent surgery and were pathologically diagnosed in Shanghai Changhai Hospital and General Hospital of Xinjiang Military Region from August 2000 to December 2013, including gender, age, medical history, clinical presentations, laboratory examinations and imaging findings and etc , were retrospectively analyzed .All cases were classified into two groups:MCN with noinvasive carcinoma ( MCN-nIC, including MCN with low-or intermediate-grade dysplasia and MCN with high-grade dysplasia ) and MCN-IC.The univariate and multivariate logistic regression was used to analyze the differences on laboratory examinations and imaging findings and the like to identify the predictors for the preoperative diagnosis of MCN-IC.Receiver operator characteristic ( ROC) curve was used to evaluate fitting performance and Hosmer-Lemeshow test was performed to evaluate goodness of fit .Results Of the 132 patients, 115 (87.12%) were MCN-nIC, 17(12.88%) were MCN-IC.Univariate analysis identified old age(≥60 years), abdominal pain, anorexia, GLU elevated, CEA≥5 ng/ml, CA19-9≥37 U/ml, unclear border of tumor , thick wall (>2 mm) , presence of mural nodules and absence of the septa as independent predictors for MCN-IC. Multivariate analysis identified old age (≥60 years), abdominal pain, CA19-9≥37 U/mL, unclear border of tumor, presence of mural nodules and absence of the septa as the predictors for MCN -IC.The maximal area under ROC ( AUC) was 0.947, which indicated that the fitting performance of the model was satisfactory and the goodness of fit was better (P=0.056).Conclusions MCN-IC had a generally low prevalence .Old age (≥60 years), abdominal pain, CA19-9≥37 U/ml, unclear border of tumor, presence of mural nodules and absence of the septa may predict the diagnosis of MCN-IC.
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OBJECTIVE To evaluate whether the IDO1 inhibitor 1- methyl- L- tryptophan (1- MT) combine calcium influx inhibitor carboxyamidotriazole (CAI) could further enhance the suppression of programmed death 1 (PD-1) in CD8 + T cells and investigate the curative effect of the combined use. METHODS CD8 +T cells were isolated from normal mice spleen by negative selection using magnetic cell separation. The isolated CD8 +T cells were cultured in RPMI 1640 medium containing 10% FBS and 100 U·mL- 1 IL-2 and activated by the addition of anti-CD3 and anti-CD28 (1 g·L- 1 each mabs). CD8 + T cells were pretreated for 48 h with drug and the fluo- 3 as a marker of intracellular calcium concentration was detected by flow cytometry. The calcineurin (CaN) levels were assayed with ELISA in CD8+T cells after 48 h incubation with 10 μm CAI. The nuclear translocations of NFAT and AHR were detected by immunofluorescent staining after 48 h of drug treatment. The expression of PD-1 in CD8+T cells was analyzed by flow cytometry. RESULTS Intracellular fluorescent intensity was markedly debase due to CAI treatment(P<0.01). Meanwhile, the changes of CaN content had a resembled correlation (P<0.01). Immunofluorescence experiment showed that after combination therapy the transfer of NFAT and AHR in nuclear substantially reduced. Flow cytometry revealed that after the combination caused a significant decrease in PD-1 expression in CD8+T cells. CONCLUSION CAI and 1-MT could inhibit markedly the expression of PD-1 in CD8 +T cells by inhibiting the nuclear translocation of NFAT and AHR, respectively and the combination of them has synergetic effect.
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Oval cells have a potential to differentiate into a variety of cell lineages including hepatocytes and biliary epithelia. Several models have been established to activate the oval cells by incorporating a variety of toxins and carcinogens, alone or combined with surgical treatment. Those models are obviously not suitable for the study on human hepatic oval cells. It is necessary to establish a new and efficient model to study the human hepatic oval cells. In this study, the hepatocyte growth factor (HGF) and epidermal growth factor (EGF) were used to induce differentiation of mouse embryonic stem (ES) cells into hepatic oval cells. We first confirmed that hepatic oval cells derived from ES cells, which are bipotential, do exist during the course of mouse ES cells' differentiation into hepatic parenchymal cells. RT-PCR and transmission electron microscopy were applied in this study. The ratio of Sca-1+/CD34+ cells sorted by FACS in the induction group was increased from day 4 and reached the maximum on the day 8, whereas that in the control group remained at a low level. The differentiation ratio of Sca-1+/CD34+ cells in the induction group was significantly higher than that in the control group. About 92.48% of the sorted Sca-1+/CD34+ cells on the day 8 were A6 positive. Highly purified A6+/Sca-1+/CD34+ hepatic oval cells derived from ES cells could be obtained by FACS. The differentiation ratio of hepatic oval cells in the induction group (up to 4.46%) was significantly higher than that in the control group. The number of hepatic oval cells could be increased significantly by HGF and EGF. The study also examined the ultrastructures of ES-derived hepatic oval cells' membrane surface by atomic force microscopy. The ES-derived hepatic oval cells cultured and sorted by our protocols may be available for the future clinical application.
Subject(s)
Animals , Mice , Antigens, CD34 , Genetics , Metabolism , Antigens, Ly , Genetics , Metabolism , Cell Differentiation , Genetics , Physiology , Cell Line , Embryonic Stem Cells , Cell Biology , Metabolism , Epidermal Growth Factor , Pharmacology , Flow Cytometry , Gene Expression Regulation, Developmental , Hepatocyte Growth Factor , Pharmacology , Liver , Cell Biology , Metabolism , Membrane Proteins , Genetics , Metabolism , Mice, Inbred BALB C , Microfilament Proteins , Metabolism , Microscopy, Atomic Force , Microscopy, Electron, Transmission , Reverse Transcriptase Polymerase Chain Reaction , Stem Cells , Cell Biology , Metabolism , Time FactorsABSTRACT
Oval cells have a potential to differentiate into a variety of cell lineages including hepatocytes and biliary epithelia. Several models have been established to activate the oval cells by incorporating a variety of toxins and carcinogens, alone or combined with surgical treatment. Those models are obviously not suitable for the study on human hepatic oval cells. It is necessary to establish a new and efficient model to study the human hepatic oval cells. In this study, the hepatocyte growth factor (HGF) and epidermal growth factor (EGF) were used to induce differentiation of mouse embryonic stem (ES) cells into hepatic oval cells. We first confirmed that hepatic oval cells derived from ES cells, which are bipotential, do exist during the course of mouse ES cells' differentiation into hepatic parenchymal cells. RT-PCR and transmission electron microscopy were applied in this study. The ratio of Sca-1+/CD34+ cells sorted by FACS in the induction group was increased from day 4 and reached the maximum on the day 8, whereas that in the control group remained at a low level. The differentiation ratio of Sca-1+/CD34+ cells in the induction group was significantly higher than that in the control group. About 92.48% of the sorted Sca-1+/CD34+ cells on the day 8 were A6 positive. Highly purified A6+/Sca-1+/CD34+ hepatic oval cells derived from ES cells could be obtained by FACS. The differentiation ratio of hepatic oval cells in the induction group (up to 4.46%) was significantly higher than that in the control group. The number of hepatic oval cells could be increased significantly by HGF and EGF. The study also examined the ultrastructures of ES-derived hepatic oval cells' membrane surface by atomic force microscopy. The ES-derived hepatic oval cells cultured and sorted by our protocols may be available for the future clinical application.
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Objective To evaluate the value of medical treatment in the management of SAP.Methods From January 2000 to December 2011,a total of 1064 cases out of 931 SAP patients were admitted and retrospectively analyzed.The etiologies,severity score,complication rates,therapies,effectiveness and costs of those SAP cases were summarized.Results There were 559 males and 372 females with a mean age of (51 ± 15)years old.The main cause was biliary tract disease (58.3%),followed by fat-rich diet (31.2%),hyperlipidemia (13.6%) and alcohol (7.1%).At the time of admission,95.5% of SAP patients presented with level D disease according to Balthazar CT severity index,26.0% had a Ranson score ≥3 and 30.1% had an APACHE Ⅱ score ≥ 8.There were 42.7% cases complicated with systemic inflammatory response syndrome (SIRS).Acute lung injury and acute respiratory distress syndrome (ARDS),acute kidney injury,shock or heart failure,acute liver dysfunction,and diffuse intravascular clotting (DIC)occurred in 24.0%,8.1%,5.4%,3.2%,and 1% of all patients,respectively.Other complications of SAP included abdominal cavity bleeding (n =17),pseudocyst bleeding (n =9),pancreatic abscess (n =78) and gastrointestinal fistula (n =33).Totally 25 (2.3%) patients died in hospital and 36 (3.4%) patients were discharged against advice,with an overall treatment success rate of 94.3%.The mean hospital stay was (23.7 ± 19.2) d,and the average cost was 52.3 thousands of RMB.Conclusions A comprehensive treatment pathway relying on medical treatment,focusing on organ function support and assisted by miniinvasive intervention may improve the treatment success rate of SAP,which is worth of further application.
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As the rapid growth of population and economic development,the types and intensity of water pollution in urban river stream are increasing,which on one hand restrict the sustainable development of the social,economic and environment,on the other hand,damage people health. Since having safety,economy,practicality,systematic and other merits,ecological remedy technology has been the main means for controlling river contamination. In this paper,the recent researches on the ecological restoration technique for the urban stream in China in recent years were reviewed.
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<p><b>OBJECTIVE</b>To observe calf thymus DNA damage induced by potassium dichromate in combination with glutathione (GSH).</p><p><b>METHODS</b>Atomic force microscope and ultraviolet spectrum (UV) were used to observe the alterations of the DNA ultrastructure and absorption spectrum.</p><p><b>RESULTS</b>Atomic force microscopy revealed no breaks of the DNA strand in response to treatment with potassium dichromate alone, but when coupled with GSH at proper concentrations, potassium dichromate induced alterations in the DNA structure and DNA fragmentation. UV examination also confirmed these findings by showing increased absorption intensity of the maximum UV peak following combined treatment of the DNA with potassium dichromate and GSH.</p><p><b>CONCLUSION</b>These morphological and spectrographic evidences verified the important role of GSH in mediating the generation of various tumor-inducing intermediate products of potassium dichromate.</p>
Subject(s)
Animals , Cattle , DNA , Chemistry , Genetics , DNA Damage , DNA Fragmentation , Glutathione , Toxicity , Microscopy, Atomic Force , Methods , Nucleic Acid Conformation , Potassium Dichromate , Toxicity , Spectrophotometry, UltravioletABSTRACT
Human CD34(+) hematopoietic cells, a distinctive cell population containing hematopoietic stem/progenitor cells (HSPC), have the capability to highly self-renewal, differentiation into all lineages of committed progenitor cells and reconstitution of both long-term hematopoiesis and immunefunctions after transplantation. CD34(+) hematopoietic cells from bone marrow (BM) recently have been employed for treating neoplastic and genetic disorders. This study was aimed to investigate membrane surface ultrastructures of bone marrow CD34(+) cell from mormal persons and leukemia patients and to compare their morphologic differences by using atomic force microscope (AFM). BM was collected from 5 normal donors and 6 leukaemia patients. All samples were layered on Ficoll-Paque gradients (specific gravity 1.077 g/ml) to separate the mononuclear cells. After that CD34(+) cells were purified by immuno-magnetic bead separation and evaluated with a FACS Calibur, these cells were detected by AFM of tapping mode inair. At lest 20 cells per samples were observed. The results showed that most of CD34(+) hematopoietic cells were like circle plate, the diameter was 10 - 14 microm. The surface of CD34(+) hematopoietic cell membrane was comparatively complex. The surface of CD34(+) hematopoietic cell membrane appeared as granular, with packed particles. With the region analysis function of IP2.1 software, the region of 2 microm x 2 microm was selected and four parameters of the surface (maximum peak-to-valley distance, average roughness, root-mean-squared roughness and mean height) were measured. Values of the 4 parameters showed that the characteristic parameters of CD34(+) HSPC from leukaemia were higher than that from normal person. It is concluded that AFM has specific advantages in analyzing cell membrane in the nanometer level and can gain more information. With the help of analysis software, AFM can be a helpful tool for fast leukaemic diagnosis and CD34(+) hematopoietic cells selection.