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BACKGROUND@#Malignant pleural effusion (MPE) is a complicated condition of patients with advanced tumors. Further dissecting the microenvironment of infiltrated immune cells and malignant cells are warranted to understand the immune-evasion mechanisms of tumor development and progression.@*METHODS@#The possible involvement of microRNAs (miRNAs) in malignant pleural fluid was investigated using small RNA sequencing. Regulatory T cell (Treg) markers (CD4, CD25, forkhead box P3), and Helios (also known as IKAROS Family Zinc Finger 2 [IKZF2]) were detected using flow cytometry. The expression levels of IKZF2 and miR-4772-3p were measured using quantitative real-time reverse transcription polymerase chain reaction. The interaction between miR-4772-3p and Helios was determined using dual-luciferase reporter assays. The effects of miR-4772-3p on Helios expression were evaluated using an in vitro system. Correlation assays between miR-4772-3p and functional molecules of Tregs were performed.@*RESULTS@#Compared with non-malignant controls, patients with non-small cell lung cancer had an increased Tregs frequency with Helios expression in the MPE and peripheral blood mononuclear cells. The verified downregulation of miR-4772-3p was inversely related to the Helios Tregs frequency and Helios expression in the MPE. Overexpression of miR-4772-3p could inhibit Helios expression in in vitro experiments. However, ectopic expression of Helios in induced Tregs reversed the effects induced by miR-4772-3p overexpression. Additionally, miR-4772-3p could regulate Helios expression by directly targeting IKZF2 mRNA.@*CONCLUSION@#Downregulation of miR-4772-3p, by targeting Helios, contributes to enhanced Tregs activities in the MPE microenvironment.
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Background@#Malignant pleural effusion (MPE) is a complicated condition of patients with advanced tumors. Further dissecting the microenvironment of infiltrated immune cells and malignant cells are warranted to understand the immune-evasion mechanisms of tumor development and progression.@*Methods@#The possible involvement of microRNAs (miRNAs) in malignant pleural fluid was investigated using small RNA sequencing. Regulatory T cell (Treg) markers (CD4, CD25, forkhead box P3), and Helios (also known as IKAROS Family Zinc Finger 2 [IKZF2]) were detected using flow cytometry. The expression levels of IKZF2 and miR-4772-3p were measured using quantitative real-time reverse transcription polymerase chain reaction. The interaction between miR-4772-3p and Helios was determined using dual-luciferase reporter assays. The effects of miR-4772-3p on Helios expression were evaluated using an in vitro system. Correlation assays between miR-4772-3p and functional molecules of Tregs were performed.@*Results@#Compared with non-malignant controls, patients with non-small cell lung cancer had an increased Tregs frequency with Helios expression in the MPE and peripheral blood mononuclear cells. The verified downregulation of miR-4772-3p was inversely related to the Helios+ Tregs frequency and Helios expression in the MPE. Overexpression of miR-4772-3p could inhibit Helios expression in in vitro experiments. However, ectopic expression of Helios in induced Tregs reversed the effects induced by miR-4772-3p overexpression. Additionally, miR-4772-3p could regulate Helios expression by directly targeting IKZF2 mRNA.@*Conclusion@#Downregulation of miR-4772-3p, by targeting Helios, contributes to enhanced Tregs activities in the MPE microenvironment.
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[ ABSTRACT] AIM:To investigate whether the increase in PTEN expression is related to apoptosis, and whether it is regulated by reactive oxygen species( ROS) .METHODS: The rat islet cells were divided into constant low glucose group ( group L) , constant high glucose group ( group H) , glucose fluctuation group ( group F) , low glucose after high glucose group (group HL) and low glucose after fluctuation group (group FL).The ROS level, apoptotic rate, intracellu-lar calcium, insulin release and PTEN protein expression were analyzed.RESULTS:Compared with groups H and L, the insulin secretion decreased, and intracellular calcium, ROS level, PTEN protein expression and apoptotic rate increased in group F ( P<0.05) .Compared with group H, the intracellular calcium, ROS level, PTEN protein expression and apoptot-ic rate in group HL decreased, but were still higher than those in group L (P<0.05).Compared with group F, the intra-cellular calcium, ROS level, PTEN protein expression and apoptotic rate in group FL decreased, but were still higher than those in group L (P<0.05).CONCLUSION:Glucose fluctuation can cause the apoptosis of islet cells more easily than constant high glucose.This may be related to the change of intracellular calcium and increase in oxidative stress which pro-motes PTEN expression.The recovery of glucose level to some extent relieves oxidative stress, decrease PTEN expression and reduce cell damage.
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OBJECTIVE To investigate the antidepressant effect of triptolide in chronically unpredictable stressed mice and its possible protective effect on brain derived neurotrophic factor(BDNF). METHODS One method was selected from 8 different stress methods each day,and the mice were treated with triptolide(20,40,80 and 160μg·kg-1)10 min before the stress method. A chroni?cally unpredictable stressed model was established and after 14 d stress experiment, the total distance in the locomotor activity and the immobility time in the force swimming test and tail test were observed respectively. Triptolide(20,40,80 and 160μg·kg-1)was given 10 min before the test. In addition, Western blot was used to analyze the expression of phosphorylated cAMP response element binding protein(p-CREB) and BDNF in the hippocampus and frontal cortex. RESULTS There was no effect on the locomotor activity in any group. Compared with the normal control group,the chronically unpre?dictable stressed group showed obvious depressive-like behavior,while the immobility time in the force swimming test decreased from(161 ± 18)s in chronically stressed group to(102 ± 14)s(P<0.05) and(83±14)s(P<0.01)when mice were ip given triptolide 80 and 160μg·kg-1, respectively,and(77± 11)s(P<0.01)in imipramine(IMI)hydrochloride group(10 mg?kg-1),and(96±9)s(P<0.01)in fluox?etine(FLU)group(10 mg?kg-1). The immobility time in the tail suspension test decreased from(128± 8)s in chronically stressed group to(93±9)s(P<0.05),(85±8)s(P<0.01)and(77±11)s(P<0.01)when mice were ip given triptolide 40,80 and 160μg · kg-1 respectively,and(64 ± 9)s(P<0.01)in IMI hydro?chloride 10 mg?kg-1 group,and(72±6)s(P<0.01)in FLU group(10 mg?kg-1). Moreover,the expression of p-CREB in the hippocampus and frontal cortex significantly increased in triptolide 80 and 160μg·kg-1 groups(P<0.05),so did the expression of BDNF in the hippocampus and frontal cortex in triptolide 80 and 160μg·kg-1 groups(P<0.05). CONCLUSION Triptolide can ameliorate the depressive-like behavior in chronically unpredictable stressed mice,which may be related to the cAMP-CREB-BDNF signal transduction cascades.
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<p><b>OBJECTIVE</b>To compare the role of radionuclide lymphoscintigraphy and dynamic magnetic resonance lymphangiography (MRL) for the diagnosis of extremity lymphedema.</p><p><b>METHODS</b>Sixteen patients with primary extremity lymphedema and two with Klippel-Trenaunay syndrome combined with lymphedema were examined by lymphoscintigraphy using the tracer 99Tc-labelled dextran, and also by MRL using gadobenate dimeglumine as contrast agent. The results of morphological abnormalities and functional state of the lymphatic system at affected limbs from the two imaging methods were compared.</p><p><b>RESULTS</b>Lymphatic vessels were imaged in 14 of 18 limbs with lymphedema using MRL, compared with one of 18 using lymphoscintigraphy. MRL detected the inguinal nodes in 16 of 17 patients, whereas lymphoscintigraphy revealed inguinal nodes in only nine cases. MRL revealed more precise information about structural and functional abnormalities of lymph vessels and nodes than lymphoscintigraphy by real-time measurement of lymph flow in vessels and nodes.</p><p><b>CONCLUSIONS</b>Dynamic MRL is more sensitive and accurate than lymphoscintigraphy in the detection of anatomical and functional abnormalities in the lymphatic system in patients with extremity lymphedema.</p>
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Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Extremities , Lymphedema , Diagnostic Imaging , Lymphography , Methods , Lymphoscintigraphy , Methods , Magnetic Resonance ImagingABSTRACT
Objective Vibrio cholera was extremely rare in Sichuan province (no cases in 2008). Any outbreak could indicate contamination through the food supply system. In July 2009, a hospital reported a cluster of 7 diarrhea patients; all attended the same banquet. One patient was confirmed to have Vibrio cholera (O139). We conducted this investigation to identify the source of this possible cholera outbreak. Methods We defined a suspect case as any banquet attendee with diarrhea ( ≥3 times/day). A confirmed case was a suspect case with a positive Vibrio cholera culture. We took stool samples or rectal swabs from all attendees for cholera culture and interviewed 272 banquet attendees about foods they ate at the banquet and kitchen workers about food preparation. Results 7.1% (24/337) of attendees developed cases within an average of 65 hours after eating. Three meals were served. All patients had the lunch whereas no patients only ate breakfast and/or dinner. Of 180 attendees who ate turtle meat 12% were case-patients, compared to 3.3% of 92 attendees who did not (RR=3.6,95%CI: 1.1-12). Of the 150 attendees who ate peanuts 13% were cases compared to 4.1%of 122 attendees who did not eat peanuts (RR=3.1,95%CI: 1.2-8.0). During preparation, the same utensil was used for fresh turtle meat and peanuts without washing in-between the process. Turtle meat and peanuts were stored for ＞ 16 hours at room temperature after cooking before consumption. All 33 turtles originated from commercial production in another province. Conclusion This outbreak was likely caused by poor food handling of commercially produced turtles. We proposed that to improve microbiologic monitoring of aquatic food animals, and raise the awareness of good handling practices at mass gathering in rural China.
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<p><b>OBJECTIVE</b>To evaluate anatomical and functional images of contrast MR lymphangiography in the diagnosis of limb lymphatic circulation disorders.</p><p><b>METHODS</b>30 patients with limb lymphedema were enrolled in the study. There were 27 patients of primary lymphedema and 3 of secondary lymphedema. Contrast enhanced lymphangiography was performed with 3.0 T MR Unit after intracutaneous injection of gadobenate dimeglumine into the interdigital webs of the dorsal foot and hand. The kinetics of enhanced lymph flow within the lymphatics were calculated using the formula: Speed (cm) = total length of visualized lymph vessel (cm)/ inspection time (minutes) and by comparing dynamic nodal enhancement and time-signal intensity curves between edematous and contralateral limbs. Morphological abnormalities of the lymphatic system were also evaluated.</p><p><b>RESULTS</b>Following injection of the contrast agent enhanced lymphatic channels were consistently visualized in all clinical lymphedematous limbs and five contralateral limbs of unilateral lymphedema cases. The speed of enhanced flow within the lymphatics of lymphedematous limbs ranged from 0.30 to 1.48 cm/min. The contrast enhancement in inguinal nodes of edematous limbs was significantly lower than that of contralateral limbs (P < 0.01). Dynamic measurement of contrast enhancement showed a remarkable lowering of peak time (P < 0.01) and peak enhancement (P < 0.01) and a delay in outflow in inguinal nodes of affected limbs compared with that of control limbs. Post-contrast MR imaging also depicted varied distribution patterns of lymphatics and abnormal lymph flow pathways within lymph nodes in the limbs with lymphatic circulation disorders.</p><p><b>CONCLUSIONS</b>Contrast MR lymphangiography with gadobenate dimeglumine was able to visualize the precise anatomy of lymphatic vessels and lymph nodes in lymphedematous limbs. It also provided comprehensive information about the functional status of lymph flow transportation in lymphatics and lymph nodes.</p>
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Adolescent , Adult , Child , Humans , Middle Aged , Young Adult , Lymph Nodes , Pathology , Lymphatic Vessels , Pathology , Lymphedema , Diagnostic Imaging , Pathology , Lymphography , MethodsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effect of manual lymph drainage on chronic extremity lymphedema.</p><p><b>METHODS</b>Fifty patients with chronic lymphedema of extremity were treated with manual lymph drainage (MLD) complex decongestion therapy. Among them, 29 had primary lymphedema, 21 had secondary lymphedema. 42 had lymphedema of lower extremity and 8 had lymphedema of upper limb. The result of treatment was evaluated with measurement of circumference of extremities and edema fluid in tissue with Multiple-frequency bioelectrical impedance analysis.</p><p><b>RESULTS</b>After 1-2 treatment courses, all 50 patients showed significant decrease of circumference of lymphomatous limbs (P < 0.05) and remarkable reduction of accumulated edema fluid in tissue (P < 0. 05). There was highly correlation between the decrease of limb circumference and edema fluid in tissue (r(s) = 0.774, P < 0.01).</p><p><b>CONCLUSIONS</b>MLD complex decongestion therapy is effective for the treatment of chronic lymphedema of extremity.</p>
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Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Chronic Disease , Drainage , Methods , Extremities , Lymphedema , General Surgery , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To study the therapeutic effect of vascular endothelial growth factor C (VEGF-C) gene for chronic obstructive lymphedema in mouse tail model which may provide a new treatment for lymphedema.</p><p><b>METHODS</b>RT-PCR and immunoabsorption were applied to detect VEGF-C gene expression in fibroblasts and secretion of VEGF-C protein in COS7 cells respectively after pCDNA3.1 (+) VEGF-C transfection. A mouse tail model of chronic obstructive lymphedema was created. Then the pcDNA3.1-VEGF-C plasmid was injected into the tail. The effect of modulating lymphangiogenesis was observed.</p><p><b>RESULTS</b>Compared with control group, overexpression of VEGF-C enhanced lymphangiogenesis in vivo and mouse tail skin suffering chronic obstructive lymphedema was improved by VEGF-C gene significantly.</p><p><b>CONCLUSIONS</b>VEGF-C can improve the lymphedema through enhancing lymphangiogenesis.</p>
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Animals , Female , Humans , Mice , COS Cells , Cells, Cultured , Chlorocebus aethiops , Disease Models, Animal , Gene Transfer Techniques , Genetic Therapy , Lymphedema , Therapeutics , Mice, Inbred BALB C , Plasmids , Transfection , Vascular Endothelial Growth Factor C , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the efficacy of gene therapy with human vascular endothelial growth factor-c (VEGF-C) on obstructive lymphedema.</p><p><b>METHODS</b>Two animal models of lymphedema were created: one in the right hind limb of adult New Zealand white rabbits and the other in SD mouse tail. Each model was randomly divided into two groups to receive intradermal injection of either VEGF-C gene (experimental group), or saline(control group). In rabbit model, the volume change of affected limb was measured. In mouse model, biopsy was performed after 3 weeks treatment to detect the expression of VEGF-C mRNA and proteins. The lymphagenesis was evaluated by immunohistochemical examination with lymphatic endothelium hyaluronan receptor antibody.</p><p><b>RESULTS</b>The volume of the affect rabbit limb decreased by (24.40 +/- 1.08) ml in experimental group, compared with (5.80 +/- 1.92) ml in control group (P = 0.0001). The expression of VEGF-C mRNA and protein increased markedly in experiment group, but not in controls. More lymphatic vessels with large caliber were seen in experiment group (P = 0.0004).</p><p><b>CONCLUSIONS</b>VEGF-C gene therapy may alleviate or treat lymphedema by inducing lyphmangiogenesis.</p>
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Animals , Humans , Rabbits , Rats , Disease Models, Animal , Gene Transfer Techniques , Genetic Therapy , Lymphedema , Therapeutics , RNA, Messenger , Genetics , Rats, Sprague-Dawley , Vascular Endothelial Growth Factor C , GeneticsABSTRACT
0.05). Conclusion Enough high-purified LECs can be isolated by collagenase digestion procedure followed by immunomagnetic beads sorting. Post-thawed endothelial cells are proved to have high vitality and growth potential in vitro without significant morphological changes. Cryopreserved LECs may serve as a cell choice for research of lymphangiogenesis and lymphatic patterning.
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<p><b>OBJECTIVE</b>To observe treatment effects of primary lower limb lymphedema using ultrasonic assisted liposuction.</p><p><b>METHODS</b>Internal ultrasonic liposculpture system combined postoperative continual elastic stockings or bandages were used for reducing lymphatic burdens of the affected limbs by partly removal of lymphedematous tissues.</p><p><b>RESULTS</b>Edema regression in the affected limbs were obvious at 2 weeks postoperative and kept to stable without recurrence during 1 year follow-up.</p><p><b>CONCLUSIONS</b>Ultrasonic assisted liposuction combined with elastic compression is safe and effective for the treatment of primary limb lymphedema.</p>
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Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Lipectomy , Lower Extremity , Lymphedema , General Surgery , Stockings, Compression , Suction , Methods , Ultrasonic TherapyABSTRACT
<p><b>OBJECTIVE</b>To investigate the possible influence of the impairment of lymph fluid on the metabolism of hyaluronic acid (HA) in the lymphedematous skin tissue.</p><p><b>METHODS</b>Tissue fluid was collected in lymphedematous limbs and the contralateral healthy limbs of 39 patients and HA content was measured with radioimmunoassay. The protein contents were also measured.</p><p><b>RESULTS</b>The HA contents in interstitial fluid of lymphedematous limb were significantly (8 fold) higher than that of normal limb. The protein concentration in the tissue fluid did not show significant differences between lymphedema and those with normal tissue.</p><p><b>CONCLUSION</b>The result suggests blockage of regional draining lymphatics may impairs breakdown of HA and the stagnation of HA in the limb may exert a deleterious effect on the interstitium.</p>
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Humans , Extracellular Fluid , Metabolism , Forearm , Hyaluronic Acid , Metabolism , Leg , Lymphedema , Metabolism , Radioimmunoassay , Skin , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of several homeobox genes during the wound healing in fetal and adult skin and their roles in fetal scarless wound healing.</p><p><b>METHODS</b>The expressions of PRX-2, HOXB13, HOX2.2 and HOX2.3 during wound healing in fetal and adult skin were determined with in situ hybridization.</p><p><b>RESULTS</b>(1) PRX-2 positive expression could be identified in normal fetal and adult skin, especially in the fetus. But there was difference in location sites of the genes. The positive expression in normal fetal skin was mainly found in the peripheral cells at the hair shafts within dermal papilla layers and was also found in the epithelium. Nevertheless, weak positive expression of PRX-2 was found in the epithelial basal layer cells in normal adult skin but not in dermal tissue. There was strong positive expression of the PRX-2 in the tissue around the wound in fetus but not of that in adults except the epithelial basal layers. (2) Positive expression of HOXB13 could be identified in both normal fetal and adult skins. And the expression was concentrated mainly in hair follicle cells in the dermis and in the basal layer cells in the epithelium. Furthermore, the expression became weak after trauma, especially in fetal skin. (3) The positive expression of HOX2.2 and HOX2.3 in normal fetal skin was observed mainly in the whole layer of the epithelium and especially in the epithelial basal layers. Weak positive expression could be found in the dermis and strong expression found in the tissue near the wound. But there was no positive expression of the HOX genes in normal adult skin and wounds.</p><p><b>CONCLUSION</b>The difference in the HOX expression in fetal and adult skin wound healing might be the key factor leading to different wound healing. Homeobox genes might be closely related with the developmental biology.</p>