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Objective To investigate the effect of U50488H on the ultrastructure and organ function in septic shock rats. Methods Forty SD male rats were randomly(random number) divided into 5 groups: sham group, septic shock group, U50488H+septic shock group, nor-BNI+U50488H+septic shock group, and nor-BNI+septic shock group, with 8 rats in each group. Cecal ligation and puncture (CLP) was performed to induce septic shock in the septic shock group. Rats in the U50488H+septic shock group were treated with U50488H injection by intravenous at the shock point, and other procedures were the same as the septic shock group. Rats in the nor-BNI+U50488H+septic shock group were treated with nor-BNI injection by intravenous 3.5 h after abdomen closed, and other procedures were the same as the U50488H+septic shock group. Except for U50488H injection, rats in the nor-BNI+septic shock group received procedures the same as the nor-BNI+U50488H+septic shock group. Albumin, cardiac troponin I (cTnI) and N terminal pro B type natriuretic peptide (NT-proBNP) in serum were measured at abdomen-closed, 3, 6, and 12 h after CLP. The changes of histology and ultramicro structure under electron microscope of lung, heart, liver and kidney of rats were observed at 12 h after CLP. Statistical analysis was performed using SPSS 19.0. Comparison among groups was carried out using ANOVA, and Student's t-test was used for multiple comparisons as post-hoc. Results At 6 and 12 h of CLP, serum albumin of the septic shock group were significantly lower than those of the sham group (P<0.01), while those in the cTnI and NT-pro BNP groups were higher at 3, 6, and 12 h of CLP (P<0.01). Compared with the septic shock group, serum albumin of the U50488H+septic shock group increased significantly (P<0.01), whereas the serum levels of cTnI and NT-pro BNP decreased remarkably at 3, 6 and 12 h of CLP (P < 0.05, P < 0.01, respectively). Compared with the sham group, the alveolar wall was severely damaged, the alveolar septum and blood vessel wall were thickened obviously; the myocardial fiber was swollen, necrotic, and the infiltration of central granulocyte was increased significantly; hepatocyte showed edema, vacuolar-like steatosis, fatty degeneration, spotty and focal necrosis; and slight edema and vacuolar degeneration were found in the glomerulus endothelial in the septic shock group. Compared with the septic shock group, the ultrastructural damage of the lung, heart, liver and kidney of the U50488H+ septic shock group was significantly improved. All the above effects of U50488H could be blocked by nor-BNI (a selective κ-opioid receptor antagonist) (P<0.01). Conclusions U50488H can promote the recovery of serum albumin, and protect organ function in septic shock rats.
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Objective To investigate the treatment efficacy,feasibility and safety of laparoscopic resection for gastric gastrointestinal stromal tumors (GISTs) larger than 5 cm.Methods The clinical data of 31 patients who underwent laparoscopic resection of large gastric GISTs between Jan 2010 and Jun 2017 at Sir Run Run Shaw Hospital and People's Hospital of Zhejiang Province were retrospectively analyzed.Results All operations were successful.Two patients were converted to open surgery.The mean tumor size was (7.2 ±0.5) cm.The mean operation time was (127 ± 12) min,intraoperative blood loss was (83 ±25) ml.The first flatus time was (2.2 ±0.8) days,the first diet resumption time was (2.4 ± 1.2) days,and the median postoperative hospital stay was (5.5 ± 1.5) days.The median follow-up time was 25 months.One patient developing liver metastasis 41 months after primary resection received oral mesylate imatinib therapy for 24 months and was still alive at the last follow-up.One patient died of lung cancer 33 months after primary operation without recurrence of gastric GIST.Conclusions Laparoscopic surgery for gastric GIST larger than 5 cm on laparoscopicaly accessible location is feasible and safe.
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Objective To obtain a non-invasive subclinical hypothyroidism ( SCH) mouse model, and to explore microRNAs profile related to lipid metabolism in the model mice. Methods C57BL/6 male mice (8 weeks) were treated with methimazole ( MMI, 0. 08 mg/kg BW/d) to construct SCH mouse model. MicroRNAs profiling analysis was performed by real-time quantitative polymerase chain reaction ( qRT-PCR) . Results Compared with the control group, the serum thyroid stimulating hormone ( TSH ) in subclinical hypothyroidism group increased significantly (P>0.01), while the serum free thyroxine(FT4) level did not show significant difference between the two groups (P>0. 05), which was in line with the diagnostic criteria of SCH. SCH mice was accompanied by dyslipidemia and liver lipid metabolism disorders. Four lipid metabolism related miRNAs, miR-33, miR-122, miR-199a-5p, and miR-375 in the liver of SCH mice were significantly decreased compared with those of control ( P>0. 05). Conclusion The noninvasive SCH model generated by MMI and miRNAs profile provide an animal model and a molecular basis for the study of SCH related lipid metabolism disorders.
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Objective To investigate the association between maternal pre-pregnant body mass index and gestational weight gain,as well as their interaction on neonatal birthweight.Methods We built a cohort in Anqing Municipal Hospital from January 2014 to March 2015,enrolling pregnant women who decided to give birth in this hospital.All women were asked to fill a questionnaire for basic information collection.Medical information of both pregnant women and their newborns were obtained through electronic medical record.Chi-square analysis,multinomial logistic regression,multiplicative and additive interaction methods were used to analyze the association between prepregnant body mass index and gestational weight gain as well as their interactions on birth weight of the neonates.Results A total of 2 881 pregnant women were included in this study.Of the 2 881 newborns,359 (12.46％) were small for gestational age (SGA) and 273 (9.48％) were large for gestational age (LGA).After adjusting the possible confounding factors,results from the multinomial logistic regression showed that pre-pregnancy underweight women were more possible to deliver SGA (aRR=1.33,95％CI:1.02-1.73).If the gestational weight gain was below the recommended criteria,the risk of SGA (aRR=1.64,95％ CI:1.23-2.19) might increase.Pre-pregnancy overweight/obese could increase the risk of being LGA (aRR=1.86,95％ CI:1.33-2.60).Maternal gestational weight gain above the recommendation level was associated with higher rates of LGA (aRR=2.03,95％CI:1.49-2.78).Results from the interaction analysis showed that there appeared no significant interaction between pre-pregnancy BMI and gestational weight on birthweight.Conclusion Pre-pregnancy body mass index and gestational weight gain were independently associated with neonatal birthweight while pre-pregnancy BMI and gestational weight gain did not present interaction on birthweight.
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Objective To investigate the association between maternal pre-pregnant body mass index and gestational weight gain,as well as their interaction on neonatal birthweight.Methods We built a cohort in Anqing Municipal Hospital from January 2014 to March 2015,enrolling pregnant women who decided to give birth in this hospital.All women were asked to fill a questionnaire for basic information collection.Medical information of both pregnant women and their newborns were obtained through electronic medical record.Chi-square analysis,multinomial logistic regression,multiplicative and additive interaction methods were used to analyze the association between prepregnant body mass index and gestational weight gain as well as their interactions on birth weight of the neonates.Results A total of 2 881 pregnant women were included in this study.Of the 2 881 newborns,359 (12.46％) were small for gestational age (SGA) and 273 (9.48％) were large for gestational age (LGA).After adjusting the possible confounding factors,results from the multinomial logistic regression showed that pre-pregnancy underweight women were more possible to deliver SGA (aRR=1.33,95％CI:1.02-1.73).If the gestational weight gain was below the recommended criteria,the risk of SGA (aRR=1.64,95％ CI:1.23-2.19) might increase.Pre-pregnancy overweight/obese could increase the risk of being LGA (aRR=1.86,95％ CI:1.33-2.60).Maternal gestational weight gain above the recommendation level was associated with higher rates of LGA (aRR=2.03,95％CI:1.49-2.78).Results from the interaction analysis showed that there appeared no significant interaction between pre-pregnancy BMI and gestational weight on birthweight.Conclusion Pre-pregnancy body mass index and gestational weight gain were independently associated with neonatal birthweight while pre-pregnancy BMI and gestational weight gain did not present interaction on birthweight.
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<p><b>OBJECTIVE</b>To evaluate the safety and feasibility of laparoscopic gastrectomy for gastric stump cancer.</p><p><b>METHODS</b>Clinical and follow-up data of 7 patients who underwent laparoscopic gastrectomy for gastric stump cancer in our department from January 2008 to July 2015 were analyzed retrospectively.</p><p><b>RESULTS</b>There were 5 male and 2 female patients, with a mean age of (62.1±10.7) years. Initial gastrectomy was performed for gastric cancer in 3 patients and peptic ulceration in 4. The initial surgery was B-II( gastrojejunostomy in 6 patients and Roux-en-Y gastrojejunostomy in 1. Duration between primary gastrectomy and occurrence of gastric stump cancer was ranged from 6-30 years for peptic ulceration, and from 11-15 years for gastric cancer. During the operation, adhesiolysis and exploration to locate the tumor were performed. Following total remnant gastrectomy and lymphadenectomy, intracorporeal anastomosis was accomplished by Roux-en-Y reconstruction. The methods of intracorporeal esophagojejunostomy were end-to-side approach using a circular stapler in 1 patient, side-to-side approach using an endoscopic linear staple in 2 patients, and hand-sewn technique in 4 patients. The operation time was (247.1±17.5) minutes and the intraoperative blood loss was (100.0±30.8) ml without transfusion. The number of retrieved lymph node was 19.1±4.8. The first flatus time, diet resumption time, postoperative hospital stay were (3.3±1.5) days, (3.7±0.8) days, (9.4±2.6) days, respectively. One patient experienced gastrointestinal bleeding that was managed conservatively and ultimately cured. Seven patients were followed up till January 2016. After follow-up from 6 to 38 months, 1 patient died of peritoneal metastasis 17 months after surgery, and 1 patient died of Alzheimer's disease 19 months after surgery. The other 5 patients were still alive without metastasis or recurrence.</p><p><b>CONCLUSION</b>Laparoscopic gastrectomy for gastric stump cancer is feasible and safe.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Anastomosis, Roux-en-Y , Blood Loss, Surgical , Gastrectomy , Gastric Bypass , Gastric Stump , Pathology , General Surgery , Laparoscopy , Length of Stay , Lymph Node Excision , Neoplasm Recurrence, Local , Operative Time , Retrospective Studies , Stomach Neoplasms , General Surgery , Surgical StaplingABSTRACT
Objective It has been reported that Ik6 overexpression is associated with poor outcome of childhood pre-B acute lymphoblastic leukemia and Sup-B15 cell line is Ik6 positive.There were fewer studies on the Sup-B15 in the country,so the aim of this research is to explore chemosensitivity and angiogenic potency of pre-B acute lymphoblastic leukemia cell line Sup-B15 associated with Ik6 positive expression through making a comparison between Sup-B15 and Nalm-6 cell line.Methods The Ik6 mRNA and protein expression were detected by nested reverse transcription polymearse chain reaction and Western blot.The effect of vincristine,daunorubicin,L-asparaginase and dexamethasone on the proliferation of cells was measured by using CCK-8 assay.The migration and invasion of cells were detected by transwell assay and mRNA expression of angiogenesis genes (VEGF,Flt-1,PlGF,IGF-1,Ang1 and Ang2) was measured by real-time PCR.Results No expression of Ik6 was detected in Nalm-6 cell line.Compared with Nalm-6,Sup-B15 cell line had a lower rate of growth.The 4 drugs could inhibit the proliferation of cells in a dose-and time-depen-dence manner.Sup-B15 was more sensitive to daunorubicin,L-asparaginase and dexamethasone,and the folds of respective IC50 value were 9.5,1.1 and 1.5.But Sup-B15 was more resistant to vincristine than Nalm-6.The result of transwell assay showed that less Sup-B15 cells transmigrated to the lower chamber.A significant lower expression level of angiogenesis genes was detected in Sup-B15 cells.Conclusion These findings suggest that Sup-B15 is more sensitive to chemotherapy and less angiogenic compared with Nalm-6.
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BACKGROUND: Placenta mesenchymal stem cells have become hot spots in stem cells study in recent years because of its advantages. OBJECTIVE:To analyze the biological characteristics of amniotic mesenchymal stem cells and amnion epithelial cells, and to explore the feasibility of amniotic mesenchymal stem cells and amnion epithelial cells applied as seed cells in three-dimensional liquid culture to construct the tissue-engineered skin. METHODS:The amniotic mesenchymal stem cells and amnion epithelial cells were obtained by using multi-step digestion with trypsin and col agenase;then the flow cytometry, reverse transcription-PCR and immunofluorescent staining techniques were adopted to identify the surface molecular markers, stem cellcharacteristics and keratinocytes similarity respectively. Based on these data, amniotic mesenchymal stem cells and amnion epithelial cells used as seed cells together with rat type Ⅰ col agen matrix were adopted for three-dimensional liquid culture. RESULTS AND CONCLUSION:Flow cytometry test showed that amniotic mesenchymal stem cells and amnion epithelial cells cultured in vitro could highly express CD90, CD73 and CD105, and could not express the hemopoietic stem cellmarker of CD34 and MHC-class Ⅱ molecular HLA-DR. Reverse transcription-PCR results detected that amniotic mesenchymal stem cells could express stem cellcharacteristic genes CMCY and NANOG;amnion epithelial cells could express stem cellcharacteristic genes CMCY and KLF4, showing that both amniotic mesenchymal stem cells and amnion epithelial cells have stem cellproperties. Reverse transcription-PCR results showed that amniotic mesenchymal stem cells could express keratinocytes characteristic genes K19,β1-integrin and K8;amnion epithelial cells could express K19,β1-integrin, K5 and K8. Immunofluorescence staining results showed amnion epithelial cells could express keratinocytes proliferation related protein K14, which revealed that there was certain similarity in the mRNA expression between keratinocytes and amnion epithelial cells, and indicating that it has the potential to differentiate into keratinocytes. Tissue-engineered skin was successful y constructed by using amniotic mesenchymal stem cells and amnion epithelial cells, hematoxylin-eosin staining section showed that it has certain skin structure, and amnion epithelial cells had a preliminary differentiation. Al these prove that it is feasible to construct human skin tissues with amniotic mesenchymal stem cells and amniotic epithelial cells through the three-dimensional culture.
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Idiopathic thrombocytopenic purpura (ITP),also called immune thrombocytopenia more frequently by the international,is the most common acquired hemorrhagic disease characterized by thrombocytopenia,presence of anti-platelet antibody in the plasma,with increased or normal megakaryocytes with abnormal maturation.About 70％ of 1TP patients recover spontaneously within 6 months,while a small proportion of patients will transform to persistent,chronic or refractory idiopathic thrombocytopenic purpura.New immunosuppressive agents and thrombopoietin for the treatment of children with refractory ITP have been developed.Dramaric progresses in the diagnostis,severity assessment and treatment of refractory ITP have been made recently,which is comprehensively illustrated in this review.
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Objective:To investigate the effect of IL-7 on differentiation of thymic T lymphocytes and dendritic cells' differentiation in vitro.Methods:Thymic lobes were apspectically removed from 15-day-old or 16-day-old mouse fetueses and were cultured by the way of fetal thymus organ cultures (FTOC) in vitro.Lobes were cultured in the dishes with or without Interleukin-7(IL-7),then collected the cultured cells respectively.The expression of CD4,CD8,CD11c,B220,Ia,and CD11b was detected by flow cytometry,and the morphology was observed under the ligt microscope.The proliferation of these cells was determined by cell counting.The cultured cells were used as stimulators for allogeneic T cells,whose proliferation were detected later by MTT method.Results:The result of cell counting indicated that the total amount of the thymocytes was decresed obviously after being cultured with IL-7,while the proportion of CD4~+ CD8~+ thymocytes decreased,the proportion of CD4~- CD8~-,and CD4~- CD8~+ thymocytes increased,and no much difference in the amount of CD4~+ CD8~- thymocytes was found.In addition,the amount of B lymphocytes,natural killer cells,and dendritic cells increased in different degree.Conclusion:IL-7 participates in differentiation and development of thymic T lymphocytes and dendritic cells.
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Objective:To optimize the culture system for the mesenchymal stem cells from the Placenta Chorion (hCMSC) in vitro by adding the selected cytokines to amplify the hCMSC effectively,with furthermore some preliminary analysis of the hCMSC performed.Methods:Immunohistochemistry was applied to show the localization of the mesenchymal stem cells within the placenta chorion,then the corresponding part of chorion was digested by collagenase Ⅳ to get the cell suspension for primary culture and subculture;Flow Cytometry was used to detect the surface marker of hCMSC,whose proliferation was detected by direct cell counting and the MTT method;then Butylated Hydroxyanisole (BHA),and DMSO was used combinedly as the inducer in the differentiation of the hCMSC to the neuron like cells,which was then detected by Immunofluorescence staining for the expression of NSE;finally RT-PCR was used to detect the expression of flt3.Results:The result of immunohistochemistry showed CD105~-,CD90~- positive hCMSC was much easier to be found around the blood vessels within the villous of chorion,which then could be obtained by mechanical dissociation and collagenase digestion.bFGF was good at amplifying the hCMSC among all selected cytokines,but FL was showed even obviously effective on the proliferation of the hCMSC.The propagating cells were found to highly express CD73,CD90,CD105;and negative for CD14,CD34,CD45 and HLA-DR.We also found that the hCMSC happened to express PDL-1 the negative co-stimulatory molecules,while their potential to different into the neuron like cells was still well maintained.Conclusion::There is low immunogenicity mesenchymal stem cells within the placenta chorion,both bFGF,FL are shown good effects on the proliferation of the hCMSC,with maintaining their differentiation potential as well.
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Objective To investigate the distribution and changes of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1) in normal heart or heart after myocardial infarction (MI),and to explore beneficial effects of bFGF on heart after MI. Methods Eleven adult minipigs were randomly divided into 2 groups,control (n=5) and MI group (n=6). MI was induced by ligating the left anterior descending coronary artery. Sham-operation was carried out on the animals of control. bFGF-like-immunoreactivity (bFGF-ir) and FGFR-1-like-immunoreactivity (FGFR-1-ir) in heart tissues were detected by immuno-histochemistry and image analysts in 4 weeks after surgery. Results In controls,bFGF-ir and FGFR-1-ir in the atrium showed a considerable high level compared with 2 ventricles (P