Congenital Hyperinsulinism ; diagnosis ; Humans ; Infant, Newborn ; Male

ABSTRACT:Objective For preparation of vascularized islets , to isolate and culture human adipose derived stem cells , investigate the role of adipose derived stem cells (ADSCs ) in promoting the proliferation and vascularization of human umbilical vein endothelial cells (HUVECs ) co‐cultured in vitro , and explore its mechanism .Methods ADSCs and HUVECs were isolated by collagenase digestion method ,then cultured ,and identified by morphology ,immunofluorescence or multi‐directional differentiation .The co‐culture system of ADSCs and HUVECs was established , HUVECs cultured alone were set up for control group . The proliferation , vascularization and concentration of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b‐FGF)in the supernatant were compared between the two groups .Results The third generational ADSCs had uniform long spindle fiberous morphology and multi‐directional differentiational function . Immunofluorescence test of surface antigens on ADSCs revealed CD44/CD49d (+ ) ,CD31/CD34 (-) ,on HUVECs CD31/vWF (+ ) . High vascular density was found when co‐cultured in Matrigel of ADSCs and HUVECs than alone of HUVECs .Growth curve shown at days 3 , 4 and 5 of the logarithmic phase , HUVECs count in co‐culture group of ADSCs and HUVECs was (4 .52 ± 0 .31) × 104 ,(7 .18 ± 0 .45) × 104 ,and (8 .23 ± 0 .36) × 104 under indirect co‐culture condition , while that in individual HUVECs group was (2 .71 ± 0 .25) × 104 ,(4 .87 ± 0 .26) × 104 ,and (6 .86 ± 0 .33) × 104 ( P<0 .01) .Population doubling time of HUVECs was shorter in co‐culture group than in individual group .Also ,the OD value of HUVECs was higher in co‐culture group than in individual group when cultured at days 1 ,3 ,5 and 7 ( P<0 .01) .When cultured at days 3 ,7 and 13 ,the concentration of VEGF and b‐FGF in the supernatant was higher in co‐culture group than in individual group ( P< 0 .01 ) . Conclusion ADSCs can promote the proliferation and vascularization of HUVECs in vitro co‐culture conditions by secreting or increasing the HUVECs secretion of VEGF and b‐FGF .

Objective To isolate and culture human adipose-derived stem cells (hADSCs),investigate the influence of hADSCs on the cellular morphology,survival rate,and function of human islet cells under the in vitro non-contact co-culture conditions,and explore its mechanism.Method hADSCs were isolated by collagenase digestion method,then cultured,and identified by morphology,immunofluorescence and multi-directional differentiation.Adult islet cells were separated and purified by Liberase enzyme and Ficoll 400,then divided into co-culture group and individual group.The cellular growth morphology of islet cells was observed by inverted phase contrast microscope.The survival rate of islet cells,insulin secretory volume,insulin stimulation index and concentration of growth factor in the supernatant were compared between the two groups.Result hADSCs of the third generation showed uniform long spindle fibrocyte-like morphology,and had multi-directional differentiational potentials of osteogenesis and adipogenesis.Immunofluorescence test of surface antigens on hADSCs revealed CD44 + and CD49d +,CD31-,CD34-and CD106-.After 14-day culture,the islet cellular morphology in co-culture group was more intact than that in individual group.The survival rate of islet cells in co-culture group was (82.83 + 2.32) ％,and that in individual group was (53.00 + 2.82) ％ (P＜0.01).Insulin secretory volumes were (23.66 + 2.11) and (7.82 +1.09) mU/L respectively in co-culture group and individual group under high glucose concentration,and 13.22 + 0.77 and 6.40 + 0.44 mU/L respectively under low glucose concentration (P＜0.01 for all).Insulin stimulation index was decreased from 1.67 + 0.10 (at 3rd day) to 1.77 + 0.13 (at 14th day) in co-culture group,and from (1.67 + 0.10) (at 3rd day) to (1.77 + 0.13) (at 14th day) in individual group (P＜0.01).After 14-day culture,the concentrations of HGF,TGF-β,VEGF and bFGF in the supernatant were higher in co-culture group than in individual group (P＜0.01).Conclusion hADSCs were isolated and cultured successfully from adult adipose tissue.They could increase the survival rate and improve the function of islet cells when co-culture with the adult islet cells in vitro through secreting HGF,TGF-β,VEGF and b-FGF.

Objective To investigate the expression and significance of nuclear transcription factor-?B(NF-?B) in childhood acute lymphoblastic leukemia(ALL) and the effect of dexamethasone(DEX) on its expression,to provide the experimental base for corresponding clinical treatment of the ALL,in which NF-?B is taken as a target.Methods 1.The biotin-streptavidin method was used to detect NF-?B P65 protein on 20 childhood ALL patients and 20 healthy children.2.The effect of DEX at clinically relevant dosage on NF-?B P65 protein were also detected by the biotin-streptavidin method.Results 1.The positive expression rate of NF-?B P65 protein in childhood ALL patients was 85.50%,obviously higher than that in normal group(10.0%)(?~2=22.56 P

Objective To evaluate the efficacy and outcomes with the use of a saline-coupled bipolar sealer during liver resection.Methods 101 patients underwent liver resection for primary hepatic carcinoma at the People's Hospital Affiliated to Zhengzhou University from August 2015 to December 2016.The patients were divided into two groups according to whether the Aquamantys(R) system was used or not.In group A (n =62) the clamp crushing technique was used for liver parenchymal transection.In group B (n =39) the Aquamantys(R) system was used.The intraoperative and postoperative complication rates were compared.Results The operation time in group B was significantly longer than group A (216.4 min vs.253.5 min,P ＜ 0.05).The intraoperaitve blood loss in group B was significantly less than group A (381.1 ml vs.257.2 ml,P ＜ 0.05),and less blood transfusion was required (211.3 ml vs.90.9 ml,P ＜ 0.05).The volume of abdominal drainage fluid in group B in the first and the 5th day was significantly less than group A (242.6 ml vs.199.2 ml,P＜0.05;84.3 ml vs.70.4 ml,P＜0.05,respectively).The drainage tube in group B was taken off earlier than in group A (8.1 d vs.7.0 d,P ＜ 0.05).The average hospitalization stay after surgery in group B was also significantly shorter (13.4 d vs.11.6 d,P ＜ 0.05).There was no significant difference in the overall postoperative complication rate (P ＞ 0.05) between the 2 groups,and no death was observed.Conclusion The use of a saline-coupled bipolar sealer (Aquamantys(R)) in liver resection for hepatocellular carcinoma was associated with less intraoperative blood loss and was better for the patients' postoperative recovery.

Objective To investigate the effects of Saccharomyces boulardii ( S. boulardii) on the col-onization of Helicobacter suis ( H. suis) in stomach and the formation of gastric mucosa associated lymphoid tissue (MALT) during H. suis infection. Methods Sixty C57BL/6 wild type mice were randomly divided into six groups. The mice in group A and group B were respectively given sterile distilled water and S. boulardii twice by gavage and then infected with H. suis for one week. The mice in group C and group E were given sterile phos-phate buffer saline by gavage for one week and then respectively given sterile distilled water and S. boulardii by gavage twice a week for 12 weeks. The mice in group D and group F were infected with H. suis for one week and then respectively given sterile distilled water and S. boulardii by gavage twice a week for 12 weeks. Serum and gastric tissue samples were collected from each mouse. Results The bacterial loads of H. suis in the stomachs of mice in group B were significantly lower than those in group A. No significant differences in the levels of se-creted IgA( sIgA) in serum and gastric tissue samples and the expression of IFN-γat mRNA level in gastric mu-cosa samples were found between the two groups. The expression of H. suis 16S RNA and the formation of gastric lymphoid follicles were detected in mice in groups D and F. The levels of sIgA in serum and gastric tissue sam-ples and the expression of IFN-γ and CXCL13 at mRNA level in gastric mucosa samples increased significantly in groups D and F as compared with groups C and E. Compared with the mice in group D, the bacterial loads of H. suis in stomachs, the numbers of MALT per unit length of gastric mucosa and the expression of IFN-γ and CXCL13 at mRNA level in gastric mucosa decreased significantly in mice from group F, but the levels of sIgA in serum and gastric tissue samples increased significantly. Conclusion S. boulardii could inhibit the colonization of H. suis in stomach and suppress the formation of gastric MALT during H. suis infection.

AIM To observe whether acute stimulation of BRL 37344, a β3-adrenergic receptor (β3-AR) agonist, has the same effects of exacerbating hemodynamics and increasing in β3-AR expression on rats with failing heart as chronic administration. METHODSRats received two doses of isoprenaline (Iso, 340 mg·kg-1, sc, with a 24-h interval) to prepare heart failure model. After 8 weeks, rats were given iv BRL 37344 0.4 nmol·kg-1·min-1 for 10 min. Hemodynamics were measured at 0, 10, 30 min, 1, 2, 3 h, 1, 2 and 7 d after BRL 37344. Levels of β-AR mRNA in myocardium were measured at 0, 1, 2 and 7 d after BRL 37344 by reverse transcription-polymerase chain reaction. RESULTSCompared with Iso group, heart rate, left ventricular end systolic pressure and +dp/dtmax were significantly higher, and left ventricular end diastolic pressure was significantly lower during 1-3 h after BRL 37344 injection in Iso+BRL group. Then, they were restored to the same level as that prior to BRL 37344 injection. Compared with normal control, the levels of β1-, β2- and β3-AR mRNA displayed no significant change in BRL group;the level of β1-AR mRNA was lower and the level of β3-AR mRNA was higher in Iso group. In Iso+BRL group, much more lower β1-AR mRNA level and much higher β3-AR mRNA level were shown on d 2 and d 7 than Iso group. CONCLUSION Acute administration of β3-AR agonist has a shorter improved hemodynamics. But it caused the same result as chronic administration in reduction of β1-AR mRNA and increment of β3-AR mRNA in failure hearts, which may aggravate the cardiac functions.

Cleidocranial Dysplasia ; pathology ; Diagnosis, Differential ; Dwarfism ; Humans ; Maxillofacial Abnormalities ; Osteosclerosis ; pathology ; Pycnodysostosis ; diagnostic imaging ; etiology ; pathology ; therapy ; Radiography ; Tooth Abnormalities

Objective To investigate the alterations of Treg cells , Th17 cells and related cyto-kines in peripheral blood of patients during the early stage of hand-foot-mouth disease ( HFMD) .Methods Flow cytometry was performed to analyze the percentages of Treg cells ( CD4+CD25+Foxp3 T cells) and Th17 cells ( CD3+CD8-IL-17+T cells) in peripheral blood samples collected from 49 patients with severe HFMD , 26 patients with common HFMD and 30 healthy children.The levels of IL-6, IL-10, IL-17, IL-23 and TGF-β1 in serum samples were measured by ELISA .Results The percentages of Treg cells , ratios of Treg/Th17 cells, serum levels of TGF-β1 and IL-10 in patients with HFMD were significantly decreased as compared with those of control group (F=5.580, 6.205, 0.000, 0.014, respectively, P<0.05).Patients with se-vere HFMD showed a significantly increased Th17 cells (F=3.189 P<0.05) and a tendency of enhanced IL-17 expression , but no significant differences with the levels of IL-17 were observed .No significant differ-ences with the expression of IL-23 in the patients among each group were detected (P>0.05).The levels of IL-6 in serum samples from severe disease group were obviously increased as compared with those of common HFMD group and control group (F=7.318, P<0.05).Conclusion The results of this study demonstrated that the levels of Treg , Th17 cells and some related cytokines were varied in peripheral blood of patients dur-ing the early stage of HFMD .Inflammatory responses were enhanced to promote anti-virus activities by sup-pressing Treg cells and stimulating Th17 cells.