Objective To explore the application of the automatic bone age assessment of BoneXpert software in Children and adolescent of China (Shanghai).Methods Left hand and wrist radiographs of 434 cases of children from trauma (264 boys and 170 girls) were obtained from Jan.2010 to Dec.2013.Bone age was assessed by BoneXpert software and 40 radiographs were randomly chosen and re-evaluated after a week.Results Seven images were rejected by BoneXpert,so the efficiency of the software was 98.4％ and it takes only 14.5 s for every image.Sample assessments by two times are exactly the same.As to BoneXpert software,there were statistical differences between bone age and chronological age in 8 groups:7,8,9,11 years old boys and 2,3,9,11 years old girls(P ＜ 0.05) out of 26 groups,and the average difference was about 1 s.The following modifications suggested that 0.57,0.79,0.93,-0.38 and-0.22,0.30,0.37,0.59 years could be added respectively for the groups of boys aged 7,8,9,11 and girls aged 2,3,9,11 years old according to their average differences between the bone age and the chronological age.Conclusions The BoneXpert method for automated determination of skeletal maturity which evaluates bone age objectively and quickly can be applied clinically after slight adjustment in some age groups.BoneXpert software is 100 percent automatic and could eliminate the subjective evaluation inaccuracy.

Cytokinesis ; Micronucleus Tests

UNLABELLEDOBJECTIVE To investigate the effect of Nepsilon-(carboxymethyl) lysine albumin (CMLs), a primary advanced glycation end products (AGEPs) isoform in diabetic body, on the function and angiogenesis of adipose tissue-derived stem cells (ADSCs) and the protective effect of Danhong Injection (DH). METHODS Human ADSCs were cultured and separated from human subcutaneous fatty tissue using enzymatic digestion and centrifugation. The morphology was observed using optical microscope and differentiation capacities assessed. Cells were exposed to 5 different interventions respectively for 24 h, i.e., PBS, 60 1 microg/mL BSA, 60 microg/mL CML-BSA, 100 microL/mL DH, and 60 micro./mL CML-BSA +100 microL/mL DH. Their effect on the proliferation, migration, apoptosis, and secretion were observed using WST-1 assay, Transwell assay, Annexin V-FITC/PI flow meter test reagent kit, human VEGF reagent kit, ELISA reagent kit, respectively. The effect on ADSCs angiogenesis was observed by in vitro angiogenesis test.

RESULTSCompared with the BSA group, the capacities of proliferation and migration could be significantly inhibited by CML-BSA, the apoptosis promoted, the secretion of VEGF reduced, and the angiogenesis of ADSCs weakened (P < 0.05). Compared with the blank control group, 100 microL/mL DH could significantly promote the proliferation and migration capacities of ADSCs, inhibit apoptosis of ADSCs, increase the secretion of VEGF, and improve the angiogenesis of ADSCs (P < 0.05). Compared with the CML-BSA group, the inhibition of CML-BSA on the proliferation and migration capacities of ADSCs could be significantly reversed, the promotion of CML-BSA on the apoptosis of ADSCs improved, the secretion of VEGF increased, and the angiogenesis of ADSCs elevated (P < 0.05).

CONCLUSIONclusion CMLs could significantly inhibit the proliferation and migration capacities of ADSCs, promote their apoptosis, and inhibit their angiogeneses, which could be improved by DH.

Adipose Tissue ; cytology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glycation End Products, Advanced ; pharmacology ; Humans ; Neovascularization, Pathologic ; drug therapy ; Stem Cells ; cytology ; drug effects

Objective In the previous study completed in Korea, early three-dimensional conformal radiotherapy concurrent with chemotherapy in rectal cancer after radical surgery demonstrated a better prognosis compared with late radiotherapy. However, with the report of MOSAIC trial, the adjuvant chemotherapy regimen had transferred from 5-Fu alone to combined 5-Fu and Oxaliplatin. We need more evidence to clarify the best radiotherapy timing during the whole period of adjuvant therapy . Methods Patients who met the following criteria were accrued in this study: 18-70 years old, lower anterior resection,pathologically proven rectal adenocarcinoma, stage T3-4 and/or N+, no evidence of distant metastases and KPS≥70. Three dimensional conformal Radiotherapy was started at the fourth week after surgery, which included 45 Gy whole pelvic radiation following by 6-10 Gy tumor bed boost. Oxaliplatin of 50 mg/m2 weekly and Xeloda 625 mg/m2 twice a day, every week from d1-5 were used concurrent with radiotherapy.Toxicity was evaluated and graded by common toxicity critera version 3. 0. The study was designed as Simon two-phase design, in the first phase, a total of 15 patients were accrued, and if more than or equal to 9 patients had grade 3 toxicity, we had 85% power to confirm the toxicity caused by early radiotherapy more than 50%. Otherwise, another 15 patients of the second phase were accrued, we would have 85% power to confirm the high toxicity of more than 50% if 18 out of 30 patients had grade 3 toxicity. Results From July 2008 to December 2008, 15 patients were treated with early radiotherapy concurrent with combined chemotherapy, Grade 3 gastrointestinal toxicity occurred in 12 patients and Grade 3 hematologic toxicity occurred in 2 patients. According to Simon design, we had 85% power to confirm the toxicity caused by early radiotherapy more than 50%. Conclusions For locally advanced rectal cancer patients, whole pelvic radiotherapy concurrent with oxaliplatin and xeloda had severe toxicities. Further studies are needed to decrease toxieities.

Objective To explore the clinical characteristics and diagnostic criteria of migraine in children.Methods The migraine cli-nical characteristics of patients that consistent with the chronic headache diagnostic criteria, excluding other diseases,were analyzed.ICHD-Ⅱ childhood migraine diagnostic criteria was used as gold standard to explore the migraine diagnostic criteria in children.Results 1.In 346 patients, 157 fitted ICHD-Ⅱchildhood migraine diagnostic criteria.2.Type of migraine: probable migraine was the most common(68.8%), the most commonly unfulfilled criterion was associated gastroenteritis symptoms; migraine without aura was the second (19.7%).3.The migraine diagnostic criteria: the most important single-variable was headache associated symptoms,with sensitivity of 70.7%,specificity of 83.6%,the positive predictive value(PPV) was 78.2%,positive likelihood ratio (PLR) was 4.31 and area under curve(AUC) was 0.771.The most important three-variables was headache duration of 1-72 hours, moderate-severe headache and headache associated symptoms, with the sensitivity of 52.2%,specificity of 96.2%,the PPV was 90.1%, the PLR was 13.7 and the AUC was 0. 657.Conclusions The most important single-variable is headache associated symptoms,the most important three-variables are headache duration 1-72 hours, moderate-severe headache and headache associated symptoms.

Objective To evaluate the effects of Guhong Injection on motor dysfunction in rats after cerebral ischemia- reperfusion. Methods The middle cerebral arteries were occluded for 2 hours and re-perfused in Sprague-Dawley rats. They were divided in sham group, model group, Aceglutamide group, Safflowere group and Guhong group, which were intravenously administrated with normal saline, Aceglutamide, Safflower or Guhong 24 hours after operation, and continued for 14 days. They were tested with the beam-walking test after treatment. Tyrosine hydroxylase (TH) immunohistochemical staining was used to investigate the viability of neurons in the substantia nigra. Results The model group spent more time in the beam-walking test than that in the sham group (P<0.01), and it decreased in the Safflower group and Guhong group compared with that in the model group (P<0.05). The TH-positive neurons decreased in the model rat compared with that in the sham group (P<0.001), and increased in both Safflower and Guhong groups compared with that in the model group (P<0.01). Conclusion Guhong administration could significantly improve the motor dysfunction in rats after cerebral ischemia- reperfusion, which might be related to provent the neurons from injury in the substantia nigra.

Objective To study the effects of Guhong injection on the expression of vascular endothelial growth factor (VEGF) in the cortex 14 days after cerebral ischemia-reperfusion. Methods 30 male Sprague-Dawley rats were divided into sham group (n=6), ischemia group (n=6), aceglutamide injection group (n=6), Honghua injection group (n=6) and Guhong injection group (n=6). The middle cerebral arteries of all the rats were occluded for 2 hours and reperfusion, except the sham group. Drugs were administered once a day 24 hours after reperfusion. The expression of VEGF in cortex was detected with enzyme-linked immunosorbent assay (ELISA) 14 days after reperfusion. Results The expression of VEGF decreased in the ischemia group compared with the sham group (P<0.001), and it increased both in the aceglutamide and Guhong injection groups compared with the ischemia group (P<0.05). Conclusion Guhong injection can significantly increase the expression of VEGF in the cortex 14 days after ischemia-reperfusion, which may be one of the ways for neuro-protection.

OBJECTIVETo explore the effect of berberine on lipid metabolism disorder and lipid deposition in liver cells of non-alcoholic fatty liver disease (NAFLD) rats induced by high fat diet.

METHODSAfter one week adaptable feeding, 45 SPF level male SD rats were randomly divided into 3 groups, the normal control group, the model group, and the berberine group, 15 in each group. Except those in the normal control group, all rats were fed with high fat diet to prepare NAFLD model. As for rats in the berberine group, Berberine Hydrochloride was administered by gastrogavage. HE staining and oil red O staining were performed to identify the model after 8 weeks. Hepatocytes were isolated, and their activities and purities were tested by Typan blue staining and flow cytometry (FCM). Serum levels of TC, TG, HDL-C, and LDL-C were detected using automatic biochemical analyzer. mRNA expression levels of LXRα and FAS in liver cells were analyzed by Real-time quantitative polymerase chain reaction (PCR). Protein levels of LXRα and FAS in liver cells were examined by Western blot.

RESULTSThe NAFLD rat model was successfully established by high fat diet. The yields of purified liver cells in each rat were (6.0-7.5) x 10(8). The viability of isolated liver cells with purity over 90% (tested by FCM analysis) was higher than 95%. Compared with the normal control group,the expression of LXRα and FAS at mRNA and protein levels was higher in the model group (P < 0.01). Compared with the model group, the expression of LXRα and FAS at mRNA and protein levels was obviously down-regulated in the berberine group (P < 0.01).

CONCLUSIONSLXRα/FAS signaling pathway was one of important signaling pathways of NAFLD lipid metabolism disorders. Berberine could recover hepatocyte fatty deposits in NAFLD rats by adjusting the LXR/FAS signaling pathway of hepatocytes, which might be one of important mechanisms for fighting against NAFLD.

Animals ; Berberine ; therapeutic use ; Diet, High-Fat ; Down-Regulation ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver ; Hepatocytes ; Lipids ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

BACKGROUNDAcute lung injury (ALI) is a serious and common condition for which there are currently no specific strategies for treatment. Recent studies have suggested that bone marrow-derived multipotent mesenchymal stem cells (MSCs) may have therapeutic applications in multiple clinical disorders. We explored the biological effects of MSCs during endotoxin-induced ALI and the mechanisms involved.

METHODSMSCs were isolated from male rat bone marrow and the ALI model was induced by intravenous endotoxin injection. Female rats were sacrificed at 6 hours, 24 hours, 4 days, 1 week and 3 weeks post-injection of MSCs or saline and the lung tissue, bronchoalveolar lavage fluid, and serum were harvested for analysis. We further evaluated the survival of the rats and examined the effects of endotoxin-induced injury on the interaction between alveolar macrophages (AMs) and MSCs in ex vivo.

RESULTSThere was a significant decrease in numbers of neutrophils in bronchoalveolar lavage fluid (P < 0.05), and myeloperoxidase activity in the lung (P < 0.01), and of TNF-α and IL-1β in serum (P < 0.05) in the MSC treated rats at 4 days. Furthermore, MSC treated rats exhibited improved survival, lower lung injury score, higher concentration of IL-10 in the serum and a reduced hydroxyproline content, but these differences were not statistically significant. Moreover, co-cultures of MSCs and AMs had significantly reduced levels of TNF-α, IL-1β and macrophage inflammatory protein (MIP)-1α and significantly increased levels of IL-10 (P < 0.05) in the culture supernatants.

CONCLUSIONSTreatment with intravenous injection of bone marrow-derived MSCs have beneficial effects on endotoxin-induced ALI in rats. The beneficial effect might be achieved through the engraftment of differentiated MSCs in the lungs and appears derive more from their capacity to secrete soluble factors that modulate immune responses.

Acute Lung Injury ; chemically induced ; metabolism ; therapy ; Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Coculture Techniques ; Endotoxins ; toxicity ; Female ; Lung ; metabolism ; pathology ; Macrophages, Alveolar ; cytology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; physiology ; Peroxidase ; metabolism ; Random Allocation ; Rats ; Rats, Wistar