Objective To investigate the effects of N-acetyl-D-glucosamine(GlcNAc)on acute pancreatitis in rats.Methods Twenty male SD rats were randomly divided into a control group,AP group,low GlcNAc + AP group and high GlcNAc + AP group,with five rats in each group.Acute pancreatitis was induced in AP group,low GlcNAc + AP group and high GlcNAc + AP group by two intraperitoneal injections of 2.5 g/kg L-arginine with a 1 hour interval.Among them,low GlcNAc + AP group and high GlcNAc + AP group were administered 50 and 200 mg/kg GlcNAc,respectively,by intraperitoneal injection at 24 hours before the first intraperitoneal injection of L-arginine.Group control and AP were intraperitoneally injected with the same volume of normal saline.After 24 h,the rats were sacrificed,and serum and pancreatic tissues were collected.Pancreatic tissue morphology was observed by HE staining,and serum levels of amylase(AMY),lipase(LPS),interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),superoxide dismutase(SOD)and malondialdehyde(MDA)were measured by enzyme-linked immunosorbent assay.Protein expression of nuclear factor E2-related factor 2(NRF2),heme oxygenase-1(HO-1),and peroxisome proliferator-activated receptor-γ(PPAR-γ)in pancreatic tissue was detected by Western Blot.Cluster of differentiation(CD)86,CD206 and macrophage markers(F4/80)were detected by immunofluorescence.Expression of CD86 and CD206 in pancreatic tissue was detected by immunohistochemistry.Results(1)Compared with control group,AMY,LPS,IL-1β,IL-6,TNF-α,and MDA levels and pancreatic CD86 expression in AP group were significantly increased(P<0.05),while SOD activity,protein expression levels of NRF2,HO-1,and PPAR-γ,and pancreatic CD206 expression were significantly decreased(P<0.05).(2)Compared with AP group,serum IL-1β,IL-6,TNF-α,MDA,and LPS and the pancreatic CD86 expression in low GlcNAc + AP group were significantly decreased(P<0.05).The PPAR-γ protein level in the pancreas was significantly increased(P<0.05).(3)Compared with AP group,serum AMY,LPS,IL-1β,IL-6,TNF-α,and MDA and pancreatic CD86 expression in high GlcNAc + AP group were significantly decreased(P<0.05),while serum SOD,and NRF2,HO-1,PPAR-γ,and pancreatic CD206 expression were significantly increased(P<0.05).(4)Compared with low GlcNAc + AP group,serum LPS,IL-1β and IL-6 in high GlcNAc + AP group were significantly decreased(P<0.05).Pancreatic expression of HO-1,PPAR-γ,and pancreatic CD206 were significantly increased(P<0.05).Conclusion GlcNAc treatment attenuates acute pancreatitis injury in AP rats,possibly by activating the NRF2/HO-1 signaling pathway to inhibit oxidative stress and promoting M2 macrophage polarization to attenuate pancreatic injury in AP rats.

OBJECTIVETo investigate a cerebral autosomal dominant arteriopathy with the subcortical infarcts and leukoencephalopathy (CADASIL) case with clinical manifestations of baldness, lumbago and Parkinson's symptoms.

METHODSClinical and imaging data of the patient were analyzed. The patient and his family members were also subjected to genetic testing.

RESULTSThe symptoms of the patient included recurrent stroke, dementia, and mood disturbance, in addition with lumbago, baldness and Parkinson's symptoms but no migraine. Cranial MRI of the patient showed bilateral symmetric leukoencephalopathy and multiple small subcortical lacunar infarcts. A point mutation in exon 11 of the NOTCH3 gene (R558C) was discovered in the proband and four asymptomatic relatives.

CONCLUSIONCADASIL is characterized by recurrent subcortical ischemic stroke, dementia, pseudobulbar palsy, and mood disturbance. Baldness, lumbago and Parkinson's symptoms may also be seen in such patients.

Alopecia ; etiology ; CADASIL ; complications ; diagnostic imaging ; genetics ; Humans ; Low Back Pain ; etiology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Mutation ; Parkinsonian Disorders ; etiology ; Receptor, Notch3 ; genetics

Objective To analyze the clinical, imaging characteristics and NOTCH3 mutations of cerebral autosomal dominant arteriopathy with the subcortical infarcts and leukoencephalopathy (CADASIL) in Henan, China.Methods CADASIL patients diagnosed by gene or biopsy in People′s Hospital of Zhengzhou University between 2012-2016 were recruited.Clinical and imaging features of these patients were analyzed retrospectively.The distribution of NOTCH3 gene mutations hotspots was described in Henan region at the same time.Results There were 37 patients from 19 families who were diagnosed as CADASIL by genetic testing or biopsy, 27 of whom had symptoms of CADASIL.Two families were confirmed by pathological examination and 17 by genetic testing.Of these 17 families, 13 mutations were found.Mutations in exon 11 were found in eight families, in exon 4 were detected in four families, and in exon 13 were found in two families.Mutation in exons 3, 8 and 20 was detected in one family respectively.Most patients presented with stroke and several presented with cognitive decline.Twelve patients had been attacked by risk factors.Magnetic resonance imaging (MRI) was performed on 22 patients.White-matter lesions were distributed in brain stem, basal ganglia, subcortical, temporal pole, external capsule.There were 19 patients with white-matter lesions in temporal pole and seven in capsula externa, showed as a high signal in T2WI.Conclusions CADASIL patients can be associated with risk factors.T2WI hyperintensities in the anterior temporal lobe were more common than that in the capsular external.Exon 11 and exon 4 were the hotspots for the NOTCH3 mutation in Henan patients.

Objective To investigate the NOTCH3 gene mutation and clinical features in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) families.Methods The clinical features of 4 CADASIL probands in Henan,China were analyzed retrospectively,and the incidences of other members in their families were investigated.The NOTCH3 gene mutations in the 3rd,4th,llth,and 18th exons were detected and the results were analyzed in the patients and some family members.Results Gene sequencing showed that 6 patients in 4 families and 1 mutant carrier had NOTCH3 gene R607C mutation in exon llth,they all met the clinical features of CADASIL.Three patients accompanied with vascular risk factors.The clinical stroke patients had unilateral limb weakness.All 5 patients with complete head MRIdata had thalamic infarction.Conclusions In the 4 CADASIL families of R607C mutation,the clinical features of 6 patients with CADASIL were similar,but there were individual differences in different family members.Imaging examination has important role in the diagnosis of CADASIL.The vascular risk factors,such as hyperte.

Objective To establish a scientific and rational nurse scheduling model for saving nursing human resources and schedule time for head nurses. Methods The nurses and the patients were divided into groups respectively. According to the principle of convention and common multiple, the mathematical logic algorithm was used to calculate working hours, rest and backup scheduling. Results ST basic scheduling model was established and the extension model 1, 2, and merge models 1, 2 was added on the basic of ST scheduling. Conclusions Based on the basic model, free combination and extension model, appropriate scheduling model was selected according to the number of patients and disease condition. It′s convenient which saves the scheduling time of the head nurses. At the same time, it standardizes the backup scheduling, fixes the backup nurses, and saves the nursing resources.

Objective To explore the genetic mechanism of Yang-deficiency constitution by detecting genomic copy number variations (CNVs). Methods Thirty cases of Yang-deficiency constitution and 30 cases of balanced constitution were included according to the standards of Classification and Determination of Constitution in Traditional Chinese Medicine. DNA was extracted from white blood cells in peripheral blood. A genome-wide association study was conducted by using Affymetrix SNP 6.0 platform. CNVs of each sample were analyzed using PennyCNV software. The Yang-deficiency constitution-specific copy number variation regions (CNVRs) of each autosome were identified. CNVR-related genes and their annotations were searched at online Human Genome Browser. Results The mean number of CNVs in balanced constitution group was 12.63±3.39, ranging from 8 to 20. After stepwise elimination of two Yang-deficiency constitution subjects, the mean number of CNVs in Yang-deficiency constitution group was 15.04±8.95, ranging from 2 to 38. A total of 26 CNVRs were identified from 28 Yang-deficiency constitution subjects, including 19 duplicated CNVRs, 6 deleted CNVRs, and 1 mixed type CNVR. Most CNVRs were shared by a few Yang-deficiency constitution subjects, and only 7 CNVRs were shared by more than 5 Yang-deficiency constitution subjects. The functions of representative genes in Yang-deficiency constitution-specific CNVRs were related with extracellular and intracellular signal transduction, metabolic regulation, and immune response, etc. Conclusion Yang-deficiency constitution subjects have some specific genomic CNVs, which might result in Yang-deficiency constitution phenotypes by influencing the expression of genes associated with extracellular and intracellular signal transduction, material metabolism (energy metabolism), and immune response, etc.

To explore the analysis methods for composite traditional Chinese medicine (TCM) constitutions.

BACKGROUND: Recent study showed that osteocalcin may elevate Insulin secretion and sensitivity, prevent the fat accumulation, play a role in the metablism of glucose and lipid. Undercarboxylated osteocalcin works as the main role. OBJECTIVE: To investigate the effect of different concentrations of glucose on osteoblast undercarboxylated osteocalcin. METHODS: The rib trabeculae were resected and broken, trypsinizated and washed completely by PBS. Bone surface and non-adhesive floating cells in cleaning fluid were observed with inverted microscope. Rib trabeculae was washed by DMEM culture medium once, and cultured in culture bottle. The culture liquid was replaced by new one once a week. The osteoblast was moved from the scledte a week later. The cells were fused monolayer and could be subcultured 4 to 6 weeks later. The active second or third generation cells were inoculated to 6-pore plate forming 5 groups. Osteoblast were stimulated by 5.6 mmol/L., 7.6 mmol/L, 9.6 mmol/L, 12.6 mmol/L, 20.6 mmol/L glucose medium respectively after the 80% cells were fused, the vitamin K_2 was added into the culture liquid until the concentration of it to be 10~(-5) mol/L. Supernatant was collected after half hour culturing, the undercarboxylated osteocalcin level were detected with RIA test kit, and corrected it as the total the undercarboxylated osteccalcin, calculated the carboxylated incomplete osteocalcin rate. RESULTS AND CONCLUSION: The rate of ostecblast carboxylated incomplete osteocalcin was different under different concentration glucose. The rate of 7.6 mmol/L, 9.6 mmol/L, 20.6 mmol/L concentration glucose groups were higher than that of 5.6 mmol/L glucose group [(0.27±0.02)%, (0.29±0.04)%, (0.12±0.02)%, P < 0.05]. It is indicated that osteoblast could sense the change of glucose concentration by regulating the secretion of the undercarboxylated osteocalcin between the concentration of 5.6mmol/L to 9.6mmol/L, while the carboxylated incomplete osteocalcin decreased as the concentration of glucose increased.

The relationship between C936T polymorphism at 3'-untranslated region of vascular endothelial growth factor (VEGF) gene and diabetic nephropathy (DN) was analysed in 194 type 2 diabetic patients. The frequencies of genotype CC and allele C were significantly higher in DN group than those in non-DN group and control group. Allele C and genotype CC of VEGF may be a genetic marker susceptible to DN.

BACKGROUND:Adiponectin possess functions of lowering blood glucose and blood lipids, and improve insulin sensitivity. But, controversy results about the effect of adiponectin on skeletal muscle have been reported.OBJECTIVE:To study the effects of eukaryon expressed adiponectin on the glycogen synthesis and glucose oxidation in skeletal muscle cell strain C2C12 myotubes by transfecting plasmids carrying mouse adiponectin.DESIGN: A controlled experiment.SETTING: The Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: PcDNA3.0 plasmid with mouse adiponectin cDNA, pcDNA3.0-mad (generously presented from Dr. Gong,University of Maryland), C2C12 cell strain (purchased from ATCC, GRL-1722), DMEM high glucose (Gibco), MEM (Hyclone), fetal bovine serum (Hanagzhou Sijiqing), equine serum (Hyclone), lipofectamine 2000 (Invitrogene), G418 (Gibco), rabbit anti-mouse adiponectin IgG (ACRP303-A, Alpha Diagnostic International), chemiluminescence kit (ECL+PLUS,Amersham), SABC instant immunohistochemistry kit (Boster), D-[U-14C] glucose (specific activity 9.25-13.32 GBq/mmol,NEC), scintillation fluid POP, POPOP (SIGMA), liquid scintillation counter (LS3801, Beckman, USA).METHODS:This study was carried out in the Central Laboratory of the Second Affiliated Hospital of Sun Yat-sen University from March to August, 2003. ① After extraction of plasmid, double digest with Xho Ⅰ and Xba Ⅰ and identification with HindⅢ digest were carried out. ② Plasmid pcDNA3.0-mad and pcDNA3.0 blank vector were transfected using liposome to C2C12 cells, and the stably transfected cells were screened by 500 mg/L G418 for 3 weeks, G418 resistant C2C12 cells were thereafter harvested, therefore stable transfected pcDNA3.0-mad and pcDNA 3.0 C2C12 cell strains were established.③ Adiponectin protein expression was determined by Western blot analysis and immunohistochemistry. ④ Glucose oxidation and glycogen synthesis detections were divided into control, vector and pcDNA3.0-mad (mad)group. Each group was further divided into 4 subgroups with 0, 0.5, 5 and 100 nmol/L insulin (n =6), respectively. Detection of glucose oxidation and glycogen synthesis was carried out with 14C-labeled glucose by counting radioactivity of 14CO2 or 14C labeled glycogen with scintillation, respectively.MAIN OUTCOME MEASURES:Changes of glycogen synthesis and glucose oxidation in skeletal muscle cell strain C2C12myotubes.RESULTS: ① Results of plasmid transfection and restrict digest: After plasmid extraction, double digest with Xba Ⅰ and Xho Ⅰ was carried out along with HindⅢ digest identification.Digest fragments were in accordance with expectation.Length of adiponectin cDNA fragment was 781 bp, plasmid fragment was 5 446 bp, adiponectin cDNA was inserted between digest sites (Xho Ⅰ and Xba Ⅰ ) of eukaryotic expression vector pcDNA3.0. ② Plasmid transfection of C2C12 cell and positive clone screening: On the 10th day of G418 media culture screening after transfection, most C2C12 cells died.Positive clone appeared at the 2nd week. G418 resistant C2C12 colonies were harvested at the 3rd week. ③ Western blot and immunohistochemical identifications: Both confirmed that adipoenctin gene was stably transfected into cells in the Mad group, with successful adipoenctin expression. ④ Effect of stably transfected adiponectin gene to myocyte glucose metabolism:The myocyte glycogen synthesis and glucose oxidation increased along with the increasing of insulin concentration. The linear regression analyses of measured myocyte glucose oxidation amount showed that the regression coefficients of the control group, blank vector group and mad group were 23.34, 2;3.23 and 26.06 respectively. This result indicated that in C2C12 cell stably transfected with adiponectin gene, when insulin concentration increased, the acceleration rate of glucose oxidation increasing was higher than other 2 groups. However, no significant difference could be observed in glycogen synthesis and glucose oxidation of C2C12 cells under basic status without insulin stimulus and treatment status with different insulin concentrations between control group, blank vector group and mad group (P＞ 0.05).CONCLUSION: ① We have successfully established stably adiponectin gene transfected C2C12 cell strain with adiponectin protein expression ability. ② Transfection with adiponectin cDNA had no significant effect on the glucose oxidation and glycogen synthesis of C2C12 myotubes.③ The glucose oxidation and glycogen synthesis of C2C12 myotubes increased with the increasing of insulin concentration. ④ Adipoenctin may coordinate with insulin in improving myocyte glucose oxidation and increasing myocyte glucose uptake.