Objective To screen melanocortin 4 receptor (Mc4R) gene mutation by direct DNA sequencing in order to explore the mutation situation of Mc4R gene in simple obese children in Nanjing.Methods One hundred and five simple obese children(obesity group) and 127 healthy children(healthy control group) were examined for mutations of Mc4R gene.Body mass index(BMI)cutoff points for overweight and obesity adopted Chinese children and adolescents,recommended by China Working Group of Obesity,and all children had no other hereditary and metabolic abnormality.Touch-down PCR was performed to amplify the full length Mc4R gene,then direct DNA sequencing was used to analyze the Mc4R gene.The differences of biochemical index levels between obesity group and healthy control group were analyzed ,including alanine transaminase,aspartate transaminas,total protein,albumin,globulin,albumin/globulin,triglyceride,cholesterol,high density lipoprotein,cortisol,insulin and C peptide.Results There were significant differences of biochemical index levels between obesity group and healthy control group,including triglyceride,insulin level after dining 2 h,C peptide and BMI(Pa

Biomarkers ; urine ; Child ; Child, Preschool ; Cystatin C ; Cystatins ; urine ; Cytomegalovirus Infections ; pathology ; urine ; Female ; Humans ; Kidney ; pathology ; Male ; TATA Box Binding Protein-Like Proteins ; urine ; alpha-Macroglobulins ; urine

[ ABSTRACT] AIM:To investigate the effects of sinapic acid ( SA) on the proliferation and apoptosis of rat vas-cular smooth muscle cells (VSMCs) induced by high glucose (HG).METHODS:Cultured A7r5 cells were randomly di-vided and treated as indicated.The cell viability was determined by MTT assay.DNA synthesis was measured by BrdU as-say.Cell cycle progression and cell apoptotic rate were determined by flow cytometry analysis.The levels of reactive oxygen species (ROS) were detected by ELISA.The protein levels of cyclin D1, P21, P27, phosphorylated protein kinase C (p-PKC), p-P38 andβ-actin were evaluated by Western blot.RESULTS:Compared with control group, the viability of A7r5 cells was significantly enhanced, the DNA synthesis was increased, the cell cycle progression was promoted, the levels of ROS were elevated, the cell apoptotic rate was reduced, the protein expression of P21 and P27 was decreased, and the pro-tein levels of cyclin D1, p-PKC and p-P38 were increased in HG group (all P<0.05).These effects were reversed by SA (0.1, 1 and 10 μmol/L) treatment in a dose-dependent manner (all P<0.05).Both P38 inhibitor SB203580 and PKC inhibitor chelerythrine significantly inhibit HG-induced PKC/P38 activation and cell viability ( P <0.05).CONCLU-SION:SA inhibits HG-induced VSMCs proliferation and promotes cell apoptosis via reducing PKC/P38 activation.

Hemorrhoids ; Humans ; Male ; Mucous Membrane

The purpose of our present work was to study the discharge of bursting-firing neurons (BFNs) in ipsilateral or contralateral hippocampus (HPC), and its relations to the reestablishment of local epileptic networks. The experiments were performed on 140 Sprague Dawley male rats (150-250 g). Acute tetanization (60 Hz, 2 s, 0.4 -0.6 mA) of the right posterior dorsal hippocampus (ATPDH) was administered to establish rat epilepsy model. The single unit discharges and the depth electrographs were simultaneously recorded from ipsilateral or contralateral HPC. In other experimental rats, acute tetanization of the right anterior dorsal HPC (ATADH) was used. Extracellular unit discharges in the CA1 region were simultaneously recorded from bilateral anterior dorsal hippocampi. Analysis of hippocampal BFN firing patterns before or after administration of the tetanization was focused on according to their location in the HPC epileptic networks in vivo. Single unit discharges of 138 hippocampal neurons were recorded from ipsilateral and/or contralateral anterior dorsal HPC. Of the 138 neurons recorded, 19 were BFNs. 13 BFNs were tetanus-evoked and the remaining 6 were spontaneous ones. The evoked reactions of the single hippocampal neuron induced by the tetanization mainly included: (1) the firing patterns of the BFNs in ipsilateral anterior dorsal HPC were obviously modulated by the ATPDH from tonic firing into rhythmic bursting. The bursting interspike intervals (BISI) decreased. (2) There were mild modulations of the firing patterns of the BFNs in contralateral anterior dorsal HPC following post-inhibition of the firing rate of single neuron induced by the ATPDH. The interspike intervals (ISI) increased obviously. (3) Post-facilitation of rhythmic bursting-firing of the BFNs in contralateral anterior dorsal HPC was induced by ATADH; both the ISI and the IBI increased. (4) Synchronous or asynchronous rhythmic bursting-firing of the BFNs and the network epileptiform events ipsilateral or contralateral anterior dorsal HPC were elicited by the ATPDH. The results obtained suggest that bursting-firing of single BFNs is produced by the ATPDH in the anterior dorsal HPC along the longitudinal axis of the ipsilateral HPC or across the hemisphere to the opposite HPC. Rhythmic activities of the BFN may be implicated in the epileptic network reestablishment of the HPC. On the other hand, synaptic modulation of the BFN temporal series might be responsible for pathophysiological information transmission in the HPC-epileptic network.
Animals ; Electric Stimulation ; Electrophysiology ; Epilepsy, Temporal Lobe ; physiopathology ; Evoked Potentials ; Hippocampus ; physiopathology ; Male ; Nerve Net ; physiopathology ; Neurons ; physiology ; Rats ; Rats, Sprague-Dawley ; Synaptic Transmission

Objective:To investigate the effects of Liuweidihuang pill on the insulin levels in sera and pancreatic islets from spontaneous mouse models of human type 2 diabetes administrated with different doses .Methods:The 6-8 week-old KK-Ay mice were randomly divided into three groups including no drug control group ,low-dose group and high-dose group,in addition C57BL/6J mice were used as a genetic control group .All the animals were given with different dose Liuweidihuang pill solutions or sterile distilled water by intragastrical administration for fifteen weeks .The fasting blood glucose ,body mass and food consumption were measured weekly .The serum insulin levels were surveyed by ELISA .And the insulin levels in the pancreas islets were detected by immunofluorescence and immunohistochemistry .Results:Decreased fasting blood glucose ,controlled body mass and food consumption ,and lower levels of insulin in the sera and pancreas islets were confirmed from the KK-Ay mice administered with Liuweidihuang pill .Furthermore,the low dose program exhibits a stronger effect .Conclusion:Liuweidihuang pill has exhibited relatively therapeutic effects in the spontaneous type 2 diabetes mice including controls of hyperglycemia and body mass and relieving insulin resistance .In addition , the low-dose regimen showed even better treatment in controlling insulin levels in the sera and pancreas islets .

BACKGROUNDHyaluronan (HA) is most likely associated with tumor invasion and metastasis. Studies have shown that HA levels are often increased in serum of patients with various malignant tumors. The purpose of this study was to determine the levels of serum hyaluronan in patients with oral cancer and evaluate the value of serum HA in adjuvant diagnosis, staging and monitoring treatment response in these patients.

METHODSEighty-four hospitalized patients with oral cancer, 65 patients with benign tumors in the oral and maxillofacial region and 67 healthy individuals were included in this investigation. Venous blood was collected from these patients and the healthy individuals before therapy. One week after therapy, venous blood was collected once again in 43 patients with oral cancer. Serum samples were obtained and serum HA levels examined.

RESULTSThe serum HA concentration was significantly higher in oral cancer patients than in patients with benign tumors and in healthy controls (P<0.05). The serum HA level in patients with stages III and IV disease was higher than in patients with stages I and II disease, but there was no significant difference in the HA level between stages I and II nor between stages III and IV (P>0.05). After a complete treatment the HA levels in patients with oral cancer became lower than before treatment, but the difference was not significant (P>0.05).

CONCLUSIONSThe results of this study suggest that the determination of HA levels may provide additional information in diagnosis of oral cancer, but its usefulness as an adjunct in clinical staging and in monitoring treatment response was limited.

Aged ; Humans ; Hyaluronic Acid ; blood ; Middle Aged ; Mouth Neoplasms ; blood ; pathology ; therapy ; Neoplasm Staging

BACKGROUNDDermatomyositis (DM) and polymyositis (PM) are common inflammatory myopathies whose immunopathogenic mechanisms remain poorly understood. The NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome is a type of cytoplasmic multiprotein inflammasome and is responsible for the activation of inflammatory reactivations. Responding to a wide range of exogenous and endogenous microbial or sterile stimuli, NLRP3 inflammasomes can cleave pro-caspase-1 into active caspase-1, which processes the pro-inflammatory cytokines pro-interleukin (IL)-1β and pro-IL-18 into active and secreted IL-1β and IL-18. The NLRP3 inflammasome is implicated in infectious and sterile inflammatory diseases. However, it remains unclear whether it is involved in the pathogenesis of DM/PM, which we aim to address in our research.

METHODSIn this study, 22 DM/PM patients and 24 controls were recruited. The protein and RNA expression of IL-1β, IL-18, NLRP3, and caspase-1 in serum and muscle samples were tested and compared between the two groups.

RESULTSThe serum IL-1β and IL-18 levels were significantly higher in DM/PM patients than those in the controls by enzyme linked immunosorbent assay (ELISA, DM vs. control, 25.02 ± 8.29 ng/ml vs. 16.49 ± 3.30 ng/ml,P < 0.001; PM vs. control, 26.49 ± 7.79 ng/ml vs. 16.49 ± 3.30 ng/ml,P < 0.001). Moreover, the real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) showed that DM/PM patients exhibited higher RNA expression of IL-1β, IL-18, and NLRP3 in the muscle (for IL-1β, DM vs. control, P= 0.0012, PM vs. control, P= 0.0021; for IL-18, DM vs. control, P= 0.0045, PM vs. control, P= 0.0031; for NLRP3, DM vs. control, P= 0.0017, PM vs. control, P= 0.0006). Moreover, the protein expression of NLRP3 and caspase-1 in muscle samples of DM/PM patients were also significantly elevated compared to that in the muscles of the controls.

CONCLUSIONSOur findings demonstrate that the NLRP3 inflammasome is implicated in the pathogenesis of DM/PM. High NLRP3 expression led to elevated levels of IL-1β and IL-18 and could be one of the factors promoting disease progress.

Adult ; Caspase 1 ; analysis ; genetics ; Dermatomyositis ; etiology ; Female ; Humans ; Inflammasomes ; physiology ; Interleukin-18 ; analysis ; genetics ; Interleukin-1beta ; analysis ; genetics ; Male ; Middle Aged ; NLR Family, Pyrin Domain-Containing 3 Protein ; analysis ; genetics ; physiology ; Polymyositis ; etiology

The rice water weevil, Lissorhoptrus oryzophilus Kuschel, has two generations in southern Zhejiang, China. To determine oogenesis in first-generation females (summer females) and its relations to temperature, females were collected from a rice field in early and mid-July and reared on young rice plants at 28, 31 and 34 degrees C in the laboratory. Percentage of females having oocytes, number of oocytes of different stages (stage-I, from early previtellogenesis to middle vitellogenesis; stage-II, late vitellogenesis; and mature-oocyte stage), and length of ovarioles were determined every 10 d of feeding. At each temperature, oogenesis took place in over 40% of females after 20~40 d of feeding, but only 0.0~3.3 stage-I, 0.0~0.8 stage-II and 0.0~1.1 mature oocytes were observed at each observation date. Temperature had significant effect on number of stage-I oocytes but not on number of stage-II and mature oocytes in early July females; temperature had no significant effect on number of oocytes of either stage in mid-July females. Conclusively, in southern Zhejiang, summer L. oryzophilus females have great potential to become reproductive on rice, but their oogenesis activity is very low, with the overall procedures little affected by temperature.
Animals ; China ; Female ; Oogenesis ; Oryza ; parasitology ; Seasons ; Temperature ; Weevils ; genetics ; pathogenicity

Aim:To investigate the effect of Hemangiopoietin (HAPO) on the hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice.Methods: Balb/c mice were underwent total body irradiation at 700 cGy 137Cs ? radiation and were treated with HAPO or recombinant human granulocyte colony stimulating factor (rhG-CSF) after irradiation. The hematopoiesis reconstitution of mice were detected. Cells from bone marrow of Balb/c mice were cultured with HAPO or rhG-CSF for 24 hours or 72 hours before or after the cells were irradiated. The viability of cells were assessed and the ability of in vitro hematopoiesis reconstitution were also detected. Result: rhG-CSF and HAPO treated mice both showed increased survival rate and increased colony forming units. The peripheral WBC number increased greatly. The HAPO group was most quickest compared with rhG-CSF group and PBS control group. The number of bone marrow cells at day 14 of rhG-CSF group was higher than that in HAPO group, but the number of bone marrow cells at day 32 of rhG-CSF was lower than that in HAPO group. The number of bone marrow cells at day 42 of rhG-CSF was below normal. The number of bone marrow cells at day 42 of HAPO group was nearly normal. The number of CFU-GEMM in HAPO group was most compared with that in rhG-CSF group and PBS control group at day 7, 14 and 21 after radiation. The survival rate of cells after radiation in HAPO group was markedly higher than that in PBS control group, but the survival rate of cells after radiation in rhG-CSF group was no notable difference compared with that in PBS control group. In MTT assay, both HAPO and rhG-CSF incubation stimulated proliferation of bone marrow cell at 72 hours after radiation. Bone marrow cells formed Hematopoietic islands in HAPO group after radiation and were positive for sca-1 and CD31. CD31 positive endothelial cells increased around the Hematopoietic islands. There was no Hematopoietic islands formation, few CD31 positive endothelial cells and no sca-1 positive cells in PBS control group. Conclusion: HAPO can promote hematopoiesis reconstitution in sub-lethally irradiated Balb/c mice. It can increase the survival rate of mice and stimulate the proliferation of hematopoietic stem cells.