Background Glial cell line derived neurotrophic factor (GDNF) is determined to have a neurotrophy effect and promoting effect to the growth of axon.GDNF has been applied in ophthalmology.Research showed that artemin,a new member of GDNF family,has a better function in protection of neuron,but seldom relevant document of distruibution of artemin in retina is found so far.Objective The aim of the present study is to investigate the distribution and expression of artemin in normal rat retinal neuron cells and retinal ganglion cells,and imitate diabetic environment to observe the expression of artemin at the condition of high glucose.Methods Retinal tissue was isolated from clean neonatal SD rats and cultured by expand culture method in DMEM/F12 containing 10% fetal bovine serum.40 mmol/L of glucose was added in medium in the seventh day after culture for 12 hours as experimental group.The expression and location of artemin in retina were tested by real-time PCR and cell immunofluorescence assay.Use of experimental animals followed the Management Regulation of experimental animals of Jiangsu Province.Results Cultured cells showed the typical cell body and processes in the seventh day.Cultured retinal ganglion cells (RGCs) presented the red fluorescence for Thy1.1 antibody,and multiple fluorescence label revealed that RGCs exhibited the green fluorescence for artemin antibody and red fluorescence for Thy1.1 antibody,indicating artemin protein was positively expressed in cultured RGCs.The numbers of positive cells for Thy1.1 antibody was (442±9)/high field in normal culture group and (263±7) /high field in 40mmol/L glucose culture group,showing a significant difference between them (P＜0.05).The expression of artemin mRNA in normal culture group and in 40 mmol/L glucose culture group,was showing a considerably difference between them(P＜0.05).Conclusion Artemin can be expressed in cultured retinal neuron cells and RGCs in rats.High glucose environment down-regulate the expression of artemin.This study proved a new idea for protecting RGCs against damage.

To improve the expression of heterologous genes using baculovirus expression system, we constructed a novel shuttle vector based on the Bm-Bacmid. In the Bm-Bacmid, partial sequences of Chitinase and Cystein Protease were replaced with a tandem cassette of Cm and egfp through homologous recombination. Bombyx mori bidensovirus (BmBDV) ns1 under the control of polyhedrin promoter was inserted into the modified Bm-bacmid by transposition. For comparison, BmBDV ns1 under the control of polyhedrin promoter was also cloned in the wild type Bm-bacmid. The resulting Bm-bacmids were transfected into the cultured BmN cells to prepare recombinant virus to infect silkworms for expression of BmBDV ns1. Total proteins of hemocyte from infected silkworms were subjected to Western blotting and ELISA analysis. The yield of BmBDV NS1 1 with the modified vector was three times as much as that with the unmodified vector. The method to improve the yield of BmBDV NS1 in silkworms will facilitate the function and three-dimensional structure study of BmBDV NS1.
Animals ; Baculoviridae ; Bombyx ; virology ; Cell Line ; Chitinases ; Cysteine Proteases ; Genetic Vectors ; Insect Viruses ; Promoter Regions, Genetic ; Transfection

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy, Fine-Needle ; adverse effects ; methods ; Child ; Diagnosis, Differential ; Diagnostic Errors ; Female ; Hematoma ; etiology ; Humans ; Male ; Middle Aged ; Thyroid Neoplasms ; diagnosis ; pathology ; Thyroid Nodule ; pathology ; Young Adult

Objective To analyze the clinical and histopathologic characteristics in children with subcutaneous panniculitis-like T-cell lymphoma(SPTCL),and explore the pathological diagnosis and differential diagnosis,in order to boost paediatricians to better understanding the disease.Methods One case was diagnosed SPTCL with over 2 years protracted course of fever and multiple skin lesions.The evolvement of clinical presentation,the misdiagnosed experience,the histopathological features,the immunohistochemical results and T cell receptor(TCR)gene cloning rearrangement were observed.The related literatures published were reviewed.Results Skin biopsy showed that the histopathologic findings were limited within subcutaneous fatty tissue,the atypical lymphocytes characteristically rimmed individual fat cells in a lace-like pattern.Immunophenotypic studies showed CD45,CD45RO,CD3,CD5,T cell intracellular antigen-1(TIA-1)and perforin usually expressed possitive,while CD10,CD20,CD56,CD68,epithelial membrane antigen(EMA)and cytokeratin(CK)were negative,implying tumor cells derived from T-cell.The results of TCR gene rearrangement as following:IgH FR2(+),FR3A(-);TCR ? JVI(-),JVII(+);TCR? JD1(-),JD2(-).Although the protocol of children's T-cell Non-Hodgkin lymphoma was administrated,the treatment outcome was poor.Conclusion SPTCL is a special primary cutaneous lymphoma with poor prognosis,skin biopsy of suffered lesion is an important method for the diagnosis of SPTCL children with unclear recurrent fever and multiple skin lesions.

OBJECTIVETo explore the effects of arecoline on hepatic insulin resistance in type 2 diabetes rats and to elucidate its possible mechanism.

METHODSForty five Wistar rats were fed with high fructose diet for 12 weeks to induce type 2 diabetic rat model. rats were randomly divided into 5 groups (n = 8): control group, model group and model group were treated with different dose (0, 0.5, 1, 5 mg/kg) of arecoline. After 4 weeks, the fasting blood glucose, blood lipid and insulin level measured , mRNA expression of liver constitutive androstane receptor (CAR), pregnane X receptor (PXR), glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by reverse transcription polymerase chain reaction (RT-PCR), the protein expression of p-AKT and glucose transporter4 (GLUT4) were detected by Western blot.

RESULTS1.5 mg/kg arecoline could significantly decrease the level of fasting blood glucose, blood lipid, blood insulin level and liver G6Pase, PEPCK, IL-6, TNF-alpha mRNA level in type 2 diabetes rats. 1.5 mg/kg arecoline also could significantly increase CAR, PXR mRNA level and p-AKT and GLUT4 protein expression.

CONCLUSIONArecoline improved hepatic insulin resistance in type 2 diabetes rats by increasing the mRNA levels of CAR and PXR leading to the creased glucose metabolism and inflammation related genes expression.

Animals ; Arecoline ; pharmacology ; Diabetes Mellitus, Experimental ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; Glucose Transporter Type 4 ; metabolism ; Glucose-6-Phosphatase ; metabolism ; Insulin Resistance ; Interleukin-6 ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Phosphoenolpyruvate Carboxykinase (GTP) ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Receptors, Steroid ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

[ ABSTRACT] AIM: To investigate the protective effect of autophagy on oxidized low density lipoprotein ( ox-LDL)-induced macrophage apoptosis and the underlying molecular mechanisms .METHODS:The RAW264.7 macropha-ges were pretreated with 3 mmol/L 3-methyladenine (3-MA), 1 μmol/L rapamycin (Rap) or 4 mmol/L 4-phenylbutyric acid ( PBA) respectively for 1 h and then treated with ox-LDL (100 mg/L) for 12 h.The cell viability and apoptosis were determined by MTT assay and flow cytometry with Annexin V-FITC/PI staining, respectively.The activities of lactate de-hydrogenase ( LDH) in the medium and caspase-3 in the cells were determined by detection kits .The protein levels of bec-lin-1 (a molecular marker of autophagy ), glucose-regulated protein 78 (GRP78, an endoplasmic reticulum stress marker) and C/EBP homologous protein ( CHOP, a key-signaling component of endoplasmic reticulum stress-induced apoptosis ) were examined by Western blot .Microtubule-associated protein 1 light chain 3 (LC3, another molecular marker of autoph-agy) was observed under laser scanning confocal microscope .RESULTS: Treatment of the RAW264.7 macrophages with ox-LDL at 100 mg/L for 12 h resulted in significant decrease in cell viability , and dramatic elevation in LDH leakage , cell apoptosis and caspase-3 activity, which were promoted by 3-MA (an autophagy inhibitor) and inhibited by Rap (an autoph-agy inducer ) .ox-LDL induced autophagy in the macrophages as assessed by beclin-1 upregulation and frequent granulation of LC3, which were inhibited by 3-MA and promoted by Rap.Interestingly, 3-MA enhanced, while Rap blocked, the CHOP upregulation induced by ox-LDL.Moreover , PBA ( endoplasmic reticulum stress inhibitor ) significantly inhibited ox-LDL-induced GRP78 upregulation and autophagy as determined by the attenuation of beclin-1 upregulation and frequent granula-tion of LC3.CONCLUSION: Endoplasmic reticulum stress mediates ox-LDL-induced autophagy in macrophages , and moderates activation of autophagy may protect macrophages from ox-LDL-induced apoptosis by inhibiting CHOP expression .

The ligators we have developed is a kind of economical and effective six-ring ligator. Endoscopic variceal ligation (EVL) was performed to treat bleeding from esophageal varices in patients with liver cirrhosis using self-made ligator and foreign multiple ligator. There are similar effects with both self-made ligator and foreign mutiple ligator in the control of variceal bleeding, variceal obliteration and rebleeding (93.8%, 87.5%, 0 in the group with self-made ligator, 94.5%, 87.1%, 2.4% in the group with foreign multiple ligator, P>0.05). In terms of the quality index, successful operation rate, hemastatic rate, variceal obliteration rate, rebleeding rate, complications and variceal recurrence rate, the self-made ligator is as good as the foreign multiple ligator, but much cheaper.
Adolescent ; Adult ; Child ; Endoscopes ; Equipment Design ; Esophageal and Gastric Varices ; therapy ; Female ; Gastrointestinal Hemorrhage ; therapy ; Humans ; Ligation ; instrumentation ; methods ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo study the effects of quercetin, a flavonoid, on the learning and memory ability of 3-day-old neonatal rats with hypoxic-ischemic brain white matter damage (WMD).

METHODSSixty 3-day-old Sprague-Dawley rats were randomly divided into four groups: control, WMD model,and quercetin treatment groups (20 and 40 mg/kg). There were 15 rats in each group. Rats in the WMD model and the two quercetin treatment groups were subjected to right common carotid artery ligation followed by 2 hrs of exposure to 8% O2 to induce periventricular white matter injury. After the operation quercetin was administered daily in the two quercetin treatment groups for 6 weeks. Six weeks later, Morris water maze and open-field tests were carried out to test memory and learning ability as well as behavior and cognition.

RESULTSFrom the second day of training, escape latency in the Morris water maze test was more prolonged in the WMD model group than in the control group (P<0.01). The escape latency in the two quercetin treatment groups was shortened significantly compared with the WMD model group (P<0.05). The WMD model group crossed the original platform fewer times compared with the control and quercetin treatment groups (P<0.05). The open-field test indicated that the number of rearings increased and time spent in the centre was extended in the WMD model group compared with the control group. Compared with the WMD model group, the number of rearings was significantly reduced (P<0.05) and time spent in the centre was significantly shortened in the quercetin treatment groups (P<0.05).

CONCLUSIONSQuercetin treatment can improve memory and learning ability as well as cognitive ability in neonates with WMD, suggesting that quercetin protects against WMD resulting from hypoxia-ischemia.

Animals ; Hypoxia-Ischemia, Brain ; drug therapy ; psychology ; Learning ; drug effects ; Maze Learning ; drug effects ; Memory ; drug effects ; Quercetin ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the effect of sodium tanshinone IIA sulfonate (STS) in preventing postoperative peritoneal adhesions in rats and explore the mechanisms.

METHODSSixty SD rats were randomized into 4 equal groups, including a blank control group, adhesion model group, and high-, moderate-, and low-dose STS-treated groups, and were subjected to injuries of the parietal peritoneum and cecum to induce peritoneal adhesions, followed by intraperitoneal administration of saline and STS at the doses of 20, 10, and 5 mg/kg for 7 consecutive days, respectively. Another 15 untreated rats served as the blank control group. The adhesion scores in each group were recorded after the treatments; the activity of tissue-type plasminogen activator (tPA) in peritoneal lavage fluid was measured, tPA/PAI-1 protein ratio in the peritoneal tissue was determined by ELISA, and the expressions of TGF-β1 and collagen I were detected by immunohistochemistry. The anastomotic healing model was used to assess the impact of STS on wound healing.

RESULTSIntraperitoneal administration of STS effectively prevented peritoneal adhesion without affecting anastomotic healing in the rats. Compared with the adhesion model group, the STS-treated groups showed increased peritoneal lavage fluid tPA activity and tPA/PAI-1 ratio in the ischemic tissues with lowered TGF-β1 and collagen I expressions in the ischemic tissues.

CONCLUSIONSIntraperitoneal administration of STS can prevent peritoneal adhesion and enhance local fibrinolysis in rats, and these effects may be mediated by TGF-β signaling pathway.

Animals ; Cecum ; injuries ; pathology ; Collagen Type I ; metabolism ; Fibrinolysis ; Injections, Intraperitoneal ; Peritoneum ; injuries ; pathology ; Phenanthrenes ; pharmacology ; Plasminogen Activator Inhibitor 1 ; metabolism ; Postoperative Complications ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Tissue Adhesions ; prevention & control ; Tissue Plasminogen Activator ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing