Objective To investigate the damage mechanism of fluetuant high glucose on the INS-1 cells (pancreatic β-cell lines).Methods The cells were divided into five groups:the control groups (A group:5.5 mmol/L of glucose),the continuing high glucose group (B group:16.7 mmoL/L of glucose),the fluctuant glucose group ( C group:16.7 mmol/L of glucose for cultivation for 2 h,then the concentration changed to 5.5 mmol/L for cultivation for 3 h,which was repeated 3 times per day;the ceils were kept in the medium containing 5.5 mmol/L of glucose during night time for 9 h),the continuing high glucose plus NAC ( 1.0 mmo/L) group ( D group),the fluctuant glucose plus NAC group ( E group).The intracellular reactive oxygen species (ROS) was observed by the flow cytometry.The activity of glucose-6-phosphate dehydrogenase (G6PD) was estimated by the tetrazolium linked cytochemical method.Results 72 h after intervention,the levels of ROSwere 37.77±2.31,86.97±7.97,124.27±10.04,60.92±2.61 and 51.47±3.36,respectively,in A～E group;the activities of G6PD were 1.25±0.03,1.09±0.02,1.03±0.01,1.12±0.02 and 1.21±0.01,respectively;the levels of NADPH were (0.123±0.003) mmol/mg prot,(0.112±0.004) mmoL/mg prot,(0.099±0.002 ) mmol/mg prot,( 0.116±0.005 ) mmol/mg prot and ( 0.120±0.002) mmol/mg prot,respectively.The level of ROS in the cells of the fluctuant glucose group were significantly higher than that in the continuing high glucose group ( P ＜ 0.01 ).The G6PD activity and NADPH was significantly lower in fluctuant high glucose group than those in the continuing high glucose group (P ＜0.01 ).NAC co-cultivation decreased the extent of cell's change.Conclusions Exposure of INS-1 to high glucose lead to increased oxidative stress, possible mechanism included decreased G6PD activity and subsequent imbalance between oxidation and reduction.

Objective To identify whether the branches of cervical nerve roots joined into the accessory nerve trunk or not.Methods In 10 adult cadavers (7 males and 3 females, including 20 laterals of brachial plexus nerves), we observe source of cervical plexus branches to the accessory nerve anatomically.In 10 clinical cases of males with brachial plexus nerve injures, in the supraclavicular approach of brachial plexus exploration, the part of the supraclavicular cutaneous nerve for histological specimen were cut off;in the posterior approach, electrical stimulation of the trunk and branches of cervical plexus were performed to observed istaltrapezius muscle contraction.After accessory nerve transfer, the residual terminal accessory nerve and branch of cervical plexus were taked for histological specimens;and observed and judged of each nerve sample by acetylcholinesterase (AchE) immunohistochemical staining.Results In 10 of 20 lateral cases, cervical plexus communicating branches were derived from the fourth cervical nerve root.The intraoperative electrical stimulation of the accessory nerve trunk, 10 cases of distal trapezius muscle were significantly shrink;electrical stimulation of the cervical plexus branch, 2 cases after stimulation of the mild distal trapezius contraction, the remaining 8 cases without trapezius muscle contraction.10 cases of supraclavicular nerve staining for AchE were negative, 10 cases of accessory nerve terminal branches of AchE staining were mixed,10 cases of branch AchE cervical plexus to the accessory nerve staining were negative.Conclusion The branches of the fourth cervical nerve root constantly joins into the accessory nerve, participating in the trapezius muscle inner vation, the fibers of the branches are sensorial fibers.

OBJECTIVE1H-NMR technology was carried out to investigate the chemical difference between 30 batches of Cibotium baronetz decoction pieces and look for new method for quality control of C. baronetz decoction pieces.

METHODSix hundreds MHz H-NMR spectroscopy and principle component analysis (PCA) were used to discriminate between 30 batches of commercially available cibotium samples based on multi-component metabolite profiles.

RESULTSaccharide is the principle component of C. baronetz decoction pieces, and steroid and triterpene were the discriminately chemical component. Protocatechuic acid, protocatechuic aldehyde, cibotiumbaroside A, cibotiumbaroside B and 4-O-caffeoyl-D-glucoside could be used as the marker for controlling the quality of commercial C. baronetz decoction pieces.

CONCLUSIONPattern-recognition techniques applied to proton nuclear magnetic resonance (1H-NMR) spectra of 80% methanol extraction of C. baronetz could correctly discriminate not only the quality, but also the chemical component for batches of commercial C. baronetz decoction pieces.

Benzaldehydes ; chemistry ; Caffeic Acids ; chemistry ; Catechols ; chemistry ; Drugs, Chinese Herbal ; chemistry ; standards ; Ferns ; chemistry ; Furans ; chemistry ; Glucose ; chemistry ; Glucosides ; chemistry ; Glycosides ; chemistry ; Hydroxybenzoates ; chemistry ; Magnetic Resonance Spectroscopy ; methods ; Maltose ; chemistry ; Quality Control ; Steroids ; chemistry ; Sucrose ; chemistry ; Triterpenes ; chemistry

OBJECTIVETo study the effect of Feiji Recipe (FR) intervening indoleamine 2,3-dioxygenase (IDO) induced immune escape on the murine model of Lewis lung carcinoma. Methods Totally 48 C57BL/6 mice inoculated with Lewis lung cancer cells transfected with human (enhanced green fluorescent protein,EGFP)-IDO gene were divided into four groups according to radom digit table, i.e., the model group (administered with normal saline by gastrogavage) , the Chinese medicine group (treated with FR Decoction at the daily dose of 100 mg/g by gastrogavage), the 1-methyl-D-trytaphan (1-MT) group (administered with 1-MT mixed liquor at the daily dose of 100 mg/kg by gastrogavage), and the Paclitaxel group (treated with Paclitaxel at the daily dose of 15 mg/kg by peritoneal injection), 12 in each group. The intervention was started from the 2nd day of modeling. The survival time was observed in 24 of them. Ratios of CD4+ CD25+ FoxP3+ regulatory T cells (Treg) in the spleen were detected in the rest 24 mice by flow cytometry respectively.

RESULTSCompared with the model group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells remarkably decreased in the Chinese medicine group, the 1-MT group, and the Paclitaxel group (P < 0. 01). Compared with the Paclitaxel group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells decreased significantly in the 1-MT group (P < 0.05).

CONCLUSIONFR could inhibit the proliferation of lung cancer cells and immune eseape, improve the immune function, and prolong the survival of tumor-bearing mice.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Carcinoma, Lewis Lung ; drug therapy ; immunology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; Lung Neoplasms ; Mice ; Mice, Inbred C57BL ; Paclitaxel ; T-Lymphocytes, Regulatory

Objective To explore the method and effect of thumb and finger reconstruction and repair by emergent primary toes transplantation. Methods Three hundred and fourteen fingers of 282 cases including finger thumb defect and complex injury with hyperthena palm radid soft tissue of forearm were reconstructed and repaired by the second toes, wrap-around flap and the second third toes wrap-around flap combined with distal flap of leg or other complex tissue flap transplantation. Results Three hundred and eight reconstructed fingers survived, the survival rate was 98%. After a follow up to 6 months to 6 years, the reconstructed thumbs and fingers have accepted nip, grasp, opponent function. The result was graded as excellent in 214 fingers, good in 63, fair in 26, poor in 5. The excellent and good rate was 88%according to the upper limb functional evaluation criteria issued by the Hand Surgery Society of the Chinese Medical Association. Conclusion Different type of toes and combined toes transplantation can obtain preferable clinical effect on emergent primary thumb and finger defect.

Objective To explore the changes of structure and function of mitochondria in liver tissue of mice with acute liver injury induced by volatile oils from ArtemisiaeArgyi Folium(VOAAF).Methods VOAAF was obtained by hydrodistillation extraction.specific pathogen free (SPF) Institute of Cancer Research (ICR) mice in half male and half female were randomized into five groups,including control group(n =10),model (carbon tetrachloride,CCl4) group (n =10),low dose (1.9 g · kg-1,n=10),middle dose (2.3 g · kg-1,n =10),and high dose(2.7 g· kg-1,n =12) VOAAF group(test group).The biological samples were obtained in 6 h after gavaging equal volume (20 mL · kg-1) drugs or solvent to mice at single dose.Aspartate transaminase (AST) in serum was detected.Morphological change of mitochondria in hver tissue was observed by transmission electron microscope.Change of mitochondrial membrane potential in liver tissue was examined by JC-1.Change of mitochondrial swelling sensibility in liver tissue was observed by spectrophotometer.Activity change of Na +-K +-ATPase,Ca2+-ATPase and Ca2+-Mg2+-ATPase in liver tissue were detected by phosphorus determination method.Results The levels of AST in control group and low,middle,high dose test group were (154 ± 27),(224 ±56),(251 ±64),(316 ± 101) U · L-1 (P ＜0.05).The mitochondria got swollen and hydropic in some degree,mitochondrial membrane potemial were 31.35 ±9.14,22.06-6.18,21.34 ±5.99,16.48 ±4.88(P ＜0.05).The mitochondrial swelhng sensibility were (1.43 ±0.69) +10-2,(0.68 ±0.28) ×10-2,(0.40 × ±0.18) + 10-2,(0.26 ±0.10) × 10-2 (P ＜ 0.05).The activity change of Na+-K+-ATPase were 4.40 ± 0.90,2.60 ± 0.60,2.20±0.80,2.10 ±0.80(P ＜0.05).The Ca2 +-ATPase were 4.20 ±0.80,2.70 ±0.50,2.50 ±0.80,2.20 ± 0.60 (P ＜ 0.05).The Ca2 +-Mg2 +-ATPase were 3.90 ± 0.70,2.80 ± 0.60,2.60 ± 0.90,2.50 ± 0.70 (P ＜ 0.05).All changes were related to dose variation trend of volatile oils.Conclusion The disorder of the structure and function of mitochondria could be the mechanism of acute liver injury induced by VOAAF in mice.

OBJECTIVETo investigate the effect of high glucose on mitochondrial respiratory chain function in INS-1 cells.

METHODSThe pancreatic beta cell line INS-1 was divided into the normal control (NC), high glucose (HG), and N-acetyl-L-cysteine (NAC) pretreatment groups, which were cultured for 72 h in the presence of 5.5 mmol/L glucose, 16.7 mmol/L glucose, and 16.7 mmol/L glucose with 1.0 mmol/L NAC, respectively. The activities of the enzyme complexes I and III of the respiratory chain in the cells were assessed with spectrophotometry, the ATP levels were examined using a luciferinluciferase kit, and insulin levels detected by radioimmunoassay.

RESULTSThe activities of the respiratory chain enzyme complexes I and III were 1.53-/+0.24 and 1.08-/+0.22 micromol.mg(-1).min(-1) in high glucose group, respectively, significantly lower than those in the normal control group (2.31-/+0.33 and 1.92-/+0.39 micromol.mg(-1).min(-1), P<0.01). ATP and insulin levels also decreased significantly in high glucose group as compared with those in the normal control group (P<0.01). The addition of NAC partially inhibited high glucose-induced decreases in the enzyme complex activities, ATP levels and insulin secretion (P<0.05).

CONCLUSIONThe respiratory chain function is positively correlated to insulin secretion in INS-1 cells, and exposure to high glucose causes impairment of the two enzyme complexes activities through oxidative stress, resulting in the mitochondrial respiratory chain dysfunction. High glucose-induced damages of the mitochondrial respiratory chain function can be partially inhibited by NAC.

Cell Respiration ; drug effects ; Cells, Cultured ; Glucose ; pharmacology ; Humans ; Insulin-Secreting Cells ; cytology ; physiology ; Mitochondria ; physiology ; Oxidative Stress ; drug effects

OBJECTIVEThe beta-thalassemia major is a common hereditary hematology disease in southern China. The combination of blood transfusion and iron chelation is now the reference treatment. The allogeneic hematopoietic stem cell transplantation is the only curative therapy for beta-thalassemia major. In this study the investigators observed and evaluated the effects of umbilical cord blood transplantation (UCBT) for patients with beta-thalassemia major.

METHODSTwelve cases of beta-thalassemia major aged from 1.3 to 8.3 years (8 male and 4 female) received UCBT. Eleven of the twelve donors were siblings and one was unrelative. Eight patients received no antigen and four patients received two antigen disparate grafts. According to the Pesaro's classification for thalassemia, 10 patients were at grade I or II, and 2 were at grade III. The HLA-identical patients accepted the conditioning regimen consisting of busulfan, cyclophosphamide and antithymocyteglobulin. The HLA-mismatched patients accepted the conditioning regimen consisting of hypertransfusions, continuous iv desferrioxamine, hydroxyurea, fludarabine, busulfan, cyclophosphamide and antithymocyteglobulin. The harvest stem cells contained 3.63 - 16.0 x 10(7)/kg of nucleated cells, 0.11 - 1.03 x 10(6)/kg of CD(34)(+) cells and 0.17 - 1.18 x 10(5)/kg of colony-forming-unit-granulocyte macrophages. Cyclosporine alone or in combination with mycophenolate mofetil (MMF) was given for acute graft-versus-host disease (aGVHD) prophylaxis.

RESULTSOf the 12 patients, 10 were engrafted. Ten patients had neutrophil recovery (> 0.5 x 10(9)/L) and seven patients had platelet recovery (> 50 x 10(9)/L). The median time was 18.1 and 57.3 days, respectively. Seven patients had disease-free survival (DFS) at a median follow up of 23 months (range 4 - 63 months). Three patients had rejection and autologous hematopoitic reconstitution. Two patients were not engrafted. One patient acquired severe aplastic anemia, another patient died of severe infection. The incidences of grade I and grade II aGVHD were 60% (6/10) and 40% (4/10), respectively. There were no long-term complications in the disease free survivors.

CONCLUSIONSGrade I-II beta-thalassemia major patients receiving sibling UCBT had high DFS. UCBT is an effective way to treat beta-thalassemia major.

Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Graft vs Host Disease ; epidemiology ; Hematopoiesis ; Humans ; Infant ; Male ; beta-Thalassemia ; mortality ; therapy

OBJECTIVEHematopoietic stem cell transplantation is currently a unique curative therapy for beta-thalassemia major. However, only 30% of patients have HLA-identical siblings to serve as donors. This study investigated the feasibility of hematopoietic stem cell transplantation from HLA mismatched related donors for beta-thalassemia major in children.

METHODSBetween November 2001 and November 2007, 10 patients with beta-thalassemia major at median ages of 4.4 years (range:1.6-9.4 years) received 11 transplantations from their haploidentical donors, either HLA mismatched sibling umbilical cord bloods (n=6) or parents marrows (n=4) or sibling marrow (n=1). The conditioning regiment included fludarabine (100 mg/m2), busulfan (16 mg/kg), cyclophosphamide (200 mg/kg) and antithymocyte globulin.

RESULTSOf the 10 patients, 6 (60%) had sustained engraftment and red blood cell transfusion independence; 2 patients showed transient engraftment but rejected the graft quickly; 1 patients had no evidence of engraftment and developed aplastic anemia; 1 patient who received two transplantations had no evidence of engraftment and developed persistent aplastic anemia. All eight engrafted patients showed grade I to III acute graft-versus-host disease (GVHD), and only one developed limited skin chronic GVHD. The probability of overall and disease-free survival was 90% and 60%, respectively, with a median follow-up duration of 57.1 months (range: 2.5 to 85.1 months).

CONCLUSIONSHaploidentical stem cell transplantation is an alternative option for children with beta-thalassemia major, particularly when a matched sibling donor is not available.

Child ; Child, Preschool ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Histocompatibility Testing ; Humans ; Infant ; Male ; beta-Thalassemia ; therapy

Continued smoking following stroke is associated with adverse outcomes including increased risk of mortality and secondary stroke.The aim of this study was to examine the long-term trends in smoking behaviors and factors associated with smoking relapse among men who survived their first-ever stroke.Data collection for this longitudinal study was conducted at baseline through face-to-face interviews and follow-up was completed every 3 months via telephone,beginning in 2010 and continuing through 2014.Cox proportional hazard regression models were used to identify predictors of smoking relapse behavior.At baseline,372 male patients were recruited into the study.Totally,155 (41.7％) of these patients stopped smoking for stroke,and 61 (39.3％) began smoking again within 57 months after discharge with an increasing trend in the number of cigarettes smoked per day.Exposure to environmental tobacco smoke at places outside of home and work (such as bars,restaurants) (HR,2.34;95％ CI,1.04-5.29,P=0.04),not having a spouse (HR,0.12;95％ CI,0.04-0.36;P=0.0002) and smoking at least 20 cigarettes per day before stroke (HR,2.42;95％ CI,1.14-5.14,P=0.02) were predictors of smoking relapse.It was concluded that environmental tobacco smoke is an important determinant of smoking relapse among men who survive their first stroke.Environmental tobacco smoke should be addressed by smoke-free policies in public places.