OBJECTIVETo investigate extracellular splitting pattern of mitochondria and the depressant effects of CsA on the process and explore the mechanism of post-traumatic SIRS and its therapeutic strategy.

METHODSTen male SD rats with 60 to 70 days age and 240 to 280 g weight were used for mitochondrial isolation. Freshly isolated mitochondria were randomly divided into two groups, which were cultured in blood plasma with or without CsA respectively for 8 h. COX and MDH were assayed by ELISA every 30 min. Meanwhile, Rat macrophage cell line NR8383 were treated as follows, control (group A): cultivation with normal medium; NR8383+CsA co-culture group (group B): culture medium was supplemented with CsA of 10 mmol/L; NR8383+intact mitochondria co-culture group (group C): culture medium was supplemented with intact mitochondria (mtDNA=5 g/ml); NR8383+intact mitochondria+CsA co-culture group (group D): culture medium was supplemented with intact mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L; NR8383+disrupted mitochondria co-culture group (group E): culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml); NR8383+disrupted mitochondria+CsA co-culture group (group F): culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L. TNF-α and IL-6 concentrations in supernatant were assessed at 1, 3, 5 h after culture.

RESULTSIn the mitochondria plasma cultures, MDH and COX levels were increased with the time and peaked at about 3 h and 3.5 h; CsA can delay the appearance of peak to 4.5 h. Among different treated groups,there was no significant difference in TNF-α and IL-6 between group A and group B; there was significant difference in TNF-α and IL-6 other groups. After 1 h culture, compared with group C, no significant difference of TNF-α and IL-6 was observed in group D, while TNF-α and IL-6 were significant higher in group E; after 3 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while TNF-α and IL-6 were significantly higher in group E; after 5 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while no significant difference of TNF-α and IL-6 were observed in group E. At each time point, there was no significant difference in TNF-α and IL-6 between group F and group E.

CONCLUSIONMitochondria can split in serum with time, which will further activate macrophages. CsA has depressant effect to mitochondrial splitting on the process and will therefore inhibit the activation of macrophages.

Animals ; Cells, Cultured ; Cyclosporine ; pharmacology ; Interleukin-6 ; secretion ; Male ; Mitochondria ; drug effects ; Prostaglandin-Endoperoxide Synthases ; analysis ; Rats ; Rats, Sprague-Dawley ; Systemic Inflammatory Response Syndrome ; drug therapy ; etiology ; Tumor Necrosis Factor-alpha ; secretion

The pertinent literature on application of traditional Chinese medicine (TCM) in hemopoietic stem cell transplantation was summarized, it indicated that the intervention of TCM could raise the mobilization effect, speed up the hemopoiesis and immunologic reconstruction after transplantation, decrease the incidence of complications, and prolong the life span of patients, showing a preliminary achievement.
Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Graft vs Host Disease ; etiology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Medicine, Chinese Traditional ; methods ; Phytotherapy ; Postoperative Complications ; etiology ; prevention & control

Objective To evaluate the value of reconstruction of three-dimentional CT in development dislocation of hip(DDH)in children.Methods Twelve cases of DDH concluded 4 bilateral and 8 unilateral cases.To sum up,16 sick hips were operated and 8 normal hips were also obtained by three-dimensional CT(Hip speed Fi/x,GE Co).Results 3D reconstruction were used to show femoral head,(acetabulum) and relationship of acetabulum and femoral head respectively.The difference between FNA measurement of sick hips and those of normal hips were significant(P

Objective To investigate the clinical epidemiologic features of transient synovitis(TS) of hip in children occurred in Shen-zhen district.Methods The medical files were reviewed and a standard questionnaire was filled according to the conditions of 705 cases such as pathogeny,clinical manifestation,therapy and prognosis.Results Transient synovitis occurred in a sporadic form all the year round.The peak age of patients with TS was 3-7 years old.The ratio of boys to girls was 2.9:1.About 19.3% patients were attacked an upper respiratory tract infection and 11.9% patients attributed the symptoms to trauma or severe activities before 1 week.A varying degree of painful limp and restriction of movement at the hip were found clinically.All of cases were cured by skin traction.The incidence of recurrence was 6.95%.Conclusions Male predominance is found in TS.It is characteristic of sporadic form in the 4 seasons and intently relation to an upper respiratory tract infection and trauma or severe activities.TS is recurrent and the prognosis is good by skin traction.

OBJECTIVETo explore the effects and mechanisms of ascorbic acid (AA) and sodium selenite (SS) on growth inhibition and redifferentiation in human gastric cancer cells.

METHODSIn the present study, trypan blue dye exclusion method was used to determine the cell growth curve and mitotic index, cell electrophoresis and colonogenic potential were used as the indexes of redifferentiation. In order to find out the mechanisms of redifferentiation, the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) were assayed, the content of malondialdehyde (MDA), reduced glutathione (GSH) and H2O2 were evaluated.

RESULTSAfter treatment with AA 3 mol/L + SS 2 mu mol/L, the growth rate and mitotic index of human gastric cancer cells (MGc-803) decreased remarkably. The indexes related with cell malignancy were alleviated. For example, cell surface charge was obviously decreased, the electrophoresis rate was dropped from 2.21 to 1.15 mu m.s-1.V-1.cm-1. The indexes related with cell redifferentiation were promoted. For example, the colonogenic potential was decreased to 93.5%. These results indicated that redifferentiation of human gastric cancer cells was successfully induced by AA + SS. The activities of SOD and GPX were significantly higher, while the activity of CAT was slower in treated group than that in the control. The content of MDA was slightly decreased, GSH was sharply decreased, and H2O2 content was dramatically increased.

CONCLUSIONThese results indicated that combination of ascorbic acid and sodium selenite may induce the redifferentiation of human gastric cancer cells and inhibit cell growth by virtue of enhancing the activities of antioxidative enzymes and inducing the formation of H2O2, and altering the cell redox status. Combination of ascorbic acid and sodium selenite may be a potent anticancer agent for human gastric cancer.

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Catalase ; pharmacology ; Cell Differentiation ; Glutathione Peroxidase ; pharmacology ; Humans ; Mitotic Index ; Sodium Selenite ; pharmacology ; Stomach Neoplasms ; pathology ; Superoxide Dismutase ; pharmacology ; Tumor Cells, Cultured

Background:The preferred treatment for uncomplicated type B dissection (thoracic endovascular aortic repair [TEVAR] or medical) is still under debate.Since 2001,our center has performed TEVAR for uncomplicated type B dissection.Based on our data,5-and 10-year survival rates among patients with uncomplicated type B dissection after TEVAR were 96.5％ and 83.0％,respectively.We,therefore,believe that TEVAR is preferable for uncomplicated type B dissections.This study analyzed the impact of a pre-operative smoking history on long-term survival after TEVAR in patients with uncomplicated type B dissections.Methods:From May 2001 to December 2013,data from 751 patients with type B dissections were collected and analyzed.Patients were divided into two groups (337 smoking patients and 414 non-smoking patients).The Kaplan-Meier method and log-rank test were used to compare survival curves of the two groups.Multivariable analyses using the Cox proportional hazards model were used to estimate the effects of smoking on survival rates.Results:The 5-and 10-year survival rates of non-smokers were 97.6％ (95％ confidence interval [CI],96.0％-99.2％) and 87.0％ (95％ CI,81.6％-92.7％),respectively,and 94.9％ (95％ CI,92.2％-97.7％) and 73.8％ (95％ CI,62.3％-87.5％) for smokers,respectively (Log-rank test,P =0.006).Multivariable analyses showed that smoking increased the risk of death during follow-up,2.1-fold when compared to non-smokers (P =0.039).Conclusion:A pre-operative smoking history increases long-term mortality rates after TEVAR in patients with uncomplicated type B dissections.

Objective: The current study was designed to evaluate the clinical application of radioimmunoimaging(RII) for lymph node metastasis in esophageal carcinoma. Methods:1)131I was used to label McAb G9(specific to cellular membrane antigen of human esophageal carcinoma) and form labeling compound 131I-G9. Administration of 131I-G9 in esophagus submucosally with a specific injector for the purpose of submucosal injection via endoscopies in preoperative patients with squamous cell carcinoma of thoracic esophagus followed by RII. 2)The samples of dissected lymph node were used for detection of radioactivity. Results: 1)The pictures at 48 h showed that small radioactivity concentrated dots appeared dispersedly in mediastinum and upper abdomen around esophagus and cardiac gastric. The lymph nodes were considered metastatic in above regions. 2)The pathological results of the lymph nodes dissected compared to the RII result. The metastatic lymph nodes were found in the regions of dispersedly concentrated radioactivity, while no metastatic lymph nodes could be found in the radioactivity free regions. 3)After counting for radioactivity, lymph node metastases showed higher antibody uptake than the non-metastases lymph nodes. The difference was statistically significant. Conclusion: 131I-G9 may be used to locate metastatic lymph nodes in patients with esophageal carcinoma.

Two plasmid constructs, pcE2 and pcE3, containing 3' fragment of open reading frame 2 (ORF2,1163 bp) of hepatitis E virus (HEV) and full-length ORF3 (369 bp), were injected into bilateral tibialis of Swiss mice respectively，for three times (0, 2nd and 4th weeks) and observed the HEV IgG by ELISA. HEV IgG was induced after the injection of pcE2 or pcE3 or both, and the percentage of seraconversion was 100% after two weeks of the third injection. Compared with injection of either construct, the antibody titers were higher in the group with combined injection of two constructs.

OBJECTIVETo investigate the splice variants of the calpain 3 gene existing in human skeletal muscle tissue and white blood cells, and to explore the feasibility of gene diagnosis using CAPN3 mRNA extracted from peripheral leukocytes.

METHODSTotal RNA was extracted from peripheral blood and skeletal muscle tissue in healthy individuals. CAPN3 cDNAs were determined by reverse transcriptase polymerase chain reaction and DNA sequencing. CAPN3 cDNAs from peripheral leukocytes were compared with sequences obtained from skeletal muscle tissue.

RESULTSRT-PCR and DNA sequencing showed that the CAPN3 cDNAs comprised 24 exons in human skeletal muscle tissue, while the number of exons was 23 in white blood cells. Exon 15 was spliced out in human white blood cells.

CONCLUSIONSplice variants exist in human skeletal muscle tissue and white blood cells. Gene diagnosis may omit the mutations of exon 15 using mRNA extracted from peripheral leukocytes. These findings suggest that mutation analysis of the CAPN3 cDNA should use skeletal muscle tissue as materials instead of peripheral blood.

Calpain ; genetics ; DNA Mutational Analysis ; DNA, Complementary ; genetics ; Exons ; genetics ; Humans ; Leukocytes ; metabolism ; Muscle Proteins ; genetics ; Muscle, Skeletal ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo express endotoxin binding peptide and its mutant in E coli DH5alpha and detect the antiendotoxin activity of the purified peptides.

METHODS(1 ) E coli DH5at containing pinpointXa3-EBP and pinpointXa3-mEBP was activated by IPTG to express biotin fusion protein. The fusion proteins were purified, and then digested by factor Xa for isolation of EBP and mEBP. The target peptide was purified with affinity chromatography and reversed-phase HPLC. Sequences of 10 amino acids at N-terminal were used for identification of mEBP. (2) PBMCs were isolated from blood of normal people, and they were stimulated with 5 mg/L FITC-LPS plus 2.0,5. 0 and 12. 5 mg/L EBP or mEBP. Then the mean fluorescent intensity was detected. PBMC was also stimulated with 1 mg/L LPS plus 2.0, 5.0 and 12.5 mg/L EBP or mEBP for 5 hours for the detection of the TNF-alpha and IL-6 level in the supernatant. (3) Thirty-five Kunming mice were randomized into normal control ( n = 5, with intraperitoneal injection of 0. 2 ml isotonic saline) , model group(n = 5, with intraperitoneal injection of LPS and 20% TBSA full-thickness burns), and treatment group (n = 15, with intraperitoneal injection of 5 mg/kg PMB or 10 mg/kg EBP or mEBP after burns). The serum contents of TNF-a and IL-6, and TNF-alpha mRNA level in hepatic tissue in each group were determined 6 hours after treatment.

RESULTS( 1 ) EBP and mEBP were obtained after Xa digestion of biotin fusion protein, with purity reaching above 98% . The sequence of 10 amino acid at N-terminal was in accord with what expected. (2) The fluorescent intensity was decreased followed by an increase in mEBP or EBP concentration. Compared with normal PMBC, IL-6 and TNF-alpha level in the supernatant were obviously lowered in 1 mg/L LPS + 12.5 mg/L EBP group and I mg/L LPS +2. O0 , 5. 0, 12. 5 mg/L mEBP groups ( P < 0.01). (3) The serum level of IL-6 and TNF-ca in the therapeutic groups were obviously lower than that in model group ( P < 0.01 ) , and the levels of these cytokines were significantly lower in 10 mg/kg mEBP group than that in 10 mg/kg EBP group ( P <0. 01) , but they were similar to that in 5 mg/kg PMB treatment group ( P >0.05). (4) Relative optical density of TNF-alpha. mRNA in control, model, 10 mg/kg mEBP, 10 mg/kg EBP and 5 mg/kg PMB groups was 0.25, 0.93, 0.51 , 0.77 and 0.43, respectively.

CONCLUSIONEndotoxin binding peptides can be obtained by procaryon expression. Both EBP and mEBP have anti-LPS activity, but mEBP is more effective.

Animals ; Endotoxins ; metabolism ; Gene Expression ; Humans ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; metabolism ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred Strains ; Mutant Proteins ; isolation & purification ; metabolism ; Peptides ; isolation & purification ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism