Child ; Clinical Trials as Topic ; ethics ; Drugs, Investigational ; Humans

Objective To investigate the percentages of Th17 and CD4+CD25+FoxP3+ regulatory T(Tr) cells and the levels of related cytokines IL-6,IL-23,IL-17 and TGF-β in serum of patients with anlylosing spondylitis(AS).Methods Forty patients with AS and 37 age-matched healthy donors were studied.Flow cytometry Was used to analyze the percentages of blood Th17 and CD4+CD25+FoxP3+Tr cells.The levels of serum IL-6,IL-23,IL-17 and TGF-β were assayed by enzyme-linked immunosorbent assay ( ELISA).Results The proportion of Th17 cells in AS group was significantly higher than those in normal group [ (1.02±0.34)％ vs (0.68±0.29)％,P＜0.05) ],and the proportion of CD4+CD25+FoxP3+ cells was lower in AS group comparing with normal group [(3.77±0.81)％ vs (4.69±1.23)％,P＜0.05)].Meanwhile,serum levels of IL-6,IL-23 and IL-17 were significantly higher in AS group than those in normal group [ (6,15±2.71) ng/L vs (3.31±1.65) ng/L; (9.44±3.12) ng/ml vs (5.82±2.61) ng/ml;(10.53±4.97) ng/L vs (6.78±3.26) ng/L,all P＜0.01 ].In contrast,TGF-β level was decreased in AS group compares with the normal group [ ( 4.76±2.15) ng/ml vs (5.16±2.02) ng/ml,P＞0.05 ],but the difference was not significant.No associations of serum eytokine levels with clinical and laboratory parameters were found in AS.Conclusion The abnormality Th17 cells and Tr cells and their related cytokines IL-6,IL-23,IL-17 and TGF-β changes in patients with AS,which may be involved in immunological pathogenesis of AS.

Objective To observe the dynamic changes of NO,NOS in serum and CSF in patients with CNS infection and lay an experimental basis for discrimination of CNS infection due to different agents.Methods The method of nitrate redutase and the method of chromometry were employed to measure NO,NOS in serum and CSF at different time.The dynamic changes of NO,NOS in serum were observed on admission,on 3,5,9,14 day after admission.The dynamic changes of NO,NOS in CSF before treatment and two weeks after treatment were ob- served,too.Results There were no difference between the the concentration of NO and the vigor of total NOS in serum and in CSF of viral meningitis,bacterial meningitis and tubercular meningitis patients due to different agents. Conclusion The changes of the concentration of NO in serum and CSF,the vigor of total NOS in serum and CSF could not he seen as laboratory basis for discrimination of CNS infection due to different agents.

AIMTo clone the gene of staphylococcal enterotoxin C2 and express it in the form of a soluble fusion protein in E. coli. Then the activation of SEC2 on mice lymphocyte and its lethal effects on tumor cells were studied.

METHODSStaphylococcus aureus SEC2 gene was cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEC2 was used to transform E. coli BL21, where the GST-SEC2 fusion protein was expressed efficiently. The rSEC2 protein was purified with Glutathione Sepharose 4B affinity column and digested with thrombin. The in vitro culture system was utilized to observe the activation of the SEC2 on mice lymphocyte and the lethal effects on tumor cells of the activated mice lymphocyte.

RESULTSThe proper gene of SEC2 was cloned and purified rSEC2 was obtained. The MTT results indicated that rSEC2 have strong ability to stimulate mice lymphocyte to proliferate with a dose-dependent manner. With the proliferation of mice splenic lymphocyte, rSEC2 has a strong lethal effect on tumor cells B16, K562 and K562-AD.

CONCLUSIONIn this study, the gene of SEC2 was cloned and the rSEC2 protein was obtained, which had strong lethal effect on tumor cells B16, K562 and K562-AD.

Animals ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cloning, Molecular ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; Glutathione Transferase ; genetics ; Lymphocyte Activation ; drug effects ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred ICR ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Spleen ; cytology ; Transfection

OBJECTIVETo discuss the optimum extraction process of compound Nanxing pain-relieving cataplasm through orthogonal design and pharmacodynamic experiment

METHODThe orthogonal experiment method was adopted to optimize the ethanol extraction process with Angelica dahurica, Ligusticum chuanxiong and other herbs. The anti-inflammatory and analgesic effects of extracts from volatile oil in such herbs as syzygium aromaticum with different extraction processes were compared by tail pain tenderness test and food-pad swelling in mice, in order to optimize the extraction process of extracts from volatile oil in such herbs as syzygium aromaticum.

RESULTThe optimum extraction process of A. dahurica, L. chuanxiong and other herbs for compound Nanxing pain-relieving cataplasm were as follows: adding 8-fold amount of 70% alcohol, extracting for 2 times with 1.5 h each time. The 95% ethnol extracts of syzygium aromaticum and other herbs had more effect in the increasing the threshold of pain and the inhibition of toe swelling of mice than volatile oil obtained from steam distillation as well as volatile oil and water decoction obtained from steam distillation.

CONCLUSIONThe method is simple and reliable that it can provide technical reference for the development of modern preparations of compound Nanxing pain-relieving cataplasm.

Analgesics ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Animals ; Chemical Fractionation ; methods ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Ethanol ; chemistry ; Mice ; Oils, Volatile ; chemistry ; Pain ; drug therapy

OBJECTIVETo evaluate the roles of plasma mast cell carboxypeptidase and chymase in the diagnosis of allergic diseases by measuring the contents of both in children.

METHODSA total of 59 children with allergic diseases and 53 healthy children were recruited into the study. Plasma levels of mast cell carboxypeptidase and chymase were measured using ELISA.

RESULTSThe plasma levels of mast cell carboxypeptidase and chymase in children with allergic children were 1.089 ± 0.752 ng/mL and 0.905(0.375-2.318) ng/mL, respectively, which were significantly higher than those in healthy children [0.593 ± 0.380 ng/mL and 0.454 (0.097-1.077) ng/mL respectively; P<0.05]. There was a significantly positive correlation between plasma mast cell carboxypeptidase and chymase levels in children with allergic diseases (r=0.684, P<0.01).

CONCLUSIONSPlasma levels of mast cell carboxypeptidase and chymase increase in children with allergic diseases, suggesting that mast cell carboxypeptidase and chymase may serve as the indexes for the diagnosis of allergic diseases.

Adolescent ; Carboxypeptidases ; blood ; Child ; Child, Preschool ; Chymases ; blood ; Female ; Humans ; Hypersensitivity ; diagnosis ; enzymology ; Infant ; Infant, Newborn ; Male ; Mast Cells ; enzymology

Child ; Child, Preschool ; Female ; Humans ; Hypertension, Portal ; diagnosis ; etiology ; therapy ; Infant ; Male ; Portal Vein ; pathology ; Venous Thrombosis ; diagnosis ; etiology ; therapy

BACKGROUNDAstrocyte swelling is an important consequence of hepatic encephalopathy, and aquaporin-4 has been reported to play a vital role in this swelling. Ammonia causes astrocyte swelling and is also known to modulate aquaporin-4 expression in the astrocyte foot processes. The purpose of this study was to explore the mechanism of ammonia-induced aquaporin-4 expression, which has been suggested to involve the p38 mitogen-activated protein kinase pathway.

METHODSWe exposed cultured astrocytes to ammonium chloride, an in vitro model of hepatic encephalopathy. The purity of cultured astrocytes was evaluated by fluorescent glial fibrillary acidic protein labeling; cell morphology was assessed by light microscopy; the expression of aquaporin-4, phospho-p38, and p38 were detected by Western blotting analysis. Statistical analysis was performed by one-way factorial analysis of variance, and the relationship between variables was calculated by linear regression using SPSS version 13.0 program for Windows (SPSS, Chicago, IL, USA).

RESULTSThe purity of cultured astrocytes was (96.6 +/- 1.4)%. Astrocytes swelled significantly when exposed to 5 mmol/L ammonium chloride for 24 hours as compared to non-exposed astrocytes. Co-treatment with 10 micromol/L SB203580 (an inhibitor of p38) attenuated the degree of ammonium chloride induced astrocyte swelling. Western blotting analysis revealed that the expression levels of phospho-p38 and aquaporin-4 in ammonium chloride treated cells were significantly increased relative to the control group (P < 0.001); SB203580 co-treatment inhibited the increased expression of phospho-p38 and aquaporin-4 relative to the ammonium chloride treated group (P = 0.002 and P = 0.015 respectively). The phosphorylation of p38 and upregulation of aquaporin-4 were highly correlated (r = 0.909). There were no significant differences in total p38 expression among the groups (P = 0.341).

CONCLUSIONSAmmonium chloride induced upregulation of aquaporin-4 in astrocytes is regulated by the p38 mitogen-activated protein kinase pathway. Inhibiting p38 activation prevented ammonium chloride induced aquaporin-4 protein upregulation.

Ammonium Chloride ; pharmacology ; Animals ; Aquaporin 4 ; genetics ; metabolism ; Astrocytes ; drug effects ; metabolism ; Blotting, Western ; Cells, Cultured ; Enzyme Inhibitors ; pharmacology ; Imidazoles ; pharmacology ; Phosphorylation ; drug effects ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

BACKGROUNDIn 2008, a sharp increase of the number of children diagnosed with urinary calculi was observed in China, 9433 children were diagnosed as having melamine-induced urinary calculi at outpatient clinic in Beijing Children's Hospital. This study examined the therapeutic efficacy of potassium sodium hydrogen citrate (PSHC) used to treat melamine-induced urinary stones in Chinese children who consumed melamine-containing infant formula.

METHODSSeventy-two infants and children (average age (18.2 +/- 7.7) months) who were diagnosed with urinary calculi were randomly divided into three treatment groups using the SAS Plan program. Group 1 was given a low dose (1 g/d) of PSHC, group 2 was given high dose of PSHC (2 g/d) and group 3 was given no PSHC (control group). The dose of drug was adjusted according to the baseline urinary pH. This study analyzed the influence of the dose of PSHC, the age of patients, stone size and position, and urinary pH on the level of efficacy of PSHC (cured, effectively treated or not cured).

RESULTSAfter 1 - 6 months of therapy, 19 patients from group 1, five patients from group 2 and six patients from group 3 were cured. Five patients from group 1, five patients from group 2 and four patients from group 3 were effectively treated. There were significant differences in therapeutic efficacy between the two treatment doses after 3 and 6 months as measured by the increase in the successful expulsion rate and time of melamine-induced urinary calculi. After 6 months the mean time of expulsion of urinary calculi in groups 1 and 2 was significantly shorter than in the control group.

CONCLUSIONSPSHC can significantly increase the successful expulsion rate and time of melamine-induced urinary calculi. The therapeutic efficacy is affected by PSHC dose, treatment duration, calculi position, and urinary pH. There is no relationship between the therapeutic efficacy and the stone size or patient age.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; Citrates ; therapeutic use ; Female ; Humans ; Hydrogen-Ion Concentration ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Treatment Outcome ; Triazines ; toxicity ; Urinary Calculi ; chemically induced ; drug therapy ; urine ; Urine ; chemistry ; Young Adult

Objective To evaluate whether the protein SP70 could be used as a serum biomarker for the diagnosis of non-small cell lung cancer (NSCLC).Methods Polyclonal antibody was prepared by immunizing New Zealand rabbit with SPC-A1 cells.Sandwich ELISA was carried out by using newly-prepared polyclonal antibody(PcMb) coating assay plates,monoclonal antibody (McAb) NJ001 and HRP goat antimouse antibody as primary antibody and labeling antibody respectively.After optimizing the experiment conditions,serum from 175 lung cancer patients [ 80 NSCLC adenocarcinoma,70 NSCLC squamous carcinoma and 25 small cell lung cancer ( SCLC) ],25 benign lung disease ( BLD) patients and 300 healthy controls (HC) were examined.CEA,NSE,CYFRA21-1 were measured by ECLIA for comparison.Results Positive rates of NSCLC adenocarcinoma,NSCLC squamous carcinoma,SCLC and BLD were 68.8％,51.4％,16.0％ and 12.0％ respectively,obviously higher than that of HC (7.3％).NSCLC (adenocarcinoma,Squamous carcinoma) had significantly higher positive rate than SCLC (60.7％ υs 16.0％,x2 =17.23,P＜0.05)and BLD(60.7％ υs 12.0％,x2 =20.41,P ＜0.05).Among 68 NSCLC patients who had definite staging,positive rates at early stage ( Ⅰ/Ⅱ,n=30) reached up to 76.7％.Meanwhile,positive rates of CEA,NSE and CYFRA21-1 (32.7％,18.0％ and 37.3％) were significantly lower than the targeting antigen to McAb NJ001 in NSCLC(60.7％ υs 32.7％,x2 =23.63,P ＜0.05;60.7％υs18.0％,x2 =57.22,P＜0.05;60.7％ υs37.3％,x2=16.34,P＜0.05).Conclusions It showed high positive rates of SP70 in the serum of NSCLC patients,which suggested thai SP70 might be a potential valuable biomarker for the diagnosis of NSCLC.