To investigate the biological role of snake venom cystatin(sv-cystatin) in tumor progression, the cDNA of sv-cystatin amplified by PCR from pUC18-cystatin plasmid was cloned into methanol-inducible expression vector pPICZ?A. The linearized recombinant plasmid pPICZ?A-cystatin was transfered into Pichia pastoris, strain GS115 by electrophoration. Transfermants with phenotype Mut+ selected were identified by PCR analysis and induced in 1.0% methanol. The reombinant sv-cystatin protein was examined by SDS-PAGE, Western blot analysis. The molecular mass of expression product was about 14 kDa and approximately 16 mg/L of recombinant sv-cystatin was produced from one of GS115-cystatin transformants. The chromatography purified protein could reduce the activity of papain. The ability of B16F1 cells treated with recombinant sv-cystatin to invade the reconstituted basement membrane decreased significantly (P

Objective:To evaluate the current situation and trend of anti-infection drug usage in 11 hospitals of Shanghai during 2003-2005.Methods:Using the cost ranking approach,we analyzed the anti-infection drug usage in 11 major hospitals of Shanghai during 2003-2005.The drug species,consumption,cost,manufacturer,etc,were analyzed by Excel 2000 software. Results:It was found that the cost of anti-infection drugs in the 11 hospitals occupied 22.65% of the total drug expenditure during 2003-2005 and the expenditure decreased at an annual rate of 4.02%.We also found that 80% of the anti-infection drugs were antibiotics.The consumption of?-lactamase inhibitor valued 44.120 5,73.696 7 and 95.163 0 million Yuan in 2003,2004 and 2005,respectively(with a yearly growth of 46.86%),ranking the 3rd in 2003 and the 2nd in both 2004 and 2005.The consumption of cephalosporins ranked the first during 2003-2005.The top 20 anti-infection drugs occupied nearly 80% of the total consumption;the consumption of sulbactam sodium/cefoperazone raised from the 17th in 2003 to the 5th in 2004 and to the first in 2005.Conclusion:The consumption of anti-infection drugs has been restrained;cephalosporins are still the major drugs in clinical practice;and the consumption of sulbactam sodium/cefoperazone is on the rise year by year.

Adult ; Biopsy, Needle ; Diagnosis, Differential ; Humans ; Kidney ; pathology ; Kidney Diseases ; pathology ; therapy ; Male ; Nephritis, Interstitial ; pathology ; Renal Dialysis ; Sarcoidosis ; pathology ; therapy ; Tuberculosis, Renal ; pathology

OBJECTIVETo investigate the role of p38 MAPK on ischemic postconditioning (IPO) attenuating pneumocyte apoptosis after lung ischemia/reperfusion injury (LIRI).

METHODSForty adult male SD rats were randomly divided into 5 groups based upon the intervention (n = 8): control group (C), LIR group (I/R), LIR + IPO group (IPO), IPO + solution control group (D), IPO + SB203580 group (SB). Left lung tissue was isolated after the 2 hours of reperfusion, the ratio of wet lung weight to dry lung weight (W/D), and total lung water content (TLW) were measured. The histological structure of the left lung was observed under light and electron transmission microscopes, and scored by alveolar damage index of quantitative assessment (IQA). Apoptosis index (AI) of lung tissue was determined by terminal deoxynuleotidyl transferase mediated dUTP nick end and labeling (TUNEL) method. The mRNA expression and protein levels of and Bax were measured by RT-PCR and quantitative immunohistochemistry (IHC).

RESULTSCompared with C group, W/D, TLW, IQA, AI and the expression of Bax of I/R were significantly increased, the expression of Bcl-2 and Bcl-2/Bax were significantly decreased (P < 0.05, P < 0.01), and was obviously morphological abnormality in lung tissue. Compared with I/R group, all the indexes of IPO except for the expression of Bcl-2 and Bcl-2/ Bax were obviously reduced, the expression of Bcl-2 and Bcl-2/Bax were increased (P < 0.05, P < 0.01). All the indexes between D and IPO were little or not significant( P > 0.05). The expression of Bcl-2 and Bcl-2/Bax of SB were significantly increased and other indexes were reduced than those of IPO (P < 0.05, P < 0.01).

CONCLUSIONIPO may attenuate pneumocyte apoptosis in LIRI by inactivation of p38 MAPK, up-regulating expression of Bcl-2/Bax ratio.

Alveolar Epithelial Cells ; cytology ; Animals ; Apoptosis ; Disease Models, Animal ; Ischemic Postconditioning ; Lung ; blood supply ; enzymology ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology ; pathology ; prevention & control ; bcl-2-Associated X Protein ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

To detect the in vitro probe microdialysis recoveries based on an HPLC-DAD method for simultaneous quantification of nine active ingredients (ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and glycyrrhizic acid) in Mahuang decoction, which provides reference for in vivo pharmacokinetic study. The concentrations of nine active ingredients in dialysate were detected by HPLC-DAD, to investigate the effect of flow rates (incremental method and subtraction method) and intraday stability of the probe recoveries and medium concentrations on the recoveries. Nine active ingredients could be well separated in 52 min. At the perfusion rate of 1.0 μL x min(-1), the relative recoveries of ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and glycyrrhizic acid were (50.95 ± 0.82)%, (52.74 ± 1.13)%, (51.29 ± 0.51)%, (32.56 ± 0.84)%, (45.36 ± 0.83)%, (70.94 ± 0.99)%, (69.98 ± 2.30)%, (71.68 ± 0.63)%, and (22.14 ± 0.48)%, respectively. And the probe kept steady in 7 hours. At the same medium concentration, the probe recoveries decreased exponentially with the increase in flow rates. The recoveries of seven ingredients detected by these two methods were similar at certain flow rates, except for amygdalin and cinnamaldehyde. At the same flow rate, the relative recoveries of cinnamyl alcohol, cinnamic acid and cinnamaldehyde changed greatly (9.55%-16.2%) and the others six ingredients had less change (3.27%-5.71%) with the changes in medium concentrations. Microdialysis method could be used to detect the in vitro recoveries of nine ingredients in Mahuang decoction. Reverse dialysis method could be used for the in vivo probe recovery calibration of ephedrine, pseudoephedrine, methylephedrine, liquiritin, cinnamyl alcohol and cinnamic acid at the flow rate of 2.0 μL x min(-1).
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ephedra sinica ; chemistry ; Microdialysis ; methods ; Molecular Structure

OBJECTIVETo investigate the role and significance of ATP-sensitive K+ channels in the pathological process of hypoxia hypercapnia-induced pulmonary vasoconstriction (HHPV) and the relationship with ERK1/2 signal pathway in rats.

METHODSWe made the third pulmonary artery rings of SD rats, used the model of pulmonary artery rings perfusion in vitro. Under acute hypoxia hypercapnia condition, and observed the effects of the three stages of HHPV incubated by glybenclamide(Gly) and the combined application of Gly and U0126. At the same time, the values of rings' tension changes were recorded via the method of hypoxia hypercapnia conditions reactivity.

RESULTSUnder the normoxia condition, the values of the third pulmonary artery rings tension were relatively stable, but under the hypoxia hypercapnia condition, we observed a biphasic pulmonary artery contractile response compared with N group (P < 0.05, P < 0.01). When the third pulmonary artery rings incubated by Gly, it's phase II persistent vasoconstriction was enhanced compared with the H group (P < 0.05, P < 0.01), and the phase I vasoconstriction was also heightened. Moreover, under the hypoxia hypercapnia condition, U0126 could significantly relieve the phase II persistent vasoconstriction compared with HD group (P < 0.05, P < 0.01) induced by Gly, but the phase I acute vasoconstriction and the phase I vasodilation had no changes (P > 0.05).

CONCLUSIONGly may mediate HHPV via activating ERK1/2 signal transduction pathway.

Animals ; Glyburide ; pharmacology ; Hypercapnia ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; In Vitro Techniques ; MAP Kinase Signaling System ; physiology ; Male ; Pulmonary Artery ; drug effects ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley ; Vasoconstriction ; drug effects

OBJECTIVETo elucidate whether the polymorphism of asthma immune regulator gene TIM-4 is associated with the risk of childhood allergic asthma in the southwest region of China.

METHODSTIM-4 gene promoter region RS6882076 and intron RS4704727 were studied. PCR-RFLP was used to test the genotypes of two polymorphism loci among 579 cases (average 7.2 years old) of asthma and 524 controls (average 7.6 years old) in a case-control study.

RESULTSThere were significant differences in the frequency of gene types at RS4704727 site between the asthma and the control groups (P<0.01). The results of PCR-RFLP showed that the polyporphisms of RS6882076 and RS4704727 in TIM-4 gene were present in this study population. The frequency of T allele at the RS4704727 site in the asthma group was significantly lower than that in the control group (OR=1.603; 95%CI 1.304-1.971; P<0.01). There were no significant differences in the frequencies of gene types and allele at RS6882076 site between the two groups (P>0.05).

CONCLUSIONSRS4704727 polymorphism of TIM-4 gene may be associated with childhood asthma, providing a better understanding of the pathogenesis of childhood asthma in the Southwest region of China.

Asthma ; etiology ; genetics ; Child ; Female ; Humans ; Male ; Membrane Proteins ; genetics ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic

Objective To investigate the prevalence and characteristics of visual hallucination among patients with Parkinson's disease (PD), and analyse the potential risk factors. Methods One hundred and twenty-eight patients with PD were administered self-prepared visual hallucination questionnaires, and prevalence of visual hallucination was surveyed. The differences in sex, age, disease duration, Mini Mental State Exam (MMSE) scores, Hoehn & Yahr stage, types of medicine used and levodopa equivalent doses (LDE) were compared between the patients with visual hallucination and those without visual hallucination. The prevalence of rapid eye movement sleep behavior disorder (RBD) was investigated in patients with visual hallucination. Results Eighteen patients (14.06%) experienced visual hallucination, among whom 10 (55.56%) experienced visual hallucination no less than one time per day, 11 (61.11%) "saw" the shadow of human figure and 15(83.33%)were complicated with RBD. There were significant differences in MMSE scores, Hoehn & Yahr stage, female proportion and usage of dopamine agonists between patients with visual hallucination and those without visual hallucination(P<0.05). Conclusion Visual hallucination is a common non-motor symptom among patients with PD, and cognitive function, disease severity and usage of dopamine agonista may be related to visual hallucination.

OBJECTIVETo detect potential mutations for probands from families affected with Duchenne/Becker muscular dystrophy (DMD/BMD), and to carry out prenatal diagnosis through identification of female carriers.

METHODSA total of 43 DMD/BMD families were recruited. Multiplex PCR was used to analyze 18 exons within hotspots for DMD gene deletions. Multiplex ligation-dependent probe amplification (MLPA) was used to detect potential deletions and duplications of DMD gene for 43 patients and 36 females from 32 families. Prenatal diagnosis was performed for 27 families.

RESULTSDeletional mutations were detected in 26 patients with multiplex PCR. In addition, MLPA has detected 3 deletions and 6 duplicational mutations, and the ranges of mutations were all determined. Among 36 female members, 18 were determined as carriers of deletional mutations, 10 were excluded as mutation carriers. The status of remaining 8 could not be determined. For prenatal diagnosis, 3 out of 18 male fetuses were diagnosed as patients and 1 female fetus was identified as carrier.

CONCLUSIONMLPA is an accurate and reliable method for detecting deletional/duplicational mutations of DMD gene as well as for prenatal diagnosis and detection of female carriers.

Adolescent ; Child ; Child, Preschool ; Dystrophin ; genetics ; Female ; Heterozygote ; Humans ; Infant ; Male ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; diagnosis ; genetics ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo study the possible role of hTERT and c-myc in endometrial carcinogenesis.

METHODSThe expression of hTERT and c-myc mRNA was examined by in situ hybridization of endometrial samples from 14 cases with simple hyperplasia, 10 with complex hyperplasia, 8 with atypical hyperplasia and 42 with endometrioid carcinoma.

RESULTSExpression of hTERT was demonstrated in samples with simple hyperplasia, complex hyperplasia, atypical hyperplasia and carcinoma at frequencies of 2/14, 4/8, 8/10 and 39/42 (92.9%), respectively. The prevalence and intensity of the hTERT signal was greater in the carcinomas and lesions with atypical hyperplasia than those with simple or complex hyperplasia (P < 0.05). The expression of c-myc was demonstrated in samples with simple hyperplasia, complex hyperplasia, atypical hyperplasia and carcinoma at frequencies of 3/14, 1/8, 5/10 and 23/42 (54.8%), respectively. The frequency of c-myc expression was higher in carcinomas and hyperplastic lesions with atypia than those in lesions with simple or complex hyperplasia without atypia (P < 0.05). The expression of hTERT was shown to be correlated with the level of differentiation (P < 0.05), while the c-myc expression appeared to be associated with the depth of myometrial invasion (P < 0.05). The expression levels of hTERT and c-myc were not found to be correlated with each other in the tissues examined (P > 0.05).

CONCLUSIONSThe expression of hTERT and c-myc may be involved in the progression from the endometrial aypical hyperplasia to invasive carcinoma. The correlation between hTERT and c-myc in endometrial hyperplasia and carcinoma are not found.

Adult ; Aged ; DNA-Binding Proteins ; Endometrial Neoplasms ; genetics ; pathology ; Female ; Genes, myc ; Humans ; Male ; Middle Aged ; Precancerous Conditions ; genetics ; pathology ; RNA, Messenger ; analysis ; Telomerase ; genetics