Objective To construct luciferase reporter plasmid containing synthetic CArG elements and to investigate their radiation-inducible property in tumor cells. Methods Insert chimeric regulation elements consisting of nine tandem-repeat copies of CArG sequence (CCATATAAGG) and CMV IE basal gene promoters into pGL3-Basic vectors to construct luciferase reporter plasmids. Tumor cells(HeLa, A549 and HepG2) were transiently transfected by reporter plasmids using lipofectamine, and transfected cells were irradiated by ?-ray with different doses. After 36 h, we assayed the level of reporter gene expression. Results The CArG elements could successfully induce the expression of a downstream reporter gene following irradiation, with maximal expression seen after 3 Gy irradiation. Conclusion The synthetic CArG elements are responsive to low dose of radiation and are able to enhance down-stream gene expression. It is expected to be the essential potential for future application of radiogenetic cancer therapy.

Objective To check serum protein of children′s Hirschsprung′s disease(HD) and sift specific protein marker which was used in constructing of HD screening and early diagnosis of serum protein fingerprint model.Methods Surface-enhanced laser desorption/ionization time of flight mass spectrometry(SELDI-TOF-MS) was applied to detect protein mass spectrometry of serum specimens in 82 cases(HD group 42 cases,20 cases of other types of obstruction,healthy control group 20 cases) and data were analyzed by bioinformatics methods(support vector machine).Results 1.For HD group and healthy control group:selected 3 M/Z in 3 221.7,5 639.2,6 884.2 protein markers were selected,HD early screening and diagnostic model was established,3 markers in HD low expression,the expressions of them in HD group and healthy control group were 378.29?273.34,295.65?159.38,444.13?254.06 and 1 428.18?1 192.61,1 039.60?785.64,1 115.72?680.48,respectively.There were significant differences in two groups(Pa0.05).Conclusions The establishment of serum protein fingerprint model of HD by SELDI-TOF-MS support vector machine could screen and diagnose HD early,which is a new method of better specificity,high sensitivity and is worthy of further research and application.

BACKGROUND: Herbal wines are an important part of the traditional Chinese medicine. The traditional medicated wine, lishizhen herbal wine,which can strengthen the immune function, has long been used for some chronic diseases. But,its mechanism remains unclear.OBJECTIVE: To explore the immune pharmacodynamics of lishizhen herbal wine and to observe its effect on the immune organs (spleen,pancreas) and lymphocyte transformation rate in mice.DESIGN: A randomized controlled experiment based on the observation of the experimental animals.SETTING: Department of integrated traditional Chinese and western medicine and the department of pharmacology of a university hospital MATERIALS: The experiment was completed at the Central Laboratory of Zhongnain Hospital of Wuhan University from September 2000 to December 2000. A total of 90 healthy Kuming mice were involved in the study.METHODS: Different doses of the herbal wine,cartinellin and the same volume of distilled water were given to the experimental animals. The drug administration was orally injected directly to the stomach of the animals once a day and 10 days consecutively. One hour after the last administration,the animals were put to death. Then,the thymus gland and the spleen were taken out and weighed to calculate the indexes of the thymus gland and the spleen. In the last three days of the administration phytohemagglutinin(PHA)was injected intramuscularly at a dose of 10 mg/kg once per day. Two hours after the last administration, the tails of the mice were cut out, the blood samples were taken to perform the Wright' s staining,and 100 lymphocytes were counted under immersion and the transformation rate was calculated.MAIN OUTCOME MEASURES: Primary results: ① The effect on the immune organs of normal mice; ② The effect of PHA on stimulating the lymphocyte transformation in mice. Secondary results: ③ The death condition of the experimental animalsRESULTS: Different doses of the herbal wine increased the spleen index in different degrees. The effect of medium dose group was obvious[(3.71± 0.78) g/kg] (P <0.05),and the thymus gland index increased a little (P>0.05). The cartinellin in the positive control group increased the spleen index[(3.79±0.91 ) g/kg] and there was no impact on thymus gland index. The transformation rates of the lymphocytes of different groups were increased to a different degree and presented a good quantity-effect relationship,especially the group administered a large dose[(45±14)%] (P<0. 01).CONCLUSION: Lishizhen herbal wine has an effect of increasing cellular immune function.

Objective To study the activity of the survivin gene promoter in several tumor cell lines and evaluate the possible application of this promoter in tumor gene therapy. Methods ①The expressions of survivin gene in A549,MDA-MB231 and HepG2 cell lines were detected by RT-PCR and Western blotting.②Tumor cells(A549,MDA-MB231,HepG2) were transiently transfected by reporter plasmids containing different length of survivin promoter using lipofectamine.And 48 h later,the level of reporter gene expression was analyzed.Results There were different levels of survivin expression in A549,MDA-MB231 and HepG2 cell lines.Transient transfection assay approved that pLuc-surP-987,pLuc-surP-596,pLuc-surP-269 and pLuc-surP-158 showed high activity and 269 bp survivin promoter demonstrated the highest activity. Conclusion In transcriptional level,survivin promoter can activate the reporter gene in several tumor cell lines.It is a potential candidate promoter in tumor gene therapy.

Objectives To investigate the changes of cystatin C in the serum of patients with acute kidney injury ( AKI) or end-stage renal disease ( ESRD), and study its significance in the early diagnosis of AKI and its correlation with the degree of renal function injury. Method The cases in Xiangya hospital were enrolled in this study according to the RIFLE criteria, including 20 cases of slight acute kidney injury, 30 cases of medium-severe acute kidney injury, 48 cases of victims of the 5. 12 wenchuan earthquake, 32 cases of end-stage renal disease and 20 healthy patients. The microparticle-enhanced immunoturbidimetry method was used to detect serum cystatin C, and the colorimetric method was used to detect urine N-acetyl-beta-D-glucosaminidase (NAGase). The enzymic method was used to detect serum creatinine. The correlation between serum cystatin C and serum creatinine was analyzed, and the sensitivity and specificity of serum cystatin C were evaluated with the receiver operator characteristic curve (ROC curve). Results Compared with healthy control group, the serum cystatin C increased obviously in acute kidney injury group and ESRD group( P <0. 05 and P <0. 01). The serum cystatin C was positively correlated with serum creati-nine( P <0.01). The serum cystatin C in the 5. 12 wenchuan earthquake injured group was also higher than that in healthy control group ( P <0.05), an the serum cystatin C had an AUC - ROC of 0.931 ( P <0. 01). Conclusion Compared to the conventional biomarkers, the earlier emergence of serum cystatin C can contribute better to early clinical diagnosis of AKI. The serum cystatin C is positively correlated with renal function, and reflect changes in renal function accurately.

Objective To modify the methods of operation and establishment of cerebral ischemia rat models made by thread occlusion. Method We randomly divided 120 male SD rats into a common group (n = 50), an improvement group (n = 60) and a sham-operated group (n - 10). Rats in the common and improvement groups were made into models using the common and improvement methods separately. All models were evaluated on the basis of physical sign indices and 2,3,5-triphenyltetrazolium chloride (TTC) staining. The TTC staining results were taken as gold standards. Then, we compared the achievement ratios of the two groups, and computed the sensitivity and specificity of every physical sign index based on these standards. The χ~2 or correction χ~2 test was used to compare the ratios. Results (1) The achievement ratio in the improvement group was significantly higher than that in the common group (71.67% vs. 52.00%, P = 0.034). (2) The sensitivity of evaluation for both common and improvement methods was 98.55%. However, the specificity of evaluation for the improvement method was significantly higher than that for the common method (100.00% vs. 40.00%, P =0.000). Conclusions The establishment achievement ratio and evaluation correctness of models are obviously elevated by modification of the operation and establishment methods.

BACKGROUNDAngiogenesis and lymphogenesis which were promoted by vascular endothelial growth factor (VEGF) and VEGF-C are important in the growth and metastasis of solid tumors. The high level of VEGF and VEGF-C were distributed in numerous types of cancers, but their distribution and expression in Wilms tumor, the most common pediatric tumor of the kidney, was unclear.

METHODSTo learn about the distribution, mass spectroscopy and immunohistochemistry were used to measure the level of VEGF and VEGF-C in serum and tissue of Wilms tumor.

RESULTSThe expression level of VEGF in serum of Wilms tumor was the same as in pre-surgery and control, so it was the same case of VEGF-C. Both of these factors were chiefly located in Wilms tumor tissue, but not in borderline and normal. In addition, the higher clinical staging and histopathologic grading were important elements in high expression of VEGF and VEGF-C. Gender, age and the size of tumor have not certainly been implicated in expression level of VEGF and VEGF-C.

CONCLUSIONSThe lymph node metastasis and growth of tumors resulted from angiogenesis and lymphogenesis which were promoted by VEGF and VEGF-C in Wilms tumor. The autocrine and paracrine process of VEGF and VEGF-C were the principal contributor to specific tissues of Wilms tumor but not to the entire body.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Immunohistochemistry ; Infant ; Male ; Mass Spectrometry ; Vascular Endothelial Growth Factor A ; blood ; metabolism ; Vascular Endothelial Growth Factor C ; blood ; metabolism ; Wilms Tumor ; blood ; metabolism

OBJECTIVETo investigate the prognostic factors in non-small cell lung cancer (NSCLC) at stage III and IV and establish a reliable model of clinical prognostic index.

METHODSKaplan-Meier and Cox regression were used to analyze the relationship between the prognostic factors and survival time in 114 cases of NSCLC. The prognostic factors included clinical-pathological features and serum levels of cytokeratin fragment 19 (Cyfra21-1), CEA, neuron-specific enolase (NSE), CA125, interleukin-2 (IL-2) and soluble interleukin-2 receptors (sIL-2R).

RESULTSKaplan-Meier analysis showed that KPS, sex, disease stage, treatment, Cyfra21-1, sIL-2R and CA125 were related to prognosis. Multivariate analysis indicated that Cyfra21-1, stage and treatment were independent prognostic factors. When Cyfra21-1 > 3.5 mg/L, stage IV and chemotherapy < 3 cycles, the relative risk (RR) was 1.691, 2.229 and 3.035, respectively. In patients given 3 or more cycles of chemotherapy, serum Cyfra21-1, sIL-2R and stage at diagnosis were significantly independent prognostic factors. Three of these prognostic factors were used to establish a prognostic index (PI) model based on a simple algorithm: PI = Cyfra21-1 + sIL-2R + stage. The median survival period of patients with 3 or more cycles of chemotherapy were 18 months if PI = 0, 8 months if PI = 1 or 2, and 5 months if PI = 3.

CONCLUSIONThe serum Cyfra21-1, sIL-2R and disease stage in unresectable NSCLC were independent prognostic factors. PI calculated on the basis of Cyfra21-1, sIL-2R and stage is recommended to predict the survival period of NSCLC.

Antigens, Neoplasm ; blood ; Biomarkers, Tumor ; blood ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; mortality ; pathology ; Female ; Follow-Up Studies ; Humans ; Keratin-19 ; Keratins ; Lung Neoplasms ; drug therapy ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Receptors, Interleukin-2 ; blood ; Survival Rate

OBJECTIVETo investigate genetic and antibiotic resistance characteristics of Campylobacter jejuni (C. jejuni) isolated from Shenzhen.

METHODSMultilocs sequence typing and agar dilution methods were used to define the genotype and antibiotic resistance of C. jejuni, respectively.

RESULTSIn total, 126 C. jejuni strains were isolated. The prevalence of C. jejuni was 5.3% in diarrheal patients. The prevalence in poultry meat (36.5%) was higher than that in cattle meat (1.1%). However, the prevalence in poultry cloacal swabs (27.0%) was lower than that in cattle stool (57.3%). Sixty-two sequence types were obtained, among which 27 of the STs and 10 alleles were previously unreported. The most frequently observed clonal complexes were ST 21 (11.9%), ST-22 (10.3%), and ST-403 (7.1%). ST-21, ST-45, ST-354, ST-403, and ST-443 complexes overlapped between isolates from patients and cattle, whereas ST-45 and ST-574 complexes overlapped between isolates from patients and poultry. All C. jejuni were resistant to at least one antibiotic. The highest resistance rate was toward ciprofloxacin (89.7%), followed by tetracycline (74.6%), and nalidixic acid (69.0%).

CONCLUSIONThis is the first report of the genotypes and antibiotic resistance of C. jejuni in Shenzhen. Overlapping clonal complexes were found between isolates from patients and cattle, and between patients and poultry.