Objective To construct pGL4.14-1uc eukaryotic expression vector for HLA-B27 promoter gene and explore the activity regulation of this promoter in Hela cells.Methods The HLA-B27 gene promot- er(-419 bp～1 bp)was amplified by polymerase chain reaction and was cloned into pGL4.14-luc vector to construct eukaryotic expression vector pGL4.14/B27 pro-luc.The purified pGL4.14/B27 pro-luc was stablely transfected into HeLa cells and the activity of HLA-B27 gene promoter was detected by luminometer.Results About 432 bp gene fragment was amplified by PCR from genomie DNA and pGL4.14/B27 pro-luc vector was constructed successfully.The activity of HLA-B27 promoter was 1.67?0.20,1.79?0.71,2.94?0.68,1.98?0.45 in Hela stable cells after treated with TNF-?,IFN-?,IFN-?and IFN-?for 48 hours.Conclusion TNF-?. IFN-?,IFN-?and IFN-?can regulate the B27 promoter activity.The high specific activity of constructed HLA-B27 promoter eukaryotic expression vector may be a good method for further research.

BACKGROUND: Serum creatinine (SCr) is the most commonly used parameter to estimate renal function impairement, but there are some shortcomings. Many factors including age, gender, drug, diet, muscle mass and metabolic rate can influence SCr, leading to an inaccurate estimation of kidney impairment. Studies have shown that cystatin C (CysC) is not affected by factors such as muscle mass, age, gender, diet, inflammation or tumor. The present study was undertaken to compare the sensitivity of CysC and SCr in evaluating renal function impairment at early stage of shock. METHODS: Seventy-one patients aged 38.3±21.4 years, who had been treated at the Emergency Medicine Department of the First Affiliated Hospital, Sun Yat-sen University between February 2006 and June 2007, were studied. They were divided into groups A, B, C, and D according to the shock time. Serum sample was drawn from each patient at 1, 2, 3, 4 hours after shock to determine SCr and CysC. CysC and SCr were determined again at 72 hours and 7 days after shock. RESULTS: CysC increased earlier than SCr in the 71 patients, and CysC decreased slower than SCr when shock was corrected. CysC increased at 1 hour after shock. There was a negative correlationship between CysC, SCr and glomerular filtration rate (GFR), especially at early stage of shock. CONCLUSIONS: There is renal injury at early stage of shock. CysC is more sensitive than SCr in assessing renal function at the early stage of shock.

Objective To investigate the clinical features,diagnosis and therapy of adrenal incidentalomas. Methods The data of 66 cases of adrenal incidentalomas were retrospectively analyzed. Results The accuracy rates of localization detected by B-Ultrasound,CT scan and MRI were 84.3%,100% and 100% respectively. The accuracy rates of qualitative diagnosis by B-Ultrasound, CT scan and MRI were 30.0%,75.5% and 79.2% respectively. There were 60 cases underwent operation and the postoperative pathological examination confirmed that there were 31 cases of adrenal cortical adenoma, 6 cases adrenal cyst, 5 myelolipoma, 5 pheochromocytoma, 3 adrenal cortical carcinoma, 3 metastatic carcinoma, 3 ganglioneuroma, 1 adrenal neurilemmoma, 1 adrenal hematoma, and 1 nodular hyperplasia of adrenal cortex. Conclusions Ultrasonography is the first choice of scanning adrenal incidentaloma. However the CT scan and MRI are satisfactory in localization of the tumour. The qualitative diagnosis of adrenal incidentaloma will depend on the diameter, imaging features of tumors, the age of patients, and endocrine examination.

Objective To evaluate the functional changes in the isometric,concentric and eccentric muscle strength of the knee extensors and flexors in patients with knee osteoarthritis,and to explore the relationship among these contractions.Methods A Biodex System-3 isokinetic test system was used to assess the isometric,concentric and eccentric strength of the knee extensors and flexors of the involved and uninvolved limbs of 54 patients suffering from osteoarthritis.Results The strength of the knee extensors and flexors of the involved limbs was significantly less than that of the uninvolved legs in the different contraction modes(P＜0.05).The difference was especially marked in concentric and eccentric contraction at low angular velocity.The hamstrings/quadriceps ratio and the dy- namic control ratio showed abnormalities in the muscle balance of the hamstrings and quadriceps.Conclusion Iso- kinetic testing should be applied and the hamstrings/quadriceps ratio and the dynamic control ratio should be analyzed in evaluating patients with knee osteoarthritis.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Intracranial Hemorrhages ; etiology ; pathology ; Magnetic Resonance Imaging ; Prognosis ; Tomography, X-Ray Computed

OBJECTIVETo detect the expressions of CCR5 and CCR7 on dendritic cells (DCs) in patients with rheumatoid arthritis (RA) in different phases of disease activity, and explore the relationship between the disease activity and the expression of chemokine receptors.

METHODSTwenty-eight patients with low, moderate and high disease activity and 10 normal control subjects were enrolled in this study. Peripheral blood was obtained from the subjects and the DCs were isolated. The expression of CCR5 and CCR7 on DCs were detected by flow cytometry, and the serum levels of rheumatoid factor (RF), C-reactive protein (CRP) and anti-CCP antibody (ACPA) were assessed. The correlation of the expressions of CCR5 and CCR7 to serum RF, CRP, and ACPA levels of the RA patients were analyzed.

RESULTSCompared to the normal control group, RA patients showed enhanced expressions of CCR5 and CCR7 on the DCs. A linear correlation was noted between CCR5 and CCR7 expressions on the DCs and the serum levels of RF and CRP, but not ACPA, in the RA patients.

CONCLUSIONThe expressions of CCR5 and CCR7 on the DCs may correlate to the disease activity of RA, and may serve as valuable indices in monitoring the disease activity and the efficacy of the treatment.

Adult ; Arthritis, Rheumatoid ; blood ; immunology ; Dendritic Cells ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Receptors, CCR5 ; metabolism ; Receptors, CCR7 ; metabolism

Objective:To study the abnormal suppressive capacity of regulatory T cells ( Tregs) to effector cells,Th17 cells and Th9 cells in patients with mild to moderate asthma attack.Methods: Recruited asthmatic patients (n=30) and healthy control (n=30),collected peripheral venous blood from healthy control and asthmatic patients in mild to moderate attack stage respectively,and then isolated CD4+CD25+CD127-/low Tregs and CD4+CD25-T effect cells with immunomagnetic beads method ( purity was above 90%).The effect cells were cultured with PHA stimulation with or without Tregs.Measured the proliferation (3H-Thymidine), expression of RORC and PU.1 ( RT-PCR) genes,production of IL-17 and IL-9 ( LUMNEX) in effect cells with or without Tregs inter-vention.Analyzed the abnormality of Th17 and Th9 cells and the supressive capacity of Tregs on Th17 and Th9 cells in patients with asthma attack,comparing with control group.Results:Tregs of asthmatic patients could suppress the proliferation of effector cells,but less effectively than healthy control (P=0.03).In asthmatic patients,RORC expression and IL-17 production increased (P<0.05), and the suppressive capacity of Tregs to RORC expression decreased, when compared with healthy group ( P<0.05 ) .In asthmatic patients,IL-9 production of effector cells increased significantly (P<0.05),but PU.1 expression was higher than healthy group only with intervention of Tregs ( P=0.04 ) .And the suppressive capacity of Tregs to the expression of PU.1 and production of IL-9 in patients with asthma attack did not decrease (P>0.05).Conclusion:The suppressive capacity and amount of Tregs decreased,but the specific gene expression and cytokines production of Th17 cells and Th9 cells were upregulated in patients with mild to moderate asthma attack.This deficiency for the suppressive capacity of Tregs to Th17 but not Th9 cells may indicate some pathogenesis of asthma devel-opment.

Background Our previous study demonstrated that kallikrein-kinin is a special protein in vitreous of the eye with proliferative vitreoretinopathy(PVR),and the expression intensity of kallikrein-kinin showed the positive correlation with the grade of PVR.Objective This study was to further explore whether kallikrein-kinin participate in the formation of PVR.Methods Rat retinal pigment epithelial cell line(RPE-J cells) was cultured in DMEM containing 4% fetal bovine serum and then prepared into suspension by PBS with the cells density of 2.5×108 cells/ml.Platelet-rich plasma was prepared by PBS with the platelet 2.5×108 /ml.RPE cell suspension(4μl) and platelet-rich plasma(6μl) was intravitreally injected in the left eyes of 30 clean Wistar rats to establish the PVR models,and 10μl sterile pyrogen-free normal saline solution was used in the same way in other matched rats as controls.The PVR was graded on Francine's criteria in 1 day,3,7,14,21,28 days after injection under the slit lamp.The serum,vitreous and retina were obtained in 28 days after injection to assess the expression of bradykinin using Western blot.The histopathology examination of rat retina was performed in the 28th day after injection.This experimental procedure followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Typical PVR was seen in 25 models with the successful rate 89.3% at 28 days after injection.PVR 1,2,3 grades were respectively exhibited in 7,14,28 days under the slit lamp.Infiltration of inflammatory cells and migration of RPE cells were found in the 7th day.In the 14th day after injection,RPE cells transformed into fibroblasts and retinal detachment occurred after that.Western blot analysis revealed that bradykinin was detected in vitreous,serum and retinal samples of rats in experimental and control rats,but the expression intensity was higher in the rats of model groups.Conclusion Intravitreal co-injection of RPE cells and platelet-rich plasma can effectively induce a model of PVR in Wistar rat.The kallikrein-kinin system probably takes part in the onset of PVR.

Literature review of the world on hospital brands, data collection in summary and comparison, and specialist consultancy were used to set the appraisal indicators, while the stratum analysis was used to calculate the weight coefficients of individual indicators, for building the appraisal indicators system for hospital brand effect. In addition, the indicators system and appraisal model were called into play to explore the use of hospital brands, offering scientific references for hospital management.

Objective To explore the role of Beclin 1 and PTEN in gastric caicinoma genesis and the effects on prognosis.Methods The expression of Beclin 1 and PTEN in 199 gastric caicinoma specimens and corresponding adjacent noncancerous tissues were examined by tissue microarrays and immunohistochemistary,and the relation with gastric cancer was analyzed.The rate of Beclin 1 and PTEN expression in 15 fresh gastric carcinoma samples and corresponding adjacent noncancerous tissues were detected by Western blot.All the samples were from Changzheng Hospital.Results The Results of immunohistochemistary showed that the rates of Beclin 1 and PTEN positive expression in cancinoma tissues were 47.2％ (94/199) and 55.8％ (111/199),both were lower than that of adjacent noncancinoma tissues (94.5％,188/199 and 92.5％,184/199; P ＜ 0.01).The lower expression of Beclin 1 and PTEN in gastric carcinoma were associated with gender,differentiation degree,depth of tumor invasion,lymph node metastases and disease stage(P＜0.05).There was a positive correlation between Beclin 1 and PTEN expression in gastric carcinoma tissues (r =0.680,P＜0.01). The survival analysis indicated that Beclin 1 and PTEN were independent factors in determining the prognosis of gastric cancinoma patients.The 5-year survival rate of Beclin 1 positive patients was 67.0％ (63/94),and of negative patients was 33.3％ (35/105).The 5-year survival rate of PTEN positive patients was 71.2％ (79/111),and of negative patients was 21.6％ (19/88) ( all P＜0.001).The Results of Western blot indicated that the expression of Beclin 1 and PTEN in gastric carcinoma tissues were significantly lower than that in the adjacent noncarcinoma tissues ( all P＜0.001).Conclusion The abnormal expression of Beclin 1 and PTEN may be related to carcinogenesis and the development of gastric carcinoma.