1.Molecular cloning of HLA-B27 gene promoter and transcriptional activity detection in HeLa cells
Li-Ke ZHAO ; Jie-Ruo GU ; Deen YU ;
Chinese Journal of Rheumatology 2003;0(08):-
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.
2.Cystatin C and serum creatinine in estimating acute kidney injury of shock patients
Qiang LI ; Jie-Yu FANG ; Wei-Ping WANG ; Jiang-Hui LIU ; Ke-Ke WANG
World Journal of Emergency Medicine 2010;1(3):185-189
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.
3.Report of 66 cases of adrenal incidentalomas
Yanhong YU ; Minhui XIAO ; Xiaohua YANG ; Shuwu QI ; Jie HUAN ; Ke ZHANG ; Jie SHEN
Journal of Endocrine Surgery 2009;3(6):412-414
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.
4.The isometric concentric and eccentric strength of patients with knee osteoarthritis
Xiao-Jie YU ; Yi WU ; Yong-San HU ; Yu-Long BAI ; Wen-Ke FAN ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(07):-
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.
5.Detection and its clinical value of CCR5 and CCR7 in dendritic cells from patients with active rheumatoid arthritis.
Journal of Southern Medical University 2010;30(9):2067-2069
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
7.Percutaneous therapy of hemostatics of injected gelatin matrix under the guidance of contrast-enhanced ultrasound for splenic trauma in canine
Jiang-ke, TIAN ; Xia, XIE ; Rong, WU ; Fa-qin, LÜ ; Yu-kun, LUO ; Jie, TANG
Chinese Journal of Medical Ultrasound (Electronic Edition) 2013;(9):65-70
Objective To evaluate the efifcacy and safety of hemostatics of injected gelatin matrix (HIGM) under the guidance of contrast-enhanced ultrasound (CEUS) for treating splenic trauma in canine model. Methods A total of 24 commercial hybrid dogs underwent celiotomy with creation of uniformly blunt splenic trauma lesion of 4.0 cm×4.0 cm×2.5 cm (length, width and depth, respectively) by hemostatic clamp. Subjects were prospectively randomized into two groups. The treatment group was treated with HIGM under the guidance of CEUS and the positive control group received thrombin solution. Conventional ultrasound and CEUS were performed to record the ascites and the splenic lesion areas at 1st, 3rd, 7th, 14th and 21st day. The ifne needle biopsy and splenectomy were performed for histopathologic examination. The weight, free intraperitoneal lfuid and injury site were compared with t test between HIGM and postive group. Results All animals in two groups survived. All dogs stopped hemorrhage after injection of HIGM under CEUS guidance. The area of injury site was (12.91±0.89) cm2, (4.45±0.75) cm2 and (1.38±0.23) cm2 at 1st, 3rd and 7th day and splenic lesions were not found at 14th and 21st day in all dogs (n=12) of HIGM group. The splenic lesion was (16.74±0.91) cm2, (11.26±0.99) cm2, (8.02±0.82) cm2 and (1.58±0.36) cm2 in the postive group at 1st, 3rd, 7th and 14th day and splenic lesions were not found at 21st day in all dogs (n=12). At 7th and 14th day post-injection, lesion areas were statistically significant between two groups (t=27.162, P=0.008;t=15.129, P=0.001). Free intraperitoneal lfuid was (0.91±0.05) cm at 1st day detected by conventional ultrasound and free intraperitoneal fluid was not found at 3rd, 7th, 14th and 21st day in all dogs (n=12) of HIGM group. The free intraperitoneal fluid in thepositive group was (1.96±0.17) cm, (1.30±0.11) cm and (0.81±0.12) cm at 1st, 3rd and 7th day and free intraperitoneal lfuid was not found at 14th and 21st day in all dogs (n=12). At 1st, 3rd and 7th day post-injection, free intraperatitoneal lfuid was statistically significant between two groups (t=20.934, P=0.003; t=41.310, P=0.000; t=22.520, P=0.000). Histopathological examination showed that there was no foreign body and foreign body granuloma and the structure of red pulp was recovered at 7th, 14th and 21st day. Gross anatomy showed that the splenic injury site was recovered completely without complications. Conclusion This study explored the value of HIGM for splenic trauma and provided a preliminary experimental evidence for clinical treatment.
8.Building and use of the appraisal indicators system of hospitals brand effect
Fei YU ; Jie CHEN ; Ke FEI ; Yingchun SHEN ; Wei JIANG ; Yuelong JIAO
Chinese Journal of Hospital Administration 2011;27(2):132-134
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.
9.Research on forecasting values of the plasma levels for TGF-β, TNF-α and IL-10 in radiation pneumonitis
Xiaofang DAI ; Gang WU ; Li LIU ; Jie YU ; Qian DING ; Yanxia ZHAO ; Yang KE
Chinese Journal of Radiological Medicine and Protection 2009;29(1):77-79
Objective To study the relationship between plasma TGF-β, TNF-α, IL-10 levels and radiation pneumonitis (RP) in patients received thoracic irradiation with 3DCRT. Methods Sixty-nine patients of lung cancer stage Ⅲ or esophageal carcinoma were evaluated prospectively by EUSA for plasma TNF-α, TGF- β, IL-10 levels and IL-10/TNF-α before 3DCRT, after 40 - 50 Gy and after 3DCRT. Results Twenty-eight patients had RP. In RP patients, the plasma TGF-β, TNF-α, IL-10 levels and IL-10/TNF-α was (15.2 ± 13.4) μg/L, (28.4 ± 13.4), (24. 1 ± 17. 1) ng/L and 1.01 ± 0.86 before 3DCRT, respectively;TNF-α increased to (36.1 ± 15.5) ng/L(t = 2.01, P = 0.040), IL-10 and IL-10/TNF-α decreased to (18.8 ± 10.8) ng/L (t =1.40, P = 0.166) and 0.62 0.55 (t = 1.90, P = 0.063)after 40-50 Gy. After 3DCRT TNF-α was higher (36.9 ± 15.5) ng/L than that before 3DCRT(t = - 2.20, P = 0.032) ,but IL-10 and IL-10/TNF-α were lower than that before 3DCRT [(13.7 ± 6.2) ng/L, t = 3.03, P = 0.005 ;0.41 ± 0.21, t = 3.60, P = 0.001]. TGF-β was not change in three times(P > 0.05) .In non-RP patients, TGF-β,TNF-α, IL-10 and IL-10/TNF-α was not yet change in three times(P > 0.05) respectively. TGF-β was not yet change between RP and non-RP patients before 3DCRT (t = 0.54, P = 0.594), and TNF-α was higher in RP group than that in non-RP group after 40-50 Gy(t = 2.02, P = 0.048), but IL-10 and IL-10/TNF-α was less in RP group than that in non-RP group after 3DCRT(t=2.50,P=0.015;t=4.63,P=0.000). Conclusions The levels of TNF-α and IL-10 are closely related to the occurrence of RP. Monitoring the changes in dynamic state could predict the generation of RP, which could be employed as a sensitive index for indicating risks for acute RP.
10.Experimental study of kallikrein-kinin system participating in proliferative vitreoretinopathy procedure
Hong-mei, ZHAO ; Jing, YU ; Min-jie, SHENG ; Ke-sheng, WANG
Chinese Journal of Experimental Ophthalmology 2011;29(7):591-595
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.