OBJECTIVETo establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen III N-terminal peptide (P III NP) in serum.

METHODSA sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of P III NP as the catalytic enzyme and the luminol as the luminescence reagent. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on. The CLEIA was compared with imported ELISA kits, by detecting clinical serum.

RESULTSThe linear range was 0.8-85 ng/ml. The detection limit was 0.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 96.2%, 91.2% and 101.1%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.99 and RSD lower than 6%. The detected results of clinical sera with CLEIA closely corresponded to those with imported ELISA.

CONCLUSIONEstablished CLEIA for quantity determination of serum P III NP has high accuracy, sensitivity and repeatability.

Adult ; Aged ; Antibodies, Monoclonal ; analysis ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Female ; Humans ; Liver Cirrhosis ; blood ; diagnosis ; Luminescence ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Peptide Fragments ; blood ; chemistry ; Procollagen ; blood ; chemistry ; Sensitivity and Specificity

OBJECTIVETo establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.

METHODSA sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on.

RESULTSThe linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.

CONCLUSIONEstablished ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.

Antibodies, Monoclonal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Membrane Proteins ; blood ; Sensitivity and Specificity

OBJECTIVETo evaluate the effects of basic fibroblast growth factor(bFGF) on proliferation of periodontal fibroblast-like cells in vivo.

METHODSA U-shaped osseous defect was produced on the buccal side of the mesial root. Four posterior teeth were conducted in four quadrants. Each quadrant included 4 groups: control, bFGF, expanded polytetrafluoroethylene(ePTFE) membrane, bFGF + ePTFE. Each time the 4 teeth sites in one quadrant were operated weekly and each dog experienced 4 times of operations. Bromodeoxyuridine(BrdU) was injected 1 hour prior to sacrificing the dogs at 4 weeks after first surgery. Immunohistochemical method was applied to count the BrdU-labeled fibroblast-like cells.

RESULTSThe number of BrdU-labeled cells reached the maximum at the 2nd week among all groups and then, decreased with time. Both bFGF and bFGF + ePTFE treated group had significantly more BrdU+ cells than remained control or ePTFE groups (P < 0.05) at 1st, 2nd weeks after surgery.

CONCLUSION2 weeks after periodontal surgery is active phase of proliferation of periodontal fibroblasts. bFGF enhances fibroblast proliferation in early periodontal wound healing, and in turn accelerate periodontal regeneration.

Animals ; Cell Division ; drug effects ; Dogs ; Fibroblast Growth Factor 2 ; pharmacology ; Fibroblasts ; cytology ; Periodontium ; cytology ; surgery ; Regeneration ; Wound Healing ; drug effects

OBJECTIVETo explore the changes of expression of uncoupling protein 2 (UCP2) in pressure overload induced failure myocardium in rats.

METHODSMale SD rats were randomized into 3 groups (n = 15 each): abdominal aorta constriction (AC) 20 weeks group (H20w group), sham operation group (SH20w group) and normal control group (N group). Twenty weeks later, myocardial function was evaluated by echocardiography and hemodynamic measurements. Mitochondria in ventricular tissue were isolated by centrifugation. Adenine nucleotide pools (ATP, ADP, AMP, PCr) in myocardium were measured by high performance liquid chromatography. The expression of UCP2 in mitochondria was detected by PT-PCR and Western blot analysis.

RESULTSMyocardial function was significantly decreased 20 weeks post-AC compared to SH20w group and N group. Myocardial ATP, ADP, AMP and PCr contents were also significantly decreased in H20w group than the other 2 control groups. The expression of UCP2 in myocardial mitochondria was significantly increased in H20w group and negatively correlated with ATP contents (r = -0.929, P < 0.01).

CONCLUSIONSThe expression of UCP2 was upregulated in pressure overload induced failure heart and might be responsible for decreased myocardial adenine nucleotide and energy metabolism disturbance in this model.

Adenine Nucleotides ; analysis ; Animals ; Echocardiography ; Heart Failure ; diagnostic imaging ; metabolism ; physiopathology ; Ion Channels ; metabolism ; Male ; Mitochondria, Heart ; metabolism ; Mitochondrial Proteins ; metabolism ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Uncoupling Protein 2

OBJECTIVEAnalyzing the relationships between peripheral blood CD4+ CD25hi regulatory T (Treg) cells and peripheral blood immune status or plasma HIV-lviral load in HIV-infected individuals,so as to determine whether Treg were related to the progression of HIV-infected disease.

METHODS116 HIV-infected patients in different stages and 21 healthy control individuals were included in this study. The CD4+ and CD8+ T cell counts were determined by a standard 4-color flow cytometry technique. The Treg cells were examined with 3-color immune staining flow cytometry. The plasma HIV-1 viral load was detected by real time PCR.

RESULTSThe frequencies of Treg cells decreased in HIV-infected individuals with high CD4+ T cell counts( > 300/microl) compared with normal controls. With the progression of disease the frequencies of Treg cells were raised gradually, until were increased in HIV-infected individuals with low levels of CD4+ T cell counts ( < 100/microl). In addition, the frequencies of Treg cells were inversely related to CD4+ T cell counts and CD4+ /CD8+ ratio, data showed a statistically significant (respectively, r = -0.564, P < 0.001; r = -0.377, P < 0.001). Furthermore, the proportions of Treg cells were closely related to plasma HIV-1 RNA viral load (r = 0.514, P < 0.001).

CONCLUSIONCD4 CD25hi Treg cells should be a kind of important cells participating the immunopathogenesis of AIDS. It may play different roles in different stages of HIV-infected disease. The exact mechanism of Treg cells in the progression of the HIV-infected disease needs to be investigate further.

Adult ; Case-Control Studies ; Cells, Cultured ; Disease Progression ; Female ; HIV Infections ; immunology ; pathology ; virology ; HIV-1 ; genetics ; immunology ; isolation & purification ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; Viral Load

OBJECTIVETo compare the efficacy of three-dimensional (3D) and two-dimensional (2D) quantitative coronary X-ray angiography (QCA) and visual estimation in the assessment of target vessels.

METHODSThe radiographic data of 60 patients (65 vessel segments) receiving coronary angiography and interventional stent placement were retrospectively analyzed. The area stenosis, diameter stenosis, lesion length, and reference diameter assessed by Medis 3D QCA, Siemens 2D QCA and visual estimation were compared.

RESULTSThree-dimensional reconstruction was successfully performed for 65 vessel segments, and 3 target vessel were excluded due to the lack of a second angiographic view for 3D reconstruction. There were significant differences in the assessments of the area stenosis [(73.87 ∓ 8.98)% vs (79.10 ∓ 8.06)% vs (83.53 ∓ 8.19)%, P<0.001], lesion length (28.95 ∓ 17.31 mm vs 26.20 ∓ 16.04 mm vs 27.21 ∓ 16.58 mm, P<0.001), reference diameter (28.95 ∓ 17.31 mm vs 26.2 ∓ 16.04 mm vs 27.21∓16.58 mm, P<0.001) by 3D QCA, 2D QCA and visual estimation; the diameter stenosis assessed by 3D [(54.21 ∓ 9.48)%] and 2D QCA [(57.84 ∓ 10.17)%] also differed significantly (P=0.016).

CONCLUSION3D QCA allows successful three-dimensional reconstruction of the target vessel and restores the actual dimensions of the vessel for a more accurate assessment of coronary artery disease than 2D QCA and visual estimation.

Aged ; Coronary Angiography ; methods ; Coronary Disease ; diagnostic imaging ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo investigate the prevalence of diabetes and prediabetes and their association with the risk for coronary heart disease (CHD) in elderly residents in Haizhu District of Guangzhou.

METHODSStratified random sampling was employed to select a total of 1800 resident aged 50 years or older in the region. The fasting fingertip blood glucose>5.6 mmol/L was used as the criterion for the initial screening. The data were collected from qualified subjects via scheduled questionnaire surveys, blood collection and testing, and physical examination. The subjects were divided into the 3 groups, namely normal blood glucose, prediabetes, and diabetes groups. The combination rates of the relevant risk factors (hypertension, hyperlipemia, obesity, and central obesity) were compared among the groups by Framingham Heart Study to predict the occurrence of CHD in 10 years.

RESULTSThe incidence was 11.00% for prediabetes and 7.56% for diabetes in the elderly residents in Haizhu District. The occurrence of hypertension, hyperlipemia, obesity, and central obesity was significant higher in the prediabetes and diabetes group than in the normal blood glucose group, and showed no significant differences between the former two groups. The 10-year risks for CHD were markedly higher in both the prediabetes and diabetes groups than in the normal blood glucose group, but similar between the former two groups.

CONCLUSIONElderly patients with prediabetes and diabetes have significantly increased 10-year risk for CHD in comparison with those with normal blood glucose, but the risk is similar between the former two groups, indicating a close association of IGR (impaired fasting glucose+ impaired fasting glucose) with CHD. Early control of blood glucose is essential to the prevention and control of CHD.

Aged ; Blood Glucose ; analysis ; China ; epidemiology ; Coronary Disease ; blood ; epidemiology ; Cross-Sectional Studies ; Diabetes Mellitus, Type 2 ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prediabetic State ; epidemiology ; Prevalence ; Risk Factors ; Sampling Studies

OBJECTIVETo evaluate the association between the polymorphisms of excision repair cross complementation group 1 (ERCC1), X-ray repair cross complementing 1 (XRCC1), glutathione S-transferase Pi 1 (GSTP1) and the survival of advanced gastric cancer patients treated with oxaliplatin-based combination chemotherapy.

METHODSEighty five patients with advanced gastric cancer accepted oxaliplatin/5-FU-based chemotherapy as first-line chemotherapy were investigated. Peripheral venous blood was taken before chemotherapy. DNA was extracted from peripheral venous blood. The genetic polymorphisms were detected by real-time PCR assay. The association between time to progression, overall survival and the polymorphisms was analyzed.

RESULTSThe median time to progression of the 85 cases was 5.3 months, and the median overall survival was 8.0 months. ERCC1-118 C/C, XRCC1-399 G/G and GSTP1-105 A/G + G/G were favorable genotypes and the number of the favorable genotypes was associated with survival of the patients. The median overall survival was 12.5 months, 10.0 months, 6.5 months and 4.5 months for patients with 3 favorable genotypes, 2 favorable genotypes, 1 favorable genotype and none favorable genotype, respectively, with a significant difference (χ(2) = 35.54, P < 0.01).

CONCLUSIONGenetic polymorphisms of ERCC1-118, XRCC1-399 and GSTP1-105 are associated with TTP and OS of advanced gastric cancer patients treated with oxaliplatin/5-Fu-based combination chemotherapy as the first-line chemotherapy.

Adenocarcinoma ; drug therapy ; genetics ; pathology ; Adenocarcinoma, Mucinous ; drug therapy ; genetics ; pathology ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; DNA-Binding Proteins ; genetics ; Disease Progression ; Endonucleases ; genetics ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Glutathione S-Transferase pi ; genetics ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Organoplatinum Compounds ; administration & dosage ; Polymorphism, Genetic ; Stomach Neoplasms ; drug therapy ; genetics ; pathology ; Survival Rate ; X-ray Repair Cross Complementing Protein 1

Purpose To investigate the association of the PD-1 and PD-L1 protein expression with K-RAS gene mutations in lung adenocarcinoma.Methods The protein expression of PD-1 and PD-L1 was detected by immunohistochemical EnVision two-step staining,the K-RAS mutation was examined by realtime fluorescent quantitative PCR.Results The positive rate of PD-1 and PD-L1 was higher in lung adenocarcinoma than benign lung disease (P ＜ 0.01).There was no relationship between PD-1 and PD-L1 protein expression with the gender,age,smoking condition,differentiation,lymph node metastasis and TNM stages (P ＞ 0.05).The K-RAS gene mutations were detectable in 8 patients (22.2％) among 36 lung adenocarcinoma,there was no association between K-RAS gene mutation with the gender,age,smoking condition,differentiation,lymph node metastasis and TNM stages (P ＞ 0.05).The correction analysis showed that there was no relationship between PD-1 and PD-L1 protein expression with K-RAS mutation (P ＞ 0.05).Conclusion The positive rate of PD-1 and PD-L1 is higher in lung adenocarcinoma than benign lung disease,but there is no relationship among PD-1 and PD-L1 protein expression with its clinal pathological characteristics and K-RAS mutation in lung adenocarcinoma.

Hip trauma has been a leading cause of death in senile patients for more than a centenary. Although the mortality decreased due to the advanced technique in medication, surgery and nursing, the increasing mortality should not be neglected in elders after orthopedic operation nowadays. Many factors are considered to influence the causes of death after trauma, such as age, gender, personal customs, comorbidities, types of fracture, timing of surgery, procedure, anesthesia, complications, medical treatment, activity of daily living, or even marriage status. This article reviews these causes from the aspects of patient's own factors, iatrogenic factors, medical treatment and other factors and provides some clues for further clinical application according to the recent foreign and domestic researches. According to the present research, it is essential for surgeons to perform a comprehensive estimation for patients suffering from hip trauma.
Cause of Death ; Hip Fractures ; surgery ; Humans ; Orthopedic Procedures ; Regression Analysis