Objective The purpose of this study was to develop a high-throughput micro-plate radiobinding assay (RBA) of glutamic acid decarboxylase antibody (GAD-Ab) and to evaluate its clinical application. Methods 35labeled GAD65 antigen was incubated with sera for 24 h on a 96-well plate, and then transferred to the Millipore plate coated with protein A, which was washed with 4℃ PBS buffer, and then counted by a liquid scintillation counter. The GAD-Ab results were expressed by WHO standard unit (U/ml). A total of 224 healthy controls, 162 patients with type 1 diabetes mellitus(T1DM) and 210 patients with newly diagnosed type 2 diabetes (T2DM) were recruited. A total of 119 TI DM and healthy cases with gradually changing GAD-Ab levels were selected to compare the consistency of micro-plate RBA with conventional radioligand assay (RLA). Blood samples were obtained from the peripheral vein and finger tip in 32 healthy controls, 35 T1DM and 24 T2DM patients, and tested with micro-plate RBA and then compared with the conventional RLA to investigate the reliability of finger tip sampling. Linear correlation,student's t-test, variance analysis and receiver operating characteristic (ROC) curve were performed using SPSS 11.5. Results (1) The optimized conditions of micro-plate RBA included 2 μl serum incubated with3 ×104 counts/min 35S-GAD for 24 h under slow vibration, antigen-antibody compounds washed 10 times by 4℃ PBS buffer, and radioactivity counted with Optiphase Supermix scintillation liquid. (2)The intra-batch CV of the micro-plate RBA was 3.8%- 10.2%, and the inter-batch CV was 5.6%- 11.9%. The linearity analysis showed a good correlation when the GAD-Ab in serum samples ranged from 40.3 to 664 U/ml and the detection limit of measurement was 3.6 U/ml. The results from Diabetes Autoantibody Standardization Program (DASP) 2005 showed that the sensitivity and specificity for GAD-Ab were 78% (39 positive among 50 new-onset T1DM) and 98% (2 positive among 100 healthy controls). The results of GAD-Ab obtained with micro-plate RBA and RLA were closely correlated (r=0.915,P<0.001) with a high concordance level of 97.5% and a Kappa value of 0.95. (3)TI DM and T2DM patients showed higher positive rates for GAD-Ab than the healthy controls(46.9% and 5.2% vs 0.89% ,X2=123.5 and 10. 1 ,P <0.001 and <0.01, respectively). (4)The consistency of GAD-Ab measurement with RBA using finger tip blood and RLA measurement using venous blood was 96.7% (r =0.946,P <0.001, Kappa value: 0.905). Conclusions The micro-plate RBA of GAD-Ab has high sensitivity, specificity and reproducibility, and can be measured with finger tip blood sampling. It might be a better alternative for clinical practice.

The content of the asarone submicro emulsion injection was determind by HPLC method, and thereby a quality evaluation method was established based on indexes of pH value, particle size, peroxide value, methoxy aniline values, free fatty acid, lysophosphatidylcholine, visible foreign substances, insoluble particle, sterility, bacterial endotoxin and impurities, etc. The results showed that the injection exhibited uniform physical appearance and all the products were in milkwhite liquid. The content of the three batches products were respectively 102.9%, 100.8%, 97.70% of the labeled amount, with mean particle size of 210-250 nm, and other indexes all met with the standards. The reserved samples showed no obvious change in terms of detection indexes and indicated good stability after the accelerated stability test and long-term stability for 12 months. The quality evaluation method established in this study could be applied to quality control and stability investigation of asarone submicron emulsion injection, which laid a basis for further clinical research and application.
Anisoles ; chemistry ; Chromatography, High Pressure Liquid ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Emulsions ; chemistry ; Particle Size ; Quality Control

Advance Care Planning ; ethics ; legislation & jurisprudence ; Aged ; Aged, 80 and over ; Cross-Sectional Studies ; Decision Making ; ethics ; Female ; Humans ; Male ; Middle Aged ; Patient Preference ; statistics & numerical data ; Risk Assessment ; Singapore ; Terminal Care ; ethics ; methods ; Third-Party Consent ; ethics ; legislation & jurisprudence

OBJECTIVE: To evaluate the long-term efficacy and safety of ultrasound-guided percutaneous nephrolithotomy (PCNL) in the treatment of patients with solitary kidney stones. METHODS: The clinical data of 22 patients with solitary kidney stones treated with PCNL in Peking University People's Hospital from September 2008 to June 2014, with the follow-up data of more than 5 years were analyzed retrospectively. Perioperative indicators, postoperative stone free rate (SFR) and incidence of complications were recorded. Ultrasonography was used to evaluate the long-term stones recurrence rate. Serum creatinine and estimated glomerular filtration rate (eGFR) were used to assess the long-term renal function. RESULTS: In this group of 22 patients, the average age was (50.3±11.8) years, with 10 cases of anatomic solitary kidneys, 12 functional solitary kidneys, and the median stone diameter was 1.65 (1.1-3.9) cm. All the patients had multiple stones, including 7 cases of staghorn stones. The median pre-operative serum creatinine was 104.5 (60.0-460.0) μmol/L, and the mean eGFR was (60.3±29.4) mL/min, showing no statistically significant difference compared with that before surgery. The mean operative time was (88.2±42.0) min, and there were 11 cases of single-channel and double-channel PCNL. The median serum creatinine on the first day after surgery was 102.0 (63.0-364.0) μmol/L, and the mean eGFR was (58.0±25.1) mL/min. The mean postoperative hospital stay was (8.7±5.2) days. In this group, 5 patients (22.7%) presented short-term complications, among which 4 patients presented postoperative infection and massive hemorrhage at the same time, which improved after conservative treatment, and 1 patient presented pleural injury and improved after closed thoracic drainage. Two patients (9.1%) developed long-term complications, and ureteral stricture occurred 3 months after operation, which improved after balloon dilatation. The median follow-up time was 6.2 (4.7-11.1) years. The median serum creatinine at the last follow-up was 104.0 (72.4-377.0) μmol/L, and the mean eGFR was (60.1±23.7) mL/min, showing no statistically significant difference compared with that before surgery. Renal function decreased in 6 patients (27.3%). Initial and final SFR were 72.7% and 100%, respectively. During the 6.2-year follow-up, 9 patients (40.9%) experienced recurrence of kidney stone. After stone recurrence, 13 lithotomy surgeries were performed, and the SFR by the latest follow-up was 63.6%. CONCLUSION This study had the longest follow-up time for patients with solitary kidney stones after PCNL reported at home and abroad. Ultrasound-guided standard PCNL was safe and effective in the treatment of solitary kidney stones. Long-term follow-up results showed that the recurrence rate of kidney stones was still high, but the long-term renal function was stable after operation, and some patients showed mild renal function decline.
Adult ; Humans ; Kidney Calculi ; Middle Aged ; Nephrolithotomy, Percutaneous ; Retrospective Studies ; Solitary Kidney/surgery* ; Treatment Outcome

Adolescent ; Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Humans ; Influenza A virus ; immunology ; Male ; Seroepidemiologic Studies ; Students, Medical ; Vietnam ; epidemiology ; Young Adult

OBJECTIVETo study the absorption, translocation and accumulation of N, P and K on Achyranthes bidentata.

METHODThe contents of N, P and K were determined by mean of sulfuric acid-hydrogen peroxide assimilating method, vanadium-ammonium molybdate colorimetric method and flame photometric method, respectively.

RESULTThe contents of N, P and K in the plant were decreasing during the growth period. The absorption rates of the three nutrients by A. Bidentata showed double-peak curves in the whole growth period, maximum absorption rate appeared in the middle ten days of October. About 8.59 kg of N, 1.36 kg of P and 7.40 kg of K were needed to produce each 100 kg root.

CONCLUSIONThe key nutrients absorption period is in the first ten days of September and in the middle ten days of October.

Achyranthes ; metabolism ; Biological Transport ; Nitrogen ; metabolism ; Phosphorus ; metabolism ; Potassium ; metabolism

OBJECTIVETo detect the expression levels of telomere binding factor 2 (TRF2) on leukemia cell lines and primary leukemia cells.

METHODSThe expression of TRF2 mRNA was detected with quantitative real-time RT-PCR in leukemia cell lines and primary leukemia cells. The Western blot analysis was used for the detection of TRF2 protein expression.

RESULTTRF2 was overexpressed in T-cell leukemia cell lines but not in myelogenous leukemia cell lines. Significant higher expression levels of TRF2 were observed in primary leukemia cells from patients with M0 and M1 subtypes of acute myelogenous leukemia (AML) compared with normal control and other subtypes of AML.

CONCLUSIONIncreased TRF2 expression levels are found in T-cell leukemia cell lines and AML patients with poor prognosis, which suggests that TRF2 expression might be related to the prognosis of leukemia.

Adolescent ; Adult ; Female ; HL-60 Cells ; Humans ; Jurkat Cells ; K562 Cells ; Leukemia, Myeloid, Acute ; metabolism ; Leukemia, T-Cell ; metabolism ; pathology ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomeric Repeat Binding Protein 2 ; metabolism ; Young Adult

Objective To determine the changes of cerebrospinal fluid(CSF) ?-endorphin(?-EP) and C-reactive protein(CRP) in children with central nervous system(CNS) infection.Methods Sixty-five children suffered from CNS infection were determined the plasma and CSF ?-EP and CRP concentration during the acute and recovering stage with radioimmunoassay, which included 48 viral encephalitis, 12 purulent meningitis and 5 tuberculou meningitis,and 24 non-CNS disease children were as control group.Results The concentrations of plasma and CSF ?-EP of every experimental group were obviously higher than those of control group during the early stage of CNS infection and these were obviously lower during the recovering stage. The serum concentration of CRP during acute stage was significantly higher than that during recovering stage. No change of serum and CSF CRP concentration was determined during either the acute or recovering stage in the other two experimental groups.Conclusions Determining the plasma and CSF ?-EP is mea-(ningful) in early diagnosis of CNS infection,and determining the serum CRP at the same time may be helpful in differentiating septic and inseptic infection.

Objective To investigate the predietors of road traffic accidents(RTAs)in middle school students.a ease-control study WaS designed to seek new clues for prevention and control on RTAs in this population.Methods Cases were 116 students who experienced RTAs from 2005/01/01 to 2005/12/31.116 students were.served as controls who were from the same clams of the cases and matched by age and sex.Information on personal behavior,family conditions,and knowledge,attitude,practice(KAB)on road traffic of these children were collected and compared.Conditionallogistic regressions were conducted to analyze the relationship between these factors and RTAs.Results Compared to the controls.the cases showed more negative attitude and risk-taking-behaviors especially in riding bicycle.However,knowledge on road safety showed less significant differences between eaSe and control groups.The negative attitude and risk-taking-behaviors were independent risk factors for RTAs by logistic regression.In addition.distance from home to school that longer than 1000 m seemed a risk factors for RTAs.while high education levels among mothers WaS a protective factor.Conclusion Road safety attitude and behavior.distance from home to school were the main influenee factors for RTAs among middle school students.It seemed more important in improving the attitude and behavior on road safety than to enhance the knowledge for prevention and eontrol of RTAs among middle school students.

OBJECTIVETo construct a eukaryotic expression vector for expressing hepatitis B virus (HBV) recombinant HBsAg-EGFP fusion protein and obtain a stable transfected Chang Liver cell line.

METHODSThe coding region of HBsAg gene of HBV was amplified by PCR and was digested by BamH I/EcoR I . This fragment was inserted into pEGFPN1 with T4 ligase and transformed E-coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into Chang Liver cell by Lipofectamine 2000 cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level HBsAg-EGFP fusion protein was obtained.

RESULTSThe eukaryotic expression vector named pEGFPN1-HBsAg was successfully constructed and the stable transfected Chang Liver cell line expressing pEGFPN1-HBsAg fusion protein was obtained.

CONCLUSIONThe stable transfected Chang Liver cell line could express pEGFPN1-HBsAg fusion protein, could be used to screen the proteins differentially expressed in HBsAg expression Chang Liver cells, which brought some new clues for studying the potential molecular mechanism of HBsAg protein.

Cell Line ; Gene Expression ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Liver ; cytology ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; methods