Objective To investigate the changes of iodide uptake and the expression of thyroidspecific genes in poorly differentiated follicular thyroid carcinoma (FTC) cells after transfection of human TSH receptor (hTSHR) gene in vitro. Methods The recombinant eukaryotic expression plasmid PcDNA3. 1/hTSHR-cDNA was transformed into DH5a bacterial for amplification and then the recombinant plasmid was extracted. The recombinant was identified with PCR amplifying, restriction enzyme digestion analysis and DNA sequencing. The recombinant plasmid pcDNA3.1/hTSHR was transfected into FTC-133 cell line by lipofectin methodin vitro. Immunofluorescence, iodide uptake studies and real time-PCR were applied to detect target protein expression. Statistical analysis was performed with t-test using SPSS 13. 0 software. Results Kpn Ⅰ and Xba Ⅰ restriction enzyme digestion, PCR amplifying and DNA sequencing confirmed that pcDNA3. 1/hTSHR was successfully constructed. After transfection of the recombinant plasmid pcDNA3. 1/hTSHR-cDNA and the stimulation of hTSH, the tumor cells displayed the expression of hTSHR protein at cell surface and cytoplasm. The iodine uptake in pcDNA3. 1/hTSHR transfected cells was 2. 9 times higher than that of control(pcDNA3.1(+) transfected cells) group(t = 28.63, P ＜0. 01). The expression of TSHR,NIS, TPO and Tg (mRNA levels) in pcDNA3. 1/hTSHR transfected cells were also significantly elevated by 1.74 (t =5.959, P＜0.01), 7.2 (t =3.807,P＜0.05), 2.88 (t=4.769,P＜0. 01) and 2.67 times (t=6.388,P ＜0.01) respectively compared to those of the control group. Conclusion The study demonstrates that iodide uptake may be reactivated by hTSHR receptor gene transfection in poorly differentiated FTC cell.

A reliable method for simultaneous determinition of eleven representative components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin) in combination of chromatographic fingerpint analysis for Reduning injection was developed by ultra high-performance liquid chromatography (UPLC). The method was performed on an Agilent ZORBAX SB-C18 anlytical column (3. 0 mm x 100 mm, 1. 8 µm) with a guard column of Agilent UPLC Guard ZORBAX SB-C18 (3.0 mm x 5 mm) at the column temperature of 30 °C. The gradient mobile phase consisted of acetonitrile (A)-0. 1% phosphoric acid (B) with a flow rate of 0. 4 mL . min-1. The injection volumn was 2 µL. The detection wavelengths were set at 324 nm and 238 nm for quantit tive analysis and 225 nm for fingerpint chromatography. Neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin were baseline seperated with good linearity relationships (r >0. 999) between concentration and peak areas over the linear ranges. The average recoverys of the investigated compounds were 103.5%, 100. 2%, 103. 3%, 102. 8%, 101. 3%, 102. 8%, 97. 36%, 99. 62%, 98. 16%, 102. 8%, 99. 27%, respectively. Reduning injection of forty-five batches was analyzed by UPLC finge print chromatography. Thirty batches were selected to generate the reference fringerprint chromatography with fourteen common peaks. The similarity values between the reference fringerprint chromatography and the remaining fifteen batches were higher than 0. 99. The developed method was fast, accurate and sensitive. It could be used as a reference for the quality control of multiple components determination and fingerprint chromatography for Reduning injection in future.
Chlorogenic Acid ; chemistry ; isolation & purification ; standards ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Glucosides ; chemistry ; isolation & purification ; standards ; Iridoids ; chemistry ; isolation & purification ; standards ; Quality Control ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity ; Time Factors

Objective To assess the prognosis of patients with bladder neck (BN) involvement in radical prostatectomy specimens and compare it with patients with seminal vesicle invasion (SVI).Methods From January 2002 to December 2008,42 patients with prostate carcinoma who underwent radical prostatectomy were analyzed retrospectively.The mean age was 70.3 years (range 59-78 years).The patients were divided into two groups according to 2002 TNM system:pT4a group (bladder neck invasion) with 17 cases and pT3b group (unilateral or bilateral seminal vesicle invasion)with 25 cases.Postoperative outpatients were followed-up on a regular basis,two consecutive postoperative serums PSA greater than 0.2 μg/L defined as a biochemical recurrence.The postoperative follow-up to the time of biochemical recurrence or follow up to the deadline without biochemical recurrence were defined as biochemical recurrence free survival time.Results All patients were followed up for 50-122 months.The patients with BN involvement 17.6％ (3/17) had biochemical recurrence,whereas seminal vesicle invasion 24.0％ (6/25),there was no statistical significance between the two groups (P＞0.05).Conclusions Postoperative adjuvant therapies can improve the biochemical recurrence free survival.In this study,the prognosis of pT4a and pT3b group is similar.A conceivable down-staging of BN involvement in the TNM staging system should be considered.

Objective To explore the clinical value of transgastric natural orifice transluminal endoscopic surgery(NOTES)in diagnosing unexplained ascites.Methods The clinical data in 12 cases of unexplained ascites diagnosed by adopting transgastric approach NOTES and performed abdominal exploration and peritoneal biopsy in our hospital from November 2015 to July 2016 were retrospectively analyzed.The operative risk and clinical application value were evaluated by statistically analyzing the postoperative complications occurrence and the diagnosis rate of disease.Results The definite diagnosis rate reached 100％ verified by pathology after abdominal exploration and peritoneal biopsy,in which 8 cases(66.7％)were tuberculous peritonitis,2 cases(16.7％)were liver cirrhosis,1 case(8.3％)was peritoneal mesothelioma,1 cases(8.3％)was peritoneal metastatic carcinoma;2 cases appeared abdominal pain after operation,including 1 case of neutrophil ratio increase,symptoms and persistent time of abnormal laboratory indexes did not exceed 24 h,the incidence rate was 8.3％;no complications of abdominal cavity infection,incision bleeding and puncture site fistula occurred.Conclusion The transgastric NOTES for conducting abdominal exploration and peritoneal biopsy in the diagnosis of unexplained ascites has the advantages of small trauma,less complications and rapid postoperative recovery,possesses an important clinical application value.

OBJECTIVETo evaluate the effects of two sample preparation methods on two-dimensional polyacrylamide gel electrophoresis (2-DE)-based serum protein separation, and produce high-resolution and reproducible 2-DE images for identifying disease-related serum protein.

METHODSDirect solubilization and hot SDS methods were used separately to extract and handle the total proteins of serum samples from patients with colorectal carcinoma. Immobilized pH gradient 2-DE was used to separate the total proteins. After image analysis of silver-stained 2-D gels, 3 differential protein spots were identified by matrix-assisted laser desorption/time-of-flight mass spectrometry.

RESULTSThe total proteins treated with hot SDS method were used to perform 2-DE. 2-DE patterns with high resolution and reproducibility were obtained for human serum samples. 2-DE was performed 3 times for the samples treated by direct solubilization and hot SDS methods, respectively, resulting in the average number of spots of 675-/+46 and 702-/+49, respectively. The average matching protein spots were 573-/+42 and 623-/+52, with average matching rate of 85.3% and 89.6%, respectively. The average position deviation of matched spots in different gels was 0.85-/+0.30 mm and 0.81-/+0.28 mm in IEF direction, and 1.02-/+0.18 mm and 0.97-/+0.12 mm in SDS-PAGE direction. Mass spectrometry of the 2-D gels treated with hot SDS method generated high-quality mass spectra, and the sample preparation method allowed detection of relatively low abundance protein.

CONCLUSIONHot SDS method is more effective for human serum protein sample preparation and well-resolved, reproducible 2-DE profiles of human serum have been established in this study.

Blood Protein Electrophoresis ; Blood Proteins ; analysis ; isolation & purification ; Electrophoresis, Gel, Two-Dimensional ; methods ; Electrophoresis, Polyacrylamide Gel ; methods ; Humans ; Peptide Mapping ; Protein Interaction Mapping ; Reproducibility of Results

Objective To identify the epidemic characteristics and risk factors of an emerging infectious disease-severe fever with thrombocytopenia syndrome (SFTS) in Hubei province.Methods Active surveillance program on SFTS was set up in monitoring sites-hospitals,at the township level or above,in Suizhou,Huanggang and Wuhan from January to December,2010.Specific surveillance program on SFTS was launched across the province in hospitals above the county level.Cases that matched the definition of surveillance case were identified and reported to Centers for Disease Control and Prevention (CDCs).Cases were interviewed and their blood samples collected and detected using PCR and virus isolation.We also conducted serum antibody surveys among healthy population and livestock and surveillance on vector ticks in those high-epidemic areas.Results 188 cases that matched the definition of surveillance case and 21 deaths were reported in 11 cities,32 countries and 100 towns in 2010,with an incidence rate of 0.33/106.The fatality rate was 11.2％.Data showed that the patients were from hilly areas at the altitude elevated between 28-940 meters.The epidemic period was between April and December with the peak from May to September.The youngest case was an 11-year old,while the eldest was 81 with median age as 56-year old.95.3 ％ of the patients were farmers.All Patients did not have the history of traveling,two weeks before the onset of SFTS.93.6％ of the patients engaged in different kind of work which was associated with agriculture.52.8％ of the patients had been exposed to ticks.22.0％ of the patients had been bitten by ticks.Skin injury was found in 64.2％ of the patients.Samples from 129 cases (68.6％) were collected and detected,with 67.4％ of them (87 cases) showed positive by Real time-PCR for SFTS virus.An elevation in antibody titer by a factor of four or evidence of sero-conversion was observed in 11 patients; SFTS virus was isolated from 2 patients.The total antibody positive rates were 3.8％,55.0％ (6/11 ),36.7％ (2/3) and 80.0％ (4/5) respectively in healthy population,dogs,sheep and cows.Ticks from grass,cattle and sheep were detected positive by Real time-PCR.Conclusion Most cases of SFTS in Hubei were infected by SFTS virus,and cases of livestock were infected by SFTS virus.Ticks might serve as an important vector.Skin injury,exposure to tick bites seemed to be the risk factors.

OBJECTIVETo study soft tissue changes observed through musculoskeletal ultrasound (MSUS) in the treatment of knee osteoarthritis with needle-knife, so as to provide MSUS basis for needle-knife in the treatment of knee osteoarthritis.

METHODSForty patients with knee osteoarthritis treated in the Third Affiliated Hospital of Beijing University of Chinese Medicine from December 2011 to December 2012 were selected according to inclusion and exclusion criteria. All the patients were treated with needle-knife release method. The VAS scores and knee joint circumference were recorded before treatment and 2 weeks after treatment. The changes of knee joint hydrops articuli and joint synovial thickness were measured through MSUS.

RESULTSThe knee pain index was 6.850 +/- 1.417 before treatment and 2.790 +/- 1.299 after treatment;the index after treatment was lower than that of before treatment. The knee joint circumference was 407.320 +/- 45.151 mm before treatment and 391.240 +/- 41.129 mm after treatment; the knee joint circumference decreased after treatment. The amount of hydrops articuli observed by musculoskeletal ultrasound showed that 47 knees were cured, 19 knees improved and 2 knees failed. The synovial membrane thickness: 43 knees cured, 17 knees improved and 8 knees failed.

CONCLUSIONThe hydrops articuli and synovial thickness of knee joint of patients with knee osteoarthritis observed under the MSUS is consistent with the main symptoms and signs, which suggests that MSUS observation on soft tissue changes before and after needle knife in the treatment of knee osteoarthritis with high reliability.

Adult ; Aged ; Female ; Humans ; Knee Joint ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Needles ; Osteoarthritis, Knee ; complications ; diagnostic imaging ; pathology ; surgery ; Pain ; complications ; Synovial Membrane ; pathology ; Treatment Outcome ; Ultrasonography

Glycoprotein (GP) Ibα ectodomain shedding has important implications for thrombosis and hemostasis. A disintegrin and metalloproteinase 17 (ADAM17) was identified to play an essential role in agonist induced GPIbα shedding. The relationship of GPIbα shedding and ADAM17 in the acute stage of atherosclerotic ischemic stroke (AIS) patients has not been thoroughly studied. A total of 306 patients and 230 controls matched for age, sex, race, history of hypertension and diabetes mellitus were enrolled in the study. GPIbα, ADAM17, glycocalicin were detected by flow cytometry, Western blotting, and enzyme-linked immunosorbent assay (ELISA) respectively. Compared with the control group, the expression of GPIbα in patients with acute ischemic stroke was significantly lower (P=0.000, P<0.01). Plasma glycocalicin and ADAM17 in AIS group were higher than those in control group (P=0.699, P=0.000). Pearson's analysis showed glycocalicin bore no correlation with GPIbα in AIS patients (r=0.095, P>0.05). GPIbα and National Institute of Health Stroke Scale (NIHSS) had negative correlation (r=-0.514, P<0.01). Our findings indicate that ADAM17 may be a risk factor for ischemic stroke in Chinese and the expression of GPIbα can serve as a measure for stroke severity.
ADAM Proteins ; blood ; ADAM17 Protein ; Biomarkers ; blood ; Blood Platelets ; metabolism ; Brain Ischemia ; blood ; diagnosis ; China ; Female ; Humans ; Intracranial Arteriosclerosis ; blood ; diagnosis ; Male ; Middle Aged ; Platelet Glycoprotein GPIb-IX Complex ; metabolism ; Reproducibility of Results ; Sensitivity and Specificity ; Severity of Illness Index ; Stroke ; blood ; diagnosis

Objecfive To establish a method for in vitro culture and identification of neural stem cells(NSCs)derived from the olfactory bulb(OB)of adult mice and test the possibility of the OB as a new source of seed cells of adult NSCs. Methads NSCs were isolated from the OB of adult mice and cultured in serum-free medium.Clonal culture and BrdU incorporation assay were performed to assess the self-renewal and proliferative activities of the NSCs.Fluorescence immunocytochemistry was carried out to examine the expression of the NSC markers nestin and SOX2,neuronal marker Tujl,astrocyte marker GFAP and oligodendroeyte marker 04. Results NSCs possessing self-renewal and proliferative capacities were obtained from the OB of adult mice,and the cells grew in the form of floating neurospheres in the medium.The neurospheres consisted of cells were positive for NSC markers nestin and SOX2,which Were able to differentiate into Tuj1-positive neurons,GFAP-positive astrocytes and 04-positive oligodendrocytes. Conclusion NSCs are present in the OB of adult mice,and the NSCs isolated from the OB can proliferate and differentiate in vitro with obvious stem cell properties.suggesting the feasibility of using OB as anew source of adult NSCs.

Objective To study the influence of age,gender and history of diabetes in maximum standardized uptake value (SUVmax) and standardized cerebral metabolic rate (SCMR) of glucose.Methods Seventy-nine healthy volunteers were recruited and evaluated by PET-CT examination.Region of interest (ROI) analysis was employed to measure the SUVmax in 13 ROIs of the brain and 13ROIs of the cerebellum.SCMR was amounted by left SUVmax/right SUVmax in the corresponding ROI point of each side in the same volunteer.The influences of age,gender and history of diabetes in SUVmax and SCMR were analyzed. Results No statistical significance of SUVmax was noted between the 2corresponding points in bilateral ROI (P＞0.05).Statistical significance of SUVmax was noted between each 2 points in the left and right ROI (left:F=14.280,right:F=14.680,P=0.000).Statistical difference of SUVmax was recorded in different age groups (P＜0.05).The female had significantly lower SUVmax in bilateral measurement points (2/26) as compared with the male (P＜0.05).Concerning the influence of history of diabetes,statistical significance of SUVmax was noted in almost all ROIs (18/26,P＜0.05).No statistical significance of SCMR in each ROI was noted in different groups of gender,age and diabetes (P＞0.05). Conclusion SCMRglu can eliminate the role of such influencing factors as age,gender and history of diabetes,which have been proved to exert some effect on the results of SUVmax; therefore,SCMRglu may provide a reliable indicator for further study.