OBJECTIVETo evaluate the effect of COX-2 silencing on the radiosensitivity of a nasopharyngeal carcinoma (NPC) cell line C666-1.

METHODSAnti-COX-2 C666-1 cell line with COX-2 gene silencing mediated by shRNAmir lentiviral vector and the control cell line Anti-GL-2 C666-1 were exposed to various radiation doses. The clonogenic survival assay and curve fitting was used to calculate the radiobiological parameters and the sensitization enhancement ratio after the radiation. Cell cycle changes were assessed after the exposure by flow cytometric analysis. In a BALB/c nude mouse model, the growth curve of the xenografts was generated and the tumor growth inhibition rate was calculated.

RESULTSCompared with the control cells, Anti-COX-2 C666-1 cells showed obviously lowered values of SF2, D0 and Dq but significantly increased α/β with a sensitivity enhancement ratio of 1.4014. COX-2 gene silencing increased the inhibition rate of the tumor xenografts after the radiation, and caused also decreased percentage of G2/M arrest resulting from the exposure.

CONCLUSIONStable COX-2 silencing in NPC cells can improve the effect of radiotherapy both in vitro and in vivo. By changing the radiobiological parameters, genetically based COX-2 inhibitor may be a potentially promising radiosensitizer of NPC.

Animals ; Carcinoma ; Cell Line, Tumor ; Cell Survival ; Cyclooxygenase 2 ; genetics ; Female ; Gene Silencing ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Nasopharyngeal Neoplasms ; radiotherapy ; RNA, Small Interfering ; Radiation Tolerance ; genetics ; Xenograft Model Antitumor Assays

OBJECTIVETo develop a follicular unit-like construct with allogeneic hair, evaluate its histocompatibility and long-term stability after transplantation, and explore the possibility of its clinical application.

METHODSHuman hair and medical polypropylene was processed according to the structure of follicular units and prepared into hair prostheses for transplantation. The histocompatibility of polypropylene and human hair in New Zealand rabbits was observed by HE staining and scanning electron microscope, and the loss rate of the hair was recorded to evaluate the long-term result of transplantation.

RESULTSMild inflammatory cell infiltration around polypropylene and human hair was observed early after the transplantation, accompanied with local epithelial cell proliferation. The prosthesis mimicking the follicular units still showed good histocompatibility one year after the transplantation without degradation of the hair. The loss rate of the hair was averaged (4.1∓4.0)% at one year after the transplantation, and the total appearance of the prosthesis remained satisfactory.

CONCLUSIONAllogeneic human hair and polypropylene in the hair prosthesis show good histocompatibility in rabbits. The prosthesis allows good cosmetic effect after transplantation with low rate of hair loss, demonstrating its potential in clinical application.

Animals ; Biocompatible Materials ; Female ; Hair ; transplantation ; Hair Follicle ; transplantation ; Humans ; Materials Testing ; Polypropylenes ; Rabbits

OBJECTIVETo construct the plasmid reference molecules for detection of transgenic tomato line Zeneca B,Da,F using quantitative real-time PCR(qPCR).

METHODSThree plasmid reference molecules pEasy-T3-APX, pEasy-T3-16A and pEasy-T3-16S were cloned based on reverse genetics, which contain the target fragments of tomato endogenous reference gene apx (ERG-apx), gene-specific sequence of pg(GS-pg) and construct-specific sequence of vectors pJR16S/pJR16A (CS-16S/CS-16A) of Zeneca B,Da,F, respectively. Primers and Taqman probes were designed by Beacon Designer 7.5.The specificity, sensitivity, reproducibility and the limit of detection(LOD) of the qualitative and quantitative PCR system based on the plasmid reference molecules were evaluated. PicoGreen was used to measure the DNA concentration of the plasmid reference molecules. Two sets of samples containing 1% or 0.1% (w/w) pEasy-T3-16A or pEasy-T3-16S mixed with pEasy-T3- APX as background DNA were prepared for evaluating the efficacy of the qPCR system.

RESULTSThe target fragments for qPCR detection were anchored, ERG-apx 108 bp, GS-pg 108 bp , CS-16S 109 bp and CS-16A 102 bp. The three plasmid reference molecules were confirmed at the expected sizes by restriction enzyme digestion. The qPCR results showed that the RSD of reproducibility were 0.2% to1.5%, LOD was 25 copies, R2 values for these standard curves were 0.994 ~0.998 and amplification efficiencies were 93.3%~102.4%.The bias between the test and true values of two sets of mixed samples ranged from -9.3% to 14.7% after adjusting by conversion factors(Cf).

CONCLUSIONThe plasmid reference molecules and qPCR system for qualitative and quantitative detection of transgenic tomato line Zeneca B,Da,F have been established successfully.

Base Sequence ; DNA, Plant ; genetics ; Lycopersicon esculentum ; genetics ; Plants, Genetically Modified ; genetics ; Plasmids ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVETo explore the optimal methods for labeling human adipose-derived stem cells (ASCs).

METHODSASCs were isolated by collagenase digestion and density gradient centrifugation, and their cell surface markers and ability to differentiate into the adipogenic, chondrogenic, and osteogenic lineages were examined in vitro. Three different cell labeling methods, namely 5 µl DiI, 10 µg/ml BrdU and 50 MOI adenovirus carrying GFP, were used for ASC labeling, and the labeling efficiency were compared at different time points and in different passages using fluorescent microscope.

RESULTSThe isolated ASCs were capable of differentiating into adipogenic, osteogenic, chondrogenic lineages with positive stem cell marker expression. At 48 h after DiI staining, 100% of the ASCs emitted red fluorescence in the cytoplasm with fluorescent-negative nuclei, but the fluorescence intensity declined quickly after cell passaging. With 10 µg/ml BrdU, 90% of the cells showed green fluorescence in the cell nuclei at 48 h after the labeling, but the positivity rate also decreased gradually after cell passaging. Cell labeling with GFP adenovirus showed more stable labeling efficiency, and green fluorescence was detected at 24 h after labeling, and even till 5 days later more than 90% of the ASCs remained positive without an obvious attenuation of the fluorescent intensity even after cell passaging.

CONCLUSIONAll the 3 techniques are applicable for labeling ASCs. Cell labeling with DiI and BrdU can be convenient and economic and well serve the purpose of short-term labeling. Adenovirus carrying GFP gene is the optimal choice for long-term ASC tracing.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; Adult ; Biomarkers ; Bromodeoxyuridine ; Cell Differentiation ; Cells, Cultured ; Female ; Green Fluorescent Proteins ; Humans ; Staining and Labeling ; methods ; Stem Cells ; cytology

OBJECTIVETo investigate the effect of fluvastatin on lysophosphatidylcholine (LPC)-induced ventricular arrhythmias and its mechanism.

METHODSTwenty male SD rats were randomly allocated into two equal groups, namely LPC treatment group and fluvastatin pretreatment group. Langendorff apparatus was used for cardiac perfusion ex vivo with 5 µmol/L LPC for 5 min followed by washing for 30 min in LPC treatment group, and in fluvastatin pretreatment group, a 30-min perfusion with 10 µmol/L fluvastatin was administered before LPC perfusion. The LPC-induced nonselective cation current (I(NSC)) in the ventricular myocytes was recorded using the whole-cell voltage-clamp method.

RESULTSFluvastatin significantly inhibited LPC-induced ventricular tachyarrhythmia/fibrillation and I(NSC). The small G-protein Rho inhibitor (C3) and Rho-kinase inhibitor (Y-27632) in the pipette solution also suppressed LPC-induced I(NSC).

CONCLUSIONFluvastatin offers cardiac protection against LPC by inhibiting LPC-induced I(NSC). LPC induces fatal arrhythmia via a Rho/Rho-kinase-mediated pathway.

Animals ; Arrhythmias, Cardiac ; chemically induced ; metabolism ; Drug Antagonism ; Fatty Acids, Monounsaturated ; pharmacology ; Indoles ; pharmacology ; Ion Channels ; drug effects ; Lysophosphatidylcholines ; adverse effects ; Male ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Sprague-Dawley ; rho-Associated Kinases ; metabolism

OBJECTIVETo study the value of basal antral follicle count (AFC) and age in predicting ovarian response and clinical outcome of in vitro fertilization-embryo transfer (IVF-ET).

METHODSA total of 1319 oocyte retrieval cycles in women with an AFC≤10 and complete IVF/ICSI cycles were analyzed retrospectively. According to the AFC, the patients were divided into groups A, B, and C with AFC≤4, of 5-7, and of 8-10, respectively, and each was further divided into <38 years old group and ≥38 years old group. The oocytes retrieved, ovarian response, implantation rate, cancellations, pregnancy, pregnancy loss, and live births were evaluated.

RESULTSAs the AFC increased, the total gonadotrophin (Gn) dose increased and the follicles aspirated and oocytes retrieved decreased significantly (P<0.001). Patients below 38 years of age had a lower total Gn dose and more follicles aspirated and oocytes retrieved than older patients. An AFC>7 and age≥38 years was associated with significantly lower total Gn dose, greater number of follicles aspirated and oocytes retrieved, and lower pregnancy rate than an AFC≤7 and age<38 years (P<0.05). Bivariate correlation and linear regression analysis identified AFC as the best single predictor of ovarian response in IVF. The pregnancy rate differed significantly between the 3 groups, and older patients (≥38 years) had higher early miscarriage rate.

CONCLUSIONAntral follicle count≤7 or age≥38 years old with AFC≤10 is the suitable threshold of diminished ovarian reserve in controlled ovarian stimulation for infertile women. Combination of AFC and age is the best predictor of ovarian response in IVF. Age has a better predictive value of pregnancy rate than AFC. AFC influences mainly the oocytes quantity, while age also affects oocyte quality.

Adult ; Age Factors ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Ovarian Follicle ; anatomy & histology ; cytology ; physiology ; Ovulation Induction ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies

OBJECTIVETo identify the genetic susceptibility to Kashin-Beck disease (KBD) and explore the interaction between low selenium (Se) and the susceptibility gene loci in KBD.

METHODSThe DNA samples collected from 23 KBD nuclear families were analyzed using PCR and GeneScan Analysis 3.7 and Genotyper3.7 software. The haplotype relative risk (HRR) and transmission disequilibrium test (TDT) were used to test the data of the genotypes. The serum selenium (Se) concentration was measured by atomic fluorescence spectrometry, and the interaction between low Se and the susceptibility loci was calculated using a binary logistic regression.

RESULTSIn the 23 nuclear families, the alleles of D2S151 (248 bp), D2S305 (320 bp), and D11S4094 (194 bp) showed significant correlation to KBD (P<0.05). Serum Se concentrations in the studied individuals was 0.037 µg/ml. No significant statistical interaction was observed between low Se exposure and the susceptibility loci (P>0.05).

CONCLUSIONThe polymorphisms in the STR loci D2S305, D2S151, and D11S4094 or the polymorphism loci near them might been related to KBD susceptibility. Low Se exposure shows no significant interaction with the susceptibility loci.

Adolescent ; Adult ; Alleles ; Child ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Kashin-Beck Disease ; blood ; etiology ; genetics ; Male ; Microsatellite Repeats ; Middle Aged ; Pedigree ; Selenium ; blood ; Young Adult

OBJECTIVETo evaluate the effect of short-term intensive therapy on blood glucose control, BETA-cell function, and blood lipid levels in newly diagnosed type 2 diabetic patients.

METHODSOut-patients with newly diagnosed type 2 diabetic patients were enrolled for intensive treatment with sulfonylureas and metformin for 12 weeks, and the therapeutic effect was evaluated.

RESULTSAfter the intensive treatment, FPG, 2 hPG, and HbA1c decreased significantly (P<0.01); HOMA-IR decreased and HOMA-B increased significantly (P<0.01), and TG, CHOL, LDL decreased significantly (P<0.01) after the treatment.

CONCLUSIONShort-term intensive treatment with glimepiride combined with metformin is safe and effective in newly diagnosed type 2 diabetic patients with HbA1c>9%.

Diabetes Mellitus, Type 2 ; drug therapy ; Drug Therapy, Combination ; Female ; Humans ; Male ; Metformin ; therapeutic use ; Sulfonylurea Compounds ; therapeutic use ; Treatment Outcome

OBJECTIVETo evaluate the effect of immunoadsorption therapy in patients with myasthenia gravis (MG) and explore the mechanism.

METHODSThis investigation involved 20 patients with MG treated with immunoadsorption combined with hormonal therapy and another 20 with only hormonal therapy, and 15 healthy subjects served as the control group. Enzyme-linked immunosorbent assay (ELISA) was used to measure the changes in serum tumor necrosis factor-α (TNF-α) and interleukin-18 (IL-18) after the treatments, and the therapeutic effect of the treatments was evaluated using clinical scores.

RESULTSThe clinical scores were significantly decreased after immunoadsorption therapy, showing a significant difference from that in the hormonal treatment group (P<0.05). The serum TNF-a and IL-18 levels were significantly higher in the two patient groups than in the control group (P<0.05), but in the former two groups, their levels were significantly lower in immunoadsorption therapy group (P<0.01).

CONCLUSIONImmunoadsorption therapy eliminates the inflammatory cytokines and free radicals as well as the circulating autoantibodies to improve the clinical symptoms of MG.

Adult ; Case-Control Studies ; Female ; Hormones ; therapeutic use ; Humans ; Immunosorbent Techniques ; Interleukin-18 ; blood ; Male ; Middle Aged ; Myasthenia Gravis ; blood ; therapy ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

Genus Wautersia is a Gram-negative bacterium that has rarely been associated with human infections abroad, and is firstly reported in home. We report a case of sepsis caused by Wautersia species, which was separated from blood and bone marrows.
Adult ; Cupriavidus ; isolation & purification ; Female ; Humans ; Sepsis ; microbiology