Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

Objective To investigate the effect of cnrcumin on the expression of pituitary tumor transforming gene (PTTG) in glioma C6 cell line. Methods Glioma C6 cells were allocated into the control group and 3 cttrcnmin treatment groups with curcumin treatment at 10, 20 and 30 μmol/L for 24 h. After the treatments, PTTG mRNA and protein expressions in the cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. Results The expression levels of PTTG mRNA in the 4 groups, showed significant differences between any two groups (P<0.01). Significant differences were also found in PTTG protein expressions between the 4 groups of C6 cells after the treatment (P<0.01). Conclusions Cureumin can down-regulate PTTG expression at both the mRNA and protein levels in glioma C6 cells in a dose-dependent manner.

Clinical Laboratory Techniques ; Hepatitis, Autoimmune ; diagnosis ; Humans

OBJECTIVETo investigate the feasibility of in vivo magnetic resonance imaging (MRI) tracking of transplanted adipose-derived stem cells (ADSCs) labeled with superparamagnetic iron oxide (SPIO) in rat heart.

METHODSADSCs were labeled with poly-L-lysine (PLL)-SPIO complexes. Intracellular iron uptake was identified by Prussian blue stain and transmission electromicroscopy. Trypan blue staining was used to test the viability of the labeled cells. In vitro MRI of labeled cells was performed. SPIO-labeled ADSCs were transplanted into normal rat hearts and were in vivo imaged with MRI. Image findings on MRI were correlated with histological findings of the rat hearts.

RESULTSThe labeling efficacy of ADSCs with PLL-SPIO was nearly 100%. Light microscopy revealed the SPIO particles were located in the cytoplasm of the ADSCs by Prussian blue staining. Transmission electromicroscopy revealed that the SPIO particles were located in the endosomes in the cytoplasm. There was no significantly deference in viability between labeled and unlabeled groups demonstrated by Trypan blue test (P > 0.05). MRI showed signal loss in gel mixed with labeled cells as compared with the unlabeled cells group and blank group. Signal void on rat hearts were demonstrated on MRI and were well correlated with histological findings where Prussian-blue-stain positive cells presented.

CONCLUSIONMRI can be used to in vivo track the transplanted ADSCs labeled with SPIO into rat hearts and facilitate to understand the conditions of the labeled cells in the transplanted areas.

Adipocytes ; cytology ; Animals ; Cell Differentiation ; Contrast Media ; administration & dosage ; Dextrans ; administration & dosage ; Feasibility Studies ; Image Enhancement ; methods ; Magnetic Resonance Imaging ; methods ; Magnetite Nanoparticles ; administration & dosage ; Male ; Myocardium ; cytology ; pathology ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; methods ; Stem Cells ; cytology

OBJECTIVETo investigate the alteration of apparent diffusion coefficient (ADC) in diffusion-weighted MR imaging (MR-DWI) of liver fibrosis and its pathological basis in rabbits.

METHODSFive rabbits in the control group and 22 with experimental liver fibrosis induced by transperitoneal injection of carbon tetrachloride (CCl4) were examined with MR-DWI. Diffusion-weighted SE EPI sequence with a relatively high b factor (b=600 s/mm2) was used to measure the ADC. The mean values of ADC were compared among the rabbits in different stages of liver fibrosis and analyzed in relation to the pathological findings.

RESULTSThe mean ADC value decreased significantly with increased severity of liver fibrosis (P<0.05). Pathologically, the amount and extension of fibrotic matrix increased, and the hepatic necroinflammation worsened with the progression of the liver fibrosis.

CONCLUSIONThe ADC value decreases with the progression of liver fibrosis possibly as the result of water diffusion limitation due to increased fibrous tissue in the liver and abnormal water diffusion within the intracellular and extracellular spaces.

Animals ; Carbon Tetrachloride ; toxicity ; Liver Cirrhosis ; chemically induced ; diagnosis ; pathology ; Magnetic Resonance Imaging ; methods ; Male ; Rabbits ; Random Allocation

Objective To explore the effectiveness of head nurse responsibility in night shift in department of pediatrics.Methods "Head nurse responsibility" was implemented in night shift since July,2011.One head nurse was entirely responsible for the treatment and nursing in patients' night shift,and ensured that highly qualified and experienced nurses were available at clinic.Self-designed questionnaire on satisfaction was used to investigate 120 patients,30 doctors and 30 nurses before head nurse responsibility from July to December 2010 and after head nurse responsibility from July to December 2011.Conventional nursing was carried out before the implementation of head nurse responsibility,and head nurse was added after the implementation.Scores in night shift nursing quality,changes and observation of patient' s condition,rescuer operating skills,health education,nurses' responsibility,guide ability,cooperation ability,attitude,overall evaluation of night shift quality,and satisfaction of patients,doctors and nurses were compare between two groups.Results The score of comprehensive night shift nursing quality was obviously better after head nurse responsibility than before [(93.45 ± 4.72) vs (86.93 ± 4.69)],and the difference was statistically significant (t =1.989,P ＜ 0.01).The score of satisfaction of parents of patients in overall evaluation of night shift quality were higher after head nurse responsibility than before [(4.70 ± 0.31) vs (3.73 ± 0.69)],and the difference was statistically significant (t =2.01,P ＜ 0.01).Satisfaction of patients,nurses and doctors improved respectively from 88.24％ to 98.51％,88.47％ to 99.43％ and 86.27％ to 98.56％ after the implementation,and the differences were statistically significant (x2 =9.64,4.32,4.28,respectively; P ＜ 0.05).Conclusions "Head nurse responsibility" can guarantee the safety of night shift nursing in department of pediatrics,as well as improve the nursing quality and patients' satisfaction,which is beneficial to improve the quality of night shift nursing in high-quality nursing service of the children' s ward.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; HIV Infections ; physiopathology ; Humans ; Liver Diseases ; physiopathology ; virology ; Male ; Middle Aged ; Risk Factors ; Young Adult

Objective To find out the status of brick-tea type fluorosis in the epidemic areas.Methods Based on "Scheme for Epidemiological Brick-tea Type Fluorosis in Sichuan Province",ten counties were selected in Sichuan brick-tea areas and ten towns were selected in every county,then the epidemicologic survey was performed in children of 8～12 year-old and adults aged above 20 years old.Results 5044 children and 4053 adults were selected from brick-tea areas.The rates of dental fluorosis in children and adults were 55.69%(2809/5044)and 60.41%(4053/6709)respectively.The dental fluorosis was mainly of mild damage.The skeletal fluorosis found in X-ray film was 44.64%(167/1241)and in clinical examination,38.94%(3883/9973).The levels of urine fluoride in children and adults were 1.88 and 2.78 mg/L.The level of urine fluoride was not differenet among children of different age,but in adults it was higher in the elder than the younger.The level of fluoride in urine was related to the severeness of skeletal fluorosis(r=0.74).The detective rates of skeletal fluorosis in agricuIture,pasturing,and agriculture-pasturing areaswere 31.70%(1369/4318),50.04%(1228/2454),and 40.17%(1286/3201),respectively.The X-ray detecting rates of skeletal fluorosis in men and wonlen were 49.57%(229/462)and 41.72%(325/779) respectively(χ2=11.72,P＜0.05).Conclusion The prevalence of brick-tea type fluorosis is very serious in the regions studied.

OBJECTIVETo compare the therapeutic effect of percutaneous cervical disc nucleoplasty (PCN group) and percutaneous cervical discectomy (PCD group) for the treatment of cervical disc herniation.

METHODSA retrospective study was carried out from July of 2002 to December of 2004, and there were 80 cervical disc herniation cases who were operated by PCN (42 cases) or PCD (38 cases). The time of operation, clinical result and the stability of cervical spine after operation were evaluated and compared between 2 groups.

RESULTSAll cases had been followed up from 6 months to 26 months, average (12 +/- 5) months on the PCN group and (12 +/- 4) months on the PCD group, and there was no significant difference on 2 groups (t = -0.06, P = 0.953). All cases had been successfully operated. There was significant difference in the operation time between 2 groups (t = -21.70, P = 0.000). There was significant difference in the pre- and post-operation scores of each group (PCN group: t = 14.05, P = 0.000; PCD group: t = -14.79, P = 0.000). There was no significant difference in 2 groups of the clinical outcomes (z = -0.377, P = 0.706, > 0.05). There was no instability of cervical spine cases in 2 groups after operation (P > 0.05), and the cervical spine stability was no significant difference in pre- and-operation in each group.

CONCLUSIONSPCN and PCD for the treatment of cervical disc herniation achieves good outcomes and no difference on the stability of cervical spine. PCN and PCD is a safe, minimally invasive, short time of operation, less traumatic operation and excellent clinical outcome.

Adult ; Catheter Ablation ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Diskectomy, Percutaneous ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Collection and rating of evidence,survey of patients' preference and value and economic evaluation of the Practice Guideline for Therapeutic Drug Monitoring of Voriconazole have been finished.The Guideline Consensus Panel reached consensus of the recommendations by 3 rounds Delphi and Grading of Recommendations Assessment,Development and Evaluation grid method.Consensus of 38 recommendations was reached,with a consensus percentage of 50.7％.Twenty seven recommendations were finalized and approved by the Guideline Steering Committee.Draft recommendations were generated.