Calcineurin ; metabolism ; Cardiovascular System ; growth & development ; metabolism ; Humans ; NFATC Transcription Factors ; metabolism ; Signal Transduction

Objective To explore the effect of Mycobacterium tuberculosis antigen (Mtb-Ag) on neutrophils apoptosis.Methods The fresh isolated neutrophils from healthy adults blood were cultured with Mtb-Ag for 24 h,with or without pretreatment of nuclear factor -?B (NF-?B) inhibitor N-tosyl-L-phenylanyl chloromethyl ketone (TPCK) for 30 minutes.Annexin V staining and Flow cytometry were used to measure cell apoptosis of neutrophils.NF-?B DNA binding was measured by gelelectrophorestic mobility shift assay (EMSA) in neutrophils after incubated with Mtb-Ag for 0,1,2,4,6,24 hours.Results Comparing to the spontaneous apoptosis (55%?6%) of neutrophils after culture in vitro for 24 h,treatment of Mtb-Ag (1.125 mg/ml) decreased the cell apoptosis of neutrophils (32%?3%).The NF-?B shift bands were detected at 1 h in neutrophils after stimulated by Mtb-Ag,and reached maximum peak at 2 hours,and then returned to basal levels within 24 h.Pretreatment of TPCK inhibited the anti-apoptosis role of Mtb-Ag in neutrophils.Conclusion Mtb-Ag prevents neutrophils apoptosis and its inhibitory role concerns NF-?B pathway.

Objective To evaluate the clinical value of the oral contrast-enhanced three-dimensional spiral CT cholangiography to display better the intrahepatic and extrahepatic bile ducts. Methods Before spiral CT scanning, the preparation was similar to the conventional X-ray oral cholecystography. The collected parameters included 3-5 mm slice, 1 or 1.5 pitch, 120 kV, 220-250 mAs, and 1 mm reconstruction image. Volume rendering was used to construct the 3D bile duct images. Results In the 20 patients without serious biliary obstruction, oral three-dimensional CT cholangiography displayed the grade Ⅰ branch of hepatic ducts clearly and the displaying rate was 100%. The displaying of the grade Ⅱ branch of hepatic ducts was not good enough and the rate was 60%. The grade Ⅲ branch of hepatic ducts could not be displayed. Conclusion As the substitute and supplement of invasive ERCP, spiral CT 3D cholangiography is safe, simple, painless, and non-invasive. Furthermore, spiral CT 3D cholangiography can display the space structure of bile duct tree more clearly than ERCP. This technique is of great clinical value.

Objective To compare the clinical value of 2 oral contrast agents for magnetic resonance cholangiopancreatography(MRCP)and to investigate the safety,potency ratio of negative gastrointestinal contrast agents and to improve MRCP image quality.Methods Before MRCP,30 patients took oral ferric ammonium citrate(FAC)solution as group A and another 30 patients took Gd-DTPA dilution as group B.Then all patients underwent MRCP with T2-haste-fs-thick-slab sequence.Image assessment was done before and after oral taking of negative gastrointestinal contrast agents.Results After oral taking of negative gastrointestinal contrast agents,the interference of liquid in stomach and duodenum was suppressed and even effectively eliminated on MRCP.MRCP image quality was improved.Though the display of bile duct and pancreatic duct was of less image artifact,it reached the quality for diagnosis.No significant difference of MRCP image quality was found between those took FAC solution or Gd-DTPA solution.Conclusion FAC and Gd-DTPA both are effective negative gastrointestinal contrast agents.Gd-DTPA solution is of better function and higher quality-price ratio.

OBJECTIVE:To provide reference for the further improvement of the drug purchasing system in Chongqing city. METHODS:30 primacy medical and health institutions in Chongqing were selected to collect the data about cost changes and busi-ness development in above-mentioned institutions from 2009 to 2013,the purchasing price,use and supply of essential medicines were collected via Chongqing Municipal Health and Family Planning Commission to evaluate and analyze the effects and existing problems of drug purchasing system,and suggestions were put forward. RESULTS:Compared with 2009,the non-essential drugs’ prices fell an average of 28.00%in 2013,prices of 307 varieties in National Essential Medicine System(2009 edition)fell an aver-age of 32.97%,205 varieties in supplementary list of local prices fell an average of 43.78%. The proportion of sales amount(not including Chinese herbal medicine)in primacy medical and health institutions increased from 76.48% in 2010 to 92.07% in 2013, the average rate of essential medicines increased from 6.84% to 28.12%. Compared with 2009,the average drug cots and inpatient drug costs decreased 16.78% and 25.85% in 2013,the proportion of drugs decreased from 56.32% in 2009 to 49.70% in 2013. CONCLUSIONS:Generally speaking,the implementation of drug purchasing system in Chongqing is successful,but also exists some problems,such as imperfect pricing mechanism,difficult supply of low-cost drugs,slow rise of average outpatient costs in primacy medical and health institutions and irrational drug use. It is suggested to improve it from aspects of perfecting drug pricing mechanism,optimizing logistics and distribution,strengthening the role of the medical insurance institutions,regulating medication overuse and improving compensation policy of medical institutions.

Adolescent ; Adult ; Child ; Child, Preschool ; Electrocoagulation ; methods ; Female ; Humans ; Hypothermia, Induced ; Male ; Middle Aged ; Prospective Studies ; Tonsillectomy ; methods ; Young Adult

0.05).(4) Significant differences between CC and CIN Ⅰ for p16,CDH1,RASSF1A and TIMP3 genes(P

OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.

METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.

RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.

CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

To establish a method for the determination of astilbin, peoniflorin, rasmarinci acid, isofraxidin and liquiritin contained in Shaolin Xiaoyin tablets, in order to lay a foundation for designing late-stage dosage forms and clinical medication schemes. In this paper, efforts were made to establish a method for the determination of the blood concentration of the five components and study the in vivo pharmacokinetics in rats. The blood concentration was determined by HPLC. Phenomenex C18 column (4.6 mm x 250 mm, 5 microm) was adopted and eluted with methanol-acetonitrile-0.05% formic acid, the flow rate was 0.8 mL x min(-1), and the wavelength was 275 nm. The samples were processed by the solid phase extraction method. After oral administration of Shaoling Xiaoyin tablets, the rat bloods were collected at different time points to determine the blood concentrations. The experimental results showed that the baseline separation could be adopted for the five components, and astilbin, peoniflorin, rasmarinci acid, isofraxidin and liquiritin showed good linear relations within ranges of 2.48-248, 0.213 6-21.36, 0.531-53.1, 0.704-70.4, 0.253-25.3 mg x L(-1). All the five components could be absorbed in blood and excreted quickly. The method established in this paper is rapid and accurate, and could be used for in vivo analysis on preparations containing similar components. The main components in Shaoling Xiaoyin tablets could be absorbed and excreted quickly, and thus suitable to be made into sustained release tablets. Common preparations are required to be taken for 4-6 times a day.
Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; Cinnamates ; blood ; pharmacokinetics ; Coumarins ; administration & dosage ; blood ; pharmacokinetics ; Depsides ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Flavanones ; administration & dosage ; blood ; pharmacokinetics ; Flavonols ; administration & dosage ; blood ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Male ; Monoterpenes ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley