Objective To observe the Pin1 expression pattern in skin and to establish an inducible skin specific Pin1 overexpression mouse model. Methods The mouse Pin1 gene was cloned into modified vector pTRE2 with C?terminal Myc tag. The linearized pTRE2?Pin1 DNA was micro?injected into one?cell embryos followed by implantation into foster mice to produce TRE?Pin1 transgenic mice. Results TRE?Pin1 transgenic founder mice were successfully created. These mice were crossed with transgenic tool mice K14?rtTA to create epithelial specific double transgenic progenies. Pin1 gene was induced by incorporating doxycycline into drinking water of the mice. Pin1 protein overexpression in the skin was con?firmed by Western blot and immunohistochemistry. The endogenous Pin1 protein was predominantly expressed in epidermal cells in the skin. Conclusions The inducible skin specific Pin1 overexpression mouse model is successfully established which may serve as a useful model for further study of Pin1 functions in the skin.

Adaptation, Psychological ; Adolescent ; Adolescent Behavior ; Humans ; Mental Disorders ; etiology ; Parents ; Psychology, Adolescent ; Turner Syndrome ; psychology

MicroRNAs (miRNAs) are small (approximately 22 nt) RNAs that exhibit a diversity in sequence,structure,abudance,and expression profile. Bioinformatic approaches and direct cloning methods have identified 3518 miRNAs from various species. MiRNAs play a pivotal role in the regulation of genes involved in diverse processes such as development,differentiation,and cellular growth control. Recently,many viral-encoded miRNAs have been discovered, most from the herpesviruses viral family. Virus-encoded miRNAs seem to evolve rapidly and regulate both the viral life cycle and the interaction between viruses and their hosts. The detailed study on the virus-encoded microRNAs and the role they play in the progress of viral infection,replication and expression will be beneficial to understand viral molecular biology,and also provide new strategies for the prevention and treatment of virus.

Objective To prepare the monodisperse,colloidal gold nanoparticles (AuNPs) with controllable sizes (50 nm,65 nm,79 nm and 102 nm) for the qualitative detection of cardiac troponin Ⅰ (cTnⅠ) by immunochromatography assay,and to evaluate the effectiveness of the detection.Methods Four kinds of monodisperse citrate-stabilized AuNPs were prepared using small AuNPs as growth centers (seeds) by a seeded growth thermal aging protocol.As controls,two conventional AuNPs (20 nm,40 nm) were prepared by the traditional citrate-reduction method.The mouse monoclonal antibody against cTnⅠ labeled AuNPs were dropped on polyester mat to make AuNPs conjugate pad.The detection line and quality control line of immunochromatography assay kits for detection of cTnⅠ were coated by mouse anti human cTnⅠ monoclonal antibody paired with antibody in AuNPs and goat anti mouse polyclonal antiboy respectively.The six kinds of AuNPs were employed as color-labels in immunochromatography assay kits for detecting cTnⅠ,and the corresponding detection effects were evaluated in signal intensity,sensitivity,specificity and stability.The assay kit with the best performance was chosen and compared with the commercialized kits for the detection of cTnⅠ in clinical samples.Results Four kinds of monodisperse AuNPs with large sizes of 50 nm,65 nm,79 nm,102 nm respectively were successfully synthesized by the seeded growth thermal aging method.The signal strength of four kinds of kits produced by the four large-sized AuNPs was superior to the kits produced by 20 nm AuNPs in detecting cTnⅠ(all P＜0.01).The signal strength of the kits produced by 65nm AuNPs showed the best performance among the six kinds of AuNPs(all P＜0.01).The lowest detectable limit was 0.50 ng/ml.To compare the agreement of results from chemiluminescent immunoassay versus the results from kits produced by 65nm AuNPs,100 serum samples have been used for detecting cTnⅠ.Their positive coincidence rate was 97.30％ and negative coincidence rate was 100％,the sensitivity and signal strength of the kits produced by 65nm AuNPs was superior to similar products which produced by ABON and Bottests company(all P＜0.01).Conclusions Monodisperse,largesized,citrate-stabilized AuNPs are controllably prepared by a seeded growth-thermal aging method.The development of large-size AuNPs-based immunochromatography assay kits is feasible.65 nm AuNPs can be a suitable candidate for cTnⅠ immunochromatography assay kit.Our findings provides a new idea for the current immunochromatography assay kits which still adopt small-sized AuNPs as color labels.

To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 μm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 μL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 μL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½β) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
Administration, Oral ; Animals ; Biological Availability ; Coumaric Acids ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Iridoids ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; administration & dosage ; blood ; pharmacokinetics

The quality of a donor liver after cardiac death is closely associated with energy metabolism during preservation. Ex vivo mechanical perfusion has broad application prospects because this technique can help energy metabolism and repair ischemia injury of donors' livers. Some core issues are presented in this review in order to provide references for propelling secure application of liver transplantation based on donation after cardiac death.
Death ; Humans ; Liver ; Liver Transplantation ; Organ Preservation ; methods ; Perfusion ; methods ; Warm Ischemia ; adverse effects

Objective To investigate the effects of small interfering RNA(siRNA)on the expression of CD147 on peripheral blood T-lymphocytes from patients with psoriasis vulgaris,and its effect on the proliferation and activation of these cells.Methods Peripheral blood monouclear cells(PBMC)were obtained from 10 patients with psoriasis vulgaris,and T lymphocytes were isolated.CD147 siRNA was chemically synthesized,then electroporated into the peripheral T-lymphocytes.Untransfected cells,blank-transfected cells and unspecifically transfected cells served as the control.After 24-,48-,72-and 96-hour incubation,RT-PCR was used to detect the mRNA expression of CD147 in these cells.MTT assay and flow cytometry were utilized to assess the proliferation of these cellas,and the expression Of CD25 at 24,48,and 72 hours after the transfection.Results Compared with untransfected cells,the mRNA expression of CD147 declined significantly in CD147 siRNA-transfected cells at 24 hours(P＜0.05),reached to the minimum at 48 hours (P＜0.01):there was no significant difierence in the expression of CD147 between the two groups of cells at 96 hours after the transfection(P＞0.05).There was a decrease of cell proliferation level by(44.5±3.13)%,(50.7±3.5)%and(53.98±4.15)%in CD147 siRNA-transfected cells 24,48 and 72 hours following the transfection,respectively;the corresponding decrease in blank-transfected cells was (37.28±3.56)%,(33.73±3.29)%,and(28.80±1.49)%,respectively,and that in unspecifically transfected cells,(31.29±2.46)%,(36.1±2.62)%and(32.08±2.78)%,respectively.A significant decrease was observed in the proliferation of CD147 siRNA-transfected cells compared with that of blank-transfected cells and unspecifically transfected cells at these three time points(P＜0.05,0.01,0.01 respectively).The expression rate of CD25 at 24,48 and 72 hours was(47.23±3.65)%,(31.50±4.22)%and(23.05±4.15)%,respectively,on CD147 siRNA-transfected cells,and,(80.2±4.8)%,(81.6±3.35)%and(83.5±4.1)%,respectively,on untransfeeted cells;the differences between the two groups at the three time points were statistically significant(all P＜0.01).Conclusion CD147 is correlated with cell proliferation and activation of peripheral T lymphoeytes from patients with psoriasis vulgaris,and may serve as a new treatment target for psoriasis.

Objective To explore the effect of hepatocyte growth factor on peripheral nerve regeneration. Methods Sciatic nerve contusion injury was made by a custom-made clamp in Wistar rats,in which human hepatocyte growth factor expressed by adenoviral vector(Ad-HGF)was injected into the muscle around the injured nerve.The results of nerve regeneration were evaluated by sciatic nerve function index(SFI),muscle wet weight,neural electrophysiology and image analysis. Results Four weeks after sciatic nerve injury,the results of sciatic nerve function index(SFI),muscle wet weight,neural electrophysiology and image analysis showed better nerve regeneration in group injected with HGF than control group(P<0.05). Conclusion Hepatocyte growth factor can promote axon regeneration and functional recovery and is an effective neurotrophic factor for peripheral nerve regeneration after injury.

OBJECTIVETo investigate the clinicopathological features of testicular malignant Leydig cell tumor (TMLCT) and improve the non-invasive diagnosis of the disease.

METHODSWe retrospectively analyzed the clinicopathological data on a case of TMLCT, detected the circulating tumor cells (CTC) in the peripheral venous blood, and reviewed the related literature.

RESULTSThe patient, a 47-year-old male, underwent radical orchidoepididymectomy under general anesthesia. Postoperative pathology confirmed the lesion to be TMLCT, which was mainly composed of Leydig cells and suspected with vessel carcinoma embolus. Immunohistochemistry showed the tumor cells to be positive for α-inhibin, Ki67, CD30, vimentin, EMA, and PLAP, but negative for CK, CK7, S100, CD10, SMA, Des, AFP, hCG, CEA, CK19, CD117, Oct-4, LCA, CD20, Pax-5, CD3, and CD43. Two CTCs were detected in the peripheral venous blood. The patient received 3 courses of chemotherapy for retroperitoneal multiple lymph nodes metastasis post-operatively. Subsequent CT imaging manifested no obvious reduction of the retroperitoneal lymph nodes and consequently the patient again underwent retroperitoneal lymphadenectomy and cryoablation. At 8 months after treatment, CT examination revealed notably enlarged retroperitoneal lymph nodes with the right adrenal gland evidently invaded.

CONCLUSIONTMLCT is an extremely rare sex-gonad stromal tumor with high malignancy and poor prognosis, and CTCs may be used for its early diagnosis and prognostic prediction.

Biomarkers, Tumor ; metabolism ; Humans ; Immunohistochemistry ; Leydig Cell Tumor ; drug therapy ; pathology ; surgery ; Lymph Node Excision ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; Prognosis ; Retrospective Studies ; Sex Cord-Gonadal Stromal Tumors ; drug therapy ; pathology ; surgery ; Testicular Neoplasms ; drug therapy ; pathology ; surgery

Objective:To explore and discuss the differences of pregnancy contents and inform formats in in-formed consent form ( ICF) for the drug clinical trial between China and foreign countries. Methods:We collected Chinese and foreign ICFs for drug clinical trial that had been audited by the Ethics Committee of the third Xiangya Hospital for the past five years. Based on the relevant domestic and foreign law, we concluded the element stand-ards and inform formats about pregnancy inform. By analyzing the integrity of the whole elements, the inform rate of every element and the using rate of every inform format, we compared the differences of pregnancy contents and in-form formats between Chinese ICFs and foreign ICFs. Results:The total number of ICFs was 177 in this study, in-cluding 107 Chinese ICFs and 70 foreign ICFs. The integrity rate of pregnancy in Chinese ICFs was statistically lower than them in foreign ICFs (19% vs. 56%, P=0. 000). Compared with foreign ICFs, the low informed ele-ments were the study of the pregnancy risk (32% vs. 73%, P=0. 000), the pregnancy test during the following-up period (33% vs. 56%, P=0. 002) and the measurements for contraception (22% vs. 53%, P=0. 000). Conclusion:The integrity level of pregnancy content in Chinese ICFs was lower than that of the foreign ICFs. And the three elements including pregnancy risk study, pregnancy test during the following-up period and measure-ments for contraception was obviously defected. Pregnancy informing forms of informed consent in China was inferi-or to abroad.