Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) ％ and (64.93±2.18) ％,respectively.Their radiochemical purity was greater than 90％.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P＜0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P＞0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.

Objective:To optimize the formula of 5-aminosalicylic acid enema in situ gel. Methods:5-Aminosalicylic acid ene-mas in situ gel was prepared using a cold dissolution method with carbomer as the gel matrix and xanthan gum as the thickener. A 32 full factorial design was used to investigate the effects of the concentrations of carbomer and xanthan gum on the viscosity before and af-ter the gelling, duration of inversion tube and sedimentation rate. Response surface methodology was used to optimize the formula. Re-sults:The quantitative relationships between the two factors and the four evaluation indices were obtained. The optimum formula was as follows:the concentration of carbopol and xanthan gum in the enema was 0. 7% and 0. 15%, respectively. The viscosity before and af-ter the gelling was 500-1 000 mPa·s and 2 200-2 700 mPa·s, respectively. The duration of inversion tube test was 40-80 min and the sedimentation rate was more than 98. 5%. Conclusion:The multi-objective simultaneous optimization of the formula of 5-aminosal-icylic acid enema in situ gel is accomplished by factorial design and response surface methodology.

Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

Objective To explore the protective effects of dipeptidyl peptidase-4 inhibitor (DPP-4I) on AD-like neurodegenerative changes and its mechanism. Methods The human neuroblastoma cell line SH-SY5Y on the logarithmic phase was divided into six groups:control group (CON group, treated with PBS contained 1‰DMSO for 12 h), wortmannin intervention group (W group, treated with 0.03 μmol/L wortmannin for 12 h), DPP-4I intervention group (DPP-4I group, treated with 10μmol/L DPP-4I for 12 h), both DPP-4I and wortmannin intervention group (DPP-4I+W group, pre-treated with 10 μmol/L DPP-4I for 2 h, then 0.03 μmol/L wortmannin for 12 h), DPP-4I, wortmannin and Ex9-39 intervention group (DPP-4I+W+Ex9-39 group, pre-treated with 10μmol/L Ex9-39 for 2 h, then 10μmol/L DPP-4I for 2 h followed by 0.03μmol/L wortmannin for 12 h), and Ex9-39 intervention group (Ex9-39 group, treated with 10μmol/L Ex9-39 for 12 h). MTT assay was used to detect the cell vitality. Western blot assay was used to detect the level of total tau protein (tau-5) and phosphorylated tau at different sites (pSpS199/202, pT231 and pS396), the level of phosphorylated neurofilaments (NF-H, NF-M) and phosphorylation of critical enzyme in PI3K/Akt/GSK-3β signaling pathway. Results (1) The cell vitality decreased, the levels of pSpS199/202, pT231, pS396 and NF-H/M increased significantly in W group than those in CON group. However, comparing with CON group, the above mentioned parameters reversed in DPP-4I group. Comparing with W group, the cell vitality increased and phosphorylated levels of above mentioned indices were decreased in DPP-4I+W group. (2) The cell vitality showed a decline trend while the levels of phosphorylation tau at three different sites and NF-H/M were higher in Ex9-39 group than those in CON group. Comparing with DPP-4I+W group, the results of the phosphorylated levels showed the same changes in DPP-4I+W+Ex9-39 group. (3) Comparing with CON group, the expression levels of phosphorylated PI3K, Akt and GSK3β increased significantly in DPP-4I group, while those decreased in W group. Additionally, the expression levels of phosphorylated PI3K, Akt and GSK3β were significantly increased in DPP-4I+W group than those in W group. Conclusion DPP-4I can enhance the level of GLP-1 and activate PI3K/Akt/GSK-3βinsulin signaling pathway to improve the hyperphosphorylated tau and NFs induced by wortmannin, and to protect AD-like neurodegeneration.

Objective The purpose of this study is to construct eukaryotic gene vector of herpes simplex virus type 1 thymidine kinase(HSV1-tk)and to observe the expression of HSV1-tk in lung adenocarcinoma AGZY cell line.Methods The full length HSV1-tk gene was amplified by PCR from plasmid pHSV 106 and was inserted into pMD18-T.The recombinant plasmid was recombined with eukaryotic vector plRES 2-EGFP u-sing gene recombinant technique .HSV1 -tk was transfected into adenocarcinoma AGZY cell line with Lipo-fectamineTM 2 000.Fluorescence microscopy was used to detect the transfection and expression of HSV 1-tk.RT-PCR was used to detect the mRNA levels of HSV 1-tk.The cell proliferation was measured by MTT assay .Re-sults A length of 1 130 bp gene sequence was obtained by PCR .The expressions of HSV 1-tk at mRNA and protein levels were displayed by RT -PCR and Western blot .MTT analysis showed that there were no significant changes cell survival on after transfection .Conclusion The eukaryotic expression vector of HSV 1 -tk report gene is successfully constructed and HSV 1-tk is effectively expressed in transfected AGZY cells .

Objective To investigate the association between Has‐miRNA‐149 polymorphism and susceptibility of digestive tract cancer .Methods A systematic search of Pubmed ,EMBASE and Web of Science ,CNKI ,Wanfang was conducted for relevant studies .The odds ratios(OR) and 95% confidence intervals(95% CI) calculated by Stata12 .0 software were used to assess the asso‐ciation between Has‐miRNA‐149 polymorphism and risk of digestive tract cancer .The Q test ,I2 value and funnel plot were conduc‐ted to examine heterogeneity and publication bias ,respectively .Results A total of thirteen eligible studies containing 4 424 cases and 5 290 controls were included in this Meta analysis .There were significant relationships between Hsa‐miRNA‐149 polymorphism and susceptibility of digestive tract cancer (Domina model CT+CC vs .TT ,OR=0 .915 ,95% CI:0 .840-0 .996 ,P=0 .040 ;Hetero‐zygous model CT vs .TT ,OR=0 .880 ,95% CI:0 .803 -0 .965 ,P=0 .007) .In the stratified analysis ,it was found that the risk of colorectal cancer decreased in patients carrying CC/CT genotypes(OR=0 .834 ,95% CI:0 .715-0 .972 ,P=0 .021) ,especially in A‐sians (CT+CC vs .TT ,OR=0 .894 ,95% CI:0 .818-0 .977 ,P=0 .013) ,while that was not found in the Caucasoids .Conclusion Has‐miRNA‐149 polymorphism is associated with the susceptibility of digestive tract cancer .The CC/CT genotypes may decrease the risk of digestive tract cancer ,especially in Asians .

AIM: To explore the localization and expression of transfo rming grow th factor-? 1,2 (TGF-? 1,2 ) and alpha-smooth muscle actin (?- ASMA) in fetal a nd adult skins. METHODS: Skins of 15 cases of fetuses with different gestational ages and 5 cases of adults were taken, embedded with paraffin wax, and sectione d. Immunohistochemistry method and pathological method were used to detect the e xpression intensity and distribution of TGF-? 1,2 and ?-ASMA. RESULTS: Positive immunohistochemical signals of TGF-? 1,2 and ?-A SMA were found in fetal and adult skins. In skins derived from young fet us, the positive signals of these three proteins were very weak. Along with the incr ement in gestational age, the positive cellular rates of TGF-? 1,2 and ?- ASMA were elevated pro gressively. In elder fetal and adult skins, TGF-? 1,2 were mostly distributed i n epidermal cells, endothelial cells and some fibroblasts, while ?-ASMA was mainly located in myofibroblasts and sweat gland epithelial cells. CONCLUSION: The endo genous TGF-? 1,2 might be involved in the cutaneous development at embryoni c stage, in the cutaneous structure maintenance at adult stage, and in the wound healing af ter injury.

Male infertility microsurgery represents the fastest growing sub-specialty in urology and clinical andrology over the past two decades. The importance of microsurgery for male infertility has risen as a part of the urologist's armamentarium in the medical and surgical management of male infertility. Despite the advances in male infertility microsurgery in China, the lack of standardized and well-organized training programs for male infertility microsurgery remains a serious problem affecting its development. In this article, Zhao and Peng have shared their experience with the learning curve of male infertility microsurgery at the Center for Male Reproductive Medicine and Microsurgery, Weill Medical College of Cornell University, which centers on how to pay attention to the details and basic principles of microsurgery. Male infertility microsurgery is physically, technically and mentally challenging, and must be first learned in the laboratory. Clinical success depends heavily upon appropriate training in a microsurgical laboratory. Good training can significantly reduce operation time and surgical errors as well as improve the quality of outcomes.
Andrology ; education ; Humans ; Infertility, Male ; surgery ; Male ; Microsurgery ; education

Objective In order to know the effect of heating infusion for premature infants to pro-mote their restoration of physiological function. Methods Divided 160 premature infants who need infu-sion into the experimental group and the control group randomly, there were 80 cases in each group. Normal temperature infusion was used in the control group, while heating infusion was used in the experimental group, compared the body temperature, pluse, respiration, blood pressure, oxygen saturation and the blood ghcose between the two groups. Results There were significant difference about the indexes which had mentioned above between the two groups. Conclusions Heating infusion can effective promote the reha-bilitation of physiological function of premature infants.

Ergosta-4,6,8(14),22-tetraen-3-one (ergone) is one of main components in many medicinal fungi. Ergone has been reported to possess the activities of diuresis, cytotoxicity, antitumor, immunosuppression, as well as treatment of chronic kidney disease. According to reported literatures, an overview of spectroscopy characteristics, content determination, pharmacological activity and pharmacokinetics, etc. for ergone is presented in this review. Furthermore, the present review can provide a certain reference value for the further study and development of ergone.
Animals ; Cholestenones ; chemistry ; pharmacokinetics ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; pharmacology ; Humans