Objective This study investigates feasibility of replacing urinary epidermal cells with foreskin epidermal cells to reconstruct engineered anterior urethra with acellular collagen matrix. Methods A cellular collagen matrices were generated from allogeneic rabbit bladder submucosa.In 6 rabbits,autologous foreskin epidermal cells isolated,in vitro expanded and labeled with Brdu before seeding onto a tubular collagen matrix of 2 cm long.In 12 male rabbits, urethral mucous defect was created and urethroplasty was performed with tubular collagen matrix seeded with epidermal cells (n=6) in experimental group or matrix without cells seeding (n=6) in control group.Urethrography was performed at 1,2,and 6 months postoperatively.Urethras grafts were harvested and analyzed grossly and histologically at the same time points. Results In control group, gross and urethrography demonstrated urethral stricture of repaired defects at all time points.Histology showed a single layer of epidermal cell with unorganized and crassitude muscle fiber bundles in submucosa layer.In contrast, urethrography showed a patency of repaired urethral defect with the maintenance of a wide urethral caliber in experimental group without sign of strictures at any time points. Histologically,the graft seeded with epidermal cell had a multi-layers epidermal cells by 1 month,which demonstrated the presence of a epidermal cells layer with a normal submucosa structure.The immunoflorescent staining of Brdu confirmed the presence of implanted epidermal cells.In addition, the implanted cells also express keratin when stained with anti-pancytokeratins AE1/AE3 antibody.At 2 month,histology showed relatively normal urethral architecture with vessel abundance in submucosa structure.The staining of Brdu showed decreased at implanted epidermal cells and epithelial phenotypes were confirmed immunocytochemically using pancytokeratins AE1/AE3.At 6 month,the graft structure show multi-layers epidermal cells without staining of Brdu. Conclusions Urethra reconstruction was better achieved with cell seeded acellular collagen matrix when compare with acellular matrices alone.Foreskin epidermal cells seems succeeded in replacing urethral epidermal cells for urethra reconstruction.

Objective To investigate the relationship between impaired fasting glucose(IFG) and the distribution of body fat in adolescents.Methods Stratified cluster sampling was used to select 3874 adolescents aged 13-18 years for this cress-sectional study.Measurements included height,weight,waist circumference,hip circumference and fasting plasma glucose (FPG).Family history of diabetes was determined by using a serf-administered questionnaire.Participants were divided into normal fasting glucose group(FPG＜5.6 mmol/L,n=3738) and impaired fasting glucose group (5.6 mmol/L≤FPG ＜ 7.0 mmoL/L,n=136) according to their FPG levels. Results (1) After adjusting for age and sex using covariance analysis,the impaired fasting glucose group showed increased levels of body mass index,weight circumference,waist/hip ratio,waist/height ratio,as compared to the normal fasting glucose group (P＜ 0.05).(2) After the age and gender were adjusted,body mass index,weight circumference,waist/hip ratio and waist/height ratio were positively correlated with FPG level (P＜0.05).Among the partial relation coefficients, that between waist/height ratio and FPG(r'=0.0925) was the highest (3) In multiple regression analyses,age (β=-0.102,P＜0.05),family history of diabetes (β=0.186,P＜0.05) and waist/height ratio (β=0.842,P＜0.05) were consistently associated with FPG.Conclusion Central obesity was an important predictor of IFG in adolescents.Waist/height ratio may be an useful index of central obesity and an important predictor of IFG in adolescents.

Objective To study the effects of tanshinone on the cultured immortalized human sebocytes and explore the mechanism of action of tanshinone in the treatment of acne. Methods MTT assay was applied to determine the effects of cryptotanshinone and tanshinone Ⅱ? at different concentrations on the proliferation of SZ95 sebocytes in 24 h, 48 h and 72 h after incubation. Lipid contents labeled with Nile red in SZ95 cells were analyzed by flow cytometry technique, and the expression of androgen receptor (AR) mRNA of SZ95 cells were detected by RT-PCR. Results Both cryptotanshinone and tanshinoneⅡ? inhibited the proliferation of SZ95 cells in a dose- and time-dependent mode, with the 50% inhibition concentration (IC50) of 7.473 ?mol/L (in 48 h) and 2.146?mol/L (in 72 h) for cryptotanshinone and 6.021 ?mol/L (in 48 h) and 2.25 ?mol/L (in 72 h) for tanshinone Ⅱ?. Additionally, as compared with the control group, the lipid content of SZ95 cells exposed to tanshinone Ⅱ? at 0.125?mol/L was decreased in 48h (P

An efficient modified route based on the targeting mechanism of antibacterial fluoroquinolones for the shift from the antibacterial activity to the antitumor one was further developed. Using a fused heterocyclic ring, s-triazolothiadiazine as a carboxyl bioisostere of ciprofloxacin, the title compounds, 1-cyclopropyl-6-fluoro-7-piperazin-1-yl-3-(6-substituted-phenyl-7H-[1, 2, 4]triazolo[3, 4-b][1, 3, 4]thiadiazin-3-yl)-quinolin-4(1H)-ones (5a-5e) and their corresponding N-acetyl products (6a-6e), were designed and synthesized, separately. Meaningfully, a ring-contraction of fused six-membered thiadiazine occurred by a sulfur extrusion reaction gave new tri-acetylated fused heterocycles related to pyrazolo[5, 1-c][1, 2, 4] triazoles (7a-7e). The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated for the synthesized fifteen heterocycles compared to parent ciprofloxacin by methylthiazole trazolium (MTT) assay. Interestingly, the results displayed that fifteen fused heterocyclic compounds showed more significant growth inhibitory activity (IC50 < 25.0 micromo x L(-1)) than that of parent ciprofloxacin (IC50 > 150.0 micromol x L(-1)), and the active order decreased from 7a-7e to 5a-5e to 6a-6e, respective.
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; CHO Cells ; Cell Line, Tumor ; Ciprofloxacin ; pharmacology ; Cricetinae ; Cricetulus ; Fluoroquinolones ; chemical synthesis ; chemistry ; pharmacology ; HL-60 Cells ; Humans ; Inhibitory Concentration 50 ; Leukemia L1210 ; pathology ; Mice ; Structure-Activity Relationship ; Thiadiazines ; chemical synthesis ; chemistry ; pharmacology ; Triazoles ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETo study the role of the PI3K/AKT signaling pathway in the diallyl disulfide (DADS)-induced apoptosis of K562 cells.

METHODSK562 cells in the logarithmic growth phase were treated with 10, 20, 40, or 80 mg/L DADS for 48 hours, then fixed and stained with acridine orange/ethidium bromide (AO/EB), and examined for cellular morphological changes under an inverted microscope. Annexin V-FITC/PI staining was used for determining the apoptotic rates, and Western blot for measuring the expression of AKT, p-AKT, and Caspase-3. Two control groups, blank and solvent, were used as references.

RESULTSK562 cells treated with DADS for 48 hours exhibited the characteristic morphological features of apoptosis including cell shrinkage, irregular cell shape, and membrane blebbing. AO/EB staining results demonstrated that the number of apoptotic cells with cell shrinkage, pyknotic or bead-like nuclei, chromatin condensation, and orange staining increased with the increasing DADS concentration, and 40 mg/L DADS had the most significant effect. The apoptotic rates of cells treated with 10, 20, 40, and 80 mg/L DADS were all significantly higher than those in the control groups (P<0.05). There were no significant differences in AKT protein expression between the K562 cells treated with different concentrations of DADS; the p-AKT protein expression decreased with the increasing DADS concentration, while the Caspases-3 protein expression increased with the increasing DADS concentration (P<0.05).

CONCLUSIONSDADS induces the apoptosis of K562 cells, probably through inhibiting the protein expression in the PI3K/AKT signaling pathway.

Allyl Compounds ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Disulfides ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; K562 Cells ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Signal Transduction ; physiology

To explore an efficient strategy for further development of anticancer fluoroquinolone candidates derived from ciprofloxacin, a heterocyclic ring as the bioisosteric replacement of C3 carboxyl group led to a key intermediate, oxadiazole thiol (5), which was further modified to the bis-oxadiazole methylsulfides (7a-7h) and the corresponding dimethylpiperazinium iodides (8a-8h), respectively. Structures were characterized by elemental analysis and spectra data, and their anticancer activities in vitro against CHO, HL60 and L1210 cancer cells were also evaluated by MTT assay. The preliminary results show that piperazinium compounds (8) possess more potent activity than that of corresponding free bases (7).
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; CHO Cells ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ciprofloxacin ; chemistry ; Cricetinae ; Cricetulus ; Drug Design ; HL-60 Cells ; Humans ; Inhibitory Concentration 50 ; Leukemia L1210 ; Molecular Structure ; Oxadiazoles ; chemical synthesis ; chemistry ; pharmacology ; Piperazines ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETo establish an animal model to replicate the blunt impact brain injury in forensic medicine.

METHODSTwenty-four New Zealand white rabbits were randomly divided into control group (n equal to 4), minor injury group (n equal to 10) and severe injury group (n equal to 10). Based on the BIM-II Horizontal Bio-impact Machine, self-designed iron bar was used to produce blunt brain injury. Two rabbits from each injury group were randomly selected to monitor the change of intracranial pressure (ICP) during the impacting process by pressure microsensors. Six hours after injury, all the rabbits were dissected to observe the injury morphology and underwent routine pathological examination.

RESULTSVarying degrees of nervous system positive signs were observed in all the injured rabbits. Within 6 hours, the mortality rate was 1/10 in the minor injury group and 6/10 in the severe injury group. Morphological changes consisted of different levels of scalp hematoma, skull fracture, epidural hematoma, subdural hematoma, subarachnoid hemo- rrhage and brain injury. At the moment of hitting, the ICP was greater in severe injury group than in mild injury group; and within the same group, the impact side showed positive pressure while the opposite side showed negative pressure.

CONCLUSIONSUnder the rigidly-controlled experimental condition, this animal model has a good reproducibility and stable results. Meanwhile, it is able to simulate the morphology of iron strike-induced injury, thus can be used to study the mechanism of blunt head injury in forensic medicine.

Animals ; Brain Injuries ; Head Injuries, Closed ; Intracranial Pressure ; Rabbits ; Reproducibility of Results ; Wounds, Nonpenetrating

To investigate the urination-reducing effect and mechanism of Zhuangyao Jianshen Wan (ZYJCW). In this study, SI rats were subcutaneously injected with 150 mg · kg(-1) dose of D-galactose to prepare the sub-acute aging model and randomly divided into the model group, the Suoquan Wan group (1.17 g · kg(-1) · d(-1)), and ZYJCW high, medium and low dose groups (2.39, 1.20, 0.60 g · kg(-1) · d(-1)) , with normal rats in the blank group. They were continuously administered with drugs for eight weeks. The metabolic cage method was adopted to measure the 24 h urine volume and 5 h water load urine volume in rats. The automatic biochemistry analyzer was adopted to detect urine concentrations of Na+, Cl-, K+. The ELISA method was used to determine serum aldosterone (ALD) and antidiuretic hormone (ADH). The changes in P2X1 and P2X3 mRNA expressions in bladder tissues of rats were detected by RT-PCR. According to the results, both ZYJCW high and medium dose groups showed significant down-regulations in 24 h urine volume and 5 h water load urine volume in (P <0.05, P <0.01), declines in Na+ and Cl- concentrations in urine (P <0.01), notable rises in plasma ALD and ADH contents (P <0.05, P <0.01) and remarkable down-regulations in the P2X1 and P2X3 mRNA expressions in bladder tissues (P <0.01). The ZYJCW low dose group revealed obvious reductions in Na+ and Cl- concentrations in urine (P <0.01). The results indicated that ZYJCW may show the urination-reducing effect by down-regulating the P2X1 and P2X3 mRNA expressions in bladder tissues of rats with diuresis caused by kidney deficiency.
Aging ; physiology ; Animals ; Diuresis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; Kidney Diseases ; drug therapy ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X1 ; genetics ; Receptors, Purinergic P2X3 ; genetics ; Urinary Bladder ; metabolism

OBJECTIVETo study the effects of four kinds of traditional Chinese herbal compound, Sini powder, Banxiahoupu soup, Ganmaidazao soup and Guizhigancaolonggumuli soup, on the experimental acute stress behaviors and the hypothalamus-pituitary-adrenal gland axis.

METHODRats were divide into 7 groups: normal group, model 1 and 2 groups, Sini powder group, Banxiahoupu soup group, Ganmaidazao soup group, and Guizhigancaolonggumuli soup group. The behaviors of the rats and their CRH of the hypothalamus, ACTH of plasma and CORT of serum (by the means of radio-immunity) were detected.

RESULTSini powder could reduce the crossing times of open-field test( P < 0.01) and the contents of CRH and CORT (P < 0.01, P < 0.05); Banxiahoupu soup could extent the still time of tail-hanging test ( P < 0.05) and the content of CRH (P < 0.05); Ganmaidazao soup could prolong the still time (P < 0.05) and reduce the struggle times (P < 0.01) of tail-hanging test and reduce the contents of CRH, ACTH and CORT (P < 0.01, P < 0.05); Guizhigancaolonggumuli soup could reduce the crossing times of open-field test (P < 0.01) , extent the still time (P < 0.05) and reduce the struggle times ( P < 0.01) of tail-hanging test of tail-hanging test, also reduce the contents of CRH, ACTH and CORT (P < 0.01, P < 0.05).

CONCLUSIONThe four kinds of traditional Chinese herbal compound can increase the ability of anti stress by affect the different taches of the hypothalamus-pituitary-adrenal gland axle and change the stress behaviors, which are based on their respective functions of regulating Qi, dispersing phlegm, tonifying Qi and warming Yang.

Adrenocorticotropic Hormone ; blood ; Animals ; Behavior, Animal ; drug effects ; Corticosterone ; blood ; Corticotropin-Releasing Hormone ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Hypothalamo-Hypophyseal System ; drug effects ; Hypothalamus ; metabolism ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Medicine, Chinese Traditional ; Ostreidae ; chemistry ; Plants, Medicinal ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; physiopathology

Objective To explore the best cut-point of waist-to-heisht ratio (WHtR) for identifying metabolic syndrome(MS)in adolescents.Methods A cross-sectional study was conducted in 4 507 adolescents aged 13-18 years in Qinhuangdao.Receiver operating characteristic (ROC) curve analysis was employed to determine the optimal WHtR cut-point for detecting MS.Results The prevalences of MS were higher among higher WHtR in both boys and girls.Odds ratios for MS significantly increased from the WHtR≥0.45 in boys(OR =13.85,95%CI 4.08-46.97)and in girls (OR=12.42,95%CI 2.62-58.96,P<0.01).The ROC curve analysis showed that the optimal WHtR cut-point was 0.45 in boys with the sensitivity of 89.4%and specificity of 81.4%.and in girls with sensitivity of 90.3%and specificity of 86.5%.Conclusion There is a significant correlation between WHtR and MS.Optimal WHtR cut-point for predicting MS is 0.45.