Adolescent ; Biopsy ; Female ; Hepatocytes ; diagnostic imaging ; pathology ; Humans ; Jaundice, Chronic Idiopathic ; pathology ; Sibling Relations ; Ultrasonography

Adolescent ; Adult ; Female ; Humans ; Hydroa Vacciniforme ; Lymphoma, T-Cell, Cutaneous ; Male ; Skin Neoplasms

Aim To study the effect of CSE ( cigarette smoke extract ) on the single-molecule interactional force between thrombomodulin and thrombin by live-cell single-molecule force spectroscopy. Methods CSE was prepared by a previously reported method. The plasmid of TM-GFP was constructed and transfect-ed in COS-7 cells. The expression of TM-GFP was de-tected by fluorescence microscopy and laser scanning confocal microscopy. The transfected COS-7 cells were grouped ( 1 ) GFP -thrombin group ( 2 ) TM-thrombin group ( 3 ) CSE-TM-thrombin group ( 4 ) CSE- GFP-thrombin group. Force measurements with the thrombin modified AFM tips on the living cell surface were car-ried out on PicoSPM II with a Pico-Scan 3000 control-ler and a larger scanner. The force curves measured in living cells were recorded by PicoScan 5 software and analyzed by MATLAB R2009aMetlab. Results The single-molecule binding force of thrombomodulin and thrombin ( TM-Thr ) was determined ( 60. 90 ± 0. 82 ) pN. The binding probability for TM-Thr was about (22. 58 ± 3. 95)%. Antibody blocking binding proba-bility for TM-Thr was ( 2. 58 ± 2. 0 )%. The binding probabilities for GFP-Thr group, CSE-TM-Thr group and CSE-GFP-Thr group were significantly decreased compared with TM-Thr group ( P<0. 05 ) . The mean value of the most probable single molecular interaction force of thrombin/TM-ECD was determined as ( 45. 30 ± 1. 37 ) pN, the binding probability of thrombin and TM-ECD was ( 23. 25 ± 7. 02 )%. When the binding was blocked with the TM-MAb solution, the binding probability decreased to ( 4. 64 ± 2. 31 )%. The bind-ing probability was ( 8. 31 ± 1. 06 )% in the CSE-TM-thr-S group. When further blocked with TM-MAb, the binding probability was ( 5. 17 ± 2. 96 )%. Conclusion CSE significantly decreases the binding probability for TM-Thr to induce intravascular thrombosis.

OBJECTIVETo demonstrate the effects and mechanisms of Qifu decoction( QFD) on renal interstitial fibrosis (RIF) in model rats with yang-deficiency syndrome.

METHODThe rats were randomly divided into 3 groups, the Sham group (Group A), the Model group (Group B), the Qifu decoction group (Group C) and the Enalapril group (Group D). The RIF model was established by adenine administrated and unilateral ureteral obstruction (UUO) of the left ureter. After the model was successfully established, the rats in Group C and D were administrated with QFD or the Enalapril suspension,while the rats in Group A and B were administrated with distilled water. All rats were administrated for 3 weeks. Before administration and at the end of week 1, 2 and 3, the rats were weighted, and 24 h urinary protein excretion (Upro), urinary β2-microglobulin (Uβ2-MG) and urinary N-acetyl-D-glucosaminidase (NAG) were examined, respectively. All rats were killed after administration for 3 weeks. Blood and renal tissues were collected, renal morphology and tubulointerstitial morphology were evaluated, respectively. Serum cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), blood urea nitrogen (BUN), serum creatinine (Scr) and uric acid (UA) were detected, respectively. The protein expressions of E-cadherin, α-smooth muscle actin(α-SMA), transforming growth factor-β1 (TGF-β1), onnective tissue growth factor (CTGF) extracellular signal-regulated protein kinase 1/2(ERK1/2) and phosphorylated-ERK1/2 (p-ERK1/2) in kidney were evaluated, respectively.

RESULTQFD ameliorated serum cAMP level and the rate of cAMP/cGMP, attenuated urinary β2-MG level, NAG level and renal tubulointerstitial fibrosis, increased E-cadherin protein expression, and reduced α-SMA, TGF-β1, CTGF and p-ERK1/2 protein expressions in the kidney. However, QFD had no influence on renal function in vivo. In addition, these effects were better than those of the model rats treated by Enalapril.

CONCLUSIONQFD could alleviate yang-deficiency parameters, as well as urinary β2-MG level and NAG level in model rats induced by adenine administration and UUO. Moreover, QFD could improve EMT and RIF by up-regulating E-cadherin protein expression, and down-regulating α-SMA, TGF-β1, CTGF and p-ERK1/2 protein expressions, the key molecular in ERK1/2 signaling pathway.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Fibrosis ; Kidney ; drug effects ; pathology ; Kidney Diseases ; drug therapy ; pathology ; MAP Kinase Signaling System ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Ureteral Obstruction ; complications ; Yang Deficiency ; complications

Adult ; Aged ; Carcinoma, Squamous Cell ; rehabilitation ; surgery ; Carotid Arteries ; Female ; Head and Neck Neoplasms ; rehabilitation ; surgery ; Humans ; Male ; Middle Aged ; Neck ; blood supply ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; blood supply

OBJECTIVETo investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro.

METHODSHistological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test.

RESULTSCompared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01].

CONCLUSIONSAbnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.

Aged ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; metabolism ; pathology ; Female ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Skin ; metabolism ; pathology ; Wound Healing

This study focused on the intestinal absorption of traditional Chinese medicines (TCM) to reveal the scientific connotation of the compatibility of TCM pairs. The single pass intestinal perfusion (SPIP) was used in rats to compare the absorption of single extracts from Puerariae Lobatae Radix, single extracts from Ginseng Radix et Rhizoma, combined extracts from Puerariae Lobatae Radix and Ginseng Radix et Rhizoma and Puerariae Lobatae Radix and Ginseng Radix et Rhizoma mixture in rats. The content of puerarin, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 in liquid were tested by HPLC. The speed constant (Ka) and apparent permeability coefficients (Papp) were calculated and compared. Specifically, the order of puerarin Ka and Papp values from high to low was Ginseng Radix et Rhizoma and Puerariae Lobatae Radix mixture > single extracts from Puerariae Lobatae Radix > combined extracts from Ginseng Radix et Rhizoma and Puerariae Lobatae Radix; the order of ginsenosides Ka and Papp values from high to low was Ginseng Radix et Rhizoma and Puerariae Lobatae Radix mixture > single extracts from Ginseng Radix et Rhizoma > combined extracts from Ginseng Radix et Rhizoma and Puerariae Lobatae Radix. The combined administration of Ginseng Radix et Rhizoma and Puerariae Lobatae Radix may improve the absorption in the intestinal tract.
Animals ; Ginsenosides ; pharmacokinetics ; Intestinal Absorption ; Isoflavones ; pharmacokinetics ; Male ; Medicine, Chinese Traditional ; Panax ; chemistry ; Plant Extracts ; pharmacokinetics ; Pueraria ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rhizome

Humans ; Osteosarcoma ; metabolism ; Signal Transduction ; physiology ; TOR Serine-Threonine Kinases ; metabolism

Endocannabinoid anandamide (AEA) has protective effect on the heart against ischemia/reperfusion injury and arrhythmia, but the electrophysiological mechanism is unclear yet. In this study, the sinoatrial node (SAN) samples from New Zealand rabbits were prepared, and intracellular recording technique was used to elucidate the effect of AEA on the action potential (AP) of SAN pacemaker cells of rabbits and the mechanism. Different concentrations of AEA (1, 10, 100, 200, 500 nmol/L) were applied cumulatively. For some SAN samples, cannabinoid type 1 (CB1) receptor antagonist AM251, cannabinoid type 2 (CB2) receptor antagonist AM630, potassium channel blocker tetraethylammonium (TEA) and nitric oxide (NO) synthase inhibitor L-nitro-arginine methylester (L-NAME) were used before AEA treatment, respectively. We found that: (1) AEA (100, 200 and 500 nmol/L) not only shortened AP duration (APD), but also decreased AP amplitude (APA) (P < 0.05). (2) AM251, but not AM630, abolished the effect of AEA on APD shortening. (3) TEA and L-NAME had no influence on the AEA effect. These findings suggest that anandamide can decrease APA and shorten APD in SAN pacemaker cells of rabbits, which may be mediated by activation of CB1 receptors, and is related to blockade of calcium channels but not potassium channels and NO.
Action Potentials ; Animals ; Arachidonic Acids ; pharmacology ; Cannabinoid Receptor Antagonists ; pharmacology ; Endocannabinoids ; pharmacology ; Indoles ; pharmacology ; Myocytes, Cardiac ; drug effects ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; metabolism ; Piperidines ; pharmacology ; Polyunsaturated Alkamides ; pharmacology ; Potassium Channel Blockers ; pharmacology ; Pyrazoles ; pharmacology ; Rabbits ; Sinoatrial Node ; cytology

AIMTo report the experience with single stage dorsal inlay buccal mucosal grafts using the Snodgrass technique for complex redo cases.

METHODSFrom May 2004 to December 2005, a total of 53 patients aged from 3 to 34 years old (average 11.62 +/- 7.18 years) with failed previous hypospadias surgery were included in the present study. Indications included urethral strictures and repair breakdown. The unhealthy urethra was unroofed from the meatus in the ventral midline, a buccal mucosal graft was inlayed between the incised urethral plate and fixed to the corpora cavernosa. The neourethra was tubularized, and covered with subcutaneous (dartos) tissue and penile skin. Glanuloplasty was also performed in all cases. Outcome analysis included clinical follow-up, and endoscopy in 2 selected cases.

RESULTSThe buccal mucosal graft was 3.0-7.5 cm in length and 0.7-2.0 cm in width. All patients required glanuloplasty, with buccal mucosal grafts extended to the tip of the glans. After a follow-up of 14-30 months (mean 22.6 months), the total complication rate was 15.1%, with five cases of fistula and three cases of stricture.

CONCLUSIONInlaying dorsal buccal mucosal grafts applying the Snodgrass technique is a reliable method for creating a substitute urethral plate for tubularization. The recurrent rate of urethral stricture and fistula is at an acceptable level for redo cases. This approach represents an effective, simple and safe option for reoperations.

Adolescent ; Adult ; Child ; Child, Preschool ; Humans ; Hypospadias ; surgery ; Male ; Mouth Mucosa ; transplantation ; Secondary Prevention ; Transplants ; Treatment Outcome ; Urethra ; surgery ; Urethral Stricture ; prevention & control ; Urinary Fistula ; prevention & control ; Urologic Surgical Procedures, Male ; adverse effects ; methods