Animals ; Benzodiazepines ; pharmacology ; Ducks ; physiology ; Growth Hormone ; blood ; Insulin-Like Growth Factor I ; metabolism ; Serum ; metabolism ; Weight Gain ; drug effects

Effect of strophanthidin (Str) on intracellular calcium concentration ([Ca2+]i) was investigated on isolated ventricular myocytes of guinea pig. Single ventricular myocytes were obtained by enzymatic dissociation technique. Fluorescent signal of [Ca2+]i was detected with confocal microscopy after incubation of cardiomycytes in Tyrode's solution with Fluo3-AM. The result showed that Str increased [Ca2+]i in a concentration-dependent manner. The ventricular myocytes began to round-up into a contracture state once the peak level of [Ca2+]i was achieved in the presence of Str (10 μmol·L-1), but remained no change in the presence of Str (1 and 100 nmol·L-1). Tetrodotoxin (TTX), nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str (1 and 100 nmol·L-1), but had no obvious effects on the action of Str (10 μmol·L-1). The elevation of [Ca2+]i caused by Str at all of the detected concentrations was partially antagonized by rynodine (10 μmol·L-1) or the removal of Ca2+ from Tyrode's solution. In Na+, K+-free Tyrode's solution, the response of cardiomycytes in [Ca2+]i elevation to Str (10 μmol·L-1) was attenuated, while remained no change to Str (1 and 100 nmol·L-1). TTX, nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str at all of the detected concentrations in Na+, K+-free Tyrode's solution. The study suggests that the elevation of [Ca2+]i by Str at the low (nomomolar) concentrations is partially mediated by the extracellular calcium influx through Ca2+ channel or a "slip mode conductance" of TTX sensitive Na+ channel. While the effect of Str at high (micromolar) concentrations was mainly due to the inhibition of Na+, K+-ATPase. Directly triggering the release of intracellular Ca2+ from sarcoplasmic reticulum (SR) by Str may be also involved in the mechanism of [Ca2+]i elevation.

Objective To evaluate the treatment of aneurysms rupture during endovascular embolization.Methods Nine aneurysms ruptured during the embolization and were treated with endovascular embolization.The reasons of aneurysms rupture during embolization,the prevention and the first aid after aneurysms rupture were analysed.Results Seven patients recovered and 2 died.Conclusions The optimal treatment of aneurysms rupture during endovascular embolization is effective,(J Intervent Radiol,2007,16: 132-134)

Objective To observe the expression of growth factors (vascular endothelial growth factor [VEGF],stromal cell-derived factor-1 [SDF-1 ],basic fibroblast growth factor [bFGF],insulin-like growth factor [IGF-1],transforming growth factor-β [TGF-β],platelet-derived growth factor [PDGF],brain derived neurotrophic factor [BDNF],glial cell line-derived neurotrophic factor [GDNF] and nerve growth factor [NGF]) in rat ischemic brain tissues after intravenous implantation of bone marrow stromal cells (BMSCs) and/or endothelial progenitor cells (EPCs). Methods Healthy adult Wistar rats were randomly divided into 4 groups:vehicle group,BMSCs transplantation group,EPCs transplantation group and BMSCs combined with EPCs transplantation group (n=20). The rats were subjected to middle cerebral artery occlusion (MCAO),and 24 h after that,they were intravenously transplanted with either 3×106 BMSCs,EPCs,BMSCs/EPCs or 1 mL physiological saline.Seven d after transplantation,real time-PCR and Western blotting were employed to detect the expressions of VEGF,SDF-1,bFGF,IGF-1,TGF-β,PDGF-BB,BDNF,GDNF and NGF. Results The mRNA expressions of bFGF,VEGF and BNDF in the BMSCs/EPCs transplantation group were significantly higher as compared with those in the other groups (P＜0.05).BMSCs transplantation group enjoyed the highest mRNA levels of NGF,GDNF and TGF-β among all the groups, significantly higher as compared with those in the other groups (P＜0.05),followed by BMSCs/EPCs transplantation group.EPCs transplantation group enjoyed the highest mRNA levels of PDGF,IGF-1 and SDF-1,significantly higher as compared with those in the other groups (P＜ 0.05), followed by BMSCs/EPCs transplantation group. Conclusion BMSCs combined with EPCs implantation can promote the functional rehabilitation in rats after focal cerebral ischemia, which provides new way for improving the transplantation success rate.

OBJECTIVETo explore the feasibility of burn denatured acellular dermal matrix (DADM) as dermal substitute in repairing wounds.

METHODS(1) Nine Wistar rats received a deep partial-thickness scald on the back. Full-thickness wounded skin was collected on post scald day (PBD) 1, 2, and 3 (with 3 rats at each time point), and it was treated with 2.5 g/L trypsin/0.5% Triton X-100 to remove cells to prepare DADM, respectively called DADM-1 d, DADM-2 d, and DADM-3 d. Another 3 rats without scald injury were treated with the same method as above to prepare acellular dermal matrix (ADM) to serve as control. Gross and histological observations and microbiological and biomechanical tests, including ultimate tensile strength, maximum tension, stretched length at breaking, stress-strain relationship, were conducted for the resulting ADM and DADM. (2) Another 64 rats were divided into ADM group and DADM-1 d, DADM-2 d, and DADM-3 d groups according to the random number table, with 16 rats in each group. A skin flap in size of 2.0 cm×1.8 cm was raised on the back of each rat. The above-mentioned ADM, DADM-1 d, DADM-2 d, and DADM-3 d were cut into pieces in the size of 1.8 cm×1.5 cm, and they were respectively implanted under the skin flaps of rats in corresponding group. At post surgery week (PSW) 1, 3, 5, or 9, 4 rats in each group were used to observe wound healing condition and change in implants with naked eye, and histological observation of the implants was conducted. Data were processed with one-way analysis of variance and t test.

RESULTS(1) The freshly prepared DADM was milky white, soft in texture with flexibility, but poor in elasticity as compared with ADM. No epithelial structure or cellular component was observed in ADM or DADM under light microscope. Collagen fibers of DADM were seen to be thickened unevenly and arranged in disorder and eosinophilic. All microbiological results of DADM were negative. There was no statistically significant difference among DADM-1 d, DADM-2 d, and DADM-3 d in levels of ultimate tensile strength, maximum tension, stretched length at breaking, and stress-strain relationship (with F values from 0.088 to 3.591, P values all above 0.05). Values of the above-mentioned four indexes were the highest in DADM-3 d, they were respectively (13.0 ± 2.4) MPa, (61 ± 4) N, (173 ± 7)%, (45.7 ± 2.0)%. Values of the four indexes of ADM were respectively (19.0 ± 2.6) MPa, (95 ± 4) N, (201 ± 5)%, (62.5 ± 2.2)%, which were higher than those of DADM-1 d, DADM-2 d, and DADM-3 d (with t values from 6.424 to 17.125, P values all below 0.01). (2) No exudate or swelling in the wounds of rats, and no contraction or curling of implants were observed in every group at PSW 1, but inflammatory cells infiltration and Fbs inward migration were observed in the wound. At PSW 3, the growth of hair was normal in the wound in DADM-1 d, DADM-2 d, and ADM groups, but few and scattered hair grew in DADM-3 d group. The inflammatory cells decreased, while Fbs increased, and new capillaries were found to grow inwardly in each group. The decrease in inflammatory cells was slightly delayed in DADM-3 d group. At PSW 5, hair growth became normal, and implants shrank and thinned with fiber membrane wrapped densely and bundles of ingrowing large caliber blood vessels in all groups. The dermal matrix in each group merged with the surrounding normal tissue. At PSW 9, ADM and DADM became white, thin, and soft tissue sheet which was closely connected with the inner side of the flap. There was no infiltration of inflammatory cells in implants in either group. The collagen fibers arranged regularly and densely, and they were integrated with normal collagen tissue.

CONCLUSIONSThe burned DADM does not have obvious immunogenicity, but with good biocompatibility. It is prospective to become as a dermal substitute in repairing wounds.

Acellular Dermis ; Animals ; Burns ; pathology ; surgery ; Female ; Male ; Rats ; Rats, Wistar ; Reconstructive Surgical Procedures ; methods ; Skin ; injuries ; Skin Transplantation ; methods ; Skin, Artificial ; Wound Healing

OBJECTIVETo verify the efficacy on lumbar disc herniation treated with Shu-needle therapy in combination with ozone injection of low concentration.

METHODSOne hundred and thirty cases of lumbar disc herniation were randomized into a Shu-needle therapy group and an acupotomy group, 65 cases in each one. In the Shu-needle therapy group, Shu-needle therapy was used in combination with ozone injection of low concentration. In the acupotomy group, the conventional acupotomy therapy was applied in combination with ozone injection of low concentration. The treatment was given once every 10 days, 3 treatments made one session. After one session treatment, the clinical efficacy of two groups was observed, scores of visual analogue scale (VAS) and Oswestry disability index (ODI) were counted before and after treatment. The long-term efficacy was followed up in half a year.

RESULTSThe clinical curative rate was 69.2% (45/65) and the total effective rate was 96.9% (63/65) in the Shu-needle therapy group. The curative rate was 43.1% (28/65) and the total effective rate was 84.6% (55/65) in the acupotomy group. In comparison, the efficacy of the Shu-needle therapy group was superior to that of the acupotomy group (P < 0.01, P < 0.05). The scores of VAS and ODI were reduced obviously after treatment as compared with those before treatment in two groups (all P < 0.05). The improvements in the Shu-needle therapy group were superior to those in the acupotomy group (both P < 0.05). In the follow-up observation, the recurrence rate in the Shu-needle therapy group was lower than that in the acupotomy group [17.8% (8/45) vs 46.4% (13/28), P < 0.05].

CONCLUSIONShu-needle therapy in combination with ozone injection of low concentration achieves the superior efficacy on lumbar disc herniation as compared with the acupotomy group.

Acupuncture Therapy ; Adult ; Combined Modality Therapy ; Female ; Humans ; Injections ; Intervertebral Disc Displacement ; drug therapy ; therapy ; Lumbar Vertebrae ; drug effects ; Male ; Middle Aged ; Ozone ; administration & dosage ; Treatment Outcome ; Young Adult

ECM is a supporting structure for stabilizing the location of cells and preserving the structure of tissues. Recently, it has been discovered that ECM and its degradation products may exert profound influences on tissues and cells, such as activities of inflammatory cells and immune cells. Angiogenesis may be stimulated or inhibited by degradation products of ECM. Matrikines, liberated by partial proteolysis of ECM macromolecules, are found to regulate cell functional activities and play a significant role in wound healing or tumor invasion. Post-burn denatured dermal matrix is being studied in burn healing now. The study of post-burn denatured or necrotic dermal matrix should be emphasized in future.
Animals ; Extracellular Matrix ; metabolism ; Humans ; Inflammation ; metabolism ; Wound Healing

OBJECTIVETo investigate the expression of PTEN and loss of heterozygosity (LOH) of its epigenetic microsatellite in gastric carcinoma and explore their roles in progression of gastric carcinoma.

METHODSLOH of epigenetic microsatellites of PTEN (D10S541, D10S583 and D10S1687) in advanced gastric cancer was detected by PCR-SSCP. Expression of PTEN mRNA and protein in normal gastric mucosa and gastric cancer was evaluated by RT-PCR and SABC immunohistochemistry, respectively. The relationship between expression of PTEN mRNA and protein and lymph node metastasis or LOH of microsatellites was discussed.

RESULTSLOH of D10S541, D10S583 and D10S1687 was found in 37.5% (21/56) of advanced gastric cancers. The positive rates of PTEN mRNA expression were 80.4% (45/56), 45.5% (5/11) and 32.1% (18/56) in normal mucosa, early and advanced gastric carcinomas, respectively, while 78.6% (44/56), 44.5% (5/11) and 28.6% (16/56) at the protein level. PTEN mRNA and protein were less frequently expressed in early and advanced gastric carcinomas than that in normal gastric mucosa (P < 0.05). There was positive correlation between PTEN mRNA expression and LOH of microsatellites in advanced gastric carcinomas. PTEN protein expression paralleled with its mRNA expression (P < 0.05). The expression of PTEN mRNA and protein was negatively correlated with lymph node metastasis of advanced gastric carcinomas (P < 0.05).

CONCLUSIONDown-regulated expression of PTEN gene is found in different stages of gastric carcinoma, and is closely correlated with LOH of its epigenetic microsatellites, which probably is its underlying molecular mechanisms. It suggests that altered PTEN gene contributes to tumorigenesis and progression of gastric carcinomas.

Gastric Mucosa ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Loss of Heterozygosity ; Lymphatic Metastasis ; genetics ; Microsatellite Repeats ; genetics ; Neoplasm Staging ; PTEN Phosphohydrolase ; Phosphoric Monoester Hydrolases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVETo evaluate the therapeutic effect of laparoendoscopic single-site surgery (LESS) for treatment of male pseudohermaphroditism.

METHODSA 17-year-old patient with male pseudohermaphroditism and a female social sex was admitted. According to the request by the patient and the relatives for a female gender, LESS vaginoplasty and cryptorchidectomy were performed using a single multilumen port inserted through a 2.5 cm incision below the umbilicus, followed by reconstruction of the perineal region by open surgery.

RESULTSThe total operative time was 7 h, and the LESS procedure lasted for about 3.5 h. No other port incision was needed. The estimated intraoperative blood loss was 400 ml. No electrolyte or metabolic acid-base balance disorders were observed perioperatively. In the follow-up examination at 6 months after the operation, the reconstructed vagina healed smoothly without obvious contraction or fixation failure, and the perineal region showed good appearance.

CONCLUSIONWith minimal invasiveness, LESS surgery produces good cosmetic effect and allows rapid postoperative recovery, thus may become a promising alternative to the management of pseudohermaphroditism.

46, XY Disorders of Sex Development ; surgery ; Adolescent ; Female ; Humans ; Laparoscopy ; methods ; Male ; Reconstructive Surgical Procedures ; methods ; Vagina ; surgery

BACKGROUNDToll like receptor (TLR) 9 has been shown to play a crucial role in the pathogenesis of systemic lupus erythematosus (SLE) in animal models. Its pathogenic role in human SLE, however, was poorly elucidated. This study was performed to investigate the role of TLR9 involved in the aberrant signaling pathway and its correlation with disease activity in SLE.

METHODSmRNA level of TLR9 and interferon (IFN) regulatory factor 5 (IRF5) in peripheral blood mononuclear cells (PBMCs) were determined by real-time polymerase chain reaction (PCR). IFN-a expression was measured in the serum of the SLE patients by enzyme-linked immunosorbent assay (ELISA).

RESULTSTLR9 expression was significantly higher in SLE patients than that in health controls (P = 0.011). SLE patients with positive anti-dsDNA antibody had significantly higher expression of TLR9 than that with negative anti-dsDNA antibody (P = 0.001). TLR9 expression was positively correlated with fever (P = 0.017), alopecia (P = 0.046), safety of estrogens in lupus erythematosus national assessment SLE disease activity index (SELENA-SLEDAI) score (r(s) = 0.385, P = 0.003), and the level of IRF5 (r(s) = 0.35, P = 0.027) and IFN-a (r(s) = 0.627, P = 0.001) in SLE patients.

CONCLUSIONTLR9 is associated with SLE disease activity and might be involved in the IFN-a pathway of SLE.

Adolescent ; Adult ; Antibodies, Antinuclear ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Interferon Regulatory Factors ; metabolism ; Interferon-alpha ; blood ; Leukocytes, Mononuclear ; metabolism ; Lupus Erythematosus, Systemic ; blood ; genetics ; metabolism ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Toll-Like Receptor 9 ; genetics ; metabolism ; Young Adult