Objective To measure the concentration of intracellular free Ca2+ ([Ca2+]i) in neuron-like cells resulted from rat bone mesenchymal stem cell (BMSCs) differentiation induced by salvia miltiorrhiza injection and provide some theoretical basis for the BMSCs transplantation. Methods The rat BMSCs were separated from rat bone marrow and cultured in vitro. After induced by basic fibroblast growth factor and 10mL/L salvia miltiorrhiza injection, the cells were identified with immunofluorescence staining against NeuN. The same procedure was performed on primarily cultured hippocampal neurons. Then, the [Ca2+]i of the differentiated neuron-like cells was determined and compared with primarily cultured hippocampal neurons. Results The BMSCs after induced by basic fibroblast growth factor and salvia miltiorrhiza injection expressed neuronal phenotypes similar to the cell appearance of neurons with NeuN. The average fluorescence intensity of the neuron-like cells derived from BMSCs was 984.75±79.51, while the average fluorescence intensity of the primarily cultured hippocampal neurons was 769.42±60.93. No significant difference was found between them (P>0.05). Conclusion The neuron-like cells from rat BMSCs differentiation induced by salvia miltiorrhiza injection possess certain neuronal properties.

OBJECTIVETo summarize the development of gene delivery vectors in peritoneal fibrosis research and discuss the feasibility and superiority of lentiviral vectors.

DATA SOURCESThe data in this article were collected from PubMed database with relevant English articles published from 1995 to 2011.

STUDY SELECTIONArticles regarding the gene therapy in peritoneal fibrosis research using non-viral vectors, adenoviral vectors, retroviral vectors, and lentiviral vectors were selected. Data were mainly extracted from 60 articles, which are listed in the reference section of this review.

RESULTSNon-viral vector-mediated gene delivery (including naked DNA for ex vivo, oligonucleotides, ultrasound- contrast agent mediated naked gene delivery, etc.) and viral vector-mediated gene delivery (including adenovirus, helper-dependant adenovirus, and retrovirus vectors) have been successfully applied both in the mechanistic investigation and the potential prevention and treatment of peritoneal fibrosis.

CONCLUSIONSPeritoneal fibrosis is a major complication of peritoneal dialysis (PD). Recently, the wide use of the gene delivery technique made it possible to access and further research peritoneal fibrosis. The use of lentiviral vector is expected to be widely used in PD research in the future due to its advantages in gene delivery.

Gene Transfer Techniques ; Genetic Vectors ; administration & dosage ; Humans ; Peritoneal Dialysis ; Peritoneal Fibrosis ; therapy

OBJECTIVETo explore the expression and significance of secretions of hypothalamus-pituitary-adrenal (HPA) axis in human hypertrophic scar.

METHODSHypertrophic scar tissues obtained from 12 patients with deep-partial thickness burn or full-thickness burn and normal skin tissues from the same 7 patients with hypertrophic scar were harvested for determination of gene expression of corticotrophin-releasing hormone (CRH), CRH receptor 1 (CRH-R1), pro-opiomelanocortin (POMC), melanocortin receptor 2 (MC-2R), and glucocorticoid receptor α (GR-α) by real-time fluorescence quantitative PCR. After addition of corresponding antibodies, distribution differences of CRH, CRH-R1, adrenocorticotropic hormone (ATCH), MC-2R, and GR-α were observed with immunohistochemical staining. Data were processed with t test.

RESULTSThe mRNA expression of CRH, CRH-R1, POMC, and GR-α in hypertrophic scar was respectively 3.1 ± 0.8, 0.05 ± 0.03, 0.020 ± 0.007, and 0.0030 ± 0.0010, which were significantly lower than those in normal skin (20.6 ± 4.7, 0.30 ± 0.12, 0.060 ± 0.020, and 0.0200 ± 0.0070, with t values from 2.10 to 4.75, P values all below 0.05). There was no statistical difference in MC-2R mRNA expression between hypertrophic scar and normal skin (t = 1.48, P = 0.15). Immunohistochemical observation showed CRH, CRH-R1, ACTH, MC-2R, and GR-α in hypertrophic scar were located in basal layer of epidermis, fibroblast of dermis, and tube wall of sweat gland. Expressions of these indexes could also be observed in sebaceous gland and hair follicle besides above-mentioned structures.

CONCLUSIONSDecreasing expression of active material of HPA axis may be related to formation of hypertrophic scar.

Adolescent ; Adrenocorticotropic Hormone ; metabolism ; Adult ; Child ; Cicatrix, Hypertrophic ; metabolism ; Female ; Glucocorticoids ; metabolism ; Humans ; Hypothalamo-Hypophyseal System ; metabolism ; Male ; Pituitary-Adrenal System ; metabolism ; Young Adult

OBJECTIVETo study the effect of melatonin on proliferation and apoptosis of fibroblasts in human hypertrophic scar and its mechanism.

METHODSFibroblasts from human hypertrophic scar were isolated and cultured with DMEM medium containing 10% FBS, and then they were divided into control (C, added with ethanol), low concentration (LC, added with 1 × 10(-5) mmol/L melatonin), middle concentration (MC, added with 1 × 10(-3) mmol/L melatonin), and high concentration (HC, added with 1 mmol/L melatonin) groups according to the random number table. After being cultured for 24 hours, cell morphologic change was observed under microscope; XTT-PMS assay was used to examine cell proliferative activity; cell cycle and apoptosis were assessed with flow cytometry after double staining of FITC and PI, and the levels of cyclin E, p53, and Fas mRNA were determined with fluorescence quantitative RT-PCR. Data were processed with analysis of variance and LSD test.

RESULTS(1) Fibroblasts in C group were spindle-shaped with growth in colonies. Along with the increase in melatonin concentration, fibroblasts in LC, MC, and HC groups gradually dispersed, deformed and atrophied, with shrunk cellular membrane, and decrease in ratio of nucleus and cytoplasm. (2) Proliferative activity of fibroblasts in LC, MC, and HC groups decreased along with an increase in melatonin concentration (1.49 ± 0.15, 1.24 ± 0.20, and 0.92 ± 0.09), which were lower that in C group (1.79 ± 0.10, F = 67.61, P < 0.05). Cell ratios of S and G2/M phases in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were all lower than those in C group [(10.6 ± 1.1)%, (6.1 ± 1.2)%, (3.2 ± 0.8)% vs.(16.9 ± 1.3)%, F = 286.10, P < 0.05; (13.5 ± 1.1)%, (9.8 ± 1.0)%, (6.0 ± 0.7)% vs. (16.7 ± 1.6)%, F = 162.69, P < 0.05]. Apoptotic rates in early and late stages of LC, MC, and HC groups increased along with an increase in melatonin concentration, all higher than those in C group (with F value respectively 424.05, 236.44, P values all below 0.05). The expressions of cyclin E mRNA in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were lower than that in C group (1.58 ± 0.21, 0.90 ± 0.20, and 0.24 ± 0.12 vs. 2.90 ± 0.30, F = 266.79, P < 0.05), while the expressions of p53 mRNA and Fas mRNA showed opposite tendency (with F value respectively 10.11, 12.03, P values all below 0.05).

CONCLUSIONSMelatonin can inhibit proliferation and induce apoptosis of fibroblasts in hypertrophic scar through regulating the gene expressions of cyclin E, p53, and Fas.

Adult ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin E ; metabolism ; Female ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans ; Male ; Melatonin ; pharmacology ; Oncogene Proteins ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo investigate the effect of liquorice in functional modulation of intestinal P-glycoprotein (P-gp) in rats.

METHODSAn in vitro diffusion chamber system (Ussing chamber) was used to examine the direct effect of liquorice decoction on rhodamine 123 (a subtrate of P-gp) transport and evaluate the permeability of rhodamine 123 or fluorescein sodium through rat jejunum membranes after oral administration of liquorice decoction.

RESULTSDirect application of liquorice decoction did not obviously affect rhodamine 123 transport across the intestinal mucosa. Oral administration of liquorice decoction (10 g/kg, twice daily for a week) significantly increased the absorption of rhodamine 123 and also enhanced rhodamine 123 secretion across the jejunum mucosa. Liquorice had no obvious effect on the transport of CF across the jejunum mucosa.

CONCLUSIONLiquorice may slightly inhibit P-gp function in the intestinal mucosa to increase the intestinal absorption of rhodamine 123.

ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; metabolism ; Animals ; Glycyrrhiza ; Intestinal Absorption ; drug effects ; Intestinal Mucosa ; drug effects ; metabolism ; Intestines ; drug effects ; metabolism ; Male ; Plant Extracts ; administration & dosage ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rhodamine 123 ; metabolism

Adult ; Aged ; Aneurysm, False ; physiopathology ; Carotid Arteries ; diagnostic imaging ; pathology ; Epistaxis ; diagnostic imaging ; etiology ; pathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Radiography

The comparative efficacy of 2 anthelmintics (ivermectin and levamisole) against Baylisascaris transfuga migrating and encapsulated larvae was studied in mice. A total of 60 BALB/c mice inoculated each with about 1,000 embryonated B. transfuga eggs were equally divided into 6 groups (A-F) randomly. Mice of groups A and B were treated with ivermectin and levamisole, respectively, on day 3 post-infection (PI). Mice of groups A-C were killed on day 13 PI. Similarly, groups D and E were treated with ivermectin and levamisole, respectively, on day 14 PI, and all mice of groups D-F were treated on day 24 PI. The groups C and F were controls. Microexamination was conducted to count the larvae recovering from each mouse. The percentages of reduction in the number of migrating larvae recovered from group A (ivermectin) and B (levamisole) were 88.3% and 81.1%, respectively. In addition, the reduction in encapsulated larvae counts achieved by ivermectin (group D) and levamisole (group E) was 75.0% and 49.2%, respectively. The results suggested that, to a certain extent, both anthelmintics appeared to be more effective against migrating larvae than encapsulated larvae. However, in the incipient stage of infection, ivermectin may be more competent than levamisole as a larvicidal drug for B. transfuga.
Animals ; Anthelmintics/*administration & dosage ; Ascaridida Infections/*drug therapy/parasitology ; Ascaridoidea/*drug effects ; Disease Models, Animal ; Female ; Ivermectin/*administration & dosage ; Larva/drug effects ; Levamisole/*administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Rodent Diseases/drug therapy/parasitology ; Treatment Outcome

OBJECTIVETo investigate the expression and its significance of melatonin receptor in human hypertrophic scarring.

METHODSThe expression of melatonin receptor GPR50 was detected with immunohistochemistry and the melatonin receptors (MT1, MT2) mRNA were assessed with RT-PCR method in 10 cases of human hypertrophic scar and normal skin. The positive production was sequenced with auto sequencing instrument.

RESULTSPositive signals of melatonin receptor could be found in the cell membrane and cytoplasm. The melatonin receptor GPR50 was located in the epithelial basal cells,sweat gland cells and hair follicle in both hypertrophic scar and normal skin. The melatonin receptor GPR50 was extensively expressed in fibroblasts of hypertrophic scar, but not in fibroblasts in normal skin. RT-PCR showed that the expression of melatonin receptor (MT1, MT2) mRNA in hypertrophic scar was significantly higher than that in normal skin (P < 0.05). In normal skin and hypertrophic scar group, the expression of MT1 mRNA was higher than MT2 mRNA (P < 0.05). In normal skin and hypertrophic scar group, the expression of MT1 mRNA was 0.99081 +/- 0.26485 and 1.16584 +/- 0.21829 copy number/microl cDNA, respectively; the expression of MT2 mRNA was 0.77083 +/- 0.15927 and 0.99550 +/- 0.14624 copy number/ microl cDNA, respectively. Sequencing results indicated that the positive product coincided with cDNA of human melatonin receptor in GeneBank.

CONCLUSIONSPositive expression of melatonin receptor can be found in human hypertrophic scar and normal skin, but it is higher in scar. The over expression of melatonin receptor in hypertrophic scar may be related to the development of hypertrophic scar.

Adult ; Cicatrix, Hypertrophic ; metabolism ; Female ; Humans ; Male ; Nerve Tissue Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; Receptors, G-Protein-Coupled ; genetics ; metabolism

OBJECTIVETo investigate the expressions of stathmin gene and its coding protein in laryngeal squamous cell carcinoma, and to explore the relationship between stathmin gene and the biological behaviors of laryngeal squamous cell carcinoma for understanding the tumorigenicity and development of laryngeal squamous cell carcinoma.

METHODSLaryngeal carcinoma tissues (studying group) in the tumors center and laryngeal normal tissues (control group) parted from 1.0 cm of the safe borderline of the tumors were took from 38 patients with laryngeal squamous cell carcinoma while they were in operation. Semiquantitative method of reverse transcriptase polymerase chain reaction (RT-PCR) was used to analyze the expression level of stathmin mRNA, and immunohistochemical staining (frozen section) was used to detect the expressions of stathmin protein, in laryngeal carcinoma tissues and laryngeal normal tissues of 38 cases, respectively.

RESULTSmRNA of stathmin gene was all positively expressed in laryngeal carcinoma tissues and in laryngeal normal tissues of 38 cases by RT-PCR. However, stathmin mRNA was obviously overexpressed in laryngeal carcinoma tissues than that in laryngeal normal tissues (t = 9.655, P < 0.05). Immunohistochemical staining showed stathmin protein was positively expressed in laryngeal carcinoma tissues of 26 cases (26/38, 68.4%), and mild-positively expressed in laryngeal normal tissues in 13 cases (13/38, 34.2%). There was significant difference between the expression rate of stathmin protein in laryngeal carcinoma tissues and in laryngeal normal tissues (chi2 = 8.901, P < 0.05). Meanwhile, the expression level of stathmin mRNA and the positive-expressed rate of stathmin protein in laryngeal carcinoma tissues of the advanced stage patients group (III stage and IV stage) were significantly higher than these in laryngeal carcinoma tissues of I and II stage patients group (t = 6.284, chi2 = 5.810, P < 0.05), and they were also significantly higher in laryngeal carcinoma tissues of the patients group with cervical lymph node metastasis than in laryngeal carcinoma tissues of the patients group without cervical lymph node metastasis (t = 9.350, chi2 = 6.923, P < 0.05).

CONCLUSIONSThe expression levels of stathmin gene and protein were significantly higher in laryngeal squamous cell carcinoma than these in laryngeal normal tissues, the levels are also significantly higher in advanced stage patients group (III stage and IV stage) than in the early stage patients group (I and II), and they are also related to the cervical lymph node metastasis of carcinoma. Stathmin gene may play an important role in the pathogenesis and development of laryngeal carcinoma and may be related to its prognosis.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Stathmin ; genetics ; metabolism

OBJECTIVETo evaluate the effect of glutamine granules on protein metabolism in severe burns and trauma patients.

METHODS120 patients with severe burns, multiple trauma and post operation who met the requirements of the protocol joined this double-blind randomized controlled, multi-center clinical trail. Patients were randomly divided into two groups: placebo control group (P group, 60 patients) and glutamine granules treatment group (GLN group, 60 patients). There was isonitrogenous and isocaloric intake in both groups, GLN and P group patents had been given glutamine granules or placebo (glycine) at 0.5 g.kg(-1).d(-1) for 7 days, respectively. The level of plasma glutamine, protein and urine nitrogen exclude were determined, wound healing rate of burn area and hospital stay were recorded, and then observed the complication and side effect.

RESULTSAfter 7 days of taking glutamine granules orally, plasma GLN concentration was significant higher than that in P group (592.50 +/- 185.23 micro mol/L vs. 407.41 +/- 190.22 micro mol/L) (P < 0.01). Plasma prealbumin and transferrin in GLN group were significant higher than those in P group (P < 0.01), but the concentration of total protein and albumin were no marked changes compare with P group (P > 0.05). The capacity of urine nitrogen exclude in GLN group were significant lower than that in P group. Additional, the wound healing rate was faster and hospital stay days was shorter than P group (P < 0.05), and the occurrence of glutamine granules side effect was seldom.

CONCLUSIONTaking glutamine could promote protein synthesis, abate protein decompose, ameliorate wound healing rate and reduce hospital stay obviously.

Adult ; Burns ; drug therapy ; metabolism ; Double-Blind Method ; Female ; Glutamine ; blood ; therapeutic use ; Humans ; Length of Stay ; Male ; Middle Aged ; Nitrogen ; urine ; Proteins ; metabolism ; Treatment Outcome ; Wounds and Injuries ; drug therapy ; metabolism