objective To investigate the differentiation of bone marrow-derived mesenchymal stem cells into neurons and transplantation of the stem ceils to repair rat hemisection spinal cord injury.Methods Adherent culture was used to isolate and culture rat bone marrow mesenchymal stem cells(MSCs).The rat spinal cord homogenate supernatant was used to induce neural differentiation of the 3rd generation ceils.The nature of ceil differentiation was identified by immunohistochemistry.The rat model of hemisection spinal cord injury was prepared and BrdU was locally injected to label the induced neurons.The distribution of living cells in the injuried spinal cord was observed at 5 weeks after cell transplantation.Results MSCs were spindle and polygonal,with 1-2 nucleoli seen under the inverted microscope.After induction with spinal cord homogenate supernatant there were a number of slender cytoplasmic projections forming interwined network and showing nestin expression,therefore,indicating the neuronal nature.MSCs at 5 weeks after transplantation into the spinal cord injury were surviving and their expression of MAP-2,NF,GFAP was significantly higher than that in the control rats(P<0.05).The rat motor function was improved than before transplantation.Conclusion MSCs induced by spinal cord homogenate supernatant can be transplanted into hemisection spinal cord injury and improve the motor function of the injuries lesions.

OBJECTIVE: To investigate the bacteriostatic effect of platelet-rich plasma (PRP) and its derivative platelet gel (PG) supernatant on Escherichia coli in vitro and its relationship with platelet factor 4 (PF4). METHODS: Apheresis platelets donated by healthy volunteers were obtained from the Blood station of Lu an Blood Center as the source of PRP. The counts of platelet, white blood cell and red blood cell in PRP and its derivative PG supernatant were detected by automatic hematology analyzer. Bacterial growth of PRP and PG supernatants co-cultured with bacteria for different time was observed by plate coating culture method, and the contents of PF4 in PRP and PG supernatants were detected by ELISA. RESULTS: Apheresis platelets were collected from 28 healthy volunteers with a median age of 33 (21-56) years old. PRP can inhibit the growth of escherichia coli, but there were individual differences in antibacterial effect within 24 hours. PRP of 13 healthy volunteers had strong antibacterial effect at 24 hours, 7 cases had weak antibacterial effect at 24 hours, and 8 cases had no antibacterial effect at 24 hours. PG supernatant showed no significant individual difference, and all of them had bacteriostatic effect within 12 hours, but no bacteriostatic effect after 12 hours. There was no statistical difference in the bacteriostatic effect of PRP at 24 hours between healthy volunteers aged ≤30 years and >30 years (P>0.05), and there was no statistical difference between the white blood cell count ≤0.1×10⁹/L and (0.1-1) ×10⁹/L groups (P>0.05). There was significant difference in the bacteriostatic effect of PRP between the two groups with platelet content ≤1 000×10⁹/L and >1 000×10⁹/L (P<0.05). The platelet count in PRP was higher than that in PG supernatant ［(911.57±160.52) ×10⁹/L vs 0］. The PF4 level in PRP was higher than that in PG supernatant (23623.34±9822.14 vs 6664.74±4065.83, P<0.05). CONCLUSION Both PRP and PG supernatant have antibacterial effects in Escherichia coli. The bacteriostatic effect of PRP was better than that of PG supernatant, and the platelet and PF4 contents in PRP were higher than those in PG supernatant, suggesting that the platelet and PF4 levels play an important role in bacteriostasis.
Adult ; Humans ; Middle Aged ; Escherichia coli ; Platelet Factor 4 ; Platelet-Rich Plasma

OBJECTIVETo study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).

METHODSWe established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.

RESULTSThe prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.

CONCLUSIONThe ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.

Animals ; Apomorphine ; Carcinogens ; Diethylstilbestrol ; Erectile Dysfunction ; etiology ; Humans ; Male ; Myocytes, Smooth Muscle ; ultrastructure ; Nitric Oxide Synthase Type I ; metabolism ; Organ Size ; Penile Erection ; Penis ; enzymology ; ultrastructure ; Pituitary Neoplasms ; chemically induced ; complications ; Prolactin ; blood ; Prolactinoma ; chemically induced ; complications ; Rats ; Rats, Wistar ; Testosterone ; blood

To block tumor cell adhesion, inhibit tumor metastasis and recurrence, the anti-adhesion peptide-trimeric beta peptide (DLYYLMDLSYSMKGGDLYYLMDLSYSMKGGDLYYLMDLSYSMK, beta3) was designed. The DNA fragment of beta3 was cloned into expression vector pET-His and the fusion protein His-beta3 was expressed in E. coli. BL21(DE3)plysS. After 1.5 hours' induction with IPTG, His-beta3 peptide was expressed significantly amounting to 10% of the insoluble proteins and 4% of the total proteins. 20mg of beta3 peptide was obtained from one litter culture medium after purification by using metal-chelating sepharose 6B FF. The purity of beta3 is 92.2% according to Gel-Pro analysis. The anti-adhesion effects of beta3 peptide, beta1 peptide (DLYYLMDLSYSMK) and GRGDS on the hepatocellular carcinoma cell line SMMC-7721 and the high metastasis hepatocellular carcinoma cell line HCCLM6 were studied. The result showed the beta3 blocked the adhesion of HCCLM6 cells and SMMC-7721 cells to fibronectin (FN) specifically. The inhibition effect was dose-dependent and time-dependent and the inhibition rate of beta3 was higher than three times concentration of beta1 and GRGDS. This suggested that pET-His-beta3/BL21(DE3)plysS was a suitable expression system for beta3, and the expressed beta3 specially inhibited the adhesion of cancer cells.
Amino Acid Sequence ; Antineoplastic Agents ; pharmacology ; Cell Adhesion ; drug effects ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Peptide Fragments ; pharmacology ; Peptides ; genetics ; metabolism ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVETo establish and evaluate a new method for measurement of dental plaque by using cellular neural network-based image segmentation.

METHODSA total of 195 subjects were selected from community population. After dental plaque staining, oral digital picture of anterior teeth area was taken by an Olympus digital camera (C-7070 Wide Zoom). At the same time, the Turesky dental plaque indices of anterior teeth were evaluated. The image analysis was conducted by cellular neural network-based image segmentation.

RESULTSThe image cutting errors between two operators were very small. The Kappa value is 0.935. Pearson's correlation coefficient is 0.988 (P < 0.001). There was high correlative consistency between traditional dental plaque index and plaque percentage obtained by using image analysis. Pearson's correlation coefficient was 0.853 (P < 0.001).

CONCLUSIONSCellular neural network-based image segmentation is a new method feasible for evaluating dental plaque.

Dental Plaque ; diagnosis ; Dental Plaque Index ; Female ; Humans ; Image Processing, Computer-Assisted ; Male ; Neural Networks (Computer) ; Photography, Dental

Objective To quantitative the changing information of estrogen receptor βgene which was in tissue and organ of sex gland during oestrus and dioestrus of Beagles, and to show the different expression situation of hypothalamus-pituitary-gonad axis during oestrus and dioestrus, and providing the basic of theory to research deeply the mechanism of heat of Beagles. Methods As the key gene in regulation reproduction, ERβgene is located in hypothalamus-pituitary-gonad axis, so using Beagles which was in oestrus and dioestrus, and extract the RNA from hypothalamus、pituitary、ovary and uterus respectively,after reverse transcription we detected the expression of ERβgene by real-time quantitative PCR.Results The expression of ERβgene mRNA from ovary、uterus、pituitary、hypothalamus of Beagles which was in dioestrus was 0.35 times, 0.17 times, 0.44 times and 0.43 times than the expression of ERβgene mRNA from ovary, uterus, pituitary, hypothalamus of Beagles which was in oestrus.Conclusion The expression of ERβgene was up-regulation in hypothalamus-pituitary-ovary axis of Beagles which was in oestrus.

Carcinoma, Hepatocellular ; pathology ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Peptide Fragments ; pharmacology ; Peptides ; pharmacology ; Tumor Cells, Cultured

The disorders of DNA and histone methylation have a close relationship with the development and progression of tumors. Epigenetic regulation is critical in maintaining the stability and integrity of the expression profiles of different cell types by modifying DNA methylation and histone methylation. However, the abnormal changes of methylation often result in the development and progression of tumors. This review summarized the theory of tumor genomic and histone methylation, detection methods of methylation and their applications, and the clinical application of methylation as biological markers and drug targets.
DNA Methylation ; Histones ; metabolism ; Humans ; Methylation ; Neoplasms ; genetics ; metabolism

As the most homologic homologue of silent information regulator 2 of yeast, Sirt1 gene is extensively expressed in mature tissues, and is rich in early embryo and reproductive cells. It is involved in the regulation of gene transcription, energy metabolism and cell aging. It promotes fat mobilization in adipocytes and glucose production in liver and regulates insulin secretion in islet beta cell. Furthermore, Sirt1 gene is an essential endogenous apoptosis inhibitor. In future, it may be used as new drug targets or applied in other disease management modalities.
Animals ; Humans ; Sirtuin 1 ; genetics ; metabolism ; physiology

BACKGROUNDAngioplasty in the unprotected left main coronary artery (LMCA) has been controversial. This study aims to evaluate the safety and clinical effectiveness of stenting, including bare metal stent and drug eluting stent (DES), for treatment of unprotected LMCA disease.

METHODSBetween September 1997 and December 2005, a total of 297 consecutive patients underwent percutanous coronary intervention (PCI) on LMCA lesions in our hospital. Their in-hospital data and clinical follow-up outcomes were analyzed and those in pre-DES "era" (group I, from September 1997 to December 2002) were compared with those in DES "era" (group II, from January 2003 to December 2004. Patients in 2005 for the time of follow-up less than one year were not included in this group).

RESULTSAltogether 368 coronary stents were successfully deployed in 295 patients. Stents failed to be implanted after balloon predilation in two patients, who received coronary artery bypass graft (CABG) successfully. Bifurcation techniques for distal LMCA executed in 206 patients (69.4%, 206/297), included crossover stenting in 156 (75.7%), T stenting in 4 (1.9%), provisional T stenting in 28 (13.6%), kissing stenting in 5 (2.4%) and stent crushing in 13 (6.3%) patients. During their hospital stay, 5 (1.7%) patients died after PCI procedure, of which 4 died from cardiac origin and one of renal failure. The total in-hospital major adverse cardiac events (MACE) were 2.0% (6/297). In the follow-up period, 19 patients (6.5%) died [15 (5.1%) of cardiac death and 4 of non-fatal myocardial infarction (MI)]. Besides, 2 (0.7%) developed subacute thrombosis (SAT) and 16 (5.4%) performed target lesion revascularization (TLR). The total follow-up MACE was 14.5% (43/297). Further analysis also showed that, compared with patients in group I, those in group II apparently had more multi-vessel involvement (14.7% vs 81.9%, P < 0.001), and more bifurcation lesions (32.4% vs 72.2%, P < 0.001). After PCI, in-hospital MACE of group II was significantly lower than that in group I (1.1% vs 9.4%, P < 0.05). And the incidences of MACE, TLR and angiographic restenosis in group II were all significantly lower than those in group I (all P < 0.05) after one year follow-up.

CONCLUSIONSAs new PCI strategies and intervention devices such as DES are developed, coronary stenting, which might have brought better in-hospital and long-term outcomes than CABG, are proved to be technically successful and can be safely applied for the treatment of LMCA lesions in the experienced center for coronary intervention.

Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Coronary Disease ; therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Stents