In the study, the effects of Panax notoginseng saponins (PNS) on alveolar epithelial to mesenchymal transition (EMT) and extracellular matrix degradation were observed in a type of human alveolar epithelial cell, A549 cells, stimulated by TGF-beta1. Firstly, MTT method was applied to evaluation of cellular proliferation and found that PNS from 12.5 mg x L(-1) to 200 mg x L(-1) dosage could not inhibit significantly cellular proliferation. Then, cells were divided into five groups, normal group, TGF-beta1 group, TGF-beta1 + 50 mg x L(-1) PNS group, TGF-beta1 + 100 mg x L(-1) PNS group and TGF-beta1 + 200 mg x L(-1) PNS group. Normal cells were not stimulatec by TGF-beta1; TGF-beta1 cells were only stimulated by TGF-beta1 and the other cells were stimulated by TGF-beta1 with different doses of PNS, respectively. After stimulation, cells and supernatants were collected for assays. Cellular roundness was applied to quantitative evaluation of morphological change. Immunocytochemistry was applied to examine E-cadherion, a-SMA and FN proteins expression in the cells. Enzyme linked-immunosorbent assay was applied to MMP-9 and TIMP-1 levels. The results showed that EMT of A549 cells was induced by TGF-beta1, showing significant change of roundness, E-cadherion, alpha-SMA and FN (P < 0.05, P < 0.01). Compared to TGF-beta1, PNS significantly inhibited the changes of roundness (P < 0.05), FN and alpha-SMA (P < 0.05, P < 0.01) and not significantly inhibited the change of E-cadherion. Furthermore, MMP-9 levels were significantly increased by TGFbeta1 stimulation (P < 0.05), without significant change of TIMP-1. Compared with TGF-beta1, PNS could significantly increase MMP-9 level (P < 0.05) and decrease TIMP-1 levels (P < 0.05, P < 0.01). In conclusion, PNS could inhibit alveolar epithelial cell EMT induced by TGF-beta1, with increase of extracellular matrix degradation ability, which showed anti-fibrosis of lung ability.
Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Panax notoginseng ; chemistry ; Pulmonary Alveoli ; cytology ; drug effects ; metabolism ; Saponins ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the inhibitory effects of OMT on TGF-β1-induced CFBs proliferation, and then explore the mechanism.

METHODThe experiment was randomly divided into 6 groups as following: control group (serum free DMEM), model group (20 μg x L(-1) TGF-β1), OMT low dose group (1.89 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT medium dose group (3.78 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT high dose group (7.56 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), SB203580 group (p38MAPK blocking agent, 1 x 10(-5) mol x L(-1) + 20 μg x L(-1) TGF-β1). Vimentin of CFBs was identified by immunocytochemical methods, α-SMA of myFBs as well. Inhibitory effects of OMT on CFBs proliferation was detected by the MTT assay. Picric acid Sirius red staining was analyzed collagen type I and collagen type III deposition. Western blot was determined the expression of p38MAPK, p-p38MAPK, collagen type I and collagen type III.

RESULTMTT results showed that OMT significantly inhibited CFBs proliferation induced by TGF-β1 (P < 0.01) α-SMA immunocytochemical experiments suggested that OMT could protect against the CFBs proliferation. OMT could significantly decrease the deposition of collagen type I and collagen type III by Western bloting and picric acid Sirius red staining. Western blot results showed that TGF-β1 enhanced p38MAPK phosphorylation, however OMT attenuated the phosphorylation of p38MAPK induced by TGF-β1 (P < 0.01).

CONCLUSIONOMT can inhibit the CFBs proliferation induced by TGF-β1, and its mechanism may be involved in inhibiting p38MAPK phosphorylation.

Alkaloids ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Collagen ; metabolism ; Down-Regulation ; Female ; Fibroblasts ; drug effects ; Heart ; drug effects ; In Vitro Techniques ; Male ; Phosphorylation ; Quinolizines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; antagonists & inhibitors ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

Standardized Manipulation of Acupuncture and Moxibustion Part 8: Intradermal Needle was compiled with the following principles. The compiling standard, technical features and clinic manipulations of intradermal needle were taken as the basic principle for compiling. Literature research, expert survey and clinic practice verification were applied as the drafting methods. The key issues were focused on the relationship between standardization and individualization, normalization and effectiveness, qualification and quantification. And the postural selection, reinforcing and reducing manipulations, fixing materials and embedding duration involved in intradermal needling were emphasized particularly. At the same time, details and the future way of thinking of intradermal needle were expounded in this article as well.
Acupuncture Therapy ; instrumentation ; standards ; China ; Humans ; Moxibustion ; standards ; Needles ; standards ; Reference Standards

It has been demonstrated that neural cell adhesion molecule (NCAM) is critical for the induction and maintenance of long term potentiation (LTP) in the CA1 region of rat hippocampus. In the present study, we investigated the changes in NCAM mRNA expression and NCAM protein level after the induction of LTP in vitro using the techniques of in situ hybridization and Western blot. The results showed that the number of NCAM mRNA positive labelled neurons significantly increased (76.6+/-11.5 neurons) 10 min after tetanus when the slope of fEPSP markedly increased. The level of NCAM protein also increased significantly (7.190+/-0.64 arbitrary unit/50 microg protein) 10 min after tetanus. The number of NCAM mRNA positive labelled neurons no longer changed (73.3+/-14.0) 1 h after tetanus, however, the NCAM protein level (9.031+/-0.71) at 1 h after tetanus was higher than that at 10 min after tetanus. Moreover, the NMDA receptor inhibitor AP-5, which blocked LTP, prevented the increase in NCAM mRNA expression and NCAM protein level. The results demonstrate that NCAM mRNA expression maintains a high level, whereas NCAM protein changes from a low level to a high level during induction and maintenance of LTP.
Animals ; Hippocampus ; metabolism ; physiology ; Long-Term Potentiation ; physiology ; Male ; Neural Cell Adhesion Molecules ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar

The present study examined the changes in 26S proteasome activity and the signal molecule mechanism regulating 26S proteasome activity in long term potentiation (LTP) in rat hippocampal slices. The results are as follows: 26S proteasome activity was 190+/-14.3 cpm/(100 microg.2 h) before tetanus, a significant increase in 26S proteasome activity (273+/-18.3 cpm/(100 microg.2 h) was found 10 min after tetanus, when the slope of fEPSP was markedly increased. Interestingly, 26S proteasome activity returned to baseline level (210+/-12.8 cpm/(100 microg.2 h) 60 min after tetanus. Moreover, the N-methyl-D-aspartate (NMDA) receptor inhibitor AP-5, which blocked LTP, prevented the increase in the 26S proteasome activity. The results suggest that NMDA receptors contribute to the transient increase in 26S proteasome activity during induction of LTP in the hippocampal CA1 region.
Animals ; Hippocampus ; enzymology ; physiology ; Long-Term Potentiation ; physiology ; Male ; Peptide Hydrolases ; metabolism ; Proteasome Endopeptidase Complex ; Rabbits ; Rats ; Receptors, N-Methyl-D-Aspartate ; metabolism

OBJECTIVETo investigate the histological features of oral leukoplakia which underwent malignant transformation and its correlation with the clinical manifestation.

METHODSA total of 1832 cases of oral leukoplakia were reviewed and the clinicopathological characteristics of malignant transformation were analyzed.

RESULTSMalignant transformation occurred in 85 cases (4.6%) of the 1832 cases. Thirty cases (2.1%) of 1404 cases with simple epithelial hyperplasia had malignant transformation. Fifty-five cases (12.9%) in 428 cases with epithelial dysplasia were transformed to malignancy, especially in the cases with moderate or severe dysplasia, in which the ratio of malignant transformation was higher than in the cases with simple epithelial hyperplasia (P < 0.005). Clinical parameters associated with an increased risk of malignant transformation were female gender and epithelial dysplasia was more often seen in non-homogenous leukoplakias than in homogenous (P < 0.005).

CONCLUSIONSNon-dysplastic leukoplakia may become malignant. Epithelial dysplasia was associated with an increased risk of malignant transformation. Leukoplakia in female may be at a higher risk for malignant transformation.

Adult ; Aged ; Cell Transformation, Neoplastic ; Female ; Humans ; Hyperplasia ; Leukoplakia, Oral ; pathology ; Male ; Middle Aged ; Mouth Mucosa ; pathology ; Mouth Neoplasms ; pathology ; Precancerous Conditions ; pathology ; Retrospective Studies ; Risk Factors ; Sex Factors

Adolescent ; Biopsy ; Cystic Adenomatoid Malformation of Lung, Congenital ; diagnosis ; diagnostic imaging ; therapy ; Diagnosis, Differential ; Diagnostic Errors ; adverse effects ; prevention & control ; Humans ; Infant ; Lung Diseases ; diagnosis ; diagnostic imaging ; therapy ; Male ; Middle Lobe Syndrome ; diagnosis ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome

Objective To detect EIA factor expression in human rectal cancer and normal tissue and to determine whether it is correlated with invasion and metastasis of human rectal cancer. Methods Real- time RT-PCR was used to detect E1AF expression in matched rectal cancers and normal tissues from g6 in- patients.Results Among the 86 rectal cancer samples tested,55 cases E1AF mRNA overexpression was ob- served. The mRNA expression of E1AF in the sample group was remarkably different from that in the control group.In carcinomas,E1AF mRNA expression correlated significantly with histological type,depth of inva- sion,lymph node and distant metastasis and advanced Duke stage.Conclusion E1AF is correlated signifi- cantly with invasion and metastasis of human rectal cancer and may be an important factor in the invasion and metastasis.

OBJECTIVESTo study the effect of Houxue Zhitong decoration on the expression of the mitochondria-initiated apoptosis pathway and transmembrane protein I an II of the epidural scar tissue.

METHODSA total of 60 New Zealand rabbits (weight: 2.5-3.0 kg) were randomly divided into four groups, sham operation group (D, n=15), control group (B, n=14), sodium hyaluronate group (C, n=15), Houxue Zhitong decoration group (D, n=15). Except for group A, 1.0 cm x 1.0 cm dura mater uncovered area laminectomy was performed at I (4) and I(5), covered with 0.5 ml sodium hyaluronate in group C, covered with same amount of saline in group B and D. First 2 weeks after operation, animals in group D were lavaged with 2.5 ml/kg Houxue Zhitong decoction by one a day for 14 days. Five rabbits of each group selected randomly were killed in the 2,4,8 weeks after laminectomy. The specimens were prepared for determination of the expression of Fas and FasL, at scar tissue by semiquantitative reserve transcription-polymerase chain reaction (RT-PCR). The degree of scar adhesion was evaluated according by Rydell method.

RESULTSThe adhesion area in group B was larger than of group C and D in the 4th and 8th week. However, the number of fibroblasts and inflammantory cells in group D was the least among the three groups in the 8th week. At 2, 4, 8 weeks after operation, as compared with group B the expression of Fas, FasL of group C and D were decreased (P < 0.05). Especially, at 2 weeks, as compared with group B the expression of this two cytokines of group D was significant decreased (P < 0.05), too. In group C and D the duramater adhesion was decreased (P < 0.05). The proliferation of fibroblast and fibroblastic function were inhibited (P < 0.05).

CONCLUSIONHuoxue Zhitong is able to down-regulated the expression of Fas, FasL, which inhibited the proliferation of fibroblast, the fibroblastic function and the synthesis of extracellular matrix in the epidural scar tissue. It is an effective way of reducing peridural scar formation and prevent the failed back surgery syndrome.

Animals ; Apoptosis ; drug effects ; Cicatrix ; genetics ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Epidural Space ; pathology ; Fas Ligand Protein ; genetics ; metabolism ; Female ; Gene Expression Regulation ; drug effects ; Rabbits ; Signal Transduction ; drug effects ; fas Receptor ; genetics ; metabolism

OBJECTIVETo study the effect of Huoxuezhitong recipe on the pathway of mitochondrial apoptosis and the expression of related gene (Caspase-3, -9, Bax, Bcl-2) in epidural scar tissue.

METHODSA total of 60 New Zealand rabbits, ranging in weight from 2.5 to 3.0 kg, were randomly divided into sham operative group (group A), control group (group B), sodium hyaluronate group (group C), Huoxuezhitong recipe group (group D, included Danggui 20 g, Chishao 20 g, Honghua 20 g, Ruxiang 15 g, Myrrh 15 g etc.) with 15 rabbits in each group. Except for group A, L4,5 vertebral plate were resected leading to 1.0 cm x 1.0 cm dura mater exposed area, which were covered with 0.5 ml sodium hyaluronate in group C, covered with same amount of saline in group B and D. At 2 weeks after operation, the rabbits of group D were administered with Huoxuezhitong recipe (2.5 ml/kg, once a day, continuous 14 d). Five rabbits of each group were selected randomly and were killed at 2, 4 and 8 weeks after operation; and the expression of Caspase-3, -9, Bax, Bcl-2 in scar tissue were surveyed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe results by RT-PCR showed significant different in expressions of Caspase-3, -9, Bax, Bcl-2 of group A and others groups at each period (P < 0.01). At 2, 4, 8 weeks after operation, compared with group B, the expressions of Caspase-3, -9, Bcl-2 in group C and D were decreased (P < 0.05), but the expression of Bax in group C and D were increased (P < 0.05). At 8 weeks after operation, the expressions of Caspase-3, -9, Bax, Bcl-2 in group D was significant decreased compared with group C (P < 0.05).

CONCLUSIONHuoxuezhitong recipe can regulate the levels of expression of Caspase-3, -9, Bax, Bcl-2 mRNA and further induce the pathway of mitochondrial apoptosis, thereby, achieve the effect of prophylaxis for the proliferation and conglutination of epidural scar tissue.

Actins ; genetics ; metabolism ; Animals ; Apoptosis ; drug effects ; genetics ; Carthamus tinctorius ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Genes, bcl-2 ; drug effects ; genetics ; Mitochondria ; drug effects ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-2-Associated X Protein ; genetics ; metabolism