OBJECTIVEThis study aimed to improve students' ability in practical and theoretical courses of oral health education and to promote students' learning interest and initiative.

METHODSFourth-year students of the oral medical profession from 2006 to 2008 at Weifang Medical University were chosen as research objects for oral health education to explore the experimental teaching reform. The students were divided into test and control groups, with the test group using the "speak out" way of teaching and the control group using the traditional teaching method. Results of after-class evaluation of the test group, as well as final examination and practice examination of the two groups, were analyzed and compared.

RESULTSAfter-class evaluation results of the test group showed that the "speak out" teaching method was recognized by the students and improved students' ability to understand oral health education. The final examination and practice examination results showed that the score of the test group was higher than that of the control group (P < 0.01).

CONCLUSION"Speak out" teaching methods can improve students' ability for oral health education, in accordance with the trend of teaching reform.

Objective To investigate the action mechanism of glutamate-mediated signaling pathway in malignant melanoma. Methods WM451LU melanoma cells in log phase were classified into 6 groups, negative control group treated with PBS (100 μl), MK801 group treated with the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (100 μmol/L), CPCCOEt group treated with non-competitive metabotropic glutamate receptor 1 (mGluR1) antagonist CPCCOEt, MAP2 group transfected with adenovirus vector containing microtubule associated protein 2a (Ad-MAP2a), MK801 + MAP2 group treated with MK801 of 100 μmol/L and transfected with Ad-MAP2a, CPCCOEt + MAP2 group treated with CPCCOEt of 10 μmol/L and transfected with Ad-MAP2a. Western blot was performed to detect the expression of an ionotropic glutamate receptor, i.e., N-methyl-D-aspartate receptor type 2A (NMDAR2A) in WM451LU cells transfected with Ad-MAP2a. Scratch motility assay and cell invasion assay were conducted in vitro to detect the changes in migration and invasion ability of WM451LU cells after treated with Ad-MAP2a, MK-801, CPCCOEt alone or in combination. In vivo study was carried out to compare the inhibitory effect of the above treatments on melanoma. Results Western blot revealed a decrease in the expression of NMDAR2A in WM451LU cells after transfected with Ad-MAP2a. The scratch motility assay showed that the number of migrating cells per high power field was 117.04 ± 2.76 in MAP2 group,107.64 ± 6.50 in MK801 group,97.36 ± 4.79 in CPCCOEt group, 43.28 ± 3.02 in MK801 + MAP2 group,30.76 ± 3.97 in CPCCOEt + MAP2 group,significantly different from that in the negative control group (152.3 ± 5.75,all P ＜ 0.01 ). Cell invasion assay demonstrated that the average number of invading cells per high power field in the negative control was significantly higher than that in MAP2 group, MK801 group, CPCCOEt group, MK801+MAP2 group and CPCCOEt + MAP2 group (170.43 ±8.72 vs. 98.26 ± 3.84, 97.22 ± 5.54, 112.23 ± 7.21, 42.89 ± 5.06, 58.25 ± 6.68, P ＜ 0.05, 0.05, 0.05, 0.01and 0.01, respectively).A significant decrease was observed in the average volume of experimental melanoma in mice of MAP2 group, MK801 group, MK801 + MAP2 group, CPCCOEt group and CPCCOEt + MAP2 group compared with the negative control group (224.02 ± 46.19 mm3, 160.33 ± 33.91 mm3, 91.49 ± 21.48 mm3,202.30 ± 52.37 mm3, 111.13 ± 69.81 mm3 vs. 342.70 ± 60.92 mm3, all P ＜ 0.01 ). Conclusions To block the glutamate signaling pathway in vitro can inhibit the invasion and migration of melanoma cells, and to block the pathway in vivo can inhibit the growth of malignant melanoma and alter the morphology of melanoma cells.

Objective To breed homozygous mice of Treg-Th17 fluorescence double labeling and to investigate the role of Treg-Th17 balance in endotoxemia. Methods Mice of Treg and Th17 fluorescence single labeling were brought from Harvard Medical School,USA and were mated and bred based on the genetic rules.The genomic DNA was extracted from the tails of second generation weaning mice for genotyping by polymerase chain reaction (PCR).Thereafter,the homozygous mice of fluorescence double labeling were selected to have successive inbreeding for three generations and their genotypes and growth status were measured.The endotoxemia model in vivo was duplicated and changes of Treg-Th17 balance were detected by flow cytometry analysis at 24 h and 48 h after endotoxemia. Results Among the seven second generation mice in the first batch,only three (two males and one female) were the wanted homozygotes.Meanwhile,the genotypes of their offsprings for the next three generations were all Foxp3 RFPKI-homo IL-17A GFPKI with normal growing status.As important component of endotoxin,lipopolysaccharide (LPS)could induce significant signal expressions of red fluorescence protein (RFP) and green fluorescence protein (GFP).The ratio of CD8a + Foxp3 +,CD8a + IL-17A +,CD4 + Foxp3 + and CD4 + IL-17A + in the lymphocytes went up significantly in the group of 24 hours of LPS treatment,as compared with the control group (P ＜ 0.01 or P ＜ 0.05 ),while the difference between the control group and the group of 48hours of LPS treatment had no statistical significance (P ＞ 0.05). Conclusion Foxp3 RFPKI-homoIL-17A GFPKI mice have identical genotypes and stable genetic characteristics and is a model animal and novel research platform that can provide real-time monitoring of the changes of Treg-Th17 balance in vivo.The role of T reg-Th17 balance works significantly at 48 hous after the subject is exposed to LPS stimulation,and the roles of CD8a + Foxp3 and CD8a + IL-17A + in Treg-Th17 balance require further investigation.

Objectives To analyze the clinical features and long-term follow-up results of neuroblastoma (NB) without chemotherapy and radiotherapy, and to provide evidence for further improvement of treatment. Methods The clinical data of children diagnosed with NB who received operation during January 2005 to December 2015 was retrospectively analyzed, and the long-term follow-up results were evaluated. Results In 57 cases of NB, 43 cases (81.1％) were in stage 1, 8 cases were in stage 2 and 2 cases were in stage 4S. The median age at diagnose was 7 months (11 days - 10 years and 11 months). There were 47/51 cases had the pathological type with a good prognosis (accounting for 92.2％). FISH was detected in 1/49 case which had the amplification at greater than 10 copies. 56/57 cases underwent surgical resection of the primary tumor, 50 cases of which were completely resected, and 3 cases had very good partial remission after tumor resection. The abdominal mass was found in the uterus in 1 case, and surgical operation was not performed and the imaging was regularly checked for follow-up , and the mass subsided completely at 7-month-old. The median follow-up time was 36 months (4 - 99 months). Five children were lost to follow-up and the median time of follow-up was 19 months (4 - 45 months). One child in stage 4S relapsed at 1 year of follow-up, 2 cases in stage 1 relapsed at 6 months of follow-up. Five years event free survival rates (EFS) in all patients were 94.6％, and overall survival rate (OS) of the 5 years was 100％. Conclusions Children younger than 18 months without MYCN amplification in the stages 1 and 2 are safe by surgical treatment alone with good prognosis. Simple surgical treatment can also be extended to all age groups of NB without MYCN amplification in the stages 1 or 2.

Objective To evaluate the outcomes of children with stage Ⅳ malignant extracranial germ cell tumors. Methods Twenty-five patients were enrolled in the retrospective analysis. Event-free survival (EFS) and overall survival (OS) rates were estimated by Kaplan-Meier method with SPSS 13.0. Results Of the 25 children, there were 13 males and 12 females. The mean age at diagnosis was 2 years old (ranged 1 to 11). Five patients receiving chemotherapy in another hospital before (n=1), or giving up treatment after confirmed diagnosis (n=1), or giving up effective treatment after received less than 2 cycles (n=3) were excluded from this analysis. Of the 20 patients, 90.0% (18/20) achieved complete remission and 5.0% (1/20) achieved partial remission after treatment. The 5-year EFS rate and 5-year OS rate were 70.0%±10.2% and 82.4%±9.2% respectively. There was no death occurred due to complications. Conclusions The effect of this treatment program is positive. The cumulative dose of the drugs is not high, compared with other schemes such as PEB, but there are more drugs involved. Whether these drugs may cause long-term adverse reactions needs further research.

Objective To explore the clinical significance and treatment regimen of sentinel lymph node(SLN) micrometastases and isolated tumor cell metastasis in breast cancer.Methods Ninety-seven breast cancer patients with sentinel lymph node micrometastases or isolated tumor cell metastasis from January 2013 to December 2015 were retrospectively studied.The patients were assigned to axillary lymph node dissection group (ALND,41 cases) and non axillary lymph node dissection group(non-ALND,56 cases) according to the final surgery mode to the axilla.Disease-free survival(DFS) and overall survival (OS) were compared between the two groups.Results Neither clinico-pathological factors,such as age,tumor size,grade,ER/PR status,HER-2 gene expression,Ki-67 expression and the size of the SLN metastasis,nor the treatment,such as breast surgery,postoperative adjuvant chemotherapy,radiotherapy and hormone therapy were found statistically different between the two groups (P ＞ 0.05).There were 96 patients evaluable with a median follow up of 24 months.The DFS of the ALND and non-ALND group was 97.5％ and 96.6％ (P ＞ 0.05),and the OS was 100％ and 98.2％ (P ＞ 0.05) respectively with no difference between the two groups.There were 2 ispilateral axillary recurrence in the non-ALND group and non in the ALND group.Conclusion Axillary lymph node dissection may be omitted for the breast cancer patients with sentinel lymph node micrometastases and isolated tumor cell metastases.But the postoperative adjuvant systemic treatment should be emphasized.

Aim To study the effect of ginsenoside F1 on the enzyme activity and expression of gene of CYP3 A4 through activation of pregnane X receptor ( PXR ) . Methods With different concentrations of ginsenoside F1 treated on LS174T cells, the expression of CYP3A4 mRNA was determined by Q-PCR, and the enzyme activity was measured by P450-GloTM CYP3A4 assay according to the manufacturer′s instructions, fur-ther PXR-CYP3 A4 stable translation HepG2 cell lines were used to test ginsenoside F1 activates PXR by re-porter gene screening assay. Results The results re-vealed that the levels of CYP3 A4 gene and protein ex-pression were significantly increased by ginsenoside F1 in a concentration-dependent manner. At the same time, reporter gene screening showed that ginsenoside F1 could also enhance the transcriptional activity of PXR. Conclusion Ginsenoside F1 can significantly up-regulate the gene expression and enzyme activity of CYP3A4 via the PXR-CYP3A4 pathway.

Objective To study and summary the treatment of talus fracture. Methods Fifteen patients with talus fracture were treated by limited cut off reduction by leverage and hollow lag screw under C arm X-Ray machine January 2008 to November 2014, of whom there were 11 males and 4 females, aged 15-61 years old. Seven patients sufferred from talus fracture because of traffic accidents, 4 patients because of falls, 2 patients because of crush injury, and 2 patients because of sprains. Hawkins typing:Ⅰtype in 1 patient, Ⅱ type in 6 patients, Ⅲ type in 6 patients of Ⅲ type, and Ⅳ type in 2 patients. Results Followed up for 6 months to 4 years (mean 1.5 years), fracture healed better. Hawkins grading standard:9 patients showed excellent (1 patient ofⅠtype, 5 patients ofⅡtype, 3 patients ofⅢtype), and two patients showed good (Ⅲtype). The excellent and good rate was 11/15. Necrosis of talus occurred in one patient, and degenerative joint disease occured in one patient. Conclusions Minimally invasive internal fixation with hollow lag screw under C arm X-Ray machine in the treatment of talus fracture has a small injury to the periosteum and the blood supply of small soft tissue. It can fix reliably and can pressurize the fracture fragments, and is conducive to fracture healing.

BACKGROUND/AIMS: The Wnt/beta-catenin signaling pathway has been reported to play an important role in liver fibrosis. This study was designed to investigate whether mesoderm-specific transcript homologue (Mest), a strong negative regulator of Wnt/beta-catenin signaling, could inhibit liver fibrosis. METHODS: pcDNA-Mest was transfected into hepatic stellate cells (HSCs) and rats. Rats were randomly divided into four groups: normal group (normal saline), treatment group (pcDNA-Mest+CCl4), control group (pcDNA-neo+CCl4), and model group (normal saline+CCl4). Changes in liver pathology were evaluated by hematoxylin and eosin and Masson's trichrome staining. The levels of alanine transaminase, aspartate transaminase, lactic dehygrogenase, hyaluronic acid, and laminin in the serum and hydroxyproline in the liver were detected by biochemical examination and radioimmunoassay, respectively. The expression and distribution of beta-catenin, alpha-smooth muscle actin (alpha-SMA), Smad3, and tissue inhibitor of metalloproteinase type I were determined, and the viability of the HSCs was tested. RESULTS: Our data demonstrate that Mest alleviated CCl4-induced collagen deposition in liver tissue and improved the condition of the liver in rats. Mest also significantly reduced the expression and distribution of beta-catenin, alpha-SMA and Smad3 both in vivo and in vitro, in addition to the viability of HSCs in vitro. CONCLUSIONS: We found that Mest attenuates liver fibrosis by repressing beta-catenin expression, which provides a new therapeutic approach for treating liver fibrosis.
Animals ; Carbon Tetrachloride/toxicity ; Cells, Cultured ; Hepatic Stellate Cells/physiology ; Liver Cirrhosis, Experimental/*physiopathology ; Male ; Proteins/*physiology ; Random Allocation ; Rats, Wistar ; Transfection ; Wnt Signaling Pathway/*physiology ; beta Catenin/metabolism

A cDNA, named Dd-ace-2, encoding an acetylcholinesterase (AChE, EC3.1.1.7), was isolated from sweet-potato-stem nematode, Ditylenchus destructor. The nucleotide and amino acid sequences among different nematode species were compared and analyzed with DNAMAN5.0, MEGA3.0 softwares. The results showed that the complete nucleotide sequence of Dd-ace-2 gene of Ditylenchus destructor contains 2425 base pairs from which deduced 734 amino acids (GenBank accession No. EF583058). The homology rates of amino acid sequences of Dd-ace-2 gene between Ditylenchus destructor and Meloidogyne incognita, Caenorhabditis elegans, Dictyocaulus viviparous were 48.0%, 42.7%, 42.1% respectively. The mature acetylcholinesterase sequences of Ditylenchus destructor may encode by the first 701 residues of deduced 734 amino acids.The conserved motifs involved in the catalytic triad, the choline binding site and 10 aromatic residues lining the catalytic gorge were present in the Dd-ace-2 deduced protein. Phylogenetic analysis based on AChEs of other nematodes and species showed that the deduced AChE formed the same cluster with ACE-2s.
Acetylcholinesterase ; genetics ; Amino Acid Sequence ; Animals ; Cloning, Molecular ; DNA, Complementary ; genetics ; Genes, Helminth ; genetics ; Ipomoea batatas ; parasitology ; Molecular Sequence Data ; Nematoda ; enzymology ; genetics ; Plant Stems ; parasitology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, Protein