Objective:To investigate the effect of human p27~(kip1) gene on the proliferation of human tongue squamous cell carcinoma(TSCC) Tca8113 cells.Methods:Human p27~(kip1) gene was cloned from normal tissue and reconstructed into eukaryotic expression vector pcDNA3, the recombinant plasmid was transfected into Tca8113 cells with liposome.The integration and expression of exogenous objective gene in the target cells were examined by PCR and flowcytometry,the growth of transfected cells and non-transfected cells was investigated by MTT assay and flowcytometry.Results:p27~(kip1) cDNA was successfully cloned and reconstructed into vector pcDNA3.The gene was trasfected into Tca8113 cells and stable transfection was achieved by screening with G418.After p27~ kip1 transfection the cell proliferation was inhibited,proliferation index decreased(P

Objective To explore the relationship between serum adiponectin (APN) concentration and insulin resistance(IR) in elderly pregnancy patients(GDM) with gestational diabetes mellitus.Methods Data of 120 cases of elderly pregnancy patients with gestational diabetes mellitus admitted from Jan.2011 to Dec.2016 were retrospective analyzed.The relationship between serum APN concentration and IR was assayed.Healthy subjects and normal pregnancy patients in our Department were taken as the control.Results The concentration of APN was lower while the CysC,BNP,IGF1,IGF2 and IGFIR were higher in elderly pregnancy patients with gestational diabetes mellitus than those in the control group with a significant difference (P＜0.05).At the same time,Spearman analysis results showed that the serum APN was negatively related with IR while CysC,BNP,IGF1,IGF2 and IGFIR were positively related with IR (P＜0.05).Conclusion Theserumconcentrationofadiponectin and visfatin were negatively related with insulin resistance in elderly pregnancy patients with GDM.

Objective To study the expression of vascular endothelial growth factor(VEGF) in carcinoma of cheek in order to investigate the relation with carcinogenesis and development of carcinoma of cheek.Methods The expression levels of VEGF were determined by S-P method of immunohistochemistry technique in 40 tissues of carcinoma of cheek,30 para-carcinoma and 10 normal mucosa.Results The expression rates of VEGF in the tissues of carcinoma of cheek,para-carcinoma and normal mucosa were 85.0%,26.7%,10.0%,respectively.There was remarkable difference in the statistic(?~2=32.26,P

This paper describes the anti-inflammatory, antipyretic and analgesic effects of Sudoxicam ( Sud) synthesized by Shenyang Pharmaceutic College.Sud is found to possess significant anti-inflammatory, antipyretic and analgesic quality in such animal models as carrayeenan-induced paw oedema ( ED50=5.26mg/kg ) , tylolinduced ear oedema in mice, adjuvant-induced arthritis in rats, yeast-induced fever in rats, acetic acid-induced arithing ( ED50= 5.0mg/kg ) and hot plate-induced pain reaction in mice.

ObjectiveTo investigate the effect of the displacement of the selected silver clip in the different respiratory state achieved by active breathing control ( ABC ) system on the displacement of the geometry constituted by all of the silver clips at the border of the cavity in external-beam partial breast irradiation (EB-PBI).MethodsTwo sets of CT images in state of moderate deep inspiratory breathing hold (mDIBH),deep expiratory breathing hold (DEBH),and free breath (FB) were acquired in the same CT simulation assisted by ABC system for each of the 27 patients after breast conservative surgery.All silver clips in the cavity were delineated based on each set of CT images.Thereafter,the irregular geometry based on the silver clips as the vertices was automatically formed.Four selected clips located at the top,bottom,lateral border and medial border of the cavity were correspondingly manually registered based on automatic registration of the CT images acquired in the same or different state of respiration including mDIBH,FB,and DEBH.The displacement of center of the geometry in the direction of right-left (RL),anterior-posterior (AP),and superior-inferior (SI) separately based on automatic registration and manual registration was evaluated.The difference of the displacement was analyzed by Kruskal-Wallis H-test and Kolmogorov-Smirnov Z-test.Results When registered between mDIBH and mDIBH,FB and FB,DEBH and DEBH,the differences of the displacement of the center of geometry were not statistically significant (H =0.00 - 1.76,P=0.184-0.954). When registered between mDIBH and DEBH,the differences were statistically significant ( Z =11.31 - 23.00,P =0.000 - 0.001 ).There were statistically significant differences in the displacement of geometry center based on the selected silver clip between different registration forms in AP and SI directions (Z=4.76-25.54,P=0.000-0.029).ConclusionsThe difference of intrafraction displacement of the geometry constituted by the clips between the same respiratory states in the three dimensional direction is not statistically significant,but the difference is statistically significant between the different respiratory states in AP and SI directions.

OBJECTIVE: To evaluate the therapeutic effects of Chinese materia medica in treating patients with different syndromes by tongue image analysis software 1.0 based on tongue colors, and to discuss the feasibility of applying this computer science-based techniques into drug evaluation. METHODS: The tongue colors and the areas of tongue fur were examined and analyzed by the tongue image analysis software 1.0 in healthy persons and the patients with different syndromes before and after treatment. The parameters of tongue colors consisted of the followings: the hue (H), the lightness (L), the saturation (S), and the values of red (R), green (G) and blue (B). RESULTS: Obvious differences could be revealed in tongue color index between the healthy persons and the patients in five groups of different syndromes. There also existed some significant differences in those index between patients before and after treatment. CONCLUSION: The tongue image analysis software 1.0 based on tongue colors is helpful to evaluate the therapeutic effects of Chinese materia medica.

OBJECTIVE: To investigate the effect of mevastatin (Mev) on the expression of peroxisome-proliferator-activated receptor-gamma (PPAR-gamma) mRNA and differentiation of Thyroid-associated ophthalmopathy (TAO) derived orbital preadipocytes in vitro. METHODS: Orbital adipose tissues were obtained from TAO patients undergoing orbital decompression surgery. The orbital preadipocytes cultured from the orbital adipose tissues were divided into Group A (a control group) and Group B (an intervention group). Group B was subdivided into Group B1-B5, all groups were stimulated to differentiate into mature adipocytes with cocktail differentiation medium.The entire course of differentiation was 10 d. The differentiation of orbital preadipocytes in Group A was induced with routine inducer,while at in Group B1,B2, and B3 was interfered with 5 micromol/L (B1), 10 micromol/L(B2),20 micromol/L (B3) mevastatin respectively during the whole process of differentiation. The differentiation of orbital preadipocytes in Group B4 and B5 was interfered with 10 micromol/L mevastatin day 4 (B4) or day 8 (B5) of the differentiation process until the entire course was over. Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining. The value of optical absorption was measured at 492 nm with enzyme-linked immunosorbent assay. The expression of PPAR-gamma mRNA was detected by reverse transcription polymerase chain reaction. RESULTS: The light absorption value (A) and PPAR-gamma mRNA expression of differentiated cells in Group A,B1,B2,and B3 decreased successively,and there was significant difference in any of the 2 groups among Group A, B1 and B2, and B3 (P<0.05). The value A and PPAR-gamma mRNA expression of differentiated cells in Group A, B4, and B2 decreased successively, and the difference in any of the 2 groups among these 3 groups was significant. However, there were no significant difference between Group A and B5. CONCLUSION Mevastatin inhibits the differentiation of TAO derived orbital preadipocytes by blocking PPAR-gamma mRNA expression. The degree of inhibition is not only concentration-dependent but also associated with the stage of differentiation. The earlier the differentiation, the stronger the inhibition.
Adipocytes ; pathology ; Adipose Tissue ; pathology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Depression, Chemical ; Graves Ophthalmopathy ; pathology ; Humans ; Lovastatin ; analogs & derivatives ; pharmacology ; Orbit ; PPAR gamma ; antagonists & inhibitors ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism

Objective: To explore the feassibility to edit human p53 and PTEN, two tumor suppressor genes, by CRISPR-Cas9 technology and to evaluate the editing efficiency in vitro, and to provide the experimental study tools for transforming primary healthy cells into malignant tumor cells and establishment of humanized mouse models with human oncogenesis in vivo Methods: The smgle-gmde RNA (sgRNA) sequences were designed to target the common exon regions of p53 and PTEN mRNA isoforms based on software analysis, that could predict their gene editing efficiency. The sgRNAs with high scores were selected and cloned into Cas9-P2A-GFP plasrmd to construct sgRNA-Cas9-P2A-GFP vector that co-expressed sgRNA, Cas9 and GFP. The 293T cells in logarithmic growth phase were transfected with the sgRNA-Cas9-P2A-GFP vector (experimental group) or PBS (control group) by Lipofectamine 2000. After two-week expansion, the GFP-positive 293T cells were purified by flow cytometric sorter, whose genomic DNA was extracted for further analysis. The DNA fragments containing the sgRNA targeting site were amplified from the extracted genomic DNA by PCR and purified by gel extraction. Then they were linked into the pEASY-Blunt Zero cloning vector and transformed into competent E. coli cells. The single colonies formed by pEASY-Blunt Zero vector transformed cells were used to extract the plasmid for DNA sequencing. And the sequencing results of control group and experimental group were compared to judge the gene editing efficiency. Results: Over 82% of the sgRNA-Cas9-P2A-GFP transfected cells were found to express GFP gene after flow sorting in experimental group, which was significantly higher than that of the pre-sorted cells (P< 0.05). Genomic DNA was extracted from the sorted cells after expansion and used as PCR template. The length of the amplified fragments containing the p53 mutation site was 612 bp, while the lengths of the amplified fragments containing the PTEN-1/PTEN-2 mutation site were 667 and 947 bp. The results of sequencing showed that the efficiency of editing induced by p53-l, p53-2 and PTEN-2 sgRNA were 54.5%, 45.5% and 33.3%, respectively. Conclusion: The sgRNAs p53-1, p53-2 and PTEN-2 designed for p53 and PTEN can successfully guide Cas9-mediated site-specific genome editing with high efficiency at the genome level.

ObjectiveTo explore the feasibility,efficacy and cosmetic effect of three-dimensional conformal external beam partial breast irradiation (EB-PBI) after breast-conserving surgery for the selected Chinese early stage breast cancer patients.MethodsFrom June 2003 to December 2010,Forty-four early stage breast cancer patients underwent EB-PBI after breast-conserving surgery.Twenty patients had CT simulation scan in moderate deep inspiration breathing hold,and twenty-four patients in free breathing.EB-PBI was planned and delivered by three-dimensional conformal radiotherapy (3DCRT)with four noncoplanar beams.The prescribed dose was 3.40 Gy per fraction in thirty-nine patients and 3.85 Gy per fraction in five patients,twice per day at an interval of at least six hours,in five consecutive days.Results The number of patients with follow up time of 2,3 and 5 years were 39,31 and 16.Grade 1 acute radiationinduced dermatitis was observed in 17 patients (39％) at three months.Cosmesis was good or excellent in all cases at six months after radiotherapy and in 95％ cases at two years after radiotherapy.The 2-,3-and 5-year local control rates were 100％,99％ and 94％,respectively.The 2-,3-,and 5-year survival rates were all 100％ and no metastases occurred.Conclusions EB-PBI delivered by 3DCRT is feasible for selected Chinese early stage breast cancer patients after breast-conserving surgery.The cosmetic effect,local control rate and long-term survival rate are satisfactory,and acute radiation toxicity is very low.

OBJECTIVE: To discuss the method and opportunity of operation for orbito-cranial fibrous dysplasia, and further to summarize our experience in repairing the bony defect with titanium after excision and assess the surgical outcomes. METHODS: A retrospective study was performed in 21 patients with visual function damage and/ or orbital malformation, who treated surgically. All patients were underwent CT examinations (coronal, horizontal, sagittal and three-dimensional scans) before surgery. CT image data of patients with serious orbital skull lesions were analyzed by computer to produce the three-dimensional, solid titanium mesh prostheses used to repair the lesions; for smaller lesions solid titanium mesh prostheses were shaped artificially during surgery according to the normal orbit and the cranium. All the patients were treated surgically via craniofacial approach. After removing the diseased tissue, the defective area was repaired by titanium plate and nets. RESULTS: Of the 21 patients, tumor removal was complete in 18 patients, and incomplete in 3 patients due to extensive invasion into the sious cavernosus. Of the 13 patients with vision impairment before surgery, vision was improved in 11 cases (range from 3 to 5 lines) following surgery. All orbital malformations were healed in these 11 patients. CONCLUSION Patients with fibrous dysplasia should undergo surgical treatment as soon as possible. Where this results in visual dysfunction or orbital-cranial deformities, repairing the bony defect with titanium material has many advantages: it is solid, easily molded, and easily fastened.
Adolescent ; Adult ; Aged ; Female ; Fibrous Dysplasia of Bone ; surgery ; Humans ; Male ; Middle Aged ; Orbit ; Prostheses and Implants ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Skull ; Titanium ; Young Adult