In order to investigate the role of Twist gene in the metastasis of hepatocellular carcinoma (HCC), total RNA was respectively extracted from three HCC cell strains with different metastatic potentials, HepG2, MHCC-97L and MHCC-97H. The first strand cDNA was synthesized by reverse transcription, which was then used as template to perform fluorescent quantitative polymerase chain reaction (FQ-PCR). The quantity of Twist gene expression was normalized by that of the housekeeping gene, GAPDH for each sample. ANOVA was used to estimate the relationship between Twist gene and metastasis potential of HCC. The results showed that the normalized initial cDNA concentrations of Twist gene in HepG2, MHCC-97L and MHCC-97H were (9.45+/-0.25)x10(-4), (1.82+/-0.41)x10(-3), (3.06+/-0.62)x10(-3), respectively. FQ-PCR revealed significant differences in the expression level of Twist among HCC cell strains with different metastatic potentials. It was concluded that high expression level of Twist was closely associated with more aggressive behaviors of HCC. Twist provides a novel indicator for HCC metastasis.
Carcinoma, Hepatocellular/*metabolism ; Cell Line, Tumor ; DNA, Complementary/metabolism ; Dose-Response Relationship, Drug ; Gene Expression Regulation, Neoplastic ; Liver Neoplasms/*metabolism ; Neoplasm Metastasis ; Nuclear Proteins/*biosynthesis ; Polymerase Chain Reaction ; RNA/metabolism ; Twist Transcription Factor/*biosynthesis

Contraindications ; Humans ; Laryngeal Neoplasms ; classification ; pathology ; surgery ; Laryngectomy ; methods ; Neoplasm Staging

Objective To assess the nutritional risk of patients with inflammatory bowel disease (IBD).Methods The nutritional status of 112 IBD patients from PUMC Hospital were evaluated by Nutritional Risk Screening 2002 (NRS 2002).Using Chinese standard,a body mass index (BMI) that was lowered than 18.5 kg/m~2 according to clinical material was regarded as malnutrition,and the score was recorded as 3.The se-verity of ulcerative colitis (UC) was evaluated using True-Love criteria as mild,moderate,and severe.Crohn's disease (CD) was evaluated using Harvey-Bradshaw Index as in remission stage (≤4),moderately active stage (4-8),and severely actively stage (≥9).Results All these 112 patients,including 70 UC cases and 42 CD cases,were evaluated by NRS2002.Forty-five patients (40.2%) were judged as at the risk of malnutrition.The incidence of malnutrition was significantly higher in CD patients than in UC patients (52.4% vs.32.9%,P<0.05).It was also significantly correlated with the severity of disease.In patients with severe IBD,the rate of mal-nutrition evaluated by BMI according to clinical material was significantly lower than the rate of nutritional risk.Con-clusions The risk of malnutrition is high in IBD patients.NRS 2002 can be used for nutritional risk screening.

BACKGROUND:Current decelularized methods have the certain damage to the extracelular matrix and reduce the biomechanical properties of acelular scaffolds. OBJECTIVE:To explore the biomechanical properties of decelularized scaffold of lyophilized bovine tendon. METHODS:Sixty lyophilized fiber bundles from fresh flexion tendon of calf toes were randomly divided into two groups: control group and experimental group. In the experimental group, serine protease inhibitors were placed asepticaly for 24 hours at room temperature, then the samples were rinsed with PBS and transferred to the low concentration of trypsin+ethanol mixed solution to remove the cel wal without destruction of the extracelular matrix at room temperature for 5 hours; after that, the fiber bundles were cultured in DNA enzyme solution for 5 hours, finaly the acelular scaffold was completed and rinsed with PBS for 48 hours and dried at room temperature in sterile room. No treatment was done in the control group. Modulus of elasticity, durability and maximum stress were determined in the two groups. RESULTS AND CONCLUSION:Similar elastic modulus and durability were found in the two groups, but the maximum stress in the experimental group was significantly lower than that in the control group (P < 0.01). These findings indicate that the lyophilized acelular tendon fibers can mimic the biological function of bovine tendon fibers to a certain extent.

Objective To investigate the effects of leptin on Th17 cells and the possible mechanism. Methods The leptin-deficient ( ob/ob) mice and their homologous wild-type mice were used in the study.The percentages of Th17 cells in peripheral blood samples, spleen tissues and lymph nodes were measured by flow cytometry ( FCM) analysis.The splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by using magnetic beads,were respectively cultured with leptin at various concentrations and with anti-leptin neu-tralization antibody to evaluate the effects of leptin on Th17 cells.The quantitative real-time PCR was performed to analyze Th17 cell-related cytokines at transcriptional levels.The levels of IL-6 and IL-17A in the supernatants of CD4+T cell culture were measured with Luminex technology.Results Compared with the wild-type mice, the ob/ob mice showed lower percentages of Th17 cells in both peripheral blood samples and spleen tissues (0.49%±0.03%vs 1.29%±0.1%, 1.56%±0.22%vs 2.47%±0.11%).There was a decrease in the percentages of Th17 cells upon the in vitro treatment of CD4+T cells from wild-type mice with anti-leptin antibody.The per-centages of Th17 cells were increased in a dose-dependent manner upon the in vitro treatment of CD4+T cells from ob/ob mice with leptin.Moreover, the levels of IL-17A and IL-6 and the transcriptional levels of RORγt, IL-17A and IL-6 in leptin deficiency group were lower than those of wild-type group, but were increased upon the treatment with leptin.No significant difference with the transcriptional levels of TGF-βand IL-23 was ob-served between the two groups with and without intervention.Conclusion Leptin deficiency seriously hampered the generation of Th17 cells in mice and resulted in a decreased expression of RORγt, IL-17A and IL-6 at mRNA level.The treatment of CD4 T cells with leptin might promote the generation of Th17 cells through up-regulating the transcription of RORγt and IL-6.

Objective To review the clinical,histological and diagnostic aspects of 12 documented cases of nodular regenerative hyperplasia of the liver(NRHL),to make this condition be understood and dealt with better. Method Twelve NRHL cases were diagnosed based on liver biopsy from 300 portal hypertension patients who had been underwent splenectomy.Imaging studies were performed as part of the diagnostic evaluation.Clinical manifestation and biochemical tests were recorded at the time of diagnosis.Management and prognosis were also reviewed.Results Most patients were complicated with autoimmune disease,6 cases were diagnosed systemic lupus erythematosus and 1 was Crohn's disease and 1 suspected ulcerative colitis.Six cases were treated by prednisone and 3 cases by immunosuppressant.Eleven cases had suffered from portal hypertension.All cases had no history of viral hepatitis.Biochemical tests showed mild increase of liver enzyme and relative normal synthetic liver function.The histological finding was nodular in the hepatic parenehyma,with mild periportal fibrosis,intraportal lymphocytic infiltration,narrow and obstruction of branch of portal vein,and lack of hepatocyte necrosis.All cases were diagnosed liver cirrhosis and portal hypertension before operation.Management was directed to portal hypertension and varices bleeding with satisfactory results.Most of them keep a stable condition during the follow-up. Conclusion The NRHL was uncommon and its cause and pathogenesis was unclear,may be related with immune and hepatic blood circulation disorder.It should be considered in patients with unexplained portal hypertension and distinguished it from liver cirrhosis.Liver biopsy confirms the diagnosis.Management directed to portal hypertension may improve clinical condition.

Objective To investigate the alteration of serotonin-producing gastric enterochromaffin (EC)cell in patients with functional dyspepsia(FD).Methods Fifteen healthy volunteers and 33 patients with FD were enrolled.Proximal gastric mucosal EC cells were countered after immunohisto- chemistry staining.The ultrastructure of EC cell was observed by electromicroscope.Results The EC cells in proximal gastric mucosa in patients with FD were significant higher than that in controls(12.5?2.1 vs 8.3?1.4,t=2.353,P＜0.05),and the staining intensity of EC cell in patients with FD was also stronger than that in controls(3.72?0.42 vs 2.61?0.57,t=2.078,P＜0.05).The more sever the gastric mucosal inflammation was,the more number of EC cells and the stronger staining intensity were.Under the electromicroscopy,more Golgi apparatus,mitochondria and endoplasmic reticulum were found in EC cells.Special secreting particles were also found in cytoplasm.Conclusions EC cells may be involved in the pathogenesis of FD.The number of EC cell is related with the severity of gastric mucosal inflammation.

Objective To retrospectively analyze the treatment outcomes and failure patterns in patients with head?and?neck cancer of unknown primary head?and?neck cancer of unknown primary, and to compare the efficacy between elective mucosal irradiation and ipsilateral neck treatment. Methods The clinical data of patients with head?and?neck cancer of unknown primary who were admitted from January 2007 to December 2013 were retrospectively collected. Thirty?one patients received elective pharyngeal mucosal irradiation and 61 patients only received ipsilateral neck treatment. The SPSS 19. 0 software was used for comparison of the survival and local control between the two groups. Results In the 92 patients, the median age was 57 years;79. 3% had metastasis to level Ⅱ lymph nodes;the median follow?up time was 36. 5 months;the 3?year overall survival, mucosal control, and neck control rates were 89. 0%, 86. 6%, and 82. 4%, respectively. Primary sites were found in 15 patients, containing nasopharynx in 4 patients, oropharynx in 3 patients, oral cavity in 3 patients, throat and hypopharyngeal part in 3 patients, maxillary sinus in 1 patient, and esophagus in 1 patient. The patients undergoing elective pharyngeal mucosal irradiation had significantly higher 3?year mucosal control and neck control rates than those undergoing ipsilateral neck treatment ( 100% vs. 74. 9%, P= 0. 040;87. 5% vs. 62. 2%, P= 0. 037 ) . There was no difference in the 3?year overall survival rate between the two groups ( 83. 5% vs. 88. 7%, P= 0. 910 ) . Conclusions For patients with head?and?neck cancer of unknown primary, elective pharyngeal mucosal irradiation can reduce the incidence of primary site and increase the neck control rate. A new standard for target volume delineation should be established as soon as possible for elective prophylactic pharyngeal mucosal irradiation.

Objective To investigate the diagnostic values of serum free fatty acids (FFA) and urine podocalyxin (PCX) in the patients with type 2 diabetic nephropathy(DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) were enrolled,and they were divided into three groups,normal albumin group(NA),microalbuminuria group(MA) and heavy albuminuria group(HA),based on the urinary albumin-creatinine ratio(UACR).In addition,40 healthy persons participated in medical examination were selected as the control group.Serum FFA levels were detected by a biochemical analyzer and urine PCX levels by ELISA.The correlation between them was analyzed by Pearson correlation,and their diagnostic values in type 2 DN were evaluated by the receiver operating characteristic curve (ROC).Results Serum FFA levels in the NA,MA,HA and control groups were (0.61 ± 0.14),(0.81 ± 0.13),(0.95 ± 0.18) and (0.49 ± 0.11) mmol/L,respectively,and urine PCX levels were (1.86 ± 0.45),(4.47 ± 1.48),(6.72 ± 1.40)and(1.38 ±0.24) ng/mL,respectively.The levels of serum FFA and urine PCX in type 2 DM patients were significantly higher than those in the control group(P ＜ 0.05),and increased with the progression of type 2 DM.Pearson correlation analysis showed that serum FFA levels were positively correlated with urine PCX levels(r =0.73,P ＜ 0.05).The sensitivity of serum FFA,urine PCX and combined detection in the diagnosis of type 2 DN were 74.1％,80.6％ and 89.5％,respectively,and the latter significantly higher than the former two (P ＜ 0.05).Conclusion Serum FFA and urine PCX levels increase significantly in diabetic patients with renal injury,and both of them have important clinical values in the diagnosis of type 2 DN.

Toresolve the problems in current physiologyexam,this paper proposed the measures in examination reform of physiology,which would provide reference for better training innovative medical personnel with a good overall quality to meet the needs of society.