Objective:To investigate the relationship between N-myc downstream-regulated gene-1 (NDRG-1) gene expression and the progression and metastasis of uterine cervical squamous cell carcinoma (UCSCC). Methods: NDRG-1 protein expression was detected by immunohistochemistry (IHC) in 42 cases of UCSCC, 30 cases of high-grade squamous intraepithelial lesion (HSIL), 28 cases of low-grade squamous intraepithelial lesion (LSIL), and 20 cases of normal squamous epithelium. NDRG-1 mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in 10 cases of UCSCC and 10 cases of normal squamous epithelium. Results: The expressions of NDRG-1 mRNA and protein in UCSCC were significantly lower than normal squamous epithelium tissues (P < 0.01). From normal squamous epithelium to LSIL, HSIL and UCSCC, NDRG1 protein was gradually decreased during tumor carcinogenesis. The difference was significant (P < 0.01). NDRG-1 mRNA and protein in metastatic UCSCC were obviously lower than non metastatic UCSCC (P < 0.05). Conclusion: Down-regulation of NDRG1 mRNA and protein in UCSCC indicate that NDRG1 gene might correlate with carcinogenesis, progression, and metastasis of cervical squamous carcinoma.

The nitrate-N content in KNO3 solution and soil was rapidly predicted using techniques of mid-infrared spectroscopy, in which 15 NO-3 and 14 NO-3 were distinguished and predicted. The results showed that the characteristic band of nitrate in solution and soil was 1200-1500 cm-1 , and compared with 14 NO-3 , the red shift of characteristic band of 15 NO-3 was about 35 cm-1 . In the characteristic band of nitrate, absorption band increased with the nitrate nitrogen concentration with less interference absorption. The linear regression was made between the first principal component of characteristic band and nitrate-N content, and correlation coefficient was more than 0 . 9840 , indicating that the technique of mid-infrared attenuated total reflectance spectroscopy could be applied for rapid monitoring of nitrate in solution and soil. Meanwhile, based on the red shift characteristic of 15 NO-3 absorption band, the method of partial least squares were involved to predict the nitrate-N of different N-isotope labeled in solution and soil, resulting that all the prediction models reached excellent levels. For 14 NO3-N and 15 NO3-N in solution, the correlation coefficients ( R2 ) were 0. 9980 and 0. 9982 respectively, and ration performance to standard deviations ( RPD ) were 6. 44 and 4. 76, respectively. While for 14 NO3-N and 15 NO3-N in soil, the correlation coefficients ( R2 ) were 0. 9794 and 0. 9679, and RPD were 5. 75 and 4. 78, respectively. Therefore, the technique of mid-infrared attenuated total reflectance spectroscopy can be applied for rapid monitoring different N-isotope labeled nitrate in solution and soil, to provide a new in situ and fast time method to study nitrification process in soil.

Objective To detect the expression level of aquaporin 8 (AQP8) in patients with functional constipation(FC) or constipated irritable bowel syndrome (IBS-C),and the correlation between the expression of AQP8 and clinical features.Methods From March to December 2014,a total of 16 patients with IBS-C and 19 patients with FC met Rome Ⅲ criteria were collected,and nine healthy individuals were assigned to control group at the same period.The ascending and decending colonic tissues mucosa of FC,IBS-C and control group were taken under endoscope.The expression of AQP8 at mRNA and protein level was detected by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC).The differences in AQP8 mRNA expression and AQP8 relative area were analyzed by Kruskal-Wallis test among groups,and Pearson correlation coefficient was performed for correlation analysis between the expression and clinical features.Results The relative expressions of AQP8 mRNA of ascending colon and descending colon of FC patients (1.38,0.61 to 4.09;2.65,0.82 to 7.52) and IBS-C patients (2.23,0.82 to 4.67;1.35,0.51 to 2.03) were higher than those of control group (0.56,0.19 to 0.97;0.38,0.21 to 1.19),and the differences were statistically significant (ZFc =-2.435,-3.149,ZIBS-C =-2.690,-2.152;all P＜0.05).AQP8 mRNA expression of descending colon in patients with FC was higher than that of patients with IBS-C,and the difference was statistically significant (Z =-2.003,P =0.045).The expression of AQP8 in patients with FC and IBS-C was positively correlated with disease course (ascending colon r=0.57 and 0.53;descending colon r=0.49 and 0.54,all P＜0.05),and was negatively correlated with frequency of defecation (ascending colon r=-0.82 and-0.61;descending colon r=-0.49 and-0.53,all P＜0.05).There was no correlation between the expression of AQP8 and age,gender,onset age,presence of abdominal symptoms of the patients (all P＞ 0.05).Most of AQP8 of FC group was expressed in cytoplasm of colonic mucosa epithelial cells,while that of IBS-C group and control group was mostly expressed at apical membrane and basal membrane of epithelial cells.The results of semi-quantification demonstrated that AQP8 relative area of descending colon of FC and IBS-C group increased compared with that of control group (3.42％ (1.24％ to 5.61％),2.45％(1.72％ to 4.27％) vs 1.18％ (0.35％ to 2.81％);Z=-2.534,-2.151,both P＜0.05).Meanwhile,AQP8 relative area of ascending colon of FC group increased compared with that of control group (2.46％(1.48％ to 4.18％) vs 1.14％(1.29％ to 2.15％) Z=-2.041,P＜0.05).Conclusion There are differences in AQP8 expression quantity and location in cells of descending colon between patients with FC and patients with IBS-C,which is a way for differentiation these two diseases.

The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

Objective To summarize clinical experience of reconstructive operation with transpositional colon behind the sternum after corrosive esophageal burns and to explore the treatment for its complications.Methods Clinical data of 65 cases with esophageal scarred stricture after corrosive burns receiving reconstructive operation with transpositional colon behind the sternum were reviewed,56 of them by end-to-end anastomosis between transpositional anterograde peristaltic colon and esophagus,seven by end-to- end anastomosis between transpositional anterograde peristaltic colon and pharyngeal fundus,and two by end- to-end anastomosis between transpositional reversed peristaltic colon and esophagus,to summarize treatment experiences in pre-operation,operation and post-operation.Results Fifty-one of this group of patients recovered and discharged form the hospital smoothly,12 with cervical anastomotic leakage after operation including two cured by re-operation and ten cured by conservative treatment,and two with necrosis of transpositional colon including one died during operation and the other cured.Conclusions Corrosive burns of esophagus can be cured by leaving scarred stricture esophagus open without resection,and the effectiveness of reconstructive operation with transpositional colon behind the sternum is satisfactory with good pre-operative preparation,correct surgical operation,and correct post-operative treatment.

Objective To understand the incidence and epidemiological characteristics of hand injuries among registered population in Jiading District of Shanghai, so as to provide evidence for scientific prevention and control. Methods Anting town, Huangdu town and Huating town were selected for investigation.Incidence of hand injuries among registered population was investigated by questionnaires. Results A total of 10 083 registered population was included in the study, with the average age being 57.1 ± 10.4 years old.Of them, 4 039 were male and 6 044 were female, 92.2% were married and 7.8% had college degree or above.The 26.8% of the subjects smoked at least one cigarette per day for more than six months, 13.1% drank at least three times per week for more than six months, and 43.7% exercised at least 10 minutes per week.The 37.9% had hypertension, 8.6% had diabetes, and 14.5% had hyperlipidemia.The incidence of hand injuries was 2.1%, of which 61.8% occurred at home and workplaces.Multivariate analysis showed that hypertension was the risk factor and smoking was the protective factor associated with hand injuries. Conclusion The incidence of hand injuries is low in Jiading District.Household hand injuries may be crucial for prevention; in addition, patients with hypertension might be a key population for prevention.

Studies have proved that microRNA-101 (miR-101) functions as a tumor suppressor and is associated with growth and apoptosis of various human cancers. However, the role of miR-101 in osteosarcoma and the possible mechanism by which miR-101 affects the tumor growth and apoptosis have not been fully elucidated. In this study, we found that the expression of miR-101 was down-regulated in osteosarcoma tissues and Saos-2 cell line as compared with that in adjacent non-neoplastic bone tissues and the osteoblastic cell line. To better characterize the role of miR-101 in osteosarcoma, we used a gain-of-function analysis by transfecting human osteosarcoma cell line Saos-2 with chemically synthesized miR-101 mimics. The results showed that overexpression of miR-101 inhibited the proliferation and promoted the apoptosis of Saos-2 cells. Meanwhile, bioinformatic analysis demonstrated that mTOR gene was a direct target of miR-101. Overexpression of miR-101 significantly decreased the expression of mTOR at both mRNA and protein levels in Saos-2 cells, consequently inhibiting Saos-2 cells proliferation and promoting cells apoptosis in an mTOR-dependent manner. Taken together, these data suggest that miR-101 may act as a tumor suppressor, which is commonly downregulated in both osteosarcoma tissues and cells. mTOR plays an important role in mediating miR-101 dependent biological functions in osteosarcoma. Reintroduction of miR-101 may be a novel therapeutic strategy by down-regulating mTOR expression.
Apoptosis ; Bone Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; MicroRNAs ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Osteosarcoma ; genetics ; metabolism ; pathology ; RNA, Neoplasm ; genetics ; metabolism ; TOR Serine-Threonine Kinases ; genetics ; metabolism

OBJECTIVETo develop an HPLC method for the determination of serum level of Crebanine (Cre) and study on the pharmacokinetics of Cre injection in rabbits.

METHODTo sample blood serum from the rabbits' ears which were injected the Cre by 2.0 mg x kg(-1) at different time and use HPLC to determine the concentration of Cre in it, the pharmacokinetic parameters were accessed by the DAS software.

RESULTCre was fitted to a two compartment open pharmacokinetic model in rabbits. There was no signifiant difference between the male and female rabbits'pharmacokinetic by t-test. The mainly pharmacokinetic parameters were: t1/2alpha = (3. 246 +/-0.222) min, t1/2beta = (36.67+/-5.52) min, Cmax = (1.401 +/- 0.062) mg x L(-1), Vd = (5.928 +/- 0.877) L x kg(-1), Cl = (0. 051 +/-0.003) L x min(-1) x kg(-1).

CONCLUSIONThis experiment can objectively show the pharmacokinetics regularity of Crebanine injection in rabbits. Crebanine injection was a speeding disposition drug (t1/2 <1 h) and disposed extensively and rapidly in rabbits.

Animals ; Aporphines ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Female ; Injections ; Male ; Metabolic Clearance Rate ; Plants, Medicinal ; chemistry ; Rabbits ; Stephania ; chemistry

BACKGROUNDThe relationship between the presence of metalloproteinases and thyroid cancer remains unknown, and many controversies still exist in this field. The objective of this study was to investigate the correlations between papillary thyroid cancer and peripheral blood levels of matrix metalloproteinase-2, matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2.

METHODSThe correlations were studied by detecting the levels of matrix metalloproteinase-2, matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2 by enzyme-linked immunosorbant assay and reverse-transcription polymerase chain reaction in the peripheral blood of 30 patients with papillary thyroid carcinoma, 27 patients with benign thyroid disease, and 25 healthy volunteers.

RESULTSThe levels of matrix metalloproteinase-2, matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2 in the peripheral blood of patients with papillary thyroid carcinoma were significantly higher than those in the peripheral blood of patients with benign thyroid disease and healthy volunteers (P < 0.05). However, there were no significant differences between patients with benign thyroid disease and healthy volunteers (P > 0.05). The accuracy of detection by both enzyme-linked immunosorbant assay and reverse-transcription polymerase chain reaction in the papillary thyroid cancer group was 83.33%.

CONCLUSIONSThe levels of matrix metalloproteinase-2, matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2 in the peripheral blood are helpful in identifying thyroid carcinoma and aid in preoperative assessment.

Adult ; Aged ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Matrix Metalloproteinase 2 ; blood ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Thyroid Neoplasms ; blood ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; blood ; Tissue Inhibitor of Metalloproteinase-2 ; blood

OBJECTIVE Gastric cancer is one of the most common malignant tumors,and the inci-dence rate is the highest in all kinds of tumors in China. However,it remains unclear that how signifi-cantly gastric cells are dependent on glycolysis,and which type of gastric cells are sensitive to glycolysis inhibition. In this study, several kind of gastric cancer cell lines were used as the research object, and the metabolic characteristics of different cell lines were systematically analyzed to provide theoretical support for the accurate treatment of gastric cancer. METHODS We examined the energy metabolism of four gastric cancer cell lines(MGC-803,SGC-7901,HGC-27 and BGC-823)by using glycolysis inhibitor, 2-deoxy-D-glucose(2-DG)and inhibitor of oxidative phosphorylation,oligomycin.Oxygen consumption rates(OCR)and L-lactate were also measured with an XF96 Analyzer(Seahorse Biosciences)to deter-mine the significance of metabolism of oxidative phosphorylation and aerobic glycolysisin gastric cells. In addition, western blot was used to detect the contribution of AMP-activated protein kinase (AMPK), and anti-apoptotic proteins(Bcl-2 and survivin)to clarify the mechanism of death or survival of gastric cancer cells treated by 2-DG or oligomycin. RESULTS In this study, it was shown that the growth of gastric cell lines were suppressed by 2-DG.However,the sensitivity to 2-DG was quite different among cell lines:IC 50 of 2-DG was from 3.28 mmol·L-1(MGC-803)to 15.57 mmol·L-1(BGC-823).MGC-803 was relatively sensitive to 2-DG (IC 50:3.28 mmol·L-1), consumed more glucose and produced more lactate (waste product of glycolysis) than the three other cell lines. Consequently, MGC-803 could be more dependent on glycolysis than other cell lines, which was further confirmed by the fact that glucose (+)FCS(-)medium showed more growth and survival than glucose(-)FCS(+)medium.Alternatively, BGC-823, most resistant to 2-DG (IC50: 15.57 mmol·L- 1), was most sensitive to oligomycin, and showed more growth and survival in glucose(-)FCS(+)medium than in glucose(+)FCS(-)medium. Thus,we had reasons to think BGC-823 cells depended on oxidative phosphorylation for energy production. In BGC-823,AMPK,which is activated when ATP becomes limiting,was rapidly phosphorylated by 2-DG, and expression of Bcl-2 was augmented,which might result in resistance to 2-DG.Interestingly,AMPK phosphorylation and augmentation of Bcl-2 expression by 2-DG were not observed in MGC-803,which is 2-DG sensitive. CONCLUSION There is a large metabolic difference between gastric cancer cell lines,which will facilitate the future gastric cancer therapy by targeting metabolic pathways.