Objective To investigate the effect of parathyroid hormone(PTH)on the proliferation and invasion of chondrosarcoma cells,and the relationship between PTH and the regulation of primary cilia expression.Methods After stimulation of the chondrosarcoma cell line SW1353 with different concentrations of PTH,induction of the expression of cilia with hypoxia and destruction of cilia structure with chloral hydrate,the cell viability was detected by CCK8 assay,the proliferation and invasion of SW1353 by Western blotting and Transwell method,the primary cilia expression by immunofluorescence assay and the GLI1,PTCH1 and IFT88 expression levels by real-time PCR.Results PTH could promote the proliferation of chondrosarcoma cells in a concentration-dependent manner and this effect was correlated with the structural integrity of the primary cilia.PTH could up-regulate the invasive ability of SW1353 cells and increase the expression levels of MMP9,which was suppressed when the primary cilia structure was damaged.Additionally,it was found that PTH could down-regulate the number of primary cilia,which was related to the structural integrity of the primary cilia.It could also regulate the expression levels of GLI1 and PTCH1,the target genes in Hedgehog pathway,and the expression levels of IFT88,the gene associated with the cilia function.Conclusion PTH can promote the proliferation and invasion of chondrosarcoma cells,down-regulate the expression of primary cilia and the downstream target genes.PTH may regulate the malignant biological features of chondrosarcoma by regulating the primary cilia expression.

Abstract: Suxamethonium chloride is a depolarizing muscle relaxant used in general anesthesia. In overdose, it causes adverse reactions such as bradycardia, arrhythmia, cardiac arrest, and death. The article reviews the progress on testing methods of suxamethonium chloride such as infrared spectroscopy, chemical color reaction, chemical titration, enzyme electrode, chromatography and mass spectrometry.
Anesthesia, General ; Arrhythmias, Cardiac/chemically induced* ; Biosensing Techniques ; Bradycardia/chemically induced* ; Chromatography ; Drug Overdose ; Heart Arrest/chemically induced* ; Humans ; Mass Spectrometry ; Neuromuscular Depolarizing Agents/analysis* ; Spectrophotometry, Infrared ; Succinylcholine/analysis*

Objective To explore the pattern of lymph node metastasis in patitsen with squamous cell carcinoma of the thoracic esophagus and its significance in lymphadenectomy.Methods The clinical data of 230 patients who received radical esophagectomy with three-field lymphadenectomy was analyzed.The metastatic sites of lymph nodes were correlated with tumor location by chi-square test.Logistic regression was used to analyze the relationship between clinic pathoingical factors and lymph node metastasis.Results Lymph node metastases were found in 133 of the 230 patients(57.8%).The average number of resected lymph nodes was 25.3? 11.4 per patient(range 11～71).The rates of lymph node metastasis were 41.6%,19.44%and 8.3%in the neck,thoracic medi- astinum and abdominal cavity for patients with upper thoracic esophageal carcinoma,33.3%,34.7%and 14%for patients with mid- die thoracic esophageal carcinoma and 36.4 %,34.1%and 43.2 %for patients with lower thoracic esophageal carcinoma.No signifi- cant difference in cervical or thoracic metastatic rate was observed among upper,middle and lower thoracic carcinoma.The difference in lymph node metastatic rate for nodes in the abdominal cavity was significant among upper,middle and lower thoracic carcinoma. The lower thoracic esophageal cancers were more likely to metastasize to the abdominal cavity.Logistic-regression showed depth of tu- mor invasion and angiolymphatic invasion were factors influencing lymph node metastasis.Conclusion Cervical and mediastinal node dissection should be performed independently from tumor location.Abdominal node dissection should be conducted more vigorously for lower thoracic esophageal cancer than of other locations.Patients with greater tumor grade,depth of tumor invasion and angiolymphatic invasion were more prone to develop lymph node metastasis.

Objective To explore the feasible approach for establishment of a liver cancer model induced by N-nitrosodiethylamine(DENA)with Sprague-Dawley rat,and to provide ideal animal model for imaging diagnosis and interventional therapy.Methods One-hundred and forty male SD rats were administrated with 0.95 g/L DENA for 10 weeks,and MRI was performed for inspecting pathological changes of rat livers on the following week.When the liver tumor was proved then the rat will be the candidate for sequential procedures,otherwise the animal continued under observation until next MRI examination after 4 weeks.DSA was done in 64 rats'for detecting blood supply of liver tumors.Animals were sacrificed with an overdose of chloral hydrate,The representative tumor tissues were fixed in 10% formalin and 2.5% glutaraldehyde for light and electron microscopy analysis respectively.Alpha fetoprotein(AFP) and hepatocyte were assaied by immunohistochemistry technique in order to identify intrinsic trait of the harvested tumors.Results The earliest induced tumor was detected on 11th week and the latest was on 20th week by MRI,and the median period was 13.9 weeks.Tumor size ranged from 2 mm to 40 mm in diameter. The rate of single and multi-induced tumor was 9.7%(7/72)and 90.3%(65/72),respectively.87.7% (57/65)" of the induced multiple tumors was with hepatocirrhosis and 18.1% of these tumors combined with extrahepatic neoplasm or metastasis.Plenty blood supply was proved by DSA in most of those tumors. Tumors not only derived from hepatocyte but also manifested positive expression of AFP.Histological types of these tumors include hepatocellular carcinoma(HCC)(92.0%,66/72),intrahepatic cholangiocarcinoma (ICC)(4.0%,3/72),and combined HCC and ICC(4.0%,3/72),respectively.Electro-microscope analysis indicated that cytoplasm and organelle of induced tumors were abnormal distinctly compared with those of non-carcinomatous cells.Conclusion DENA can induce ideal rat liver cancer as a feasible animal model,MRI is the best approach for scrutinizing pathological changes of rat livers during induced period.

Objective To summarize our surgical experience in repair and functional reconstruction for compound defects of proximal phalanx and dorsal skin at multiple digits,Methods Six patients with multiple digital defects were treated in our department between June 1996 and March 2005.At the first stage,free iliac bone grafts were used to repair defects of proximal phalanges and temporary syndactyly between adjacent affected fingers was created through digital palmar skin sutures.The defects were covered with free flap transfer finally.Dorsalis pedis flaps were used in four patients,a lateral arm flap in one and a lateral thoracic flap in one respectively.At the second stage,a partial debulking procedure and division of syndaetyty followed three to six months later.Additional procedures were performed in three eases to reconstruct the digital extensor function through tendon transfer.The follow-ups ranged from six months to nine years.Results The flaps survived uneventfully in the six patients postoperatively.The dorsal aspects of reconstructed fingers demonstrated an aesthetically pleasing effect after the flap debulking procedures and division of syndactyly.Follow-up X-Ray examinations showed good lilac bone union and nearly normal structure of digital bone.The distal interphalangeal extension restored to normal in the three cases after extensor reconstruction.Conclusions Iliac bone graft to repair phalangeal defects and free flap transplantation to cover skin defects can be a good treatment for compound defects of proximal phalanx and dorsal skin at multiple digits.Secondary plastic procedure may greatly improve the appearance of a reconstructed digit,and extensor re- construction the function of distal interphalangeal extension too.

Objective To evaluate the association between recurrent miscarriages and insulin resistance.Methods The case-control studies on the association between recurrent spontaneous abortion and insulin resistance from June 1996 to April 2012 were collected from Medline,Elsevier,Chinese Journal Fulltext Database,Chinese Biological Medicine Database,data base of Wanfang,Springer link and EMBASE.RevMan 5.1 software was used for Meta analysis.Results According to the included criteria,7 clinical trials were finally selected.Total 467 cases with recurrent pregnancy loss were enrolled in study group,while 413 women with no history of abnormal pregnancies were enrolled in control group.No significant difference was found in average age and body mass index between the two groups (P ＞ 0.05).Meta analysis results showed that the level of fasting glucose was no statistical difference between study group and control group (WMD =2.27,95％ CI:-1.11 to 5.65,P ＞0.05); fasting insulin level was higher 2.05 mU/L in study group than that of in control group,the difference was statistically significant (WMD =2.05,95％ CI:1.03 to 3.08,P ＜ 0.01).Case number of study group on Homa-insulin resistance ＞ 4.5 was more than that of control group (OR =3.36,95％ CI:1.72 to 6.57,P ＜ 0.01).Case number of study group on glucose/insulin ratio ＜ 4.5 was more than that of the control group,statistical difference was found (OR =3.37,95％ CI:1.90 to 5.99,P ＜ 0.01).Conclusion Insulin resistance is associated with the susceptibility to recurrent miscarriages,and it may contribute to the occurrence of recurrent miscarriages.

Adult ; Air Pollutants ; adverse effects ; Humans ; Male ; Military Medicine ; Monitoring, Physiologic ; Noise, Occupational ; adverse effects

The nuclear transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) plays a crucial role in maintaining cellular redox homeostasis. The aberrant NRF2 signaling confers enhanced antioxidant capacity, which is linked to tumor progression and therapeutic resistance. The current study investigates the biological effects and molecular mechanism of tribbles homolog 3 (TRIB3), a stress-induced protein, in regulating cell survival and apoptosis in lung cancer. This study first performed the RNA sequencing data analysis with 576 lung adenocarcinoma patients from the cancer genome atlas (TCGA) database. The NRF2- antioxidant response element (ARE) signature was enriched in patients with high TRIB3 expression. Dual-luciferase reporter assay and real-time quantitative polymerase chain reaction (PCR) were used to confirm the effect of TRIB3 on the kelch-like ECH-associated protein-1 (KEAP1)-NRF2 pathway. Abrogation of TRIB3 impaired NRF2 transcriptional activity and reduced the expression of its target genes. Moreover, TRIB3 enhanced NRF2 stability via blocking KEAP1-NRF2 interaction. TRIB3-depletion promoted reactive oxygen species (ROS) production, restrained cell proliferation, and enhanced carboplatin-induced apoptosis. In addition, NRF2 overexpression recovered the tumor inhibition effect of TRIB3-depletion. Consistently, TRIB3 failed to modulate apoptosis in NRF2 depletion cells. In summary, this study shows that TRIB3 inhibits the KEAP1-NRF2 interaction and upregulates the transcriptional activity of NRF2, thereby promoting lung cancer cell proliferation and reducing the sensitivity to chemotherapy. Targeting the TRIB3-NRF2 signal axis may become a new strategy for ROS homeostasis and lung cancer treatment.

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology

Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.