Mitochondrial fission protein 1 determines mitochondrial fission and cisplatin sensitivity in tongue squamous cell carcinoma / 口腔疾病防治

Objective @# To investigate the effect of mitochondrial fission protein 1 (FIS1) on apoptosis and cisplatin resistance in tongue squamous cell carcinoma (TSCC) cells.@*Methods @#The squamous cell carcinoma cell lines SCC9 and CAL27 were used to detect the mRNA and protein levels of FIS1 after cisplatin treatment, the knockdown and overexpression of FIS1 of SCC9 and CAL27 with or without cisplatin treatment were accomplished through small interfering RNA (siRNA) and plasmid, respectively. The mitochondrial division state in cells was detected by mitochondrial staining, and the apoptosis state of cells was detected by TUNEL, flow cytometry and Caspase 3/7.@*Results@#FIS1 protein expression in tongue squamous carcinoma cells treated with cisplatin was increased, but the mRNA level did not change. Silencing of FIS1 expression reduced mitochondrial division and apoptosis in squamous cell carcinoma cells treated with cisplatin, whereas overexpression of FIS1 exhibited the opposite effects. The percentage of dividing mitochondria, the number of apoptotic cells and the activity of Caspase 3/7 in SCC9 and CAL27 cells were significantly different before and after modulation of FIS1 expression (P < 0.05). @*Conclusion@#FIS1 is involved in the regulation of cisplatin chemotherapy sensitivity in tongue squamous cell carcinoma and can be used as a new target for improving the sensitivity of cisplatin chemotherapy in oral squamous cell carcinoma.

Expression and change of ERRα inoxaliplatin?resistant colon cancer cells after the downregulation of tetrasoanin / 中国综合临床

Objective To observe the proliferation and apoptosis of oxaliplatin-resistant colon cancer cell lines and the expression of estrogen receptor related receptor(ERR)α when tetrasoanin was down-regulated. Methods Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)were used to detect the expression of tetrasoanin and ERRα of colon cancer cells and oxaliplatin resistant cells in mRNA and protein levels. ERRα inhibitor XCT790 was used to down-regulate ERRα expression, The expression of ERRα was down regulated by ERR α inhibitor XCT790,and the level of tetrasoanin,apoptosis and proliferation of L-OHP-SW620 cells were detected by Western blot, flow cytometry and MTT.Tetrasoanin expression was down regulated by siRNA, the expression, apoptosis and proliferation of L-OHP-SW620 cells AKT, p-AKT, tetrasoanin and ERRα were detected by Western blot,qRT-PCR,flow cytometry and MTT assay.Results The expression of tetrasoanin and ERRα protein in L-OHP-SW620 cell lines were higher than those in SW620 cells (t=6.127,P<0.01,t=12.579,P<0.01),The expression of tetrasoanin mRNA in L-OHP-SW620 cell line was higher than that in SW620 cell line(t=9.085,P< 0.01). The early apoptosis rate of L-OHP-SW620 cells in XCT790 group after XCT790 inhibited ERR -αexpression was higher than that in NC group(t=3.297, P< 0.01). The survival rate of XCT790 group after 72 h culture was(45.264±6.249)%,lower than that of NC group((63.364 ± 9.472)%)(t=4.537, P<0.01). Compared with NC group,p-AKT protein was up-regulated(t=8.139,P<0.01),ERRα protein was down-regulated(t=6.452,P<0.01),the apoptosis rate was(17.541±2.317)%,lower than that in the sitetrasoanin group((32.892±3.296)%)(t=4.526,P<0.01), the survival in sitetrasoanin group after 72 h culture was(49.653 ± 5.945)%, lower than that in NC group ((67.376±7.934)%)(t=3.109,P<0.05).Conclusion Tetrasoanin down-regulation and p-AKT protein up-regulation decreases ERRα protein and OHP-resistant colon cell proliferation is decreased, apoptosis is increased and drug resistance is decreased.

Inhibidores adquiridos de la coagulación: enfoque diagnóstico y casos especiales / Acquired inhibitors of blood coagulation: diagnostic perpective and especial cases / Inibidores adquiridos da coagulação: abordagem diagnóstica e casos especiais

Los inhibidores adquiridos son defectos raros. Se asocian a diferentes manifestaciones clínicas con morbimortalidad significativa. Su detección es importante para implementar el tratamiento sin demora. Hay inhibidores específicos (bloquean función), contra todos los factores de la coagulación y además VWF, o de interferencia (afectan una o varias vías de la coagulación). Los inhibidores específicos son alo-anticuerpos desarrollados en pacientes deficitarios (complicación terapéutica) o auto-anticuerpos presentes en individuos sin alteraciones previas. Hay anticuerpos específicos que pueden afectar la depuración pero no inhiben función. Inhibidores de interferencia: inmunoglobulinas u otras sustancias (heparina/heparinoides, PDF/pdf, PIVKAS, moléculas anómalas, etc.) asociadas a diferentes situaciones clínicas (asintomáticos, sangrados, trombosis y/o complicaciones obstétricas). El laboratorio es fundamental para el diagnóstico. Las pruebas globales detectan el defecto, que no corrigen por el agregado de plasma normal; se caracteriza luego el tipo de inhibidor y eventualmente se titula. Esto es complejo; hay variabilidad en los resultados y posibilidad de falsos positivos o negativos, además las pruebas no son estrictamente específicas. Los algoritmos diagnósticos son útiles, pero no contemplan la posibilidad de defectos combinados. Es crítico caracterizar al inhibidor y descartar posibles interferencias o defectos concomitantes; ello requiere aplicar las pruebas adecuadas e interpretarlas correctamente.

Acquired inhibitors are rare disorders. They are associated with different clinical behaviours and significant morbi-mortality. Detection is important in order to start treatment urgently. There are either specific inhibitors, which block function, against all coagulation factors, and VWF, or with interference effects, on one or more coagulation pathways. Specific inhibitors are either allo-antibodies developed in deficient patients, which give rise to therapeutic complication; or auto-antibodies, which are present in individuals without previous defects. There are specific antibodies that can affect clearance but which cannot block the function. Inhibitors with interference effects are immunoglobulins or other substances (heparin/heparinoids, FDP/fdp, PIVKAS, abnormal molecules, etc.) associated with different clinical settings (asymptomatic, bleeding, thrombosis and/or obstetric complications). Laboratory results are fundamental for the diagnosis. Global tests are able to detect the defect, which is not corrected by the addition of normal plasma; the type of inhibitor is then characterized and titration of the inhibitor is eventually performed. This is complex; there is variability in the results and there is likelihood of false positive or negative results; moreover, the tests are not strictly specific. Diagnostic algorithms are useful tools but they do not consider combined defects. It is a critical point to characterize the inhibitor and exclude possible interferences or concomitant defects; this demands application of the correct tests without misinterpretations.

Os inibidores adquiridos são defeitos raros. Associam-se a diferentes manifestações clínicas com morbimortalidade significativa. Sua detecção é importante para implementar o tratamento sem demora. Há inibidores específicos (bloqueiam função), contra todos os fatores da coagulação e também VWF, ou de interferência (afetam uma ou várias vias da coagulação). Inibidores específicos: - aloanticorpos desenvolvidos em pacientes deficitários (complicação terapêutica); - autoanticorpos, em indivíduos sem alterações prévias. Existem anticorpos específicos que podem afetar a depuração, mas não inibem função. Inibidores de interferência: imunoglobulinas ou outras substâncias (heparina/heparinoides, PDF/pdf, PIVKAS, moléculas anômalas, etc.) associadas a diferentes situações clínicas (assintomáticos, sangramentos, tromboses e/ou complicações obstétricas). O laboratório é fundamental para o diagnóstico. Os testes globais detectam o defeito, que não corrige pelo acréscimo de plasma normal; caracteriza-se depois o tipo de inibidor e eventualmente é titulado. Isto é complexo; há variabilidade nos resultados e possibilidade de falsos positivos ou negativos, além disso os testes não são rigorosamente específicos. Os algoritmos diagnósticos são úteis, mas não consideram a possibilidade de defeitos combinados. É crítico caracterizar o inibidor e descartar possíveis interferências ou defeitos concomitantes; isso requer aplicar os testes adequados e interpretá-los corretamente.

miRNA library and thermodynamic stability of Astragali Radix decoction pieces / 中草药

Objective: To establish a small RNA library of Astragalus membranaceus, in order to understand the intake and absorption of miRNAs in human body, and to lay the foundation for pharmacological research of Astragali Radix decoction pieces (ARP). Methods: The decoction of A. membranaceus after desiccation, high temperature and microwave treatment was used as sample, and using high-throughput sequencing technology, the unknown miRNA sequence was obtained. Results: Totally 9931049 small RNA sequences were identified from ARP by the high-throughput sequencing technology. After compared with all hairpin-forming precursors in miRBase 21 database, the miRNA library of ARP decoction was established, and one conserved miRNA and 9 novel miRNA sequences were confirmed. Twelve class I small interfering RNAs (siRNAs) were obtained by further bioinformatic analysis and we found that the 5' terminal was unstable while the 3' terminal was stable thermodynamically. Conclusion: The miRNAs expression profiles of ARP is first revealed by thermodynamic treatment. Our study suggests that class I siRNA can be used as a potential biochemistry components of phytomedicine, which might contribute to the epigenetic studies on Chinese materia medica.

Decreasing effect of RNA interference vector targeting the EphB4 gene on the expression of EphrinB2 in human colon cancer cells / 医学研究生学报

Objective Studies show that the role of EphB 4 in the development and progression of cancer is correlated to its ligand EphrinB2.The present study was to observe the effect of the changes in EphB4 on the expression of EphrinB2 by constructing and identifying microRNA ( miRNA) interference vectors targeting the EphB4 gene in colon cancer cells . Mte hods According to the EphB4 gene sequence , 3 pairs of oligo DNA sequences of miRNA were designed .The single strand of oligo DNA was annealed to form double-strand DNA, and then connected with the plasmid pcDNA 6.2-GW/EmGFP-miRNA.The expression vector pcDNA6.2-GW/EmGFP-miR-EphB4 was linked to pDONR221 and pLenti6/V5-DEST to construct the lentiviral expression vector pLenti 6/V5-DEST-EphB4, which was cotransfected with packaging mix (pLP1, pLP2 and pLP/VSVG) into 293FT cells by lipofectamine 2000 transfec-tion to produce lentivirus , and the lentivirus titer was measured by infection of HEK 293 cells.The stable cell lines were selected and cultured.The expression levels of EphB4 and EphrinB2 were examined by qPCR. Results Three miRNA interference vectors SR-1, SR-2, and SR-3 targeting the EphB4 gene were successfully constructed , with SR-3 exhibiting the most significant interference efficien-cy.The constructed lentiviral vector pLenti 6/V5-DEST-EphB4 was successfully packaged in 293FT cells.The virus titer was 7 ×108 Caco-2 cells. Conclusion The exogenous EphB4 expression could be significantly inhibited by treatment with specific miRNA in co-lon cancer cells .The correlation of EphB4 and EphrinB2 may be effected by many factors and need further studies .

Study on Determination Method for Bacterial Endotoxins in Nedaplatin for Injection / 中国药师

Objective:To establish a detection method for bacterial endotoxins in nedaplatin for injection. Methods:The experi-ment was carried out and the result was evaluated according to the appendix XIE of Chinese Pharmacopoeia 2010 Vol. II. Results:There was no interference from nedaplatin for injection to bacterial endotoxin in TAL reaction. Conclusion:The method is feasible to detect bacterial endotoxin in nedaplatin for injection, which can replace the rabbit pyrogen test.

O pesquisar em ação / Research in action

O objetivo deste texto é fazer uma resenha do livro intitulado Pesquisar na Diferença, uma publicação coletiva, inspirada pelo Abecedário de Gilles Deleuze. O livro é composto por verbetes, cada um dos quais ligados a uma ação que se faz presente no pesquisar, tal como é entendido pelo grupo de pesquisadores que compõe o volume. Observar, desejar, escrever, afetar, tatear são alguns dos verbetes presentes neste livro. Os autores que se reúnem nesta publicação definem a pesquisa como uma ação encarnada e situada, como uma prática de composição de mundo e que por isso mesmo, é eminentemente política. É a partir deste ponto que os verbetes são definidos.

The aim of this paper is to present a review of the book titled Pesquisar na Diferença, a publishing collective, inspired by "Deleuze's Abecedary". The book consists of entries, each of which is linked to an action that is imanent to research, as understood by the group of researchers who make up the volume. Observe, wish, write, affect, groping are some of the entries in this book. The authors Who gather in this publication defines research as an action embodied and situated, as a practice of composing world which is eminently political. It is from this point that the entries are defined.

Building and operation of the pharmacovigilance system in the hospital / 中华医院管理杂志

Objective Effective intervention on the setbacks and safety loopholes of clinical medication, in an effort to explore, establish and improve the ADR watchfulness system in the hospital step by step. MethodsCollection and compilation of ADRs and drug compatibility cases found in 2010 in the hospital. Intervention on drugs of safety risk exposure or medication factors by means of the ADR prewarning mechanism, medical rounds by hospital's directors regarding drug issues, communication between pharmacists and clinicians, ADR monitoring leadership team, and pharmaceutical affairs council. Results 143 cases of ADR were interfered with among all the 393 ADR cases reported by clinical departments in 2010, accounting for 36.4％ of all the reported cases. ConclusionHospitals need to pay more attention to the monitoring, analyzing and evaluation of ADRs, and take efficient intervention means to make drug use more rational and ensure safe medication for the people.

Lentivirus-mediated RNAi against xenoantigen in mouse vascular endothelial cells / 中华器官移植杂志

Objective To investigate the feasibility of inhibiting Galα (1,3)-Gal expression in mouse vascular endothelial cells by lentivirus-mediated RNAi.Methods The shRNA specified to α1,3-GT mRNA was designed and synthesized in vitro and cloned into the lentivirus vector.EOMA cells were infected by recombinant lentivirus.Real-time RT-PCR was used to detect mRNA transcriptional levels of αl,3-GT as well as immunofluorescence and flow cytometry were applied to detect Galα(1,3)-Gal antigen level after gene transfection.Co-culture of infected EOMA and serum of human was done and the survival rate was measured by MTT.Results The αl,3-GT shRNA sequences were cloned into the recombinant lentivirus vector correctly and the lentivirus was produced successfully.The transfection efficiency to EOMA was 75 %.Real-time PCR revealed that the mRNA transcription of α1,3-GT was obviously inhibited by α1,3-GT shRNA recombinant lentivirus with the rate of 88 % (P＜0.05),while there were no obvious differences among control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＞ 0.05).Immunofluorescence and flow cytometry demonstrated the same results that Galα(1,3)-Gal antigen expression in EOMA transfected by α1,3-GT shRNA lentivirus was less than that of control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＜0.05),but there were no obvious differences among the later three groups (P＞0.05).After co-culture with serum of human,MTT showed the survival rate of EOMA infected by α1,3-GT shRNA lentivirus was obviously increased (P＜ 0.05).Conclusion Recombinant α2,3-GT shRNA 1entivirus is constructed successfully,which can inhibit the expression of α1,3-GT and Galα1,3-Gal in EOMA by RNAi and control hyperacute rejection in vitro.

Prevention of the Antelocation of Nursing Intervention to Constipation in Patients with Acute Myocardial Infarction / 中国康复理论与实践

@#Objective To investigate the prevention of the antelocation of nursing intervention to constipation in patients with acute myocardial infarction(AMI).Methods 70 AMI patients were divided randomly into the test group and control group with 35 patients in each group.The control group received the AMI usual care and drugs;the test group received the antelocation of nursing intervention to constipation besides the usual care and drugs.Results The death rate and the incidence of abdominal distension,abdominal pain,anxiety,and complication of the patients in the test group were lower than those of the patients in the control group(P<0.05).Conclusion The antelocation of nursing intervention to constipation can decrease the death rate of AMI patients.

Construction and Identification of NBS1-targeting microRNA Expressing Eukaryotic Vector / 中国生物工程杂志

Objective:To construct NBS1 microRNA expressing eukaryotic recombinants,and identify biological activity of recombinants in Hela cell after transfection.Methods:According to sequence of NBS1mRNA,the NBS1 pre-microRNA was designed and synthesized,then cloned into the GFP reporter pcDNA6.2-GW/EmGFP-miR vector and transfected into Hela cell line.To detect integrity of inset fragment through colony PCR and sequencing analysis.The biological activity of recombinants through identify interference efficiency of NBS1 microRNA recombinants by way of Real-Time PCR and Western blot were determined.Results:Sequences of inset fragment in four microRNA expressing recombinants were correct.NBS1 mRNA and protein expression of four microRNA recombinants were decrease,which is the lowest in the NBS1mi-2 group.Conclusion:Four NBS1-targeting microRNA expressing recombinants all have biological activity in Hela cell line,and NBS1mi-2 recombinant has the most interference efficiency.The microRNA expressing plasmid which were successfully constructed and lay foundation for the studies on the tumor gene therapy of microrna targeting NBS1.

The deficiency and complement on the law of the exemption of informed consent in China / 中国医学伦理学

Doctors must carry out informed consent in medical treatment.However,to ensure patients' rights and interests mostly,this obligation needs exempted in some special cases.This article studies the insufficiency in the existing regulation on doctors' obligations exemption.It also lists some constructive suggestions based on the existing theory to solve this problem.

Progress in surgery intervention for craniocervical unstability / 第二军医大学学报

Craniocervical unstability may cause severe motor and respiratory compromise due to recurrent upper spinal cord and/or brain stem impingement.In recent years,by improving internal fixation materials and surgical techniques,the methods of surgical treatment of craniocervical unstability has made great progress.With the appearance of these new surgery and internal fixation methods,the successful rate and safety of the operation obviously increased and the complications was reduced greatly.We reviewed the surgery methods for craniocervical unstability in this article.