Advances in research on anti-tumor drug-related renal injury / 医学研究生学报

Novel antineoplastic drugs have significantly prolonged the survival time of cancer patients. Meanwhile, nephrotoxicity of antineoplastic drugs and its adverse effects on the prognosis of cancer patients have received increasing attention. Conventional chemotherapy causes kidney injury mainly through direct renal toxicity, while new anti-tumor drugs can induce a number of kidney damage, including acute renal tubular injury, thrombotic microangiopathy, interstitial nephritis, and glomerular diseases through multiple mechanisms. Clinicians must be knowledgeable in the renal toxicity of antineoplastic drugs to minimize the nephrotoxicity of the drugs and diagnose early, especially in patients with underlying kidney disease. This article focuses on the risk factors, clinical and histological patterns, pathogenesis, prevention and treatment of renal injury associated with the antineoplastic drugs, especially novel targeted antineoplastic drugs.

Molecular features and expression of DAZAP2 in human multiple myeloma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In our previous study, we found that DAZAP2 was the most significantly down regulated gene when differential screening of complementary DNA (cDNA) chips were used to analyze mRNA isolated from bone marrow mononuclear cells from newly diagnosed multiple myeloma (MM) patients without anticancer treatment. In this study, we observed DAZAP2 mRNA and protein expression in the mononuclear cells from MM bone marrow and investigated its role in the pathogenesis of MM.</p><p><b>METHODS</b>The full-length cDNA of DAZAP2 was cloned and sequenced from mononuclear cells from human bone marrow. The nucleotide and amino acid sequences of DAZAP2 were analyzed using the ClustalW program. A dendrogram was constructed by multiple sequence alignment using ClustalW and amino acid sequence identity/similarity was derived based on comparisons attained using the MegAlign software. The recombinant pEGFP expression vector was constructed and the confocal microscopy was used for the localization of the DAZAP2 protein in transfected COS7 cells. The expression of DAZAP2 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and the expression level of DAZAP2 protein was detected by Western blotting analysis in MM samples.</p><p><b>RESULTS</b>DAZAP2 proteins of vertebrates is highly conserved in evolution. It contains a proline-rich region, several potential SH2 and SH3 domain-binding motifs and a possible protein kinase C (PKC) phosphorylation site. We showed by confocal microscopy that the DAZAP2 protein predominantly resides in the cytoplasm with a discrete pattern of punctuated distribution. The expression of DAZAP2 was not detected in 24 of 36 MM samples by semi-quantitative RT-PCR. In contrast, DAZAP2 expression was detected in all 30 normal controls. The expression level of DAZAP2 protein was assayed by Western blotting analysis, showing a robust down-regulation in MM patients (P < 0.001) that matched with the results of the RT-PCR.</p><p><b>CONCLUSIONS</b>DAZAP2 is downregulated in MM samples and it may be a signal molecule in MM cells. DAZAP2 is involved in the pathogenesis of MM and could be used as a genetic marker for MM.</p>

Analysis of current trauma epidemiology in Zhengzhou during 2004-2006 / 中华急诊医学杂志

Objective To investigate the present status of casualty epidemiology in Zhengzhou and the effects of different trauma care model.Method Statistic study of the classification of emergency disease and the number of ambulance responses to the call in Zhengzhou emergency rescue center from January,2004 through December 2006 was carried out and the efficiency between the trauma care model of an inclusive emergency rescue survices station and that of a simple emergency survices station was analyzed.Results The percentage of ambulance departure responses by Zbengzhou emergency rescue center for trauma care was 45.3 %,44.7 % and 45.8% of in 2004,2005 and 2006,respectively.There were 26 emergency rescue service stationa in Zhengzhou, including one independent station,eight internal medicine dominated semi-independent stations and seventeen simple model stations.The indicated surgical intervention can lye performed on the patients with severe multi- trauma in the independent emergency station in order to win the optimal operation time and reduce the mortality. Conclusions The trauma is the major reason for the emergency call.Emergency rescue service stations properly distrihtted,can offer quick and efficient pro-hospital first aid.The independent emergency rescue service station can increase successful resuscitation rate of serious casualties.

Cloning of distinguishing DNA sequences of Gastrodia elata Blume and application of them in identifying gastrodia tuber / 生物工程学报

Gastrodia elata Bl. is a famous and costful traditional Chinese medicine. Their genomic DNA fingerprints were investigated using a modified Randomly Amplified Polymorphic DNA method. DNA fragments common to all or to fine populations were identified and recovered. Five DNA fragments were proven not to be reported through DNA cloning, PCR identifying, nucleotide sequencing and bioinformatics analyses and were received in and recorded by NCBI GenBank. Gastrodine contents of the Gastrodia tuber samples were determined using high performance liquid chromatography technique. The distribution of the five DNA fragments in 9 Gastrodia elata Blue populations and the correlation with gastromedicine content were studied. The results show the distribution of these DNA sequences varied greatly among the populations whereby DNA Sequence 1 was the common and distinguishing molecular marker for all the populations studied and DNA Sequence 2 may relate to higher gastrodine content. In conclusion, these DNA marker sequences can be employed to identify genuine gastrodia tubers, better varieties and optimize their selection and cultivating.

Detection of MSH2 gene mutation by PCR / 中南大学学报(医学版)

OBJECTIVE@#To establish a genetic diagnosis method for a novel MSH2 mutation.@*METHODS@#A specific primer on the mutated site of MSH2 was synthesized and PCR was conducted using the specific primer and another downstream primer. PCR products were electrophoresed and then the carriers with the novel gene mutation of the carriers or non-carriers were identified.@*RESULTS@#MSH2 in a hereditary nonpolyposis colorectal cancer family were successfully found.@*CONCLUSION@#The method is effective and simple for genetic diagnosis of the novel mutation in MSH2.

The construction of recombinant AAV vector expressing HSVtk gene controlled by Tet-On and the detection of its activity / 生物工程学报

In order to investigate the application of recombinant adeno-associated virus (rAAV) vector containing Tet regulation system and HSVtk gene in cancer gene therapy, pAAV/TRE/HSVtk/Tet-On was constructed and identified with PCR and restriction enzyme digestion. Packaging cells HEK293 were cotransfected with plasmids pAAV/TRE/HSVtk/Tet-On, pAAV-RC and pAAV-helper to produce infectious rAAV, and CsCl2 densitygradient centrifugation method was performed for purification and concentration of rAAV. The viruses were then transduced into MCF-7 cells. The results of dot blot hybridization indicate that the rAAV can transfer the target gene into MCF-7 cells. MTT assay showed that GCV could kill AAV-infected MCF-7 cells under the induction of Dox. The data demonstrated that rAAV containing Tet regulation system and HSVtk gene was successfully obtained, and could be used for further investigation of in vivo and in vitro experiments.

Suicide gene therapy of human breast cancer in SCID mice model by the regulation of Tet-On / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>RevTet-On gene expression system was used to deliver the suicide gene tk to human breast cancer cell line MCF-7 and control the tk gene expression level. The animal model of human breast cancer on severe combined immune deficiency (SCID) mice was set up to explore the suicide gene therapy by the regulation of Tet-On.</p><p><b>METHODS</b>Herpes simplex virus-thymidine kinase (HSVtk) gene was inserted into the plasmid pRevTRE and the recombinant retroviral vector pRevTRE/HSVtk was constructed. Using modified calcium phosphate co-precipitation method, two transfections, pRevTRE/HSVtk and pRevTet-On were performed for MCF-7 cell line and selected by hygromycin B and G418. MCF-7 cell line that stably expressed Tet-regulated tk gene was established. HSVtk gene expression in the MCF/TRE/tk/Tet-On cell line was under the control of Doxycycline (Dox). Cell viability was also determined by MTT assay, whereas HSVtk gene expression was analyzed by reverse transcription-PCR (RT-PCR).</p><p><b>RESULTS</b>MCF/TRE/tk/Tet-On cell survival rate was decreased from 100% to less than 20% when ganciclovir (GCV) concentration was increased from 0 to 1000 microg/ml at 1 microg/ml of Dox after 72 hours of GCV administration. At 1 microg/ml of GCV concentration, the cell numbers decreased from 7 x 10(4) cells/ml to 2 x 10(4) cells/ml when Dox concentration was increased from 0 to 1500 ng/ml after 72 hours culture. In addition, bystander effects were generated in vitro when 10% - 25% of transduced MCF-7 cells were mixed in untransduced MCF-7 cells. On the other hand, the human breast cancer models in SCID mice were set up. The tk gene was expressed with the regulated character after MCF/TRE/tk/Tet-On cells were implanted into the female SCID mice 7 days after Dox induction followed by intraperitoneally administration of GCV for 23 days. Subcutaneous tumors in SCID mice that were implanted with MCF/TRE/tk/Tet-On cells shrank remarkably after Dox and GCV administration as compared with the control.</p><p><b>CONCLUSION</b>The human breast tumor cells (MCF-7) expressing HSVtk gene can be eradicated by administration of GCV and induced with tetracycline or its derivative Dox in vitro and in vivo.</p>

Retroviral vector-mediated HSVtk gene expression and acquisition of high titer recombinant virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the HSVtk gene expression mediated by the retroviral vector and to obtain high titer recombinant retroviral virus.</p><p><b>METHODS</b>The recombinant vector pRevTRE/HSVtk was constructed by inserting HSVtk gene into pRevTRE. The recombinant retrovirus, which was produced from cloned PA317 cells screened by hygromycin B after "micro-pingpong" technique transferring with pRevTRE/HSVtk plasmids DNA by using modified calcium phosphate precipitation method. HSVtk gene expression was performed on target cells and virus titers were detected in different cultured temper, time and sodium butyrate concentration.</p><p><b>RESULTS</b>The recombinant retroviral vector pRevTRE/HSVtk was constructed and HSVtk gene expression was detected on target cells after they were infected with the recombinant retrovirus.</p><p><b>CONCLUSION</b>High titer of retroviruses could be obtained in the culture medium of PA317 cell line through "micro-pingpong" technique at 30 hours and 10 mmol/L sodium butyrate concentration followed by frozen ultrafiltration.</p>