Preliminary Study on the Gene Characteristics of Oidiomycetes Mutant Strains Like Bacterial Morphology / 现代检验医学杂志

Objective To further explore the genetic characteristics of oidiomycetes mutant strains like bacterial morphology on the basis of the study on morphology and structure of mutated candida.Methods The standard strains of candida albicans were induced by low temperature and under the condition of low temperature and nutrient deficiency.Variation of standard strains of Candida albicans were induced by clinical antifungal drugs such as fluconazole with different concentration gradient.Fungal gene template was prepared by boiling method,sequences of 16SRNA and 18SRNA were amplified using bacteria conservative gene sequence of 16SRNA and fungal conserved gene sequence of 18SRNA,and observed and recorded the results agarose gel electrophoresis.At the same time,the amplified fragment of bacterial conservative gene 16SRNA was sequenced,and the sequence was analyzed by BLAST comparison.Results the 16SRNA sequences of candida variant were amplified positive,while the standard strain of candida albicans did not show the corresponding amplification band.Except 2 strains which showed a faint band,the other variants of the 18SRNA sequences did not amplified the target band,while the standard strains of candida albicans showed a corresponding amplification bands.Suggested that proportion of 18SRNA sequences in the genome of oidiomycetes mutant strains like bacterial morphology was not much even lack.The 16SRNA fragments amplified of oidiomycetes mutant strains like bacterial morphology did determination of DNA sequence after purification.BLAST comparison analysis,it was found that sequence of oidiomycetes mutant strains like bacterial morphology had higher similarity with bacterial sequences in the database.Conclusion Oidiomycetes mutant strains like bacterial morphology contained bacterial and a small amount of fungus conservative gene.Oidiomycetes mutant strains like bacterial morphology with original nuclear biological character are ones from eukaryotes.This study is great significance in biological evolution,especially in the evolution of prokaryotic cells and eukaryotic cells.

In vitro drug resistance of clinical isolated Brucella against antimicrobial agents

OBJECTIVE@#To explore the antibiotic resistance of Brucella melitensis and instruct rational use of antimicrobial agents in clinical treatment of Brucella infection.@*METHODS@#Bacteria were cultured and identified by BACTEC9120 and VITEK II automicrobic system. E-test was used to detect the minimal inhibitory concentration (MIC) of antimicrobial agents in the drug susceptivity experiment.@*RESULTS@#A total of 19 brucella strains (all Brucella melitensis) were isolated from 19 patients, who had fever between January 2010 and June 2012, and 17 samples were blood, one was bone marrow, the other sample was cerebrospinal fluid. The MIC range of ceftazidime was 2.0-8.0 mg/L, rifampicin was 0.06-2.0 mg/L, amikacin was 4.0-12.0 mg/L, levofloxacin was 2.0-8.0 mg/L, doxycycline was 8.0-32.0 mg/L, sulfamethoxazole-trimethoprim was 4.0-16.0 mg/L, ampicillin was 1.5-2.0 mg/L and gentamicin was 0.50-0.75 mg/L.@*CONCLUSIONS@#The drugs used in this experiment cover common drugs for treating Brucella. Meanwhile, the results are consistent with clinical efficacy. It is suggested personalized regimen according to patients' status in treatment of Brucella.

Transcriptional activities of tumor-specific survivin promoter and PSMA promoter and enhancer in human prostate cancer: evaluation and comparison / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect and compare the transcriptional activities of prostate-specific membrane antigen (PSMA) promoter and enhancer and survivin promoter in different human prostate cancer cell lines, and to search for some evidence for the targeting gene therapy of human prostate cancer.</p><p><b>METHODS</b>The fragments of the PSMA promoter and enhancer and survivin promoter were amplified by PCR and inserted into pGL3-Basic. The recombinant plasmids were transiently transfected into human prostate cancer cell lines and normal Chang liver cells, and, their transcriptional activities in various cells were determined by measuring the expression of luciferase.</p><p><b>RESULTS</b>The survivin promoter exhibited a higher transcriptional activity than PSMA promoter and enhancer in tumor cell lines, and the S2pro promoter showed the highest activity, reaching one third of that of the CMV promoter.</p><p><b>CONCLUSION</b>The survivin promoter is highly activated in prostate cancer cell lines and may serve as a new tool for the transcriptional targeting gene therapy of prostate cancer.</p>

Detection of transcriptional activities of tumor-specific survivin promoter in human prostatic carcinoma / 中华男科学杂志

<p><b>OBJECTIVE</b>To clone DNA sequence of the survivin promoter and study is transcriptional activities in human prostate cancer cells and normal Chang liver cells.</p><p><b>METHODS</b>The fragment of the survivin promoter was acquired by PCR amplification and inserted into pPRIME vectors to reconstruct a recombinant plasmid named pPRIME-S1pro and pPRIME-S2pro. Then the reconstructed plasmid was transiently transfected into human prostate cancer cells lines LNCaP and normal Chang liver cells. The transcriptional activities of the survivin promoter in various cells was determined by measuring the expression of green fluorescent protein (GFP).</p><p><b>RESULTS</b>The survivin promoter had transcriptional activities in LNCaP cells and the transcriptional activity of the S2pro was much higher that of the S1pro, reaching a level of 39% of the transcriptional activity of the CMV promoter.</p><p><b>CONCLUSION</b>The survivin promoter cloned in the therapy for prostate cancer.</p>

Inhibition of survivin gene expression in PC-3 cells by RNAi / 中华男科学杂志

<p><b>OBJECTIVE</b>To construct eukaryotic expression vectors by using the pSilencer3. 1-H1 neo vector for inhibiting human survivin gene by RNA interference, and to detect the effect of the silenced survivin gene on PC-3 cells.</p><p><b>METHODS</b>Three target gene segments were synthesized and cloned into the pSilencer3. 1-H1 neo vector respectively to construct three recombinant eukaryotic expression vectors: pSilencer3. 1-SVV1, pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3, which were identified by enzyme digestion analysis and DNA sequencing. Then the PC-3 cells were transfected with the recombinant vectors and the interference effect detected by RT-PCR, Western blot and immunohistochemical staining. The apoptosis index of the PC-3 cells was detected by flow cytometry and their proliferation detected by MTT method.</p><p><b>RESULTS</b>Enzyme digestion analysis and DNA sequencing showed that three target segments were cloned into pSilencer3. 1-H1-neo vectors. The results of RT-PCR, Western blot and immunohistochemical staining indicated that pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors could knock down the transcription and expression of survivin gene. After transfected with pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors, the apoptosis index of the PC-3 cells was increased by 10% - 15% and their growth obviously slowly down.</p><p><b>CONCLUSION</b>The transcription and expression of survivin gene were inhibited effectively by the recombinant eukaryotic expression vectors (pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3) in the prostate cancer cell line PC-3. After transfected with pSilencer3. 1-SVV2 and pSilencer3. 1-SVV3 vectors, the apoptosis index of the PC-3 cells was increased and their growth inhibited.</p>

Effects of tanguticum maxim polysaccharide on ulcerative colitis induced by TNBS in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the effects of Tanguticum Maxim polysaccharide (TMP-1) on TNBS-induced colitis in rats.</p><p><b>METHOD</b>Rats with TNBS/ethanol-induced colitis were used and treated with TMP-1 and dexamethasone (DX). Seventy-two rats, including animals with TNBS-induced colitis, were treated with saline, TMP-1 (100, 200, 400 mg.kg-1) and DX. White blood cells were counted on the fifth day and the rats were killed by ether on the sixth day. SOD activity in serum, MPO and SOD activity of colonic tissue were measured.</p><p><b>RESULT</b>The remarkable effects of TMP-1 at dosage of 200, 400 mg.kg-1 on TNBS-induced colitis were observed. The ulcerative area was diminished and weight of colon was reduced. White blood cell population was reduced, SOD activity in serum and SOD activity of colon tissue were remarkably increased, and, MPO activity of colonic tissue was reduced.</p><p><b>CONCLUSION</b>TMP-1 has significant effects on TNBS-induced colitis in rats with lower side effects, which suggests the effective component of rhubarb on colitis perhaps is TMP. The mechanism of the actions of TMP may relate to its antiflammation, antioxidation and immunoloregulation.</p>

Bacterial biofilms and its clinical significance / 中华检验医学杂志

Biofilms are microbial communities which are enclosed within a matrix of exopolysaccharides produced by the bacteria,fungi and protozoa growing intimately on a living or inert surfaces.Further researches on bcterial biofilms formation and molecular machines,pathogenic mechanisms (especially drug resistance),detection and treatment maybe provide novel pathways to refractory infections caused by bcterial biofilms.