Analysis of the Influence of Different MCV Level on the Platelet False Increase Counts / 现代检验医学杂志

Objective To evaluate the influence of the mean corpuscular volume (MCV) in different ranges on PLT counts,provide the theoretical basis for making PLT counts review rules in laboratory.Methods From March 2016 to August 2016 in Xijing Hospital department of outpatient who perform complete blood cell count were randomly divided into 3 groups,MCV≤65 fl in A group,65 fl＜MCV≤70 fl in B group and 70 fl＜MCV≤75 fl in C group.The reference method (Coulter principle) compared with the fluorescence method,accuracy analysis of different monitoring methods of counting PLT.Results PLT I and PLT F count values in A group was (322.8± 109.1) × 109/L and (282.60± 100.5) × 109/L respectively,and there was significant differences between the two groups (t=6.799,P＜0.05).In B group,the count values was (305.7 ± 111.7)× 109/L and (304.8 ± 112.3)× 109/L respectively,and there was no differences between two groups.In C group,the count values was (292.2±84.4) × 109/L and (291.6±84.4) × 109/L respectively,and there was no differcnces between two groups neither.Conclusion When small red blood cells or blood cell debris present in blood circulation,MCV≤65 fl,the reference testing (Coulter principle)of platelets causes a false increase in platelet count.

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>

Sexual hormone levels in semen and germ cell apoptosis / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the relationship between sexual hormones in semen and germ cell apoptosis in male population.</p><p><b>METHODS</b>Sixty-six infertile patients and thirty fertile males were selected randomly. The levels of folicle stimulating hormone ( FSH), prolactin (PRL), luteinizing hormone (LH), and testosterone (T) in semen were measured by ELISA. Terminal deoxynucleotidyl transferase mediated UTP nick end labeling (TUNEL) was used for the detection of germ cell apoptosis.</p><p><b>RESULTS</b>The levels of FSH, LH, PRL, T in thirty fertile men were (1.63 +/- 0.15) U/L, (2.18 +/- 0.21) U/L, (6.34 +/- 0.30) nmol/L, (1.85 +/- 0.11) nmol/L, respectively, and germ cell apoptosis rate was (4.61 +/- 1.23)%. FSH, LH, PRL, T levels in infertile group were (1.25 +/- 0.18) U/L, (1.76 +/- 0.32) U/L, (5.86 +/- 0.13) nmol/l, (1.45 +/- 0.13) nmol/, respectively, and germ cell apoptosis rate was (18.36 +/- 2.04)%. There were significant differences in all parameters between infertile group and fertile group. The levels of FSH, LH, PRL, T were negatively correlated with germ cell apoptosis rates( r = -0.88, -0.93, -0.90, -0.98). The volume of apoptotic germ cell decreased, and chromatin was compacted to form cell-membrane blebs and apoptotic bodies.</p><p><b>CONCLUSION</b>Low concentration of sexual hormones may increase the apoptosis of germ cells, which can induce male infertility.</p>

Sodium nitroprusside facilitates human sperm capacitation and acrosome reaction / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effect of nitric oxide (NO) on human sperm capacitation and acrosome reaction (AR).</p><p><b>METHODS</b>Different concentrations of sodium nitroprusside (SNP) were added to the sperm suspension from 48 healthy fertile men, and the suspension was incubated in 1 x Earle at 37 degrees C for 1 hour. Progesterone was used to induce AR for 15, 30, 45 and 60 min, and then acid phosphatase (ACP) activity in the suspension before and after capacitation and at different time of AR was measured by p-nitrophenyl sodium phosphate assay. In the meantime, sperm motile parameters were assayed by CASA to observe sperm capacitation and AR.</p><p><b>RESULTS</b>ACP activity and sperm motile parameters increased in the 50 approximately 100 nmol/L NO concentration group, showed no significant variation in the 150 approximately 200 nmol/L group, and decreased in the 250 approximately 300 nmol/L group.</p><p><b>CONCLUSION</b>NO can facilitate sperm capacitation, AR and sperm motile parameters in low concentration and suppress them in high concentration. ACP activity assay of sperm is an objective and reliable method to evaluate sperm capacitation and AR in whole sperm population.</p>

Study on the nucleic acid of E. coli bacteriophage with broad host range and its sterilization effect to sewage samples from the environment / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the change of nucleic acid sequence and the germicidal effect of an E. coli bacteriophage with broad host range isolated from hospital sewage as well as to study the mechanism of phage host specificity and the effect of killed bacteria by phage-disinfectant to the samples from sewage water.</p><p><b>METHODS</b>To extract the nucleic acid from phage f(2) and phage with broad host range using anti-serum-carbamidine hydrochloride assay. Purity with agarose gel electrophoresis was then evaluated. Differences of nucleic acid sequence between phage f(2) and phage with broad host range with reverse transcription-polymerase chain reaction (RT-PCR) and random amplified polymorphic DNA (RAPD)-PCR were also comparing and analysed. Through observing the germicidal test of phage f(2) and phage with broad host range to samples from environment, different sterilization effects between the two phages were compared.</p><p><b>RESULTS</b>Analystic test for nucleic acid revealed that the two phages both belonged to 6000 bp, single-stranded RNA bacteriophage. Significant differences in their specificity of RAPD-PCR and RT-PCR were found during the changed of host range; with 26 RAPD-cDNA differential fragments found that in two phages RAPD-PCR products. The RT-PCR product of phage f(2) was 450 bp cDNA fragment, but the phage with broad host range did not show PCR product. Treating the sewage water with phage under broad host range, the germicidal test showed that the cleaning rate of E. coli bacteria and phage f(2) in water samples from environment could reach 36.75% - 56.28%, 30.84% - 47.96%, 19.19% - 35.06% and 13.05% - 27.85%, respectively.</p><p><b>CONCLUSION</b>The cleaning rates to E. coli and bacteria by phage with broad host range were obviously higher than phage f(2) (P = 0.000). Analytic test for nucleic acid indicated that host-specific lytic effect of phage with broad host range had been changed at genetic level.</p>

Research on the correlation between lupus erythematosus cell and diseases activity in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To observe the morphologic changes of peripheral blood lupus cell in patients with systemic lupus erythematosus and investigate its relationship with disease activity in systemic lupus ery- thematosus.Methods Modified classical blood clotting method to observe the morphological changes of pe- ripheral blood lupus cells in 80 cases with systemic lupus erythematosus.Fifty cases were in active stage,and 30 cases were in stable stage.Comparison of serum autoantibody,complement and SLEDAI were also carried out.Results There was significant association between lupus cells in special morphous and autoantibody, such as anti-double-stranded DNA antibody,anti-nucleosome antibodies,complement C3、C4 and SLEDAI(r= 0.588,P=0.056:r=0.759,P=0.135;r=-0.648,P=0.058;r=-0.589,P=0.057,r=0.686,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.