Clinical study of different sedative methods in critically ill patients after heart surgery / 天津医药

Objective To study the effect of light sedation and traditional sedation (moderate sedation with daily sedation interruption) on hemodynamic indexes and prognosis in critically ill patients after cardiac surgery. Methods A total of 134 patients who were ventilated delay after heart surgery in our hospital from January to June 2017 were enrolled in this study. The patients were randomly divided into light sedation group (RASS score-1-1, n=65) and traditional sedation group (RASS score -3--2, n=69). All patients received sufentanil for postoperative analgesia. The light sedation group received propofol and/or dexmedetomidine as sedative drugs after operation, and the conventional sedation group used midazolam for postoperative sedation. The hemodynamic indexes, the first time of weaning off the ventilator, the duration of mechanical ventilation and ICU stay were compared between the two groups. Patients with low cardiac output syndrome after surgery were analyzed in subgroups. Results (1) There were no significant differences in heart function, operative complications and other indicators between the two groups after surgery (all P＞0.05). The low cardiac output syndrome was found in 12 patients in the light sedation group and 10 cases in the traditional sedation group. (2) Hemodynamic monitoring results displayed that the sedation/central venous oxygen saturation (SvO2/ScvO2) and cardiac index (CI) were higher after sedation than before sedation in both groups (all P＜0.05), but there was no significant difference between the two groups (all P＞0.05). Subgroup analysis showed that the SvO2/ScvO2index was higher in patients with low cardiac output syndrome in the traditional sedative group than that in the light sedation group (P＜0.05). There was no difference in the SvO2/ScvO2 index in patients with non-low cardiac output syndrome between two groups. (3) Compared with the traditional sedation group, the first off-line time, the total mechanical ventilation after surgery and the ICU stay time were significantly shortened, and the incidence of postoperative delirium was decreased in the light sedation group (all P＜0.05). Subgroup analysis showed that in patients with non-low cardiac output syndrome, the first off-line time, total postoperative mechanical ventilation time and total ICU stay were significantly shorter in the light sedation group than those in the traditional sedation group (all P＜0.05). There was no significant difference in patients with low cardiac output syndrome between the two groups (P＞0.05). Conclusion Patients with non-low cardiac output syndrome after cardiac surgery benefit significantly from the superficial sedative strategy, and the postoperative mechanical ventilation time and ICU residence time are reduced. The moderate sedation may contribute to the early cardiac function recovery in patients with low cardiac output syndrome.

Values of different biomarkers for diagnosis of Henoch-Schönlein purpura in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To screen biomarkers which can be used as an auxiliary method in the diagnosis of Henoch-Schönlein purpura (HSP) and to evaluate their diagnostic values by receiver operating characteristic (ROC) curve analysis.</p><p><b>METHODS</b>A total of 127 children diagnosed with HSP between April 2012 and March 2014 were included in the HSP group and an equal number of healthy children were included in the control group. Twelve parameters, i.e., serum amyloid protein A (SAA), interleukin-6 (IL-6), immunoglobulins (IgA, IgG, IgM, and IgE), C-reactive protein (CRP), white blood cell (WBC) count, complements C3 and C4, anti-streptolysin O, and ferritin, were analyzed. The values of the screened biomarkers for diagnosis of HSP were assessed by ROC curve analysis.</p><p><b>RESULTS</b>The HSP group had significantly higher levels of SAA, IL-6, CRP, WBC, IgA, and IgM than the control group (P<0.05). The areas under the ROC curve of SAA, IL-6, WBC, IgA, and IgM for the diagnosis of HSP were higher than 0.7 (P<0.05). The optimal cut-off values of SAA, IgA, IgM, WBC, and IL-6 for the diagnosis of HSP were 3.035 μg/mL, 1579.5 mg/L, 922.5 mg/L, 8.850 × 10⁹/L, and 7.035 pg/mL, respectively; the corresponding sensitivities of the optimal cut-off values for the diagnosis of HSP were 95.1%, 75.6%, 72.3%, 78.0%, and 63.4%, respectively, and the corresponding specificities were 90.2%, 85.4%, 82.4%, 70.7%, and 80.5%, respectively.</p><p><b>CONCLUSIONS</b>SAA, IgA, IgM, WBC, and IL-6 are valuable biomarkers for clinical diagnosis of HSP and among them SAA seems to be the best one.</p>

Construction and identification of a eukaryotic expression vector containing human homeobox gene / 中国实验血液学杂志

In order to investigate the special role of HOXB4 in expansion and self renewal of hematopoietic stem cells, the cDNA of HOXB4 was extracted and cloned from umbilical cord blood mononuclear cells by using RT-PCR. Then the eukaryotic expression bicistronic plasmid vector pIRES2-EGFP/HOXB4 was designed and constructed after cutting HOXB4 and pIRES2-EGFP respectively by restriction enzyme EcoRI and BamHI. The recombinant plasmid was delivered into competent cells of Escherichia coli. The successful construction of plasmid was confirmed by the identification of endonuclease cutting and sequencing. The results showed that the HOXB4 cDNA was cloned successfully from umbilical cord blood mononuclear cells and the recombinant eukaryotic expression bicistronic plasmid vector was constructed, and then introduced it into 293T cells successfully. It is concluded that a pIRES2-EGFP/HoxB1 eukaryotic expression bicistronic plasmid vector has been constructed successfully, which results provide a useful material basis for exploration of HoxB4 function in the proliferation and differentiation of hematopoietic cells.

Exploration on the establishment of animal models for gouty nephropathy complicated with chronic renal failure / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the method for establishing animal models of gouty nephropathy complicated with chronic renal failure.</p><p><b>METHODS</b>Six-eight weeks old male Wistar rats were fed with 10% fodder yeast. The adenine at the daily dose of 100, 150, 200, 250, and 300 mg/kg was administrated to them by gastrogavage. The serum levels of blood urea nitrogen (BUN), creatinine (Cr), and uric acid (UA) were dynamically monitored. Meanwhile, the pathological changes of rat kidney were observed.</p><p><b>RESULTS</b>Compared with the normal control group, serum BUN, Cr, and UA obviously increased in rats administered with 100 mg/kg for 7 days (P<0.05). Meanwhile, pathological changes as gouty nephropathy occurred. Along with the prolongation of the modeling time, the aforesaid biochemical indices and pathohistological changes of the kidney were more obvious. The blood Cr level just reached the chronic renal failure level on the 26th day of the administration (about the 4th week), and obviously exceeded the renal failure level on the 41st day (about the 6th week). The blood UA level increased to a higher level on the 7th day of modeling, and maintained at a higher level for a long time. It decreased rapidly from the 41st day to the 48th day. The renal pathological examination showed aggravated infiltration of lymphocytes and stromal fibrous proliferation. On the 48th day of modeling, the proliferation of the fibrous tissue and the interstitial fibrosis were obvious on the bases of the aforesaid changes. The serum BUN, Cr, and blood UA obviously increased in the rats administered with 150, 200, 250, and 300 mg/kg when compared with the normal control group, reaching the level of chronic renal failure (P<0.05). These levels obviously decreased 17 days after restoring to normal fodder feeding, and approached the normal levels till the 35th day.</p><p><b>CONCLUSION</b>Ideal experimental animal models of gouty nephropathy complicated with chronic renal failure could be established in male Wistar rats by feeding with 10% fodder yeast and 100 mg/kg adenine by gastrogavage for 5 weeks.</p>

Association of ABCG2 gene C421A polymorphism and susceptibility of primary gout in Han Chinese males / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association between a C421A single nucleotide polymorphism (SNP) in exon 5 of ATP-binding cassette, sub-family G (WHITE), member 2 (ABCG2) gene and susceptibility of primary gout in Han Chinese males.</p><p><b>METHODS</b>For 200 male patients with primary gout and 235 controls, the genotype of C421A locus was analyzed by PCR and direct sequencing. Blood glucose, uric acid, total cholesterol, triglycerides, creatinine and urea nitrogen was measured by an automatic biochemical analyzer.</p><p><b>RESULTS</b>Compared with the controls, there was a higher frequency for AA genotype and A allele of the rs2231142 SNP in gout patients (22.5% vs. 8.5% by genotype; 44.9% vs. 32.3% by allele). The association with gout reached significance (chi-square =15.91, P< 0.001, crude OR=3.02, 95% CI:1.36-4.90 and OR (adjusted by age)=1.80, 95% CI: 1.32-2.45 by dominant mode; chi-square=6.82, P=0.009, OR=1.67, 95% CI: 1.54-2.27 by recessive mode). Blood glucose, uric acid, triglycerides, creatinine and urea nitrogen levels in gout patients were significantly higher than those of controls (P< 0.001).</p><p><b>CONCLUSION</b>The C421A SNP, in particular AA phenotype, may be associated with susceptibility of primary gout in Han Chinese males.</p>

The detection and clinical significance of hepatic fibrosis index / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the relationship between levels of serum HA, LN, IV-C, PC III of chronic hepatitis and indexes of hepatic fibrosis.</p><p><b>METHODS</b>The levels of serum HA, LN, IV-C and PC III of chronic hepatitis of 124 cases and health 18 cases were measured by radio immunoassay, combined with clinical characteristics and 33 cases pathologic slice etc. The diagnostic of the indexes of serum was analyzed with statistics.</p><p><b>RESULTS</b>HA and IV-C are parallel in chronic hepatitis periods. LN and PC III are concert in the same pathologic periods. In G4 period PC III is nearly closed with comparative group. The value of HA, LN, NV-C and PC III in the chronic hepatitis group was significantly higher than that in the normal comparative group. Conclusion The levels of serum HA LN IV-C and PC III are in concert with the degree of hepatic fibrosis, and these indexes are valuable for chronic hepatitis diagnoses combined with the clinic. LN and PC III are coincidence with hepatic fibrosis degree before G4 period.</p>

Association of the exon 8 and intron 8 polymorphisms of the human urate transporter 1 gene with primary hyperuricemia in Chinese Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of the exon 8 and intron 8 polymorphisms of the human urate transporter 1 gene SLC22A12 with primary hyperuricemia (HUA) in Chinese Han population.</p><p><b>METHODS</b>Genomic DNA from 215 individuals with HUA and 323 controls was extracted. The exon 8 and intron 8 of the SLC22A12 gene was amplified by polymerase chain reaction (PCR). PCR product was sequenced directly. Single nucleotide polymorphisms (SNPs) were detected and the association of the SNPs with primary HUA was assessed.</p><p><b>RESULTS</b>(1) Two SNPs were identified, they were T1309C located in exon 8 (rs7932775) and -103A to G located in intron 8. Pairwise linkage disequilibrium analysis displayed an absolute linkage disequilibrium between the two SNPs (D'= 1). (2) The minor allele frequencies for both SNPs were 51.9% in HUA patients, which were significantly different from that of controls (42.4%)(P< 0.01). (3) The genotype frequencies of GG+ GA and CC+ CT in HUA patients were significantly higher than that in controls (80.0% vs. 69.0%, P< 0.01). (4) Individuals of both GG+ GA and CC+ CT genotypes had 1.79 fold increase of HUA risk (OR= 1.794, 95%CI: 1.19-2.70).</p><p><b>CONCLUSION</b>These findings indicated that T1309C and -103A to G polymorphisms of the SLC22A12 gene were associated with primary HUA in Chinese Han population.</p>

Effect of dendritic cells on expansion and function of autologous natural killer cells in vitro / 中国实验血液学杂志

The objective of this study was to investigate the effect of dendritic cells (DCs) on expansion and function of autologous natural killer (NK) cells and its mechanism in vitro. NK cells were expanded from peripheral blood mononuclear cells (PBMNCs) of healthy volunteers in stem cell growth medium (SCGM) supplemented with rhIL-2 (control group) in 24-well culture plates at 37 degrees C in a humidified CO(2)-containing atmosphere. NK cells were cultured with autologous DCs in the ratio of 5 to 1 (group 5:1) or 1 to 1 (group 1:1) from day 10 after expansion. Total cells of every group were counted and the expression of CD3, CD16/56 on the surface of NK cells was assayed by flow cytometry on days 7, 14 and 21 to calculate the expansion of NK cells. Cytotoxicity of expanded NK cells against K562 cells was assayed by MTT method. TNF-alpha and IL-12p70 were detected in culture supernatants by sandwich ELISA. The results indicated that the expansion and cytotoxicity of NK cells were improved after mixed with autologous DCs. Furthermore, when DCs were mixed with NK cells, the ratio of DCs to NK cells was higher, the expansion and cytotoxicity NK cells were higher. On day 14, the expansion multiple in control, group 5:1 and group 1:1 were 16.26 +/- 1.58, 29.25 +/- 4.01 and 21.23 +/- 2.91 respectively. The expansion multiple of group 5:1 was much higher than that of the other two groups (p < 0.05). The expressions of CD3(-), CD56/16(+) on surface of NK cells in control, group 5:1, group 1:1 were (34.8 +/- 5.1)%, (64.6 +/- 7.8)% and (50.6 +/- 8.7)% respectively and that of group 5:1 was the highest (p < 0.05). The cytotoxicities against K562 cells in control, group 5:1 and group 1:1 were (63.7 +/- 3.8)%, (87.4 +/- 6.8)% and (75.4 +/- 6.3)% respectively. The cytotoxicity of group 5:1 was higher than that in the other two groups also (p < 0.05). TNF-alpha and IL-12p70 levels in culture supernatants when DCs and NK cells were mixed in the ratio of 5 to 1 were much higher than those in culture supernatants of DCs and NK cells alone or in culture supernatants when DCs and NK cells were mixed in the ratio of 1 to 1 (p < 0.05). It is concluded that the expansion and cytotoxicity of NK cells can be improved by DCs and it depended on the mixed ratio of DCs to NK cells. The elevated expansion of NK cells by DCs bears relation to IL-12 produced by DCs. The enhanced cytotoxicity of NK cells is associated with TNF-alpha secreted by NK cells.

Research Progress of microRNAs and human hematological diseases - review / 中国实验血液学杂志

The microRNAs (miRNAs) are an evolutionarily conserved class of 22 nucleotide (19 - 25 nt) non-coding RNAs. The miRNAs are partially complementary to 3' untranslated region of target mRNA, resulting in the repression of gene expression at post-transcriptional level. The miRNAs have been associated with diverse biological processes. This review summarizes recent progress of research on the characteristics and function of miRNAs, and the role of miRNAs in hematological malignancy development.

Effect of pericardial suction blood re-transfusion in off-pump coronary artery bypass grafting on inflammatory cytokines, myocardial injury and pulmonary function / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of pericardial suction blood re-transfusion in off-pump coronary artery bypass grafting (CABG) on inflammatory cytokines, myocardial injury and lung function.</p><p><b>METHODS</b>31 patients of off-pump CABG were divided into two study groups (OPCABG1 group and OPCABG2 group) according to the amount of pericardial suction blood re-transfusion beyond or less than 600 ml. 13 patients of on-pump CABG were control group. Serum samples from vein were collected for measurement of IL-6, IL-8, IL-10 and TNF-alpha pre-operation and 1, 4, 24, 48 hours post-operation respectively. The results of CK-MB, TnI, AaDO2 and PaO2/FiO2 were recorded.</p><p><b>RESULTS</b>Patients of the three groups had no significant difference in terms of gender, age, bodyweight, history of hypertension and cardiac infarction and diabetes, EF and left ventricular end diastolic of pre-operation, the amount of bypass graft and shed blood. Of the three groups, IL-6, IL-8 and IL-10 reached peak level one hour after the operation, and dropped to the pre-operation level 72 hours after the operation. One hour after the operation, the level of IL-6 and IL-8 in OPCABG1 group was higher than in OPCABG2 group (P < 0.05) and about the same in CABG group (P > 0.05). Four hours after the operation, the level of CK-MB in OPCABG1 group was lower than that of CABG group (P < 0.05) and about the same in OPCABG2 group (P >0.05). 4 and 24 hours after the operation, the level of TnI in OPCABG1 group was lower than that of CABG group (P < 0.05) and about the same in OPCABG2 group (P > 0.05). Among the three groups, there was no significant difference in AaDO2 and PaO2/FiO2.</p><p><b>CONCLUSIONS</b>Re-transfusion of large amount of pericardial suction blood can increase serum level of IL-6, IL-8, but it can not cause myocardial injury and affect the gas exchange function of lung significantly.</p>

PFGE of Shigella flexneri 4c isolates from food-poisoning outbreaks and sporadic diarrhea patients / 中华预防医学杂志

<p><b>OBJECTIVE</b>To know the molecular characteristic of Shigella flexneri 4c isolates from patients in two food-poisoning outbreaks and one sporadic diarrhea case in Hangzhou, China.</p><p><b>METHODS</b>S. flexneri isolates from patients in two food-poisoning outbreaks (outbreak 1 and outbreak 2, n = 13 and n = 12, respectively) and one sporadic diarrhea patient (n = 1) in Hangzhou during 2003 and 2005 were serotyped. Antibiotic resistances of these isolates were measured by the Kirby-Bauer method. Invasive plasmid antigen gene ipaH was examined by PCR. Pulse field gel electrophoresis (PFGE) was performed for molecular typing.</p><p><b>RESULTS</b>In outbreak 1, all 13 isolates were S. flexneri 4c, of them 6 isolates tested were quite different in PFGE patterns with dice coefficient from 0.78 to 0.92. In outbreak 2, 10 isolates were S. flexneri 4c and 2 isolates were S. flexneri X, however their PFGE patterns were almost identical (dice coefficient > 0.8). Compared to the two outbreaks isolates, the sporadic isolate was demonstrated with a distinct PFGE pattern (dice coefficient < 0.8). The antibiotic resistance patterns with 14 kinds of antibiotics had a little difference among the isolates from outbreak 1, outbreak 2 and sporadic diarrhea patient, but the same pattern was found among 10 isolates of S. flexneri 4c and 2 isolates of S. flexneri X from outbreak 2.</p><p><b>CONCLUSIONS</b>PFGE might distinguish the isolates from these two outbreaks and the sporadic diarrhea patient. Some differences in PFGE patterns, serotypes and antibiotic resistance patterns might occur among S. flexneri 4c isolates during an outbreak.</p>

Multiplex real-time PCR detecting Salmonella, Shigella and diarrheagenic Escherichia coli / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop a multiplex real-time PCR for the detection of Salmonella invasion protein A gene (invA), enterotoxigenic Escherichia coli (ETEC) heat-labile I enterotoxin gene (elt), and Shigella or enteroinvasive E. coli (EIEC) invasive plasmid antigen H gene (ipaH).</p><p><b>METHODS</b>Under the optimized reaction conditions of the multiplex real-time PCR, invA, elt, and ipaH were determined in 10-fold series of dilution of DNA extracted from Salmonella enterica serovar Typhimurium, ETEC 44815 strain and Shigella F301 strain. The three genes were examined in 90 fecal samples from diarrhea patients using the multiplex real-time PCR. When PCR-positive samples were found, the target strains were isolated and identified.</p><p><b>RESULTS</b>The detectable concentration for this multiplex real-time PCR was 10 CFU/microl for Shigella F301 strain, 10(2) CFU/microl for S. enterica serovar Typhimurium and ETEC 44815 strain, respectively. Out of 90 fecal samples from diarrhea patients, thirteen were found positive for elt gene (14.4%), and five were found positive for ipaH gene (5.6%). Three E. coli strains positive for elt gene and four E. coli strains positive for ipaH gene were isolated successfully from the PCR-positive samples mentioned above. The detection of invA, elt and ipaH genes was completed in 10 h, which included an enrichment period of 6 h.</p><p><b>CONCLUSION</b>The multiplex real-time PCR assay can detect invA, elt, ipaH simultaneously in a single reaction, moreover, it can detect for virulence genes in strains of Salmonella, ETEC, and Shigella or EIEC and screen these pathogens in fecal specimens from patients with diarrhea with a high specificity.</p>

Molecular typing on Vibrio parahaemolyticus isolates from Hangzhou, China, during 2000-2002 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the molecular epidemiology of Vibrio parahaemolyticus isolates from clinical and environmental samples collected in Hangzhou area during 2000 and 2002.</p><p><b>METHODS</b>V. parahaemolyticus isolates from food-poisoning, sporadic diarrhea patients and seafood in market in Hangzhou during 2000 and 2002 were serotyped. From clinical isolates of serotype O3:K6 and environmental isolates of serotype O3:KUT, virulence genes, tdh and trh, were examined by PCR. Molecular typings [including ribotyping, random amplified polymorphic DNA analysis (RAPD), enterobacterial repetitive intergenic consensus PCR (ERIC-PCR)], were also performed.</p><p><b>RESULTS</b>Among 13 food-poisoning outbreaks caused by V. parahaemolyticus, O3:K6 strains with tdh positive and trh negative were detected in 11 episodes (84.6%), and O4:K8 strains with tdh positive and trh negative were responsible for other 2 outbreaks (15.4%). In 34 V. parahaemolyticus isolates from sporadic diarrhea patients, O3:K6, O4:K8, and O1:KUT strains with tdh positive and trh negative were detected with proportions of 26.5% (9/34), 17.6% (6/34), and 38.2% (13/34), respectively, and other 6 strains were not able to be serotyped. Of 64 isolates from seafood in which both tdh and trh were negative except trh was positive in one O1:KUT strain, 37 belonged to 7 serotypes except O3:K6 or O4:K8, and 9 were not able to be serotyped. The fingprintings of ribotyping, ERIC-PCR, and RAPD showed that almost all of O3:K6 isolates from food-poisoning and sporadic diarrhea patients were genetically close to each other and there were obviously genetical difference between O3:K6 isolates from clinic and O3:KUT isolates from environment.</p><p><b>CONCLUSION</b>There was a distinct serotype distribution between V. parahaemolyticus isolates from clinic patients and seafood. One group of close related V. parahaemolyticus O3:K6 strains with tdh positive and trh negative seemed to be prevailing in Hangzhou in those years.</p>

Study on rapid generation of dendritic cells from K562 cell line induced by A23187 alone / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore a simple, rapid and efficient way to generate dendritic cells from leukemic cells.</p><p><b>METHODS</b>K562 cells were cultured with calcium ionosphere A23187 alone, A23187 plus GM-CSF, or a DC differentiation cocktail consisting of GM-CSF, IL-4 and TNF-alpha, respectively. The expression of surface markers of induced DCs was analyzed by flow cytometry. The K562-DCs stimulating the proliferation of allo-genetic naive T cells and inducing cytotoxicity of T cells were determined by MTT assay.</p><p><b>RESULTS</b>Microscopic examination revealed that under all the three culture conditions, K562 cells became displaying DC morphology. At 72 hours in the two culture systems containing A23187, there were higher proportions of cells with dendritic morphology [(69.5 +/- 17.2)% and (73.1 +/- 13.9)%, respectively] than that in the cocktail system [(28.5 +/- 12.3)%] (P < 0.05). And the same did when cultured for 7 days [(69.5 +/- 17.2)%, (73.1 +/- 13.9)% respectively vs (51.2 +/- 10.7)%, P < 0.05]. In the 7-day cultures, the percentage of CD1a expressing cells was lower [(8.2 +/- 2.3)% and (10.3 +/- 5.1)% vs (17.2 +/- 1.6)%, respectively] while the CD83 expressing cells was higher [(85.6 +/- 8.8)% and (82.4 +/- 9.1)% vs (77.4 +/- 12.9)%, respectively] compared with that in the cocktail system (P < 0.05). No significant difference was found in the allogeneic T cell proliferation response and induced T cell cytotoxicity between A23187 containing and cocktail groups (P > 0.05).</p><p><b>CONCLUSIONS</b>A23187 treatment is a simple, rapid and efficient in vitro strategy for inducing dendritic cell from leukemic cells.</p>

Characteristics of virulence gene in Vibrio parahaemolyticus strains isolated from clinical patients and environment in Hangzhou, China / 中华预防医学杂志

<p><b>OBJECTIVES</b>To investigate the characteristics of virulence gene in Vibrio parahaemolyticus strains isolated from clinical patients and environment in Hangzhou, China.</p><p><b>METHODS</b>Thermostable direct hemolysin gene (tdh) and thermostable direct hemolysin-related hemolysin gene (trh) were determined in a total of 174 strains of V. parahaemolyticus isolated from patients and environment (seafood) in Hangzhou area by PCR.</p><p><b>RESULTS</b>The tdh was found in 92 out of 94 V. parahaemolyticus strains from food poisoning patients and in 33 out of 34 strains from sporadic diarrhea patients, and trh was not detected in all above clinical strains. Meanwhile the tdh was negative in all V. parahaemolyticus strains from environment, and the trh was also negative except one strain with urease activity. All strains with trh negative had no the activity of urease.</p><p><b>CONCLUSIONS</b>The V. parahaemolyticus strains from food poisoning patients and sporadic diarrhea patients are tdh positive and trh negative. The V. parahaemolyticus strains with tdh negative and almost trh positive in environment might be a potential pathogen in Hangzhou.</p>

The induction and function study on dendritic cells derived from blasts from patients with acute myelogenous leukemia / 中国实验血液学杂志

To investigate the induction method and function of dendritic cells (DC) derived from acute myelogenous leukemia (AML) blasts in vitro, cytokine-supplemented suspension cultures of leukemia blasts in 25 AML patients were performed. Mononuclear cells were cultured for 8 to 12 days using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF), recombinant human interleukin-4 (rhIL-4) and recombinant human tumor necrosis factor-alpha (rhTNF-alpha). Morphology, phenotype, cytogenetics, and function of induced cells were studied. The results showed that after culture for 3 days, cells in 20/25 AML cases demonstrated an increase in size with dendritic morphology. After culture for 8 - 9 days, the percentage of such cells reached peak. When cultured for 12 days, the total number of cells and the number of cells with DC morphology decreased greatly. Phenotypic analyses of cells (11/20 cases) were measured by flow cytometry before and after culture. Before culture, cells did not express CD1a, CD80 and CD83, while expressed CD54, CD86 and HLA-DR with low frequency. After culture, cells upregulated CD1a, CD80, CD83, CD54, CD86 and HLA-DR significantly. A marked increase of the T-cell stimulatory capacity could be generated in Allo-MLRs. FISH confirmed the leukemic origin of generated cells. In conclusion, leukemia-derived DC can be generated from AML blasts using cytokine combination (GM-CSF, IL-4, and TNF-alpha) in vitro.

Study on cryopreservative methods for dendritic cells derived from K562 cell line / 中国实验血液学杂志

To investigate the biological properties of cryopreserved dendritic cell (DC) derived from K562 cell line, thus to provide a simple, quick and efficient preservative method of DC for infusion of DC to patients with leukemia after complete remission, fresh DC induced from K562 cell line (K562-DC) was frozen in -196 degrees C liquid nitrogen and -80 degrees C mechanical freezer by method of steps (RPMI 1640 with 10% DMSO, 20% FCS as cryopreservatives), and thawed in different time, respectively. Survivals of cryopreserved and fresh K562-DC, expression of surface antigens, stimulating index (SI) and cytotoxic eliminating rate were detected. The results of fresh induced cells were compared with that of cryopreserved ones. The results showed that before and after frozen in liquid nitrogen, the morphological characteristics of K562-DC had no distinct change; and both their expression rates of surface molecular and capacity to stimulate allogeneic lymphocyte had no statistic significance (P > 0.05). In addition, there were no differences in terms of viability, stimulatory capacity and cytotoxicity of K562-DC from two ways for less than one month (P > 0.05), but there were differences when frozen for more than one month (P < 0.05). It is concluded that there is no significant difference when frozen less than one month between liquid nitrogen and -80 degrees C freezer; but when time is more than one month, K562-DC frozen in -196 degrees C liquid nitrogen is better than that in -80 degrees C freezer.

Development of Multiplex Real-time PCR for Detection of Toxigenic Vibrio cholerae and Virbio parahaemolyticus / 微生物学通报

A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.