Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

Objective To discuss effective nursing methods for 18F-FDG PET/CT test of colorectal cancer,in order to improve the understanding and nursing level of retention enema.Methods Totals of 130 patients who had suffered from conventional 18F-FDG PET/CT test for high FDG in colorectum were chosen,and performed with partial water enema in 18F-FDG PET/CT scans.Positive results of 18F-FDG PET/CT and pathological mechanism after enteroscope or surgery were compared.Results The sensitivity,specificity and accuracy for detecting colorectal cancer were respectively 80.8％,58.5％ and 81.5％ with conventional 18FFDG PET/CT scan,and 100.0％,97.5％ and 99.2％ with retention enema,and the difference was statistically significant (x2 =21.04,P ＜ 0.05).Conclusions Retention enema is helpful to diagnose and detect colorectal cancer,and the nursing has significance in improving image quality and accuracy of diagnosis.

OBJECTIVETo explore the effects of serum of asthmatic patients, dexamethasone, interleukin-4 (IL-4), interferon-gamma (IFN-γ) and transforming growth factor-β (TGF-β) on the expression of interleukin-22 receptor 1 (IL-22R1) mRNA and protein in HASMCs in vitro.

METHODSIL-22R1 mRNA and protein expressions in HASMCs treated with different stimulating agents were measured by real-time PCR and Western blotting, respectively.

RESULTSIL-22R1 mRNA and protein expressions in HASMCs were significantly increased after stimulation by serum from asthmatic patients, but decreased after co-stimulation with dexamethasone. IL-22R1 mRNA and protein expressions in the cells both increased after stimulation by IL-4, IFN-γ and TGF-β.

CONCLUSIONIL-22R1 in HASMCs might be involved in the pathogenesis of asthma, and the therapeutic effect of dexamethasone on asthma is mediated, at least partially, by IL-22R1. The effects of IFN-γ, IL-4, and TGF-β on asthma may also be attributed to their actions on HASMCs.

Asthma ; blood ; Cell Line ; Humans ; Interferon-gamma ; pharmacology ; Interleukin-4 ; pharmacology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; RNA, Messenger ; genetics ; Receptors, Interleukin ; metabolism ; Transforming Growth Factor beta ; pharmacology

OBJECTIVESTo investigate the effects of newborn bull serum(NBS), vitamin C and vitamin E on cryopreservation of mouse seminiferous epithelial cells.

METHODSThe seminiferous epithelial cells from 7-day-old mice were cryopreserved in different freezing solutions. The cell recoveries were examined by Trypan blue exclusive staining after thawing. The freezing solutions composed of DMEM, 10% dimethylsulphoxide(DMSO), and 0, 5%, 10%, or 20% NBS, respectively, or composed of DMEM, 10% DMSO, 10% NBS, and 150 micrograms/ml vitamin C or 50 micrograms/ml vitamin E, respectively.

RESULTSThe cell recoveries in freezing solution containing 0, 5%, 10%, or 20% NBS were 83.4%, 84.7%, 85.7% and 83.6%, respectively. There were no significant differences between them. The cell recoveries in freezing solution containing vitamin C or vitamin E were 88.0% and 82.9%, respectively. There was no significant differences compared with that in freezing solution containing 10% DMSO and 10% NBS.

CONCLUSIONSNBS, vitamin C and vitamin E have no significant protecting effects on mouse seminiferous epithelial cells, and can not significantly improve the cell recoveries.

Animals ; Ascorbic Acid ; pharmacology ; Cattle ; Cryopreservation ; Epithelial Cells ; physiology ; Fetal Blood ; physiology ; Male ; Mice ; Seminiferous Epithelium ; cytology ; Vitamin K ; pharmacology

OBJECTIVETo observe the effects of inhibiting stromal cell derived factor-1 (SDF-1) expression by RNA interference (RNAi) on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells.

METHODSSDF-1 specific short hairpin RNA (shRNA) expressing plasmid was transferred into cultured human acute leukemic bone marrow stromal cells, positive clones were isolated by screening G418 resistance (Group A) , SDF-1 protein level in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The adhesion rates to bone marrow stromal cells layer and the drug sensitivity to doxorubicin of co-cultured Jurkat cells were detected by cell counting and MTT assay, respectively. The un-transfected bone marrow stromal cells of acute leukemia patient (Group B) or normal subject (Group C) were taken as control.

RESULTSThe level of secreted SDF-1 protein (pg/10(5) cells/week) in the supernatants of Group A, B and C were 1920 +/- 205, 12,370 +/- 1355 and 6620 +/- 770, respectively. Of co-cultured Jurkat cells in Group A, B and C, the adhesion rates after 24 h co-culturing were (28.8 +/- 2.6)%, (57.4 +/- 3.8)% and (45.2 +/- 4.0)%, respectively, and the IC50 values of doxorubicin were 585, 6162 and 1758 nmol/L, respectively.

CONCLUSIONDown-regulating SDF-1 expression of bone marrow stromal cells by RNAi reduces adhesion rates and enhances drug sensitivity to doxorubicin of their co-cultured Jurkat cells.

Bone Marrow Cells ; metabolism ; Cell Adhesion ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; metabolism ; Coculture Techniques ; Drug Resistance, Neoplasm ; Gene Expression ; Humans ; Jurkat Cells ; RNA Interference ; Stromal Cells ; metabolism

OBJECTIVETo study the effects of RNA interference inhibiting stromal cell derived factor-1 (SDF-1) expression on the proliferation and apoptosis of co-cultured Jurkat cells.

METHODInhibition of SDF-1 expression by RNA interference (RNAi) was achieved by transferring SDF-1 specific short hairpin RNA (shRNA) expressing plasmid into cultured human acute leukemic bone marrow stromal cells. Resistant clones were obtained by G418 selection (group A). The concentration of SDF-1 protein in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The population double time (PDT), cell cycles, apoptosis rates and the expressions of PCNA, Bcl-2/Bax, Fas/FasL of co-cultured Jurkat cells were detected by cells counting, flow cytometry. TdT-mediated dUTP nick-end labelling (TUNEL) and immunocytochemistry (ICC), respectively. The un-transfected acute leukemic (group B) and normal (group C) bone marrow stromal cells were taken as controls.

RESULTSThe content of SDF-1 protein in supernatant of group A\[(384 +/- 41) pg/ml] was significantly lower than that in group B[(2474 +/- 271) pg/ml] or group C[(1324 +/- 154) pg/ml]. As group A compared with group B and group C, the PDT of co-cultured Jurkat cells was prolonged (group A: 42 h, vs group B: 29 h, group C: 33 h), and G(0)/G(1) stage cells increased [group A: (28.47 +/- 2.39)%, vs group B: (19.43 +/- 2.80)%, group C: (27.15 +/- 2.07)%], S stage cells decreased [group A: (25.57 +/- 1.90)%, vs group B: (74.48 +/- 3.23)%, group C: (60.99 +/- 2.33)%], G(2)/M stage cells increased [group A: (45.96 +/- 3.24)%, vs group B: (6.09 +/- 1.96)%, group C: (11.86 +/- 1.98)%], the apoptosis rate increased [group A: (15.2 +/- 0.8)%, vs group B: (5.4 +/- 0.7)%, group C: (9.5 +/- 0.4)%], and the expressions of PCNA, Bcl-2, Fas decreased; whereas the expressions of Bax and FasL were increased.

CONCLUSIONThe inhibition of SDF-1 expression in bone marrow stromal cells inhibits the proliferation and promotes the apoptosis of co-cultured Jurkat cells.

Apoptosis ; genetics ; Bone Marrow Cells ; metabolism ; Cell Proliferation ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; Coculture Techniques ; Gene Expression ; Humans ; Jurkat Cells ; RNA Interference ; Stromal Cells ; metabolism ; Transfection

OBJECTIVETo prepare and characterize polyelectrolyte multilayer film coated microbubbles for use as ultrasound contrast agent (UCA) and evaluate its effects in ultrasonic imaging on normal rabbit's liver parenchyma.

METHODSPerfluorocarbon (PFC)-containing microbubbles (ST68-PFC) were prepared by sonication based on surfactant (Span 60 and Tween 80). Subsequently, the resulting ST68-PFC microbubbles were coated using oppositely charged polyelectrolytes by microbubble-templated layer-by-layer self-assembly technique via electrostatic interaction. The enhancement effects in ultrasonic imaging on normal rabbit's liver parenchyma were assessed.

RESULTSThe obtained microbubbles exhibited a narrow size distribution. The polyelectrolytes were successfully assembled onto the surface of ST68-PFC microbubbles. In vivo experiment showed that polyelectrolyte multilayer film coated UCA effectively enhanced the imaging of rabbit's liver parenchyma.

CONCLUSIONSThe novel microbubbles UCA coated with polyelectrolyte multilayer, when enabled more function, has no obvious difference in enhancement effects compared with the pre-modified microbubbles. The polymers with chemically active groups (such as amino group and carboxyl group) can be used as the outermost layer for attachment of targeting ligands onto microbubbles, allowing selective targeting of the microbubbles to combine with desired sites.

Animals ; Contrast Media ; chemistry ; Electrolytes ; chemistry ; Fluorocarbons ; chemistry ; Liver ; diagnostic imaging ; Microbubbles ; Polymers ; chemistry ; Rabbits ; Surface Properties ; Surface-Active Agents ; chemistry ; Ultrasonics ; Ultrasonography

OBJECTIVETo prepare polyelectrolyte multilayer film-coated microbubble ultrasound contrast agent (UCA) and evaluate its effects in contrast imaging on normal rabbit's liver parenchyma.

METHODSPerfluorocarbon (PFC) -containing microbubble UCA (ST68-PFC) were prepared by sonication-based on surfactants (Span 60 and Tween 80). Subsequently, the resulting ST68-PFC microbubbles were coated using oppositely charged polylysine (PLL) and alginate (Alg) by microbubble-templated layer-by-layer self-assembly technique via electrostatic interaction. The enhancement effects in contrast imaging on normal rabbit's liver parenchyma were assessed.

RESULTSThe obtained microbubble UCA exhibited a narrow size distribution. The polyelectrolytes were successfully assembled onto the surface of ST68-PFC microbubbles. In vivo experiment showed that polyelectrolyte multilayer film-coated UCA effectively enhanced the imaging of rabbit's liver parenchyma.

CONCLUSIONSThe novel microbubble UCA obtained via layer-by-layer self-assembly, when enabling more functions, has no obvious difference in enhancement effects compared with the premodified microbubbles. The polymers with chemically active groups (such as amino group and carboxyl group) can be used as the outermost layer for the attachment of targeting ligands to microbubbles, which allows the selective targeting of the microbubbles to desired sites.

Alginates ; chemistry ; Animals ; Contrast Media ; administration & dosage ; chemistry ; Fluorocarbons ; chemistry ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Liver ; diagnostic imaging ; Microbubbles ; Polylysine ; chemistry ; Rabbits ; Ultrasonography

BACKGROUND & OBJECTIVE: Postoperative tissue adherence, scarring and radiotherapy often lead to extrinsic compression and stricture in the distal ureter of the patients who had history of pelvic malignancies. Our aim was to evaluate the efficacy and safety of endourologic techniques in treating this kind of ureteral obstruction. METHODS: From Jan. 1998 to Mar. 2007, 46 patients with obstruction in the distal ureter and had history of pelvic malignancies underwent endoscopic treatments at the Third Affiliated Hospital of Sun Yat-sen University for relief of the obstruction. Perioperative and follow-up data were analyzed. RESULTS: Of the 46 patients, 25 underwent laparoscopic ureterolysis and ureteroneocystostomy, 18 underwent placement of ureter stent under ureteroscope, 3 underwent percutaneous nephrostomy. No severe complication was recorded. The mean operating time was 82.5 min (range, 30-140 min). The mean blood loss was 45.5 ml (range, 5-180 ml). No blood transfusion was needed. The median follow-up time was 18.2 months (range, 3 months to 6.5 years). Three months after operation, B-ultrasonography and intravenous urography (IVU) showed that 39 (84.8%) patients had recovered normal renal function, the other 7 (15.2%) had hydronephrosis relief and renal function improvement. Nuclear renal scanning showed that the mean postoperative glomerular filtration rate (GFR) in the obstructive kidney was higher than the preoperative level (37.6 ml/min vs. 21.3 ml/min, P＜0.05). No stricture in the uretero-bladder anastomotic stoma was recorded. CONCLUSION: Endoscopic operation is an effective and feasible option for managing some selected kinds of distal ureteral obstruction caused by postoperative tissue adherence and radiotherapy in the patients with history of pelvic malignancies.

The traditional experimental teaching mode of forensic science and biological evidence is mostly confined to experimental operation, which is not capable of cultivating students' comprehensive quality and post competency. Therefore, it is urgent to seek an innovative teaching and training mode. At present, the experimental teaching of forensic science and biological evidence is dominated by teachers. There are some problems, such as insufficient training of students' scientific thinking and innovation ability, single teaching and evaluation model, and disconnection from the practical application. This paper proposes an experimental teaching design scheme of forensic science and biological evidence based on post competency training. The course is implanted in the framework of simulated cases, and the virtual simulation experiment platform and group discussion learning method are used to achieve a training model oriented by social needs and centered on students. In the preliminary study on the students who were trained in this mode of selected sections, we found that, compared with traditional teaching, the time for students to complete the prescribed experimental operation in this teaching mode was shortened by 4 minutes on average, and the average score of theoretical course test case analysis questions was increased by 1.5 points. In conclusion, the instructional design of the experiment teaching forensic science and biological evidence can effectively improve students' post-competency, and it deserves further exploration and application.