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Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.
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Objective:To investigate the safety and effect of transcatheter arterial chemoembolization (TACE) combined with radiofrequency ablation (RFA) and sorafenib on large hepatocellular carcinoma (HCC) patients treatment.Methods:From Jan 2012 to Dec 2017, 36 patients (Male: 33, Female: 3, average age: 51.8) with large HCC lesions(5-7 cm) received TACE plus with RFA and sorafenib in the Third Affiliated Hospital of Sun Yat-sen University. Efficacy was evaluated after TACE. Each patient was received follow-up after RFA procedure. The occurrence rate of complications and overall survival (OS) were recorded. Log-rank univariate analysis was used to analyze the OS data.Results:The median TACE time was 4, and the RFA time was (1.7±0.7) . Mean duration time of sorafenib administration was (37.7±28.8) months. Adverse events of sorafenib: 26(72.2%) hand-foot skin reaction, 6(16.7%) hypertension, 22(61.1%) diarrhea, 17(47.2%) alopecia, 3(8.3%) oral ulcer and 1(2.8%) gastrointestinal hemorrhage. Median OS was 63.0 months, and 1-year, 3-year and 5-year survival rate was 100%, 72.7% and 52.6%. The cumulative survival rate of patients taking whole course of sorafenib ( n=21) was better than that of patients taking remedial ( n=15); the cumulative survival rate of patients with alpha fetal protein (AFP) <200 μg/L ( n=26) before treatment was better than ≥200 μg/L ( n=10); the cumulative survival rate of patients with good TACE response ( n=19) was better than that of patients with no response ( n=17), and the differences were statistically significant (all P<0.05). Conclusions:TACE plus with RFA and sorafenib are safe and effective for large HCC patients with 5-7 cm lesions and this treatment might improve OS. The whole-course sorafenib, lower base AFP value (<200 μg/L) and good TACE response were considered as the good factors for the combination therapy in large HCC patients.
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Objective To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P4502C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology. Methods Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results. Results The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4%and 10.9%, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results. Conclusion A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.
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Objective@#To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P450 2C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology.@*Methods@#Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results.@*Results@#The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4% and 10.9%, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results.@*Conclusion@#A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.
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Objective To study the treatment outcomes using transcatheter arterial chemoemboliza tion (TACE) followed by hepatic resection (HR) in patients with BCLC stage B hepatocellular carcinoma (HCC).Methods The clinical data of 46 patients with intermediate stage HCC in the Third Affiliated Hospital of Sun Yat-sen University from Jan.2004 to Jul.2013 were analyzed retrospectively.The baseline characteristics of these 46 patients were recorded.The levels of serum aspartate transaminase (AST),ala nine aminotransferase (ALT),total bilirubin (TBil),and albumin (Alb),and the prothrombin time (PT) at 1 week and 1 month after HR were compared with those at the baseline.The overall survival (OS) and progression-free survival (PFS) were analyzed by the Kaplan-Meier method.The effect on PFS by the changes in the alpha-fetal protein level and the modified Response Evaluation Criteria In Solid Tumors (mRECIST) after TACE were analyzed using the log-rank test.Results The number of cycles of TACE given before HR was 1.7 ± 1.1.The mean time interval between TACE and HR was 3.0 ± 3.2 months.The levels of serum AST,ALT,TBil,Alb and PT at 1 week after HR were significantly different from the baseline levels (P < 0.05),while those levels at 1 months after HR showed no significant difference from the baseline levels (P>0.05).The mean OS was 72.1 ±6.0 (95% CI 60.36 ~83.86) months.The median PFS was 46.0 (95% CI:42.60~49.40) months.Using the log-rank test,the decrease in AFP (P < 0.001) and the improvement in mRECIST (CR + PR) (P < 0.001) after TACE were significantly associated with better PFS.Conclusions HR after TACE was safe for patients with BCLC stage B HCC and might prolong the PFS and OS.Decreases in AFP level and improvement in mRECIST (CR + PR) after TACE were factors they could be used to predict the survival outcomes of HR.
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Objective To improve the diagnostic accuracy of primary renal rare benign tumors by exploring and analyzing the CT/MRI and clinical pathologic features.Methods 9 patients with primary renal rare benign tumors pathologically proven lesions after operation who had CT or MRI exams with contrast were enrolled in our hospital.The radiological and clinical pathological features of all tumors were analyzed respectively.Results The cases were the mixed epithelial and stromal tumor of kidney (MESTK),cystic nephroma(CN),renal leiomyoma(RL)and renal oncocytoma(RO)respectively.Location:6 cases were in left kidney and 3 in right kidney.Size:long diameter 2.5-8.9 cm,mean 5.7 cm;short diameter 2.5-8.4 cm,mean 4.9 cm.The tumor shape included oval(n=7) and irregular(n=2).9 cases protruded from the renal surface.MESTK showed polycystic lesions on preconstrast,various degrees of cystic wall and septum enhancement were detected on enhancement scans.CN showed cystic lesions and calcification on preconstrast, slight septum enhancement were detected on enhancement scans.RL showed heterogeneous density or signal with patchy necrosis and clear border on precontrast,the obvious enhancement was found on cortex and medullary phase and slight washout enhancement was detected on excretory phase.RO showed heterogeneous hypointense on T1 WI,hyperintense on T2 WI and mixed isodensity and hypodensity on CT precontrast.Slight delayed enhancement was found in central scar and washout enhancement was detected in parenchyma on three phases.Conclusion There are some special CT/MRI characteristics for primary renal rare benign tumors,which could improve their diagnosis and differential diagnosis combined with the clinical pathological features.
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Objective To compare magnetic resonance spectroscopy (MRS) characteristics and changes before and after 5-Fu interventional thermochemotherapy (ITC) as well as to analyze the correlation of Cho/Lip and apoptosis index (AI) in hepatic VX2 carcinoma animal model.Methods 20 rabbit models of hepatic VX2 carcinoma were implanted successfully.Ten days later,as tumor diameter≥ 10 mm,routine scan and 1H-MRS were performed before one day and after 14 days on all of rabbits by Philips 1.5T MR scanner.The changes of Cho peak,Lip peak and the ratio of Cho/Lip before and after the treatment were analyzed.Specimens of AI were examined and compared with the ratio of Cho/Lip.Results The Cho peak and Lip peak were increased before ITC.After ITC,the Cho peak and the ratio of Cho/Lip decreased than before.The Lip peak was further increased than that of before.AI was correlated negatively with the ratio of Cho/Lip (r =-0.56,P =0.02).Conclusions The metabolites of different ROI are matched with the levels and scopes of AI.1H-MRS can be used to evaluate tumor cell apoptosis and energy metabolism changes of the rabbit models of hepatic VX2 carcinoma with the treatment.
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Objective To explore a promising system for tumor CD 44 receptor-targeted imaging and to investigate their physic-chemical properties and targeting effect on CD 44 abundant cancer cells in vitro.Methods The superparamagnetic iron oxide ( SPIO) nanoparticles were prepared by a coprecipitation in alkaline media starting from a mixed of the ferrous and ferric solution.And then the surface of the SPIO nanoparticles were modified with APTMS by a reaction with the hydroxyl groups.Finally, the hyaluronan-modified SPIO ( SPIO-HA) nanoparticles were prepared.Control and experimental groups were established after adding SPIO or SPIO-HA as agents respectively.Transmission electron microscopy ( TEM) and particle size analyzer were used to measure these nanoparticle sizes and the hydrodynamic diameters.Thermogravimetric analysis ( TGA) was carried out to evaluate the HA-content on the surface of SPIO-HA.The MRI T2 ralaxivities (1/T2 ) of the two groups at different Fe concentrations (0.09, 0.18, 0.27, 0.36, 0.45 mmol/L ) were measured on a 3.0T MR system.HepG2 cells and HL7702 cells were used for assessment of cells viability by methyl thiazolyl tetrazolium ( MTT ) assay.Prussian blue staining , immunoassay fluorescence image and flow cytometry were carried out to determine the targeted cellular uptake of SPIO-HA nanoparticles.MRI were performed to show the MR T 2 value changes after incubating with HepG2 cancer cells by using T 2 WI sequences at a clinical 3.0 T MR system.One-way analysis of variance was performed to determine significant changes in MR T 2 values of blank control , SPIO-HA and SPIO groups.Results The SPIO-HA and SPIO NPs were fairly homogeneous with an average core size of 18.2 and 22.4 nm, hydrodynamic diameter of 91.1 and 103.2 nm, Zeta potential of (-45.00 ±0.86) mV and (-18.50 ±0.73) mV, and magnetic relaxivity of 0.212 ×106 M-1 · s-1 and 0.191 ×106 M-1 · s-1.Based on the TGA data , HA accounted for 24%weight of each SPIO-HA.The internalization of the SPIO-HA was confirmed by prussian blue staining , while the cells showed no obvious blue stains with SPIO , incubation of SPIO-HA with tumor cells led to blue color inside the cells.After that, we examined cancer cell binding of FITC-SPIO-HA by immunoassay fluorescence image and flow cytometry.The green fluorescence resulting from FITC-SPIO-HA was observed inside the cells in both the cytoplasm and the plasmalemma.Tumor cells treated with SPIO-HA exhibited higher fluorescence signals with 7.97-fold enhancement observed for HepG 2 cells over control particles.In vitro MR, mean T2 values of blank control , SPIO and SPIO-HA groups were ( 115.20 ±0.36 ), ( 115.07 ±0.81 ) and ( 21.67 ±0.21 ) ms, respectively.There was significant difference among those three groups (F=31 703.339,P<0.01), MR T2 values of HepG2 cells treated with the SPIO-HA NPs were lower than blank and SPIO group.In comparison, SPIO did not generate any MRI signal changes compared with blank group.Conclusion The tumor CD44 receptor-targeted MR molecular probe SPIO-HA had a good physic-chemical property and well targeted HepG2 cells.
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Objective To investigate the nursing of traumatic carotid-cavernous fistula(TCCF)by intervention.Methods 18 patients with TCCF by intervention were given nursing measures including psychological preparation,eye care,disease observation and complications care.Results All patients gained successful embolization,and the symptoms of proptosis and vascular murmur in patients were alleviated.One case received spring embolism because of balloon rupture.The symptoms of vascular spasm in two patients were alleviated with timely treatment.Hyperperfusion syndrome was relieved after lowering blood pressure in two patients.Conclusions The intervention embolization is an effective treatment of TCCF.Good nursing could improve the efficacy and reduce the incidence of complications.