Gabexate Mesylate-Poloxamer 407 Conjugate Alleviates Sodium Taurocholate-Induced Severe Acute Pancreatitis in an Optimized Rat Model.

BACKGROUND AND AIMS: We have previously shown that gabexate mesylate-poloxamer 407 conjugate (GMTI) alleviates traumatic pancreatitis in rats. In this study, we evaluated the therapeutic effect of GMTI on sodium taurocholate-induced severe acute pancreatitis (SAP) in an optimized rat model. METHODS: An SAP rat model was established via microinjection of 3.5% sodium taurocholate and retention in the bile duct for 1 min. SAP rats were administered GMTI via tail vein injection (i.v.) or tail vein injection + intraperitoneal injection (i.v. + i.p.). All rats were sacrificed at 12 h after treatment. Biochemical approach and enzyme-linked immunosorbent assay were performed to measure the serum levels of amylase (AMY), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). Hematoxylin and eosin staining and TUNEL assay were conducted to examine histopathology and acinar cell apoptosis in the rat pancreas. RESULTS: SAP was successfully induced in all model rats, as evidenced by progressively aggravating SAP symptoms and signs, pancreatic histopathological abnormalities, as well as elevated serum levels of TNF-α, IL-6, and AMY. The mortality rates at 1 h, 6 h, and 12 h were 0%, 0%, and 25%, respectively. GMTI therapy via i.v. or i.v. + i.p. significantly reduced pancreatic wet weights, ascites amounts, pathological scores, and circulating levels of TNF-α and IL-6 while promoting acinar cell apoptosis in SAP rats. GMTI therapy via i.v. + i.p. outperformed i.v. in improving pancreatic histology and reducing TNF-α and IL-6 serum levels in SAP rats. CONCLUSIONS: Our optimized SAP rat model is reliable and reproducible. GMTI therapy is a promising approach against SAP.

Factors associated with health-related quality of life in papillary thyroid microcarcinoma patients undergoing radiofrequency ablation: a cross-sectional prevalence study.

PURPOSE: To explore the association of demographic characteristics, clinical symptoms and the fear of the disease progression factors with the physical and mental summary components of the health-related of life (HRQoL) of the papillary thyroid microcarcinoma (PTMC) patients undergoing radiofrequency ablation (RFA). METHODS: 123 PTMC survivors undergoing RFA were enrolled in this study from October 2019 to March 2020. Demographic, clinical symptoms and the fear of the disease progression data were collected. SF-36, THYCA-QoL and FoP-Q-SF were used to evaluate the HRQoL of patients, clinical symptoms and the fear of disease progression. A multivariate regression model was performed to evaluate the association between the independent variable and the HRQoL variable. RESULTS: The average self-reported HRQoL score was 81.17 ± 15.48 for the PCS and 73.40 ± 18.03 for the MCS. The multivariate linear regression model shows that the factors related to a poorer PCS were dependent for the female patients, the symptoms of neuromuscular and the throat/mouth, the fear of disease progression; the psychological disorder, symptoms of throat/mouth, inability to concentrate were related to worse scores for the MCS. The condition that was most strongly related to a poorer HRQoL (in both PCS and MCS) was the fear of their physical health. CONCLUSIONS: The factors related to significantly worse HRQoL scores across PCS and MCS for PTMC survivors include the female gender, the symptoms of neuromuscular and the throat/mouth, the psychological disorder, inability to concentrate, and the fear of their own physical health. Identification, management, and prevention of these factors are critical to improving the HRQoL of patients.

[Factors Affecting Quality of Life of Patients Undergoing Radiofrequency Ablation for Papillary Thyroid Microcarcinoma].

Objective To evaluate the factors affecting health-related quality of life (HRQoL) of patients with thyroid papillary microcarcinoma (PTMC) after ultrasound-guided radiofrequency ablation (RFA).Methods The clinical data of 100 patients with PTMC who underwent reexamination after RFA in the Ultrasound Department of our center from October to December 2019 were retrospectively analyzed.Demographic information was collected.SF-36 and Thyroid Cancer-specific Health-related Quality of Life Questionnaire scales were used to assess patients' quality of life and thyroid-related specific symptoms.The SF-36 scale includes two general domains including physical component summary (PCS) and mental component summary (MCS).The impacts of demographic characteristics and thyroid-related symptoms after RFA on PCS and MCS scores were further analyzed.Results Univariate analysis and correlation analysis showed that the PCS scores in quality of life of PTMC patients were related to sex, neuromuscular, voice, concentration, sympathetic nerve, and throat/mouth complaints, psychological state, sensory symptoms, scar, chills, tingling, and headache (all P<0.1);and the MCS scores were associated with education level, residence, neuromuscular, voice, concentration, sympathetic nerve, and throat/mouth complains, psychological state, sensory symptoms, scar, chills, tingling, and headache (all P<0.1).Multivariate regression analysis showed that the PCS scores were only associated with sex and the neuromuscular and throat/mouth complains and the psychological state.The regression equation was:PCS=110.367-8.025×sex-0.213×psychological state-0.280×neuromuscular complain-0.278×throat/mouth complain.In contrast, the MCS scores were only associated with the psychological state and the throat/mouth and concentration complains, with the regression equation being:MCS=91.323-0.237×psychological state-0.437×throat/mouth-0.304×concentration.Conclusions The main risk factors affecting the quality of life of PTMC patients after ultrasound-guided RFA were female gender, psychological burden, lack of attention, and symptoms in neuromuscular system and throat/mouth.Therefore, preoperative explanations should be made according to the relevant symptoms that the patients may report, and psychological interventions should be offered after RFA to improve the quality of life of PTMC patients after treatment.

Percutaneous injection of hemostatic agents for severe blunt hepatic trauma: an experimental study.

This study was designed to evaluate whether percutaneous injection of hemostatic agents under the guidance of contrast-enhanced ultrasound (CEUS) can stop hemorrhage from severe hepatic trauma. Eighteen dogs were impacted by a miniature impactor to create blunt hepatic trauma. Fourteen with appropriate liver lesions were divided into two groups: the treatment group (n = 7) and the control group (n = 7). In the treatment group, hemocoagulase atrox and alpha-cyanoacrylate were respectively injected into the injury sites and transected micro-vessels under the guidance of CEUS. In the control group, normal saline was injected into the injury sites. CEUS and CT were performed at 3, 7, 14, and 21 days after the focal injection. Surviving animals were killed on the 21st day for pathologic examination. All animals of the treatment group survived. Three dogs of the control group died in the first 24 h. In the treatment group, CEUS and CT demonstrated that hepatic lesions became smaller gradually from the 3rd to the 21st day after injection. The focal injection of hemostatic agents under the guidance of CEUS can stop hemorrhage from hepatic trauma of grade III~IV or IV. During the period of 3 weeks, no side effect was found.

Application of ultrasonic gas-filled liposomes in enhancing transfer for breast cancer-related antisense oligonucleotides: an experimental study.

The aim of this study was to investigate the application of ultrasonic gas-filled liposomes in enhancing transfer for breast cancer-related antisense oligonucleotides in vitro. An antisense oligodeoxynucleotide (AS-ODN) sequence, HA2741, modified with luciferase reporter plasmid, was used in evaluating the enhancing effect of gas-filled liposomes for gene transfer in breast cancer cells. Some important factors on HA 2741 transfection efficiency, such as wave intensity, ultrasound duration, gas-filled liposome concentration, and HA2741 concentration, were tested, respectively. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by propidium iodide assay. Reverse-transcriptase polymerase chain reaction and immunocytochemistry were used to detect the inhibitory effect of HA2741 on HER-2 expression. All the four factors (wave intensity, ultrasound duration, gas-filled liposome concentration, and HA2741 concentration) showed a positive effect on AS-ODN transfection efficiency. However, these factors had a negative effect on cell viability. Considering all the factors investigated, the maximum transfection efficiency with minimum cell viability achieved under 2% gas-filled liposome mixed with 80 nmol/L HA2741 for 30-second ultrasound exposure at -3.0 dB wave intensity, which gave an overall transfection efficiency exceeding 90% and a cell viability near 90%. Under controlled conditions, ultrasound-mediated AS-ODN transfer, enhanced by gas-filled liposomes, may represent an effective, safe avenue for cancer-related gene delivery.

Paclitaxel-loaded and A10-3.2 aptamer-targeted poly(lactide-co-glycolic acid) nanobubbles for ultrasound imaging and therapy of prostate cancer.

In the current study, we synthesized prostate cancer-targeting poly(lactide-co-glycolic acid) (PLGA) nanobubbles (NBs) modified using A10-3.2 aptamers targeted to prostate-specific membrane antigen (PSMA) and encapsulated paclitaxel (PTX). We also investigated their impact on ultrasound (US) imaging and therapy of prostate cancer. PTX-A10-3.2-PLGA NBs were developed using water-in-oil-in-water (water/oil/water) double emulsion and carbodiimide chemistry approaches. Fluorescence imaging together with flow cytometry verified that the PTX-A10-3.2-PLGA NBs were successfully fabricated and could specifically bond to PSMA-positive LNCaP cells. We speculated that, in vivo, the PTX-A10-3.2-PLGA NBs would travel for a long time, efficiently aim at prostate cancer cells, and sustainably release the loaded PTX due to the improved permeability together with the retention impact and US-triggered drug delivery. The results demonstrated that the combination of PTX-A10-3.2-PLGA NBs with low-frequency US achieved high drug release, a low 50% inhibition concentration, and significant cell apoptosis in vitro. For mouse prostate tumor xenografts, the use of PTX-A10-3.2-PLGA NBs along with low-frequency US achieved the highest tumor inhibition rate, prolonging the survival of tumor-bearing nude mice without obvious systemic toxicity. Moreover, LNCaP xenografts in mice were utilized to observe modifications in the parameters of PTX-A10-3.2-PLGA and PTX-PLGA NBs in the contrast mode and the allocation of fluorescence-labeled PTX-A10-3.2-PLGA and PTX-PLGA NBs in live small animals and laser confocal scanning microscopy fluorescence imaging. These results demonstrated that PTX-A10-3.2-PLGA NBs showed high gray-scale intensity and aggregation ability and showed a notable signal intensity in contrast mode as well as aggregation ability in fluorescence imaging. In conclusion, we successfully developed an A10-3.2 aptamer and loaded PTX-PLGA multifunctional theranostic agent for the purpose of obtaining US images of prostate cancer and providing low-frequency US-triggered therapy of prostate cancer that was likely to constitute a strategy for both prostate cancer imaging and chemotherapy.

Comparing transfection efficiency and safety for antisense oligodeoxyribonucleotide between phospholipids-based microbubbles and liposomes.

OBJECTIVE: To compare transfection efficiency and safety for antisense oligodeoxynucleotides (AS-ODNs) between two type of phospholipids-based vectors. METHODS: An AS-ODNs sequence HA824 combined with luciferase reporter plasmid was used. Under low intensity ultrasound (US), a breast cancer cell line SK-BR-3 was exposed to different concentration of microbubbles and liposomes. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by propidium iodide assay. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the inhibitory effect of HA824 on HER-2 expression at mRNA level. Atomic force microscopy (AFM) scanning techniques was employed to observe the change of membrane pore size. RESULTS: AS-ODNs transfection efficiency showed an increasing tend with microbubble concentration, but not with liposome concentration. Maximum transfection efficiency with minimum cell viability was achieved under 2% microbubble concentration. Too strong sonoporation activity would enlarge membrane pores significantly and cause low cell viability. CONCLUSION: US-mediated AS-ODNs transfection enhanced by phospholipids-based microbubbles represents an effective and safe avenue.

[Echogenic phospholipids-based gas-filled microbubbles as delivery system of antisense oligodeoxynucleotides].

AIM: To investigate the feasibility of transfer antisense oligodeoxynucleotides (AS-ODNs) by the phospholipids-based gas-filled microbubbles (PGM) under ultrasound activation. METHODS: An antisense oligodeoxynucleotides sequence ZL combined with luciferase reporter plasmid was used. A breast cancer cell line SK-BR-3 was exposed to different conditions to investigate the effects of such factors as ZL concentration, PGM concentration, mechanical index (MI) and ultrasound exposure duration on transfection efficiency and cell viability. The transfection efficiency and cell viability by other lipid vectors such as lipofectamine and liposome were also tested, whose results were comparied with that of PGM. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by PI (propidium iodide) assay. RESULTS: Among the factors tested, ultrasound exposure duration, MI and PGM concentration had obvious impacts on transfection efficiency and cell viability. The results showed that the optimal ultrasound condition was the exposure to ultrasound at MI 1.0 for 30 s with 2% PGM concentration, which gave an overall transfection efficiency of 78% +/- 10%, increased nearly 18 folds over the transfection by PGM (4.0%) or lipofectamine (4.3%) without ultrasound. Under same ultrasound conditions, different vectors showed significant difference in transfection efficiency while there are similar results in cell viability. CONCLUSION: Under proper ultrasound conditions, PGM can markedly enhance AS-ODNs transfection efficiency.

[Comparison of sonoporation effect of liposomes and phospholipids-based microbubbles on cultured cell membrane].

AIM: To compare sonoporation effect of two phospholipids-based vectors-liposomes and microbubbles on cultured cell membrane. METHODS: A breast cancer cell line SK-BR-3 was exposed to ultrasound alone, 2% or 5% liposome + ultrasound and 2% or 5% microbubble + ultrasound, separately. Immediately after the experiment and 24 h after ultrasound exposure, atomic-force microscopy (AFM) scanning was used to observe the membrane change of SK-BR-3 cells. RESULTS: After ultrasound exposure, normal SK-BR-3 cells more or less lost their natural shape, showing elliptic outline with obtuse curved boundary. In groups added with phospholipids-based microbubbles, more obtuse curved boundary of cells was observed. The membrane pores of SK-BR-3 cells had apparent changes after ultrasound exposure. With AFM technique, membrane pores under ultrasound alone or ultrasound with liposomes conditions were enlarged, the diameter of some pores exceeding 1 microm. But all the membrane pores in these conditions returned to normal appearance after 24 hours. In ultrasound with 2% microbubble condition, most membrane pores were about 1 - 3 microm in size and returned to normal appearance after 24 h. In ultrasound with 5% microbubble condition, however, pores of most cell membrane porosity was about 2 - 4 pm and did not totally return to normal appearance after 24 h. CONCLUSION: At 2% concentration, phospholipids-based microbubble could enhance ultrasonic sonoporation effect and produce reparable membrane pores on SK-BR-3 cells, which appeared to be a promising vehicle for drug and gene delivery.

A novel thermosensitive in-situ gel of gabexate mesilate for treatment of traumatic pancreatitis: An experimental study.

Gabexate mesilate (GM) is a trypsin inhibitor, and mainly used for treatment of various acute pancreatitis, including traumatic pancreatitis (TP), edematous pancreatitis, and acute necrotizing pancreatitis. However, due to the characteristics of pharmacokinetics, the clinical application of GM still needs frequently intravenous administration to keep the blood drug concentration, which is difficult to manage. Specially, when the blood supply of pancreas is directly damaged, intravenous administration is difficult to exert the optimum therapy effect. To address it, a novel thermosensitive in-situ gel of gabexate mesilate (GMTI) was developed, and the optimum formulation of GMTI containing 20.6% (w/w) P-407 and 5.79% (w/w) P188 with different concentrations of GM was used as a gelling solvent. The effective drug concentration on trypsin inhibition was examined after treatment with different concentrations of GMTI in vitro, and GM served as a positive control. The security of GMTI was evaluated by hematoxylin-eosin (HE) staining, and its curative effect on grade II pancreas injury was also evaluated by testing amylase (AMS), C-reactive protein (CRP) and trypsinogen activation peptide (TAP), and pathological analysis of the pancreas. The trypsin activity was slightly inhibited at 1.0 and 5.0 mg/mL in GM group and GMTI group, respectively (P<0.05 vs. P-407), and completely inhibited at 10.0 and 20.0 mg/mL (P<0.01 vs. P-407). After local injection of 10 mg/mL GMTI to rat leg muscular tissue, muscle fiber texture was normal, and there were no obvious red blood cells and infiltration of inflammatory cells. Furthermore, the expression of AMS, CRP and TAP was significantly increased in TP group as compared with control group (P<0.01), and significantly decreased in GM group as compared with TP group (P<0.01), and also slightly inhibited after 1.0 and 5.0 mg/mL GMTI treatment as compared with TP group (P<0.05), and significantly inhibited after 10.0 and 20.0 mg/mL GMTI treatment as compared with TP group (P<0.01). HE staining results demonstrated that pancreas cells were uniformly distributed in control group, and they were loosely arranged, partially dissolved, with deeply stained nuclei in TP group. Expectedly, after gradient GMTI treatment, pancreas cells were gradually restored to tight distribution, with slightly stained nuclei. This preliminary study indicated that GMTI could effectively inhibit pancreatic enzymes, and alleviate the severity of trauma-induced pancreatitis, and had a potential drug developing and clinic application value.

Percutaneous injection therapy for blunt splenic trauma guided by contrast-enhanced ultrasonography.

OBJECTIVE: The purpose of this study was to investigate the application of contrast-enhanced ultrasonography (CEUS) in managing blunt splenic trauma and the effectiveness of CEUS-guided percutaneous injection therapy. METHODS: Six patients with grade 3 or 4 splenic injuries as determined by CEUS and contrast-enhanced computed tomography were given hemocoagulase atrox and absorbable cyanoacrylate percutaneously, which were injected into the injury region and active bleeding site, respectively, under CEUS guidance. Immediately after the procedure and 1 and 3 days, 1 and 2 weeks, and 1 and 6 months after the procedure, follow-up CEUS up was performed in all patients. RESULTS: Among the 6 patients, 4 cases of CEUS-guided hemostatic injection were successful without complications. Rehemorrhage occurred in 1 patient, and a traumatic arteriovenous fistula occurred in another; repeated injection therapy in these 2 patients was effective. During the follow-up, there were no complications, and spleen perfusion recovered gradually. CONCLUSIONS: Contrast-enhanced ultrasonography can be used to guide percutaneous injection therapy and therefore achieve the goal of using interventional ultrasonography in managing splenic trauma.