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Objective:To explore the clinical feasibility and application method of speckle-tracking echocardiography (STE) in assessing left ventricular longitudinal strain in infants.Methods:One hundred and ten infants within one week of birth were randomly selected in the Third Affiliated Hospital of Guangzhou Medical University from June 2019 to February 2020, and the basic data were collected. STE was performed by two physicians with more than 5 years′ experience.Images of the same infant were analyzed by physicians with 5 years′ experience and 2 years′ experience, respectively, and the difference and correlation of the results between the two doctors were analyzed. The images of the same infant were analyzed twice by the doctor with 5 years′ experience, and the differences and correlations between the results were analyzed. The infants were divided into different groups according to weight to compare the correlations between inter-physicians and intra-physician. STE analysis indicators included global left ventricular longitudinal peak strain (Glps-avg), four-chamber left ventricular longitudinal strain (Glps-A4C), three-chamber left ventricular longitudinal strain (Glps-A3C), two-chamber left ventricular longitudinal strain(Glps-A2C), left ventricular anterior wall longitudinal strain (Glps-a), anterior interventricular septum longitudinal strain(Glps-ais), posterior interventricular septum longitudinal strain (Glps-pis), inferior wall of left ventricle longitudinal strain (Glps-i), posterior wall of left ventricle longitudinal strain (Glps-p), lateral wall of left ventricle longitudinal strain (Glps-l), left ventricular basal section longitudinal strain (Glps-bs), left ventricular middle section longitudinal strain (Glps-ms) and left ventricular apex section longitudinal strain (Glps-as), a total of 13 indicators. The differences were analyzed by paired t test, and the correlations were determined by intra-group correlation coefficient (ICC). Results:According to the inclusion and exclusion criteria, 95 patients were included. For the 13 STE indicators, inter-physicians comparison: the differences between the two physicians were statistically significant (all P<0.05) except for Glps-pis, and the ICC were 0.38-0.72 (all P<0.01). Intra-physician comparison: the differences of these indicators between two measurements were not statistically significant (all P>0.05) except for Glps-ais and Glps-I, ICC were 0.31-0.76 (all P<0.05). Among them, inter-physicians and Intra-physician ICC of Glps-avg, Glps-bs, Glps-ms were 0.68/0.75, 0.72/0.66 and 0.65/0.76 respectively. The group comparison by infants′ weight showed that: In very low weight infants group, the ICC of inter-physicians and intra-physician ranged 0.82-0.93(all P<0.05) and 0.80-0.95(all P<0.05). In low weight infants group, the ICC of inter-physicians and intra-physician ranged 0.65-0.94 (all P<0.05) and 0.69-0.93 (all P<0.01). In the normal weight infants group, ICC of inter-physicians ranged 0.06-0.68, with statistical significance except for Glps-A3C (all P<0.05); ICC of intra-physician ranged 0.36-0.59 (all P<0.05). In overweight infants group, there was no statistical significance in ICC of inter-physicians (all P>0.05), the ICC of intra-physician, Glps-bs and Glps-ms groups were 0.63 and 0.77 (all P<0.05), with no statistical significance in other indicators (all P>0.05). Conclusions:Left ventricular longitudinal strain by STE in infants has better repeatability, and the consistency of intra-physician is higher than inter-physicians, among which the Glps-avg consistency is the best, followed by Glps-bs and Glps-ms. STE is more reproducible in low and very low weight infants than in normal and overweight infants.
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Objective To investigate the quantitative value of the signal intensity ratio of extraocular muscle and ipsilateral white matter measured by MRI for the evaluation of activity in thyroid-associated ophthalmopathy. Methods A total of 129 patients and 245 eyeballs with thyroid-associated ophthalmopathy were enrolled in this study and this 245 eyeballs were set as thyroid-associated ophthalmopathy group(TAO group). There were 10 patients with newly diagnosed Graves'disease and in the same period and these 20 eyeballs were set as graves'disease group(GD group). 32 normal people from annual physical test excluded thyroid and eye diseases and their 64 eyes were selected randomly for the normal control group(NC group). The signal intensity of the extraocular muscle and the ipsilateral white matter on the MRI images were measured, while did exophthalmos and the width of the inner fat of eyeballs (FWs)measurements in the same time. Results SIR,FWs,and exophthalmos of TAO group were higher than those of the other 2 groups[SIRs:1.71(1.40,2.10)vs 1.26(1.22,1.34)and 1.23(1.14,1.32);FWs:8.04(6.70, 8.71)mm vs 6.16(4.86,7.08)mm and 6.93(6.41,7.65)mm,exophthalmos:20.10(18.56,22.15)mm vs 15.40(14.87,16.60)mm and 14.73(13.40,16.07)mm,all P<0.05]. The reference value of SIR establishing based on SIRs of NC group is less than 1.37. In total 129 TAO patients,55 patients(with 106 eyeballs)have a clinical activity score(CAS). Then,these eyeballs were grouped to activity and non-activity(grouped by CAS≥3),and the baseline group difference of these 2 groups was not statistically significant. The SIRs and exophthalmos of activity group were higher than the non-activity group[SIRs:1.70(1.45,2.33)vs 1.41(1.25,1.75); exophthalmos:(20.38 ± 2.40)mm vs(19.05 ± 3.70)mm,all P<0.05]. But the difference of FWs of these two groups was not statistically significant(P>0.05). The SIRs and CAS had a positive correlation(r=0.580,P=0.000),through the receiver operating characteristic curve(ROC)we get the best diagnostic performance of TAO activity when the SIR≥1.56(sensibility=65.6%,specificity=89.1%,AUC=0.815,P=0.000). Conclusion The signal intensity ratio of extraocular muscle and ipsilateral white matter may discriminate the activity of TAO early as a quantitative indicator, reflecting its efficacy,and is worth clinically generalizing.
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Objective To explore the correlation between injection peak pressure and tubal patency during real-time three-dimensional hysterosalpingo-contrast sonography(RT 3D-HyCoSy).Methods One hundred and seven patients who were scheduled to undergo transvaginal RT 3D-HyCoSy with SonoVue contrast agent were included in the study.They were injected with contrast agent by an YLD-YZ-800 auto contrast agent injection device. The patients were divided into tubal patency included bilateral patency group,one side patency group and bilateral positive group according to imaging results. During the examination,the injection peak pressure was recorded to analyze whether there were significant differences among the three groups.Results The contrast agent injection peak pressures of bilateral patency group,one side patency group and bilateral positive group were (34.58 ± 8.25)kPa,(44.85 ± 10.05)kPa and (54.26 ± 11.65)kPa,respectively. The differences of injection pressure among the three groups were statistically significant ( F = 38.732; P = 0.000,0.000,0.033). The peak pressure was negatively correlated with tubal patency ( r = -0.653,P =0.000).Conclusions Contrast agent injection peak pressure is associated with tubal patency in RT 3D-HyCoSy,and can be quantitatively measured to help for assessing fallopian tube patency.
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Objective To compare the efficacy of lung ultrasound and chest X-ray for diagnosis of neonatal transient respiration syndrome (TTN) and respiratory distress syndrome (RDS).Methods Totally 120 infants with respiratory distress underwent lung ultrasound and chest X-ray within 6 h after birth.Taking clinical diagnosis as the standard,the sensitivity,specificity and accuracy of lung ultrasound and X-ray for diagnosis of TTN and RDS were calculated.Results According to clinical diagnosis,there were 75 infants of TTN and 34 of RDS.The sensitivity,specificity and accuracy of lung ultrasound in diagnosis of TTN was 96.00％ (72/75),88.89％ (40/45) and 93.33％ (112/120),respectively,higher than those of chest X-ray (85.33％ [64/75],84.44％ [38/45] and 85.00％ [102/120]).Lung ultrasound showed 85.29％ (29/34) of sensitivity,95.35％ (82/86) of specificity and 92.50％ (111/120) of accuracy for diagnosis of RDS,whereas chest X-ray showed 88.23％ (30/34) of sensitivity,89.53％ (77/86) of specificity and 89.17％ (107/120) of accuracy.Conclusion Lung ultrasound is accurate for the diagnosis of TTN and RDS.
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Objective To investigate the clinical value of combination of transvaginal fourdimensional hysterosalpingo-contrast sonography(TVS 4D-HyCoSy) with uterine tubal liquid poking in assessment tubal fimbria patency.Methods Sixty-two female infertile patients with obstruction at tubal fimbria,or partial obstruction with pelvic adhesion were included.All of them were underwent 4D-HyCoSy as well as uterine tubal liquid poking,and were followed with laparoscopic chromopertubation (LPC) using Methylene blue in three months.Results Comparing with laparoscopy,the total coincidence rate of 4D-HyCoSy to assess the tubal fimbria patency was 94.3％,with the sensitivity of 90.1％ and specificity of 94.1 ％.Imaging results showed that the obstruction at tubal fimbria,uncircle-like wrapping contrast medium can be seen around ovary accounted for 80％.The patent fallopian tube fimbria,circle-like wrapping of contrast medium can be observed around ovary accounted for 85.5 ％.Conclusions The combination of TVS 4D-HyCoSy with uterine tubal liquid poking is highly in accordance with LPC by providing real-time dynamic visualization on overflowing of contrast medium from tubal fimbria,as well as the pelvic adhesion surrounding ovaries.The 4D-HyCoSy can be the preferred method for assessment of tubal fimbria patency and pelvic adhesion surrounding ovaries with its advantages of accuracy,non invasion and a good safety profile.
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Objective To evaluate the inhibitory effect of phase change doxorubicin loaded carboxymethyl hexanoyl chitosan nanodroplet on ovarian cancer cells,and the effect of its ultrasound image in vitro.Methods The carboxymethyl hexanoyl chitosan synthesized through the acylationreaction with carboxymethyl chitosan and hexanoic anhydride.The drug loaded carboxymethyl hexanoyl chitosan nanodroplets were prepared by ultrasonic emulsification.The surface morphology,particle diameter and electric potential were characterized.Ultrasound imaging of the nanodroplet was evaluated in vitro.The encapsulation efficiency was determined by ultraviolet spectrophotometry.The survival rate of ovarian cancer cell was detected using CCK-8 reagent.The statistical analysis was performed.Results The drug loaded carboxymethyl hexanoyl chitosan nanodroplet was successfully prepared which showed regular morphology in microscope,the mean diameter of (458.33± 43.50)nm.The encapsulation efficiency was (52.06 ± 10.14)％.The nanodroplet could enhance ultrasonic imaging.The survival rate of ultrasound combined with drug loaded nanodroplet group ([62.54± 3.60]％) was lower than those of the free drug group ([75.55±7.21]％) and drug loaded nanodroplet group ([76.18±4.94]％),ultrasound group ([89.90±0.83]％;P＜0.05).Conclusion Ultrasound-mediated drug loaded nanodroplet can inhibit ovarian cancer cells,and has the potential for application in the clinical diagnosis and treatment.
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<p><b>OBJECTIVE</b>To evaluate the feasibility, safety, radicality and short-term outcome of preserving left colic artery (LCA) during laparoscopic total mesorectal excision (TME) for the treatment of rectal cancer.</p><p><b>METHODS</b>From January 2013 to December 2016,136 patients with mid-lower rectal cancer received laparoscopic TME in the Gastrointestinal Surgery Department of Taizhou People's Hospital of Jiangsu Province. Patients with rectal tumor within 10 cm to the anal verge were enrolled into the study. All the enrolled patients had complete data of pathology and follow-up. Those receiving neoadjuvant chemoradiotherapy, with severe base diseases, multifocal tumor, tumor invasion of surrounding tissues, fixation of tumor, recurrent tumor, complications such as acute ileus, bleeding, perforation were excluded. In this study, 72 patients did not undergo preservation of LCA (high ligation group) and 64 patients underwent preservation of LCA (low ligation group). Operative parameters, clinicopathological data and short-term outcome were collected and compared between two groups.</p><p><b>RESULTS</b>The baseline data including gender, age, body mass index, tumor stage, and distance of tumor from anal verge of two groups were comparable (P>0.05). The differences between two groups about the mean time of operation and the operative blood loss were not significant [(164.0±12.6) min vs. (167.3±9.4) min, (30.0±3.6) ml vs. (30.1±3.0) ml, all P>0.05]. There was no operative death in both groups. Differences in the lymph node dissection (13.7±2.6 vs. 13.3±2.1) and the specimen length of proximal resection margin [(16.4±1.9) cm vs. (16.7±2.1) cm] or distal resection margins [(3.9±0.6) cm vs. (4.1±0.9) cm] between high and low ligation groups were not significant (all P>0.05). Compared with high ligation group, the low ligation group had higher rate of sphincter preservation [92.2% (59/64) vs. 79.2% (57/72), χ=4.580, P=0.032], lower rate of anastomotic leakage [1.6% (1/64) vs. 9.7% (7/72), χ=4.075, P=0.044], anastomotic stenosis [3.1% (2/64) vs. 12.5%(9/72), χ=4.006, P=0.045], and voiding and sexual dysfunction [6.3%(4/64) vs. 18.1%(13/72), χ=4.317, P=0.038]. Mean time of follow-up was 19 months. In high ligation group, the local recurrent rate was 5.56%, distant metastasis rate was 13.89%, overall survival rate was 90.28%, disease-free survival rate was 80.56%, while in low ligation group, the local recurrence rate was 4.69%, distant metastasis rate was 12.50%, overall survival rate was 90.63%, disease-free survival rate was 82.81%, whose differences between two groups were not significant (all P>0.05).</p><p><b>CONCLUSION</b>Preservation of LCA during laparoscopic TME for the treatment of rectal cancer is safe and feasible, which can reduce the incidence of anastomotic leakage and stenosis, and voiding and sexual dysfunction.</p>
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<p><b>OBJECTIVE</b>To explore the feasibility, safety and efficacy of intraoperative regional infusion chemotherapy by celiac trunk in advanced gastric cancer patients.</p><p><b>METHODS</b>One hundred and twenty-six patients with advanced gastric cancer(stageII(-III() were screened from database of Gastrointestinal Surgery Department of Taizhou People's Hospital between January 2008 and December 2010 who underwent R0 resection and D2 lymphadenectomy, received postoperative chemotherapy(XELOX or FOLFOX), and had complete follow-up data. They were divided into infusion chemotherapy group (65 cases) and control group (61 cases) according to regional infusion chemotherapy or not (fluorine 1 000 mg and cisplatin 60 mg). The side effects of chemotherapy, parameters related to the operation, long-term survival and relapse rate were compared between the two groups.</p><p><b>RESULTS</b>The baseline data between the two groups were comparable(all P>0.05). Postoperative III( and IIII( adverse reaction of chemotherapy was not significantly different between the two groups (P>0.05). The time of postoperative intestinal function recovery [(67.9±14.8) hours vs. (68.9±15.0) hours, t=-0.380, P=0.705), volume of postoperative 1-week drainage [(66.1±17.1) ml vs.(61.9±18.2) ml, t=1.478, P=0.142], recent morbidity of complications[55.4%(36/65) vs. 49.2%(30/61), χ=0.256, P=0.613], and the long-term morbidity of complications [16.9% (11/65) vs. 14.8% (9/61), χ=0.111, P=0.739] were all not significantly different between the two groups. The 3-year survival rate and 3-year relapse-free survival rate in infusion chemotherapy group were significantly higher than those in control group(58.4% vs. 37.7%, χ=5.382, P=0.020; 58.4% vs. 34.4%, χ=6.636, P=0.010).</p><p><b>CONCLUSION</b>Regional infusion chemotherapy by celiac trunk during operation for advanced gastric cancer patients is safe and feasible, and can reduce the risk of local recurrence and improve survival rate.</p>
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Humanos , Protocolos de Quimioterapia Combinada Antineoplásica , Usos Terapéuticos , Arteria Celíaca , Quimioterapia del Cáncer por Perfusión Regional , Métodos , Mortalidad , Cisplatino , Usos Terapéuticos , Desoxicitidina , Usos Terapéuticos , Supervivencia sin Enfermedad , Flúor , Usos Terapéuticos , Fluorouracilo , Usos Terapéuticos , Gastrectomía , Leucovorina , Usos Terapéuticos , Escisión del Ganglio Linfático , Recurrencia Local de Neoplasia , Compuestos Organoplatinos , Usos Terapéuticos , Complicaciones Posoperatorias , Recuperación de la Función , Neoplasias Gástricas , Quimioterapia , Mortalidad , Cirugía General , Tasa de SupervivenciaRESUMEN
Objective To analyze the occurrence rate of gastrointestinal and pulmonary infection and hospital acquired pneumonia (HAP)in severe craniocerebral injuries in my hospital using proton pump inhibitors (PPIs) ,discuss the right way to use PPIs for severe craniocerebral injuries .Methods The case histories of 141 severe craniocerebral injuries in my hospital from 2011 to 2012 were looked up ,and divided into three groups as follows :the group that never used PPIs(17 cases) ,the group that used PPIs for a long time (59 cases) ,and the group that used PPIs for a short time (65 cases) ,and the risks of getting infected using PPIs were analyzed in those three groups above .Results The patients in PPIs group had a higher risk of gastrointestinal tract infection and HAP ,especially who were treated with PPIs more than 9 days .Adjusted by GCS score ,using PPIs for a long time was a risk factor in different multivariate logistic .Conclusion Using PPIs for a long time might be a risk factor of gastro‐intestinal tract infection and HAP .Therefore ,apart from considering the state of illness ,more attention should be paid to the u‐sing time w hen use PPIs .
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Objective To evaluate the influence of ultrasound-mediated contrast agent microbubbles carrying kallidinogenase targeted therapy on neurogenesis and angiogenesis after experimental acute cerebral infarction.Methods Kallidinogenase-loaded microbubbles were prepared using mechanical shaking method.Middle cerebral artery occlusion (MCAO) model was established in male Wistar rats by sutureoccluded method.MCAO rats (n =120) were randomly divided into 5 groups:ultrasound-mediated kallidinogenase-loaded microbubbles group (group 1),ultrasound-mediated microbubbles group (group 2),ultrasound-mediated saline group (group 3),kallidinogenase group (group 4),saline group (group 5).Medication was given through tail vein daily for 2-6 consecutive days starting 24 h after MCAO.Ultrasound groups (groups 1,2 and 3) were given 2 MHz pulse ultrasonic irradiation on the ischemia lateral skull for 10 min after injection.Cell proliferation was examined using 5'-bromo-2'-deoxyuridine (BrdU,50 mg/kg).Infarction volume and neurological function were evaluated on the 3rd,7th day after MCAO respectively.Doublecortin (DCX) + cells in the subventricular zone and laminin + in the peri-infarction region were observed at the same time.Results The number of DCX + cells in the subventricular zone (under 20 × ocular) of groups 1-5 was 251.8 ± 13.1,125.7 ± 11.6,130.2 ± 13.7,234.5 ± 12.4 and 123.7 ± 10.0 respectively.The percentage of laminin + cells in the peri-infarction region (× 10 objective) of groups 1-5 was 10.0％ ± 0.8％,5.2％ ± 0.7％,5.0％ ± 1.0％,8.0％ ± 1.8％ and 5.0％ ± 0.9％ respectively.The number of DCX + cells in the subventricular zone and laminin + cells in the peri-infarction region of the group 1 and the group 4 was significantly more on the 7th day after MCAO compared with those of the other three groups (DCX:t values of group 1 vs groups 2,3,5 were 17.88,17.17 and 18.16,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 15.42,14.78 and 15.70,all P ＜0.01.Laminin:t values of group 1 vs groups 2,3,5 were 7.01,6.71 and 7.11,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 4.23,3.94 and 4.33,all P ＜0.01).Moreover,the number of DCX+ cells in the subventricular zone (t =2.46,P ＜ 0.05) and laminin + cells in the peri-infarction region (t =2.78,P ＜ 0.05) of the group 1 was much more than those of the group 4.Neurologic scores on the 7th day of groups 1-5 were 1.00 (0.75,1.25),2.0 (2.00,3.00),2.0 (1.00,2.00),1.5 (0.75,2.00),2.0 (2.00,2.50).Compared with other four groups,group 1 showed better functional improvement after stroke (U values of group 1 vs groups 2-5 were 2.0,4.0,7.5 and 2.5,all P ＜ 0.05).While there was no significant difference in infarction volume among five groups at all the time points after MCAO.Conclusions Our study demonstrates ultrasound-mediated kallidinogenase-loaded contrast agent microbubbles targeted therapy promotes neuroblasts proliferation and vascular regeneration compared with mediated and non-medicine-loaded microbubbles therapy,which attributes to functional improvement after MCAO.Therefore,ultrasound-mediated ultrasound contrast agent microbubbles carrying drugs targeted therapy may have a perspective on ischemic stroke.
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Due to its effectiveness and specificity,RNA interference (RNAi) technology is widely used in the treatment of tumor,viral infectious and metabolic diseases.Noninvasively tracing the delivery of therapeutic RNA molecules in vivo and evaluating the therapeutic effects are important to promote the clinical application of RNAi technology.Molecular imaging allows noninvasive visualization of biological processes at the cellular and molecular level in vivo.It can be used to monitor the biodistribution of therapeutic RNA molecules and to assess the effects of the RNAi.This article reviews the advance of molecular imaging in monitoring the delivery of RNAi molecules and evaluating the therapeutic effects of RNAi.
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Purpose To investigate the clinical value of three-dimensional hysterosalpingo-contrast-sonography (3D-HyCoSy) in the assessment of fallopian tubal patency.Materials and Methods Fallopian tubal patency of 33 infertile women was assessed using transvaginal 3D-HyCoSy, and the results were compared with laparoscopically salpingoraphy, the sensitivity, specificity, positive likelihood ratio and negative likelihood ratio of 3D-HyCoSy were evaluated.Results 3D-HyCoSy showed 20 passable fallopian tubes, 10 partially passable fallopian tubes and 36 obstructed tubes, while lapascopically salpingoraphy showed 23 passable fallopian tubes, 7 partially passable fallopian tubes and 36 obstructed tubes. The total coincidence rate of 3D-HyCoSy to assess the fallopian tubal patency was 86.4% (57/66), with the sensitivity of 93.0% (40/43), specificityof 73.9% (17/23), positive likelihood ratio of 7.15, and negative likelihood ratio of 0.20.Conclusion Tubal patency can be efficiently and accurately assessed using 3D-HyCoSy, but patients with obstructed or partially passable fallopian tubes should undergo laparoscopy to clarify.
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Objective To investigate the gene silencing,apoptosis induction and the suppression of proliferation in vivo transfected by UTMD techniques associated with shRNA techniques. Methods The survivin-shRNA expression vector was constructed. Nude mice were randomly arranged into 3 groups:control group, plasmid injection and ultrasound (P + US), P + UTMD group. Histological examination were evaluated. Protein expressions of Survivin and proliferating cell nuclear antigen (PCNA), Bcl-2, Bax,Caspase-3, Ki-67, nucleostemin (NS), p53 were investigated by immunohistochemistry. Results In transplanted tumors experiment, comparing with those in C and P + US groups, protein expressions of PCNA,Ki-67,Bcl-2, Survivin, NS were down-regulated markedly, while those of Bax, Caspase-3 and P53 were up-regulated significantly ( P ＜ 0.05). Conclusions UTMD combined with shRNA technique can induce apoptosis and inhibit proliferation significantly, without causing any apparently adverse effect,representing a new,promising technology that can be used in the tumor gene therapy and research.
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Objective:To transfect genes using ultrasound targeted microbubble destruction (UTMD) techniques and observe the effects of RNA interference on cervical cancer (HeLa) cell line in silencing survivin gene and inducing apoptosis. Methods: Recombinant expression plasmid of short hairpin RNA (shRNA) targeting survivin gene was constructed. It was co-treated with microbubbles and transfected to cultured HeLa cells followed by exposure to ultrasound (P+UTMD group). Moreover, blank control group (C), plasmid group (P), ultrasound exposure group (US), plasmid and ultrasound exposure group (P+US), plasmid+ Lipofectamine group (P+L) were used as controls, respectively. Transfection efficacy was evaluated by observing the red fluorescence in the cells by fluorescent microscopy and flow cytometry(FCM). Ultrasound intensity and exposure time were optimized. Cell apoptosis was investigated using flow cytometry analysis, Hoechst staining, and DNA ladder method. Expression of survivin mRNA was assessed by RT-PCR. Results: Restrictive enzyme digestion and sequencing analysis verified that the recombinant plasmid was successfully constructed. UTMD significantly increased gene transfection efficacy in cultured HeLa cells (P<0.01). Gene transfer was the most prominent at ultrasound intensity of 1.0 W/cm2 and exposure time of 3 min (P<0.01). RT-PCR showed that the expression of survivin mRNA in P+UTMD group was inhibited by (83.33±2.73)%. The differences were significant compared with any other groups (P<0.01). FCM analysis showed that the apoptosis ratio in P+UTMD group was significantly increased as compared with other groups (P<0.01). Hoechst staining and DNA ladder showed that apparent apoptosis and DNA ladder were detected only in P+UTMD and P+L groups. Conclusions:UTMD effectively enhances the transfection efficacy of expression plasmid. It is a novel and effective non-viral gene transfer system and has promising foreground. UTMD mediates RNA interference silenced survivin gene and induces significant cell apoptosis, which provides a new method for tumor research and gene therapy.
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Objective To study the transfeetion efficiency and safety of liposome microbubble(LM)on red fluorescent protein(RFP)in vitro and in vivo under ultrasound mediated gene transfection(USMGT)conditions.Methods Plasmids containing RFP were added to cultured Hela cells followed by ultrasound (US)exposure with LM.Different concentration of LM,US intensity and exposure time were optimized.Transfection efficiency was evaluated by fluorescent microscopy and FACS.Cell viability was verified by propidium iodide assay.In transplanted tumors in vivo study,LM and plasmid(P)were injected into the nude mice followed by US exposure(P+LM+US group).Nude mice undergoing plasmid injection alone(P group),plasmid injection and US exposure(P+US group)and plasmid and LM injection(P+LM group)were used as controls.Frozen section and histological examination were conducted and RFP expression was evaluated.Results LM and US exposure significantly increased transfeetion efficiency in cultured Hela cells (P＜ 0.01).Transfection efficiency was the most prominent under the condition of US intensity of 1.0 W/cm2 with 6%LM,duration 3 min.No apparent cell damage was found in the all groups.In transplanted tumors,strong RFP was seen in P+LM+US group.It was significantly higher than in any other groups(P＜0.0 1).No tissue damage was seen histologically.Conclusions LM could enhance USMGT effectively without causing any apparently adverse effect in vitro and in vivo.This method would be a novel,effective,safe non-viral gene transfection method and provide an alternative to current clinical gene therapy.
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Objective To investigate different pulsed ultrasound (PUS) parameters and culture conditionsthat would affect cell viability and sonoporation on cell membrane of human cervical cancer cells (HeLa). MethodsHeLa cells were cultured in two different conditions ( in suspension or in monolayer). Cells were exposed to differentPUS intensity (0.4 W/cm2, 1.0 W/cm2, 1.6 W/cm2, 2.2 W/cm2), duty cycle (10%, 20%, 50%) and expo-sure time ( 1 min or 3 min). Cell viability was analyzed by flow cytometry. Using microscope and scanning electronmicroscopy (SEM) , the changes of shape and the sonoporation on cell membrane induced by PUS were observed.Results Low intensity and duty cycle did not exert a great impact on the cell viability. Cell injury was found to in-crease progressively with high intensity ( 1.6 W/cm2 , 2.2 W/cm2 ) and duty cycle ( 50% ) ( P < 0. 01 ) , and celldetachment was significantly accompanied by PUS exposure in adherent HeLa cells. Results of factorial design showedthat the culture conditions and the PUS parameters had significant interaction ( P < 0.01 ). SEM demonstrated insome detail the phenomenon of transient pores in the cell membrane under suitable PUS irradiation. The ideal sonopo-ration conditions that cell viability was above 80% and more membrane holes were noted to be at 1.0 W/cm2 expo-sure for 3 min with a duty cycle of 20% in cell suspension. Conclusion The optimized conditions of the PUS pa-rameters and the culture conditions could lower the cell injury and exert a great impact on the sonoporation. It couldproduce remarkable membrane pores on cells and enhance cell membrane permeability, which facilitate transportationof macromolecules into cells.
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Objective To estimate the diagnostic value of the patients of intrauterine pregnancy abortion (IUPA) of pregnancies of unknown location (PUL) with transvaginal power Doppler ultrasound (TV-PDU) combining Logistic regression model and receiver operating characteristics (ROC) curve.Methods Endometria of PUL were detected by TV-PDU during the early pregnancy.Serum β-HCG and progesterone were analyzed meanwhile.All variables were researched by multiple factors Logistic regression.Logistic regression equationg were established.ROC curve was constructed and the cut-off value of endometrial mnographic findings and clinical data to diagnose intrauterine pregnancy abortion were studied.The accuracy of every univariate and multivariable were evaluated and the validity and stability of the Logistic regression equation were tested.Results The RI value and PSV were included in the multivariate model 1 (M1),while the serum progesterone,type of endometrial resonance and gestation age were included in the multivariate model 2(M2).The area under the curve of M1,M2 were 0.887,0.976,respectivdy.Youden index and positive predictive value were 90.8%,87.5% and 12.213,8.000,respectively.Specificity and positive predictive value of M2 was lower than M1 which had a hisher accuracy.Diagnostic value were increased obviously by the use of M1 and M2 comparing with univariate and the stability was all righL Conclusions Possibility of IUPA could be predicted by TV-PDU simply,atraumaticly and efficiently.A close rehaion is found between the hemodypnmic parameter of endometria and the IUPA.
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Objective To study the optimized condition of transfection efficiency for MCF-7 cells enhanced by ultrasound(US) irradiation and contrast agent combined with polyethyleneimine(PEI) and observe whether the combination can have a synergistic effect to increase DNA transfection. Methods MCF-7 cells were transfected with the compounds prepared by the vector of plasmid DNA encoding luciferase (pCMV-luciferase-GL3) and PEI.SonoVue microbubble was added to the cell suspension to serve as nucleation sites for aeoustic cavitation before US irradiation. The DNA expression of luciferase plasmid and viability of cells were evaluated. The strategy of US irradiation was optimized. Furthermore, the influencing factor, such as the concentration of plasmid, incubation time, serum, the type of solvent and the volume of culture media, were examined. Results The viability of cells and US-induced enhancement of luciferase activity were influenced by the US intensity,exposure time and duty cycle.US irradiation under an appropriate condition enables ceils to accelerate the permeation of the PEI/DNA complex through the cell membrane, resulted in enhanced transfection efficiency of plasmid DNA. Optimal US condition for the enhancement was determined to be 1 W/cm2,10% DC for 3 min. In contrast to the PEI/DNA complex alone without US irradiation or US irradiation alone, the combination of US irradiation with contrast agent and PEI had a significantly enhanced luciferase activity (P<0.01). The 2 h pre-irradiation incubation with PEI/DNA complex for MCF-7 ceils exhibited a significantly enhanced lueiferase activity (P<0.01). Besides,serum,type of solvent and the volume of culture media did affect the transfection efficiency. Conclusions The optimized parameters of US and transfection provide efficient gene delivery in MCF-7 cancer cells. The combination of US irradiation with contrast agent and PEI has a synergistic effect to increase DNA transfection. This is a simple and promising method to enhance the gene expression of plasmid DNA.
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Objective To investigate different ultrasound parameters and transfection conditions that would affect transfection rate of red fluorescent protein(RFP)and cell viability of cancer cells.Methods In this study,Hela cells were cultured using two different protocols:(A)24 h culture for complete adherence;(S)suspension.Subsequently,cells were transfected following different ultrasound exposure protocols[1.0W/cm2;duty cycle(DC):10%,20%and 50%;exposure 1min or 3 min].Gene transfection and cell viability were evaluated.Treatment parameters optimized in Hela cells were applied for delivery RFP in 4 other cell lines(HepG2,Ishikawa,MCF-7 and B16-F10).Results Cell injury were found to increase progressively with DC and exposure time in group A.Cell detachment was significantly accompanied by ultrasound exposure in adherent HeLa cells.Cells in group S were found more prone to be transfected than group A with the same ultrasound parameters,while the survival rate was not decreased apparently.The ideal ultrasound conditions were noted to be at 1.0 W/cm2 irradiated 3 min with 20%DC using suspended protocol,producing maximum efficiency[transfection=(28.04±2.27)%]in gene delivery with minimum cell toxicity[cell viability=(81.20±1.73)%].These experiments also revealed different response to ultrasound treatment,but for all tested cell lines,dead and transfected cells in the treated groups were significantly different from the non-irradiated groups.Conclusions Ultrasound parameters and transfection conditions have a great impact on the gene delivery and cell viability.Gene delivery of ultrasound-mediated microbubble enhance should be optimized to improve the efficiency.
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Objective To determine whether it could enhance gene delivery and tumor transfection in vivo by combination of ultrasound-targeted microbubble destruction(UTMD)with polyethylenimine(PEI)in tumor xenografts.Methods Two different reporter plasmid[lueiferase(pCMV-LUC)and red fluorescent protein(RFP)]were incubated with PEI to prepare cationic compound(PEI/DNA)in various nitrogen:phosphate ratios(N/P ratios,nmol of nitrogen in the PEI/nmol of phosphate in DNA).Formation of PEI/DNA complexes were confirmed by the gel retardation assay.Human cervical carcinoma(Hela)tumors were planted subcutaneously in both flanks of female nude mice.Tumor-bearing mice were administered by tail vein with PBS,plasmid,plasmid and Sono Vue microbubble,PEI/DNA and Sono Vue microbubble.One tumor was exposed to ultrasound irradiation (3 MHz,2 W/cm2,2 min exposure,duty cycle 20%),while the other served as control.The feasibility of targeted delivery and tissue specificity facilitated by UTMD and PEI was investigated.The mice were sacrificed 3 days after ultrasound exposure.Tissue specimens were viewed with microscopy to determine the presence of RFP expression.The efficiencies of luciferase transgene expression were determined.Histology analysis was detected.Results Electrophoresis experiment revealed that PEI was mixed with plasmid to condense DNA efficiently.The application of UTMD significantly increases the tissue transfection in vivo compared to plasmid alone.RFP expression was present in all sections of tumors that received ultrasound exposure but not in control tumors.Results of luciferase activity showed that the expression of luciferase was to be 14 times greater in ultrasound-exposed tumors(P＜0.01).More importantly,the increase in transgene expresgion was related to UTMD with the presence of PEI dramatically.At least 10-fold increase of luciferase gene transfer was obtained in irradiated tumors compared to non-irradiated controls(P＜0.01),111-fold increase compared to UTMD alone(P＜0.01).There was not significantly gene expression in other organs or tissues regardless of US exposure(P＞0.05).No tissue damage was seen histologically.Conclusions The combination of UTMD with PEI can enhance targeted delivery and expression of reporter gene to tumors at intravenous administration.It is a promising new and safe method for gene delivery in vivo.