Objective To study the effects of ultrasound mediated microbubbles destruction on endothelial cells transfected with lipofectamine DNA complexes.Methods One hour after addition of pIRES2 EGFP hVEGF gene and lipofectamine,endothelial cells on 6 well plates were exposed to ultrasound for 30 s,60 s,or 90 s in the presence(10 ?l,100 ?l,500 ?l) or absence of ultrasound contrast agent.The transducer (Sonos 5500,S3, 1.3 MHz,MI 1.0 ) was submerged in water and held at a distance of 1 cm.Expression of the marker gene EGFP was observed by fluorescent microscopy 2 days later.Results Ultrasound treatment of endothelial cells on plates for 30 s and 60 s yielded significant increase in transfection rates without damaging the cells,but 90 s treatment killed most of the cells.Ultrasound contrast agent was able to significantly increase endothelial cells transfection rate by enhancing cavitation effects at a concentration of 100 ?l,and damaged most cells when applied at a concentration of 500 ?l.Conclusions Ultrasound mediated destruction of microbubbles enhances gene expression after lipofectamine mediated transfection of endothelial cells in vitro; and it may represent an improved avenue for therapeutic gene delivery in vivo.

Objective To analyze the correlation between the levels of urinary ptasminogen activator receptor (uPAR) and the clinicopathological characteristics of patients with cervical carcinoma and it's clinical significance. Methods SABC immunohistochemistry was employed to detect the expression of uPAR in 50 cases of cervical carcinoma tissue and 50 cases of normal cervical tissue. ELISA method was used to determine the serum uPAR levels for the patients with cervical carcinoma. Results There was no expressionfor uPAR in normal tissues, The positive expression rate of uPAR was 66 % (33/50). The uPAR level of cervix cancer tissues [(70.92±28.55) ng/100 mg protein]was significantly higher than those in normal tissues [(11.01±5.40) ng/100 mg protein], (P ＜0.001). and uPAR levels were closely related to clinical stages,lymphatic metastasis and differentiation degree (P ＜0.05), but not related to deep myometrial invasion and vascular embolization (P ＞0.05) (however, they were not related to patient's age, tumor growth type and the size of tumor. Significant difference of uPAR level was observed between the patients with cervical carcinoma [(2.38±0.29) ng/ml]and in healthy controls [(0.50±0.16) ng/ml](P ＜0.001). Single factor analysis indicated that, before the treatment, the serum uPAR levels were closely related to clinical stages, lymphatic metastasis, vascular embolization, and deep myometrial invasion (P ＜0.05-P ＜0.01). However, they were not related to differentiation degree (P ＞0.05). Multifactor regression analysis showed that the pretreatment serum uPAR levels of patients were related to clinical stages (P =0.000), cavum pelvis lymphatic metastasis (P =0.000) and deep myometrial invasion. Patients with cervical carcinoma showed a dramatic drop in serum uPAR levels after treatment, which were significantly different compared to their pretreatment uPAR levels (P ＜0.001). Linear correlation analysis showed that there was a positive correlation between serum and tissue uPAR levels, (r =0.801, P ＜0.001). Conclusion There were high expression of uPAR in serum and tissues with cervical carcinoma. Pretreatment serum uPAR levels were closely related to patients' clinical stages,cavum pelvis lymphatic metastasis and deep myometrial invasion, serum uPAR levels may be an important tmnor marker for the diagnosis, detection, prognosis of cervical carcinoma.

BACKGROUND:Although drug treatment.physics-behavior therapy,and postoperative therapy have been commonly used to treat stress urinary incontinence(SUI),there is still no satisfactory treatment at present.OBJECTlVE:To build a stable animal model simulating stress urinary incontinence(SUI)by bilateral transaction of pudendal nerve and nerves innervating pelvic floor muscles,including iliococcygeous muscle and pubococcygeous muscle.METHODS:A total of 18 6-week-old female SD rats weighing(1 99.44±8.41)g were randomly divided into 3 groups:normal,model,and sham-surgery groups,with 6 rats in each group.Rats in the model group underwent bilateral transaction of pudendal nerves and nerves innervating iliococcygeous/pubococcygeous muscles,while rats in the sham-surgery group had same procedures except nerve transaction.The normal group did not undergo any operation.Each rat was subjected to measure leak point pressure(LPP)at 2 weeks after the operation.After the measurement of LPP,cross sections of connection area of bladder and urethra were sent to histology.RESULTS AND CONCLUSION:One rat in the sham-surgery group died at 1 week after the operation.The LPP of model group decreased significantly by approximately 33%compared with the normal group(P＜0 05):however,there was no significant difference in LPP between sham-surgery and normal groups(P＞0.05).The results of histology showed loosely arrangement and atrophy of urethral sttriated muscle fibers in rats of the model group.Bilateral transaction of pudendal nerves and nerves innervating to iliococcygeous/pubococcygeous muscles resulted in SUI in rats stably.

ObjectiveTo explore the effects of myoblast formation around the urethra of stress urinary incontinence (SUI) rats after treated with bone marrow mesenchymal stem cells(BMSCs) or musclelike cells/calcium alginate composite gel injection therapy.MethodsIsolation,cultivation and identification of Sprague-Dawley rat bone marrow mesenchymal stem cell were performed.5-azacytidine was introduced to induce muscle-like cells.Calcium alginate gel was initially prepared by 2％ sodium alginate and 1％ calcium chloride solution at a volume ratio of 5∶1.Compounds of stem cells or muscle-like cells were mixed with gel,respectively,and were prepared for microinjection.SUI was produced in 72 6-week-old female Sprague-Dawley rats.The rats were then divided into 4 groups:Gel group,stem cell-gel group,muscle-like cell-gel group and mock control group.Each group was further divided into 3 groups.Submucosal injection of gel was performed at urethra and bladder neck.After preparation of cross sections of rat urinary tract at 4 weeks and 8 weeks after injection,HE staining,fluorescent tracing,staining of Desmin and α-skeletal muscle actin (α-SMA) were performed.OD values of positive rates were compared.ResultsAt 4 weeks and 8 weeks after injection in stem cell-gel group and muscle-like cell-gel group,growth of blood vessels gradually increased at gel edge,BMSCs and muscle-like cells gathered around the new blood vessels observed by fl(u)orescence tracer,muscle-like cells grew into elongated spindle-like cells.Desmin and α-SMA staining were positive in these groups,and the OD values in the stem cell-gel group and muscle-like cell-gel group was significantly higher than that from the gel only group and control group,but no difference was found between stem cell-gel group and muscle-like cell-gel group.ConclusionsCompound of BMSCs,muscle-like cells and calcium alginate composite gel has the potential to differentiate into muscle cells in the microenvironment of SUI rat model.In short term,the myoblast formation potential is the same whether the BMSCs was introduced into the micro-environment in vivo directly,or the BMSCs was implanted into microenvironment after the formation of the muscles cells induced by 5-azacytidine in vitro.

Objective To evaluate the clinical effects of percutaneous micro-balloon compression (PBC) trigeminal ganglion for the treatment refractory trigeminal neuralgia. Methods Surgical results of 452 patients with trigeminal neuralgia treated by PBC from October 2009 to May 2013 were analyzed retrospectively. 125 cases aged over 80 years old and 70 cases′ pain belongs to the first branch neuralgia. Such procedures as Meckle′s cave cannulated with No.4 Forgarty catheter and the balloon inflated and compressed the gasserian ganglion monitored by X-ray were observed by PBC. Follow-up interview and curative effects were recorded. Results The average hospitalization was 6.1 days. Among them, 432(95.6%) cases had immediate relief from pain. The overall pain relief rate was 97.8% in our group without serious surgical complications. Postoperative complications include hemifacial numbness in 385 patients (85.2%), mild masseter muscle weakness in 248 patients (54.9%), diplopia in 2 patients. All symptoms relived or disappeared within 1 ~ 6 months. The average follow-up intervier period is 23.5 months. The recurrence rate is 10.2% (46 cases). Conclusion PBC is a safe and effective method with high pain relief rate in the treatment of refractory trigeminal neuralgia , especially for the treatment of the high risk patients , patients with recurrent symptoms or the patients suffered from the first branch neuralgia.

To evaluate the clinical effect of digital assisted chimeric deep circumflex iliac artery perforator flap (DCIAPF) in the reconstruction of mandibular composite defects. Methods From January, 2018 to January, 2019, 6 cases of mandibular tumor patients with postoperative defect within side were treated. Preoperative CTA was used to evaluate the deep branches of spin iliac artery.Digital simulation software and 3D printing technolo-gy was taken, vascularized iliac flap of the design guide of bone was made, and the rebuilding effect was simulated. DCIAPF was used to repair the defect of lower jawbone. The donor sites were sutured directly. The patients were fol-lowed-up in outpatient department for 3-6 months to evaluate the recovery of the patient′s shape, jaw height and oc-clusal function, as well as the complications in the donor area. Results Postoperation pathological examination re-sults: ameloblastoma in 2 cases, 4 cases of gingival cancer. The length of cut out ilium was 6.0-13.0 cm, carrying the flap area of 3.0 cm×1.0 cm-6.0 cm×5.0 cm.Six cases of DCIAPF and iliac bone flap survived.The shape, mandibular height and occlusal function were satisfactory.And no obvious complications were found in the donor area. Conclu-sion The blood supply of DCIAPF is rich with enough bone mass and height. The position of terminal skin perfora-tors is invariant. The complications of donor sites is less. With the help of digital technology, the accuracy of mandibular defect repair and the 3-dimensional wound repair can be realized, and provides an advantage condition for subsequent dental implant.It is one of the ideal method of reconstruction of mandibular defect.

Objective:To investigate the clinical application of 3-dimensional CT angiography(3D-CTA) in the repair of mandibular defects with deep circumflex iliac arterial flaps.Methods:From January 2019 to January 2020, 5 patients with postoperative defects of mandibular tumors were treated in the Department of Stomatology of the First Affiliated Hospital of Bengbu Medical College, including 2 males and 3 females. The ages of the patients ranged from 30 to 70 years, with an average of 52 years. CT angiography of both lower limbs was performed before operation. Data were processed using the CT workstation. According to the size and shape of the defect in the recipient site, the 3D model of the deep circumflex iliac arterial flap(DCIA) was constructed. the contour of the DCIA was observed and measured, and the data was recorded, including the diameter of the vassel at the starting point, the angle between the line from the starting point to the anterior superior iliac spine and the line between the bilateral anterior superior iliac spine, and the distance from the starting point to the anterior superior iliac spine. According to the 3D model, a composite tissue flap pedicled with DCIA was designed and prepared for mandibular defect repair. The actual situation during the procedure was compared with the preoperative 3D model simultaneously.Results:The diameter of the DCIA at the starting point was (2.30±0.24) mm. The angle between the line from the starting point to the anterior superior iliac spine and the line between the bilateral anterior superior iliac spine was 39.3°±6.1°, and the distance from the starting point to the anterior superior iliac spine was (5.83±0.24) cm. The shape and position of the DCIA were basically consistent with the three-dimensional model. All the flaps survived well after operation. All the patients were followed up for 3 to 6 months, the bilateral maxillofacial region were basically symmetrical. The postoperative imaging examination showed that the iliac osteocutaneous flap healed well, and the height and width of mandibular alveolar ridge were restored satisfactorily. There were no complications such as numbness in the donor site, abdominal hernia and gait abnormality.Conclusions:The 3D model of the DCIA produced by 3D-CTA can accurately locate the position and route of the vessel, clearly display the three-dimensional spatial relationship between the skin, muscles and bones around the target blood vessel, and significantly reduce the complications and operative time.

Objective:To investigate the value of contrast-enhanced ultrasound (CEUS) as a preoperative planning strategy for lymphaticovenous anastomosis (LVA) in improving the quality of LVA and the outcome of short-term limb volume reduction in patients with secondary upper limb lymphedema.Methods:Patients with breast cancer-related upper extremity lymphedema who underwent LVA at the Department of General Surgery Cancer Center Division of Breast Surgery of Zhejiang Provincial People’s Hospital from August 2021 to August 2023 were enrolled retrospectively. According to whether preoperative ultrasound lymphography was performed, the patients were divided into CEUS assisted group and control group. In the CEUS assisted group, preoperative CEUS lymphography combined with high-frequency ultrasound color Doppler imaging was utilized for precise localization of lymphatic vessels and recipient veins, as well as surgical target planning for LVA. In the control group, preoperative indocyanine green lymphography was employed to guide surgical exploration. Mann-Whitney U test was used to compare the number of LVA surgical exploration incisions per limb and the number of successful anastomoses per limb between the two groups. The success rate of anastomosis (total number of successful anastomoses/total number of surgical exploration incisions) was compared by the chi-square test. The duration of single anastomosis, mean arm circumference, and the difference between preoperative and postoperative mean arm circumference were compared by independent sample t-test. Paired-sample t-test was used to compare the improvement of the mean arm circumference of the operated limb of the two groups after 3 months of follow-up. P < 0.05 was considered statistically significant. Results:A total of 47 female patients were enrolled, including 27 patients in the CEUS assisted group, with an average age of (57.1±9.0) years and a median edema course of 2 years. There were 20 cases in the control group, with an average age of (58.1±9.6) years and a median duration of edema of 2 years. The CEUS group, compared with the control group, exhibited a higher number of surgical exploration incisions per limb [6.0 (4.0, 7.0) cases vs. 5.0 (3.0, 6.0) cases], a greater number of successful anastomoses per limb [5.0 (3.0, 6v0) cases vs. 3.0 (2.0, 3.0) cases], and a significantly increased overall success rate of anastomosis [82.8% (125/151) vs. 61.4% (54/88)]. Additionally, there was a significant increase in the preoperative and postoperative mean arm circumference difference [(6.2±3.3) cm vs. (3.9±1.9) cm]. The duration of single anastomosis was significantly shortened [(57.4±16.0) min vs. (92.8±18.5) min], with statistically significant differences observed in all comparisons (all P < 0.05). The preoperative and postoperative mean arm circumference were compared between the CEUS group [(31.4±4.6) cm vs. (25.3±4.7) cm] and the control group [(31.3±4.3) cm vs. (27.5±3.8) cm], respectively, with statistically significant differences observed in both groups (both P < 0.01). Conclusion:CEUS lymphography, as a preoperative planning strategy for LVA, can significantly increase the number and success rate of LVA anastomosis in patients with secondary upper limb lymphedema, shorten the duration of single anastomosis, and improve the short-term effect of limb volume reduction after LVA.

Objective:To investigate the value of contrast-enhanced ultrasound (CEUS) as a preoperative planning strategy for lymphaticovenous anastomosis (LVA) in improving the quality of LVA and the outcome of short-term limb volume reduction in patients with secondary upper limb lymphedema.Methods:Patients with breast cancer-related upper extremity lymphedema who underwent LVA at the Department of General Surgery Cancer Center Division of Breast Surgery of Zhejiang Provincial People’s Hospital from August 2021 to August 2023 were enrolled retrospectively. According to whether preoperative ultrasound lymphography was performed, the patients were divided into CEUS assisted group and control group. In the CEUS assisted group, preoperative CEUS lymphography combined with high-frequency ultrasound color Doppler imaging was utilized for precise localization of lymphatic vessels and recipient veins, as well as surgical target planning for LVA. In the control group, preoperative indocyanine green lymphography was employed to guide surgical exploration. Mann-Whitney U test was used to compare the number of LVA surgical exploration incisions per limb and the number of successful anastomoses per limb between the two groups. The success rate of anastomosis (total number of successful anastomoses/total number of surgical exploration incisions) was compared by the chi-square test. The duration of single anastomosis, mean arm circumference, and the difference between preoperative and postoperative mean arm circumference were compared by independent sample t-test. Paired-sample t-test was used to compare the improvement of the mean arm circumference of the operated limb of the two groups after 3 months of follow-up. P < 0.05 was considered statistically significant. Results:A total of 47 female patients were enrolled, including 27 patients in the CEUS assisted group, with an average age of (57.1±9.0) years and a median edema course of 2 years. There were 20 cases in the control group, with an average age of (58.1±9.6) years and a median duration of edema of 2 years. The CEUS group, compared with the control group, exhibited a higher number of surgical exploration incisions per limb [6.0 (4.0, 7.0) cases vs. 5.0 (3.0, 6.0) cases], a greater number of successful anastomoses per limb [5.0 (3.0, 6v0) cases vs. 3.0 (2.0, 3.0) cases], and a significantly increased overall success rate of anastomosis [82.8% (125/151) vs. 61.4% (54/88)]. Additionally, there was a significant increase in the preoperative and postoperative mean arm circumference difference [(6.2±3.3) cm vs. (3.9±1.9) cm]. The duration of single anastomosis was significantly shortened [(57.4±16.0) min vs. (92.8±18.5) min], with statistically significant differences observed in all comparisons (all P < 0.05). The preoperative and postoperative mean arm circumference were compared between the CEUS group [(31.4±4.6) cm vs. (25.3±4.7) cm] and the control group [(31.3±4.3) cm vs. (27.5±3.8) cm], respectively, with statistically significant differences observed in both groups (both P < 0.01). Conclusion:CEUS lymphography, as a preoperative planning strategy for LVA, can significantly increase the number and success rate of LVA anastomosis in patients with secondary upper limb lymphedema, shorten the duration of single anastomosis, and improve the short-term effect of limb volume reduction after LVA.

Objective:To investigate the clinical application of 3-dimensional CT angiography(3D-CTA) in the repair of mandibular defects with deep circumflex iliac arterial flaps.Methods:From January 2019 to January 2020, 5 patients with postoperative defects of mandibular tumors were treated in the Department of Stomatology of the First Affiliated Hospital of Bengbu Medical College, including 2 males and 3 females. The ages of the patients ranged from 30 to 70 years, with an average of 52 years. CT angiography of both lower limbs was performed before operation. Data were processed using the CT workstation. According to the size and shape of the defect in the recipient site, the 3D model of the deep circumflex iliac arterial flap(DCIA) was constructed. the contour of the DCIA was observed and measured, and the data was recorded, including the diameter of the vassel at the starting point, the angle between the line from the starting point to the anterior superior iliac spine and the line between the bilateral anterior superior iliac spine, and the distance from the starting point to the anterior superior iliac spine. According to the 3D model, a composite tissue flap pedicled with DCIA was designed and prepared for mandibular defect repair. The actual situation during the procedure was compared with the preoperative 3D model simultaneously.Results:The diameter of the DCIA at the starting point was (2.30±0.24) mm. The angle between the line from the starting point to the anterior superior iliac spine and the line between the bilateral anterior superior iliac spine was 39.3°±6.1°, and the distance from the starting point to the anterior superior iliac spine was (5.83±0.24) cm. The shape and position of the DCIA were basically consistent with the three-dimensional model. All the flaps survived well after operation. All the patients were followed up for 3 to 6 months, the bilateral maxillofacial region were basically symmetrical. The postoperative imaging examination showed that the iliac osteocutaneous flap healed well, and the height and width of mandibular alveolar ridge were restored satisfactorily. There were no complications such as numbness in the donor site, abdominal hernia and gait abnormality.Conclusions:The 3D model of the DCIA produced by 3D-CTA can accurately locate the position and route of the vessel, clearly display the three-dimensional spatial relationship between the skin, muscles and bones around the target blood vessel, and significantly reduce the complications and operative time.