ObjectiveTo investigate the effect of folic acid-modified crebanine polyethylene glycol-polylactic acid hydroxyacetic acid copolymer（PEG-PLGA） nanoparticles（FA-Cre@PEG-PLGA NPs， hereinafter referred to as NPs） combined with ultrasonic irradiation on subcutaneous tumor of liver cancer in Kunming（KM） mice. MethodsEighty-four healthy male KM mice were utilized to establish a subcutaneous tumor model of mouse hepatocellular carcinoma with H22 cells， then mice were randomly divided into model group， placebo group， hydroxycamptothecin group（8 mg∙kg^-1）， low， medium and high dose crebanine raw material groups（2， 2.5， 3 mg∙kg^-1， hereinafter referred to as the low， medium and high dose crebanine groups， respectively）， low， medium and high dose NPs groups（2， 2.5， 3 mg∙kg^-1）， and low， medium and high dose NPs combined with ultrasonic irradiation groups（2， 2.5， 3 mg∙kg^-1， hereinafter referred to as the low， medium and high dose combination groups， respectively）. The corresponding doses of drugs were administered via tail vein injection， the model group received no treatment， while the placebo group was injected with an equivalent amount of normal saline. Dosing was conducted for a total of 10 times on alternate days. The body mass of the mice was monitored， and parameters such as body mass change rate， thymus index， spleen index， tumor volume， tumor weight， relative tumor growth rate（T/C）， and tumor inhibition rate（TGI） were calculated. Pathological changes in liver and kidney tissues as well as the tumor were observed by hematoxylin-eosin（HE） staining. Additionally， the levels of aspartate aminotransferase（AST）， alanine aminotransferase（ALT）， blood urea nitrogen（BUN） and creatinine（CREA） in serum of mice were detected by biochemical method. Furthermore， the effect of ultrasound on the distribution of NPs in subcutaneous tumors of mouse hepatocellular carcinoma was observed by in vivo imaging technique. ResultsAmong different treatment methods， the combination of NPs and ultrasound irradiation had the best therapeutic effect. Compared with the model group， the body mass growth rates of mice in the medium and high combination groups decreased， while the thymus index and spleen index increased， but there was no statistically significant difference in serum AST， ALT， BUN and CREA levels， indicating that NPs combined with ultrasound irradiation had little effect on the normal physiological state of the body， oth groups had TGI>40% and T/C<60%， indicating a clear anti-tumor effect. Pathological analysis showed that compared with the NPs groups， the combination groups exhibited varying degrees of necrosis in tumor cells， accompanied by less damage to the liver and kidneys. In vivo imaging of small animals showed that compared with the high dose NPs group， the high dose combination group had stronger tumor targeting ability（P<0.01）. ConclusionNPs combined with ultrasonic irradiation can not only effectively targeted the drug to the tumor site， inhibit the subcutaneous tumor growth of mouse liver cancer， but also decrease damage to liver and kidney tissues.

OBJECTIVE To investigate the targeting effect of folic acid-modified crebanine nanoparticles （FA-Cre@PEG- PLGA NPs， hereinafter referred to as “NPs”） combined with ultrasound irradiation on M109 cells in vitro and in vivo after administration， and explore the anti-tumor mechanism. METHODS CCK-8 assay was used to detect the inhibitory effect of NPs combined with ultrasound irradiation on the proliferation of M109 cells， and the best ultrasound time was selected. Using human lung cancer A549 cells as a control， the targeting of NPs combined with ultrasound irradiation to M109 cells was evaluated by free folic acid blocking assay and cell uptake assay. The effects of NPs combined with ultrasound irradiation on the migration， invasion， apoptosis， cell cycle and reactive oxygen species （ROS） levels of M109 cells were detected by cell scratch test， Transwell chamber test and flow cytometry at 1 h after 958401536@qq.com administration； the changes of mitochondrial membrane potential （MMP） were observed by fluorescence inverted microscope. A mouse subcutaneous tumor model of M109 cells was constructed， and the in vivo tumor targeting of NPs combined with ultrasound irradiation was investigated by small animal in vivo imaging technology. RESULTS NPs combined with ultrasound irradiation could significantly inhibit the proliferation of M109 cells， and the optimal ultrasound time was 1 h after administration. The free folic acid could antagonize the inhibitory effect of NPs on the proliferation of M109 cells， and combined with ultrasound irradiation could partially reverse this antagonism. Compared with A549 cells， the uptake rate of NPs in M109 cells was significantly higher （P＜0.01）， and ultrasound irradiation could promote cellular uptake. NPs combined with ultrasound irradiation could inhibit the migration and invasion of M109 cells and block the cell cycle in the G0/G1 and G2/M phases. Compared with control group， the apoptosis rate of M109 cells and ROS level were increased significantly （P＜0.01）， while the MMP decreased significantly （P＜0.01） in the different concentration （100， 200， 300 μg/mL） groups of M109 cells. Compared with the mice in non-ultrasound group， the fluorescence intensity and tumor-targeting index of the tumor site in the 0 h ultrasound group were significantly enhanced （P＜0.05 or P＜0.01）. CONCLUSIONS NPs combined with ultrasound irradiation have a strong targeting effect on M109 cells in vitro and in vivo， the anti-tumor mechanism includes inhibiting cell migration and invasion， blocking cell cycle， and inducing apoptosis.

Objective:To analyze the imaging features of primary leiomyosarcoma of the inferior vena cava (PLV).Methods:This study was a case series analysis. The clinical and imaging data of 17 patients with PLV confirmed by pathology who underwent CT, MRI and/or PET-CT examinations in the Second Affiliated Hospital of Zhejiang University School of Medicine from June 2012 to September 2022 were retrospectively collected. The CT or MRI findings, including the number, location, size, shape, margin, density (signal), enhancement characteristics, surrounding invasion, and metastasis of the lesion were analyzed.Results:Among the 17 cases there were 7 males and 10 females, aged 32 to 78 years. All patients had single tumors, the maximum diameter of the mass was 3.8 to 19.0 cm, and in 12 cases it was consistent with the inferior venacava.There were 11 cases of "petal-like" necrosis cystic lesion, and 6 cases of "fissure" necrosis cystic lesion. Collateral circulation was found around the mass in 15 cases. There were 3 cases of intraluminal type, 2 cases of extraluminal type, and 12 cases of junctional type. The tumors of intraluminal type presented as "worm-like" appearance, while the extraluminal and junctional type tumors presented as lobulated shape, with local "umbilical depression sign". The margins were clear in 3 cases and unclear in 14 cases. Vascular invasion by the mass was observed in 7 cases. On CT plain scan, the solid component of PLV presented as isodense or slightly hypodense. On MRI T1-weighted images, the solid component presented as isodense or slightly hypodense, and on fat-suppressed T2-weighted images, it presented as isodense or slightly hyperdense. On MRI diffusion-weighted imaging, 12 cases showed high signal intensity, and the corresponding apparent diffusion coefficient map showed low signal intensity. On CT enhanced scan, 8 cases showed mild to moderate enhancement of the solid part of PLV in the arterial phase and 9 cases showed obvious enhancement. In the venous phase and delayed phase, 10 cases showed continuous enhancement and 7 cases showed progressive enhancement. On MRI enhanced scan, 8 cases showed moderate enhancement and 6 cases showed obvious enhancement of the solid part of PLV in the arterial phase. In the venous phase and delayed phase, continuous progressive enhancement was observed; no enhancement was seen in the necrotic and cystic areas.Conclusion:The imaging characteristics of PLV show that the maximum diameter of the masses are consistent with the direction of inferior vena cava, most masses exhibit a "petal-like" necrotic cyst morphology with collateral circulation, and a localized "umbilical concave sign", and some cases possess hemophilic tube features.

Objective:To explore the clinical effects and value for application of the tibial flap of the second toe with the vascular pedicle including a plantar vein on the reconstruction of fingertip soft tissue defects.Methods:From October 2020 to August 2022, retrospective analysis of 12 patients (12 digits) were treated at the Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital for small soft tissue defects of fingertip. The patients were 9 males and 3 females and aged 29-54 years, at 38 years in average. The fingertip defects were measured at approximately 1.2 cm × 0.8 cm to 1.2 cm × 1.2 cm. In the surgery, a tibial flap of second toe was designed to reconstruct the defect in fingertip. The flap were designed on the tibial of second toe without an extended incision in dorsal foot. The pedicle of the flap carried with the lateral proper digital artery, a nerve and the plantar vein of the second toe. At the recipient site, the artery, nerve and vein carried by the pedicle of the flap were end-to-end anastomosed with the digital artery, digital nerve and subcutaneous vein of the finger. The flaps were measured at 1.5 cm×1.0 cm - 1.5 cm×1.5 cm in size. All donor sites were reconstructed with skin grafts from the ipsilateral calf. Scheduled postoperative follow-ups were conducted at the outpatient clinic. The Michigan Hand Function Questionnaire (MHQ) evaluation criteria was employed to assess the recovery of hand function, and Total Active Movement (TAM) was used to evaluate the recovery of range of motion of the interphalangeal joints of the affected fingers.Results:All 12 flaps in the fingertips survived. Postoperative follow-ups lasted from 6 months to 2 years, with an average of 11 months. One flap was slightly bloated and a flap aesthetic surgery was followed at 3 months after the primary reconstructive surgery, and the rest of flaps were all in good appearance. TPD was found at 12 -14 mm for all flaps at 9 months after surgery. All donor sites in the feet and calfs had primary healing, without a contracture or rupture of skin graft or an obvious dysfunction at the donor sites. According to the evaluation criteria of the MHQ, 8 patients were very satisfied with the overall appearance of the hand, and 4 were satisfied. Finger movement was evaluated according to TAM criteria, all 12 fingers were rated excellent.Conclusion:Reconstruction of a fingertip defect with a tibial flap of the second toe with the vascular pedicle including a plantar vein of the second toe has a good clinical efficacy. It has advantages in flap harvest, avoids an extended incision on dorsal foot, and makes a minimal damage to the donor site.

Objective:To analyze the imaging features of primary leiomyosarcoma of the inferior vena cava (PLV).Methods:This study was a case series analysis. The clinical and imaging data of 17 patients with PLV confirmed by pathology who underwent CT, MRI and/or PET-CT examinations in the Second Affiliated Hospital of Zhejiang University School of Medicine from June 2012 to September 2022 were retrospectively collected. The CT or MRI findings, including the number, location, size, shape, margin, density (signal), enhancement characteristics, surrounding invasion, and metastasis of the lesion were analyzed.Results:Among the 17 cases there were 7 males and 10 females, aged 32 to 78 years. All patients had single tumors, the maximum diameter of the mass was 3.8 to 19.0 cm, and in 12 cases it was consistent with the inferior venacava.There were 11 cases of "petal-like" necrosis cystic lesion, and 6 cases of "fissure" necrosis cystic lesion. Collateral circulation was found around the mass in 15 cases. There were 3 cases of intraluminal type, 2 cases of extraluminal type, and 12 cases of junctional type. The tumors of intraluminal type presented as "worm-like" appearance, while the extraluminal and junctional type tumors presented as lobulated shape, with local "umbilical depression sign". The margins were clear in 3 cases and unclear in 14 cases. Vascular invasion by the mass was observed in 7 cases. On CT plain scan, the solid component of PLV presented as isodense or slightly hypodense. On MRI T1-weighted images, the solid component presented as isodense or slightly hypodense, and on fat-suppressed T2-weighted images, it presented as isodense or slightly hyperdense. On MRI diffusion-weighted imaging, 12 cases showed high signal intensity, and the corresponding apparent diffusion coefficient map showed low signal intensity. On CT enhanced scan, 8 cases showed mild to moderate enhancement of the solid part of PLV in the arterial phase and 9 cases showed obvious enhancement. In the venous phase and delayed phase, 10 cases showed continuous enhancement and 7 cases showed progressive enhancement. On MRI enhanced scan, 8 cases showed moderate enhancement and 6 cases showed obvious enhancement of the solid part of PLV in the arterial phase. In the venous phase and delayed phase, continuous progressive enhancement was observed; no enhancement was seen in the necrotic and cystic areas.Conclusion:The imaging characteristics of PLV show that the maximum diameter of the masses are consistent with the direction of inferior vena cava, most masses exhibit a "petal-like" necrotic cyst morphology with collateral circulation, and a localized "umbilical concave sign", and some cases possess hemophilic tube features.

Objective:To explore the clinical effects and value for application of the tibial flap of the second toe with the vascular pedicle including a plantar vein on the reconstruction of fingertip soft tissue defects.Methods:From October 2020 to August 2022, retrospective analysis of 12 patients (12 digits) were treated at the Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital for small soft tissue defects of fingertip. The patients were 9 males and 3 females and aged 29-54 years, at 38 years in average. The fingertip defects were measured at approximately 1.2 cm × 0.8 cm to 1.2 cm × 1.2 cm. In the surgery, a tibial flap of second toe was designed to reconstruct the defect in fingertip. The flap were designed on the tibial of second toe without an extended incision in dorsal foot. The pedicle of the flap carried with the lateral proper digital artery, a nerve and the plantar vein of the second toe. At the recipient site, the artery, nerve and vein carried by the pedicle of the flap were end-to-end anastomosed with the digital artery, digital nerve and subcutaneous vein of the finger. The flaps were measured at 1.5 cm×1.0 cm - 1.5 cm×1.5 cm in size. All donor sites were reconstructed with skin grafts from the ipsilateral calf. Scheduled postoperative follow-ups were conducted at the outpatient clinic. The Michigan Hand Function Questionnaire (MHQ) evaluation criteria was employed to assess the recovery of hand function, and Total Active Movement (TAM) was used to evaluate the recovery of range of motion of the interphalangeal joints of the affected fingers.Results:All 12 flaps in the fingertips survived. Postoperative follow-ups lasted from 6 months to 2 years, with an average of 11 months. One flap was slightly bloated and a flap aesthetic surgery was followed at 3 months after the primary reconstructive surgery, and the rest of flaps were all in good appearance. TPD was found at 12 -14 mm for all flaps at 9 months after surgery. All donor sites in the feet and calfs had primary healing, without a contracture or rupture of skin graft or an obvious dysfunction at the donor sites. According to the evaluation criteria of the MHQ, 8 patients were very satisfied with the overall appearance of the hand, and 4 were satisfied. Finger movement was evaluated according to TAM criteria, all 12 fingers were rated excellent.Conclusion:Reconstruction of a fingertip defect with a tibial flap of the second toe with the vascular pedicle including a plantar vein of the second toe has a good clinical efficacy. It has advantages in flap harvest, avoids an extended incision on dorsal foot, and makes a minimal damage to the donor site.

Objective To explore the otherness of orthotopic injection of cell suspensions and transplantation of tumor tissue blocks to establish orthotopic implantation models of hepatocellular carcinoma in mice,and to provide a technical reference for the establishment of an orthotopic implantation model.Methods Healthy KM mice were divided into four groups:group A,direct injection of H22 cells;group B,direct injection of H22 ascitic cells;group C,transplantation of tissues;and group D,direct injection of saline.Activity and weight changes were observed regularly in each group and survival times were recorded.Liver tumor formation,tumor size,abdominal organ adhesion degree,and metastasis were observed in all groups.B-ultrasound imaging was performed,concentrations of alpha fetoprotein(AFP)and abnormal prothrombin(DCP)were detected,and liver histopathological changes were detected by hematoxylin and eosin staining.Results Mice molding operation time in groups A,B,and C were(3.36±0.44)min,(3.30±0.41)min,and(5.68±0.65)min,respectively.After modeling for 25 days,the rates of model formation in groups A,B,and C were all 100.0%.Severe abdominal adhesions occurred in 40.0%of mice in group A and 60.0%in group B,but in no mice in group C or D.Ascites occurred in 40.0%,100.0%,and 0.0%and abdominal wall tumors in 30.0%,60.0%,and 0.0%of mice in groups A,B,and C,respectively,while 40.0%of mice in group B also had liver metastasis.B-ultrasound imaging,detection of serum AFP and DCP levels,and histopathological result showed smooth liver margins,uneven echo and slightly lower echo mass,maintained high AFP and DCP secretion,and large numbers of inflammatory cells and tumor cells in mice in groups A,B,and C.Conclusions At day 25,all three methods can thus be used to establish orthotopic transplantation models of HCC.Among these,inj ection of cell suspensions demonstrated the advantage of simplicity in operation and the presence of multiple metastatic nodules within the liver,compared to transplantation of tumor tissue.Conversely,transplantation of tumor tissue showed the advantage of causing less impact on the abdomen and other organs when compared to inj ection of cell suspensions.

Objective:To explore the application value and treatment effects of multiple free anterolateral perforator flaps of calf for reconstruction of multiple digital wounds in single hand.Methods:From August 2020 to March 2022, 12 patients with soft tissue defects in 35 digits were treated in Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital. Ten patients were male and 2 were female, aged 25 to 58 years old. Of the patients, 1 had soft tissue defects in 5 digits, 3 in 4 digits, 2 in 3 digits and 6 in 2 digits. The size of defects was from 1.2 cm ×1.2 cm to 7.0 cm×3.5 cm after debridement. The vascular perforators discovered from intraoperative explorations were found originating from the superficial peroneal artery in 24 flaps, from the peroneal artery in 7 flaps and from the anterior tibial artery in 4 flaps. During surgery, the perforator artery and accompanying veins of the flaps were anastomosed with the proper digital artery and palmar or dorsal subcutaneous veins in the recipient site, respectively. The size of the flaps was from 1.5 cm×1.5 cm to 7.5 cm×4.0 cm. No nerve was affected in the surgery. The wound at donor sites in the calf was sutured directly. Regular postoperative follow-ups were conducted at outpatient clinics. The comprehensive evaluation scale of flap was used to assess the conditions of the donor and recipient sites.Results:In this study, all 35 soft tissue defects of digits in 12 patients were reconstructed by the anterolateral perforator flaps of calf. All the 35 flaps survived after surgery, with a 100% of survival rate. The patients were instructed to carry out early functional training after surgery. Follow-up lasted 6 to 24 months, with an average of 11 months. Twenty-five flaps were found in slightly swollen, and further flap thinning surgery were carried out 3 months after the primary surgery, while the rest of the flaps had good appearance and texture. At 6 months after surgery, all flaps recovered a partial deep and shallow sensory and sense of touch. All wound at donor sites in calf had one-stage healing without dysfunction. The comprehensive evaluation scale was excellent in 28 flaps and good in 7 flaps. The excellent and good rate was 100%.Conclusion:It is an effective method to use multiple free anterolateral perforator flaps of calf to reconstruct multiple digit defects in single hand. The flaps can be conveniently harvested and the multiple digital defects in single hand can be reconstructed in primary surgery with small damages to the donor sites and together with satisfactory clinical outcomes.

Objective:To establish an animal model of pneumonia for research on clinical prevention and treatment of bacterial pneumonia by infecting immunocompromised rats with drug-resistant Acinetobacter baumannii ( Ab) strains. Methods:Drug-resistant Ab strains were selected. Forty-eight SD rats were randomly divided into four groups: normal group, cyclophosphamide control group (intraperitoneal injection of 45 mg/kg cyclophosphamide), bacterial infection group (intratracheal instillation of 1.5×10 8 CFU Ab suspension), and bacterial infection+ immunosuppression group (intraperitoneal injection of 45 mg/kg cyclophosphamide+ intratracheal instillation of 1.5×10 8 CFU Ab suspension). Flow cytometry analysis was used to detect the proportion of CD4 + , CD8 + and NK cells in rat peripheral blood before as well as 3 d and 7 d after infection. A lung function meter was used to detect peak inspiratory flow (PIF), peak expiratory flow (PEF), tidal volume (Vt ) and forced expiratory volume in the second second/forced vital capacity (FEV 200/FVC) at 3 d and 7 d after modeling. ELISA was used to detect the levels of IL-6, TNF-α and IL-10 in the alveolar lavage fluid. HE staining was used to observe the morphology of rat lung tissues in each group. Bacterial loads in rat lung tissues were counted by bacterial culturing. Results:A decrease in voluntary activity was observed in rats in the cyclophosphamide control group, bacterial infection group and bacterial infection+ immunosuppression group after modeling. Lung rales could be heard in the bacterial infection group and bacterial infection+ immunosuppression group. Compared with the normal group, the cyclophosphamide control group showed decreased proportion of CD4 + and CD11b + NK cells and increased CD8 + cells in peripheral blood; the bacterial infection group showed decreased PIF, PEF, Vt and FEV 200/FVC, increased IL-6 and TNF-α levels and decreased IL-10 level in the alveolar lavage fluid, and higher bacterial load in lung tissues with mild widening of alveolar walls and inflammatory cell infiltration ( P<0.05, P<0.01). Compared with the cyclophosphamide control group and the bacterial infection group, the bacterial infection+ immunosuppression group showed a lower proportion of CD4 + cells and a higher proportion of CD8 + cells in rat peripheral blood, decreased PIF, PEF, Vt and FEV 200/FVC, increased IL-6 and TNF-α levels and decreased IL-10 level in alveolar lavage fluid, higher bacterial load in lung tissues, and aggravated widening of alveolar walls and inflammatory cell infiltration ( P<0.05, P<0.01). The proportion of CD11b + NK cells in peripheral blood of rats in the bacterial infection+ immunosuppression group was significantly lower than that in the bacterial infection group ( P<0.05, P<0.01). Conclusions:A bacterial pneumonia model was successfully constructed by infecting rats with Ab alone or in combination with cyclophosphamide immunosuppression. In the model constructed with Ab and cyclophosphamide immunosuppression, the rats had more severe pneumonia, which might be related to the reduced cellular immune function and the aggravated bacterial infection in rat lung tissues by cyclophosphamide.

Objective:To evaluate the effects of the changes in respiratory status on gated stereotactic radiotherapy under free breathing guided by real-time position management (RPM).Methods:This study simulated the baseline shift, change in respiratory rate, end-expiratory delay, end-inspiratory delay, and irregular breathing using an in-house developed motion phantom. Moreover, this study analyzed the correlation between the changes in the above states of three plans, three-dimensional conformal radiotherapy(3D-CRT), intensity modulated radiotherapy(IMRT), and volumetric modulated arc therapy(VMAT) and the position of the ball at the center of the motion phantom (L) and the exposed dose of the phantom in the ionization chamber (the dose).Results:The in-house developed phantom presented high setup repeatability and measurement stability. There was a positive correlation between L and the baseline shift ( r = 0.99, P < 0.01). The change in the dose was less than 4% when the baseline shift was less than the setup error, while the dose declined rapidly and was negatively correlated with the baseline shift otherwise ( r= -0.95, P < 0.01). Moreover, there was statistically significant difference in dose when the baseline shift exceeded the setup error or not ( Z = -3.06, P < 0.01). There was no significant difference in the rate of the dose affected by baseline shift in the three plans ( P > 0.05). The changes in respiratory rate had little effect on L and the dose. Both end-inspiratory delay and end-expiratory delay reduced the planned dose of the three plans, with a maximum decrease of up to -1.74%. Furthermore, the end-inspiratory delay has greater effects on the planned dose than the end-expiratory delay( Z = -2.67, P< 0.01). However, there was no significant correlation between the dose and the delay duration ( P > 0.05), and no significant difference in the rate of the planned dose of the three plans affected by respiratory waveform change ( P > 0.05). Irregular breathing had greater effects on the dose. Specifically, the dose from six repeated measurements of 3D-CR, IMRT, VMAT was (709.68±180.00), (751.40±127.16), and (750.00±185.60) cGy, respectively, all less than the prescribed dose with a poor consistency. Conclusions:The changes in the patients′ respiratory status will reduce the dose, especially when the baseline shift exceeds the setup error threshold or large respiratory waveform variation corresponding to irregular breathing occurs. Moreover, there is no correlation between the decrease in the dose and the radiotherapy technology.