Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG^@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

Objective:To construct a finite element analysis model for the application of sustentaculum tail sustentacular screw in the treatment of calcaneal fracture of Sanders type Ⅱ. The finite element analysis method was used to study the effectiveness of the simulation treatment of Sanders type Ⅱ calcaneal fracture.Methods:A complete adult calcaneal specimen that was not affecting the normal quality of bone after teaching in the Department of Human Anatomy, Kangda College of Nanjing Medical University from 2021 to 2022 second semester was selected. A micro CT scan was performed to obtain data. The three-dimensional model of the calcaneus was reconstructed by the software of Mimics and Geomagic Wrap. The internal fixation and fracutre model were drawn using SolidWorks software. The obtained data were imported into the finite element analysis software for material assignment and meshing to obtain a three-dimensional finite element model. The load and boundary constraints were applied to each model to perform finite element analysis and calculation, and the stress and displacement of the model were extracted. Finally, the experimental screw placement of the calcaneus loading process was verified.Results:The three-dimensional model of calcaneal fracture had a high degree of appearance simulation, complete morphological structure, no defect, and was consistent with the size of the solid calcaneus. After surgical treatment, the anatomical reduction of each fracture model was good, the steel plate fit to the bone surface, and the screw did not protrude outside the model. There was no deformation or damage to the whole internal fixation, the grid division was clear, and the number was relatively uniform and consistent. The maximum stress in each type of calcaneal fracture model was in the internal fixation, and the overall stress was uniformly distributed in each component. The maximum displacement distance of each type of calcaneal fracture was less than 0.13 mm, and the angles and lengths of nails were accurate. The tip of the Kirschner wires was located in the sustentaculum tail. There was no rupture of the sustentaculum tali and no protrusion of the Kirschner wires.Conclusions:The internal fixation is an effective method to treat the Sanders type Ⅱ calcaneal fractures by using sustentaculum tail sustentacular screws.

Objective:To explore the clinical efficacy of targeted interventional embolization therapy by a multidisciplinary team for the hemorrhagic shock caused by acute pelvic fracture or/and acetabular fracture.Methods:A retrospective study was conducted to analyze the data of 63 patients with hemorrhagic shock caused by pelvic fracture or/and acetabular fracture who had been admitted to Department of Orthopaedics, The Second Hospital of Shanxi Medical University from January 2015 to July 2022. There were 44 males and 19 females with an age of (39.6±15.6) years, and 23 pelvic fractures, 35 acetabular fractures, and 5 pelvic and acetabular fractures. The time from injury to targeted interventional embolization therapy was 2.67 (2.00, 3.33) hours. All the patients were treated with targeted interventional embolization therapy by a multidisciplinary team involving orthopedics, interventional medicine, general surgery, and urology. The shock index and lactate level within 12 hours after therapy, 24-hour urine output, and incidence of complications 3 weeks after therapy were recorded.Results:No bleeding was found again in the 63 patients after embolization. Within 12 hours after therapy, the shock index was ≤1.0, indicating the shock was corrected. Within 12 hours after targeted interventional embolization therapy, the shock index (0.70±0.46) and lactate value [(2.03±1.35) mmol/L] in the 63 patients were significantly lower than those before therapy [(1.76±0.56) and (4.53±1.74) mmol/L] ( P<0.05). The 24-hour urine output [(50.26±20.38) mL/h] was significantly higher than that before therapy [(21.56±1.27) mL/h] ( P<0.05). Two patients experienced poor blood circulation in the distal skin of the great toe, which was relieved after treatment with blood circulation promotion and anticoagulation. Three patients developed necrosis of the hip soft tissue, which was cured after multiple times of debridement and anti-infection treatments. One patient with severe injury died from multiple organ dysfunction. Conclusions:The targeted interventional embolization therapy can not only diagnose the bleeding location in patients with hemorrhagic shock caused by pelvic fracture or/and acetabular fracture, but also timely and accurately carry out hemostatic treatment to correct shock. Moreover, a multidisciplinary team can help patients avoid multiple surgeries and decrease their pain and financial loss.

The rise of nanotechnology has opened new horizons for cancer immunotherapy. However, most nanovaccines fabricated with nanomaterials suffer from carrier-related concerns, including low drug loading capacity, unpredictable metabolism, and potential systemic toxicity, which bring obstacles for their clinical translation. Herein, we developed an antigen self-assembled nanovaccine, which was resulted from a simple acryloyl modification of the antigen to induce self-assembly. Furthermore, a dendritic cell targeting head mannose monomer and a mevalonate pathway inhibitor zoledronic acid (Zol) were integrated or absorbed onto the nanoparticles (denoted as MEAO-Z) to intensify the immune response. The synthesized nanovaccine with a diameter of around 70 nm showed successful lymph node transportation, high dendritic cell internalization, promoted costimulatory molecule expression, and preferable antigen cross-presentation. In virtue of the above superiorities, MEAO-Z induced remarkably higher titers of serum antibody, stronger cytotoxic T lymphocyte immune responses and IFN-γ secretion than free antigen and adjuvants. In vivo, MEAO-Z significantly suppressed EG7-OVA tumor growth and prolonged the survival time of tumor-bearing mice. These results indicated the translation promise of our self-assembled nanovaccine for immune potentiation and cancer immunotherapy.

Due to the insufficient long-term protection and significant efficacy reduction to new variants of current COVID-19 vaccines, the epidemic prevention and control are still challenging. Here, we employ a capsid and antigen structure engineering (CASE) strategy to manufacture an adeno-associated viral serotype 6-based vaccine (S663V-RBD), which expresses trimeric receptor binding domain (RBD) of spike protein fused with a biological adjuvant RS09. Impressively, the engineered S663V-RBD could rapidly induce a satisfactory RBD-specific IgG titer within 2 weeks and maintain the titer for more than 4 months. Compared to the licensed BBIBP-CorV (Sinopharm, China), a single-dose S663V-RBD induced more endurable and robust immune responses in mice and elicited superior neutralizing antibodies against three typical SARS-CoV-2 pseudoviruses including wild type, C.37 (Lambda) and B.1.617.2 (Delta). More interestingly, the intramuscular injection of S663V-RBD could overcome pre-existing immunity against the capsid. Given its effectiveness, the CASE-based S663V-RBD may provide a new solution for the current and next pandemic.

Mucosal vaccines that stimulate both mucosal and systemic immune responses are desirable, as they could prevent the invading pathogens at their initial infection sites in a convenient and user-friendly way. Nanovaccines are receiving increasing attention for mucosal vaccination due to their merits in overcoming mucosal immune barriers and in enhancing immunogenicity of the encapsulated antigens. Herein, we summarized several nanovaccine strategies that have been reported for enhancing mucosal immune responses, including designing nanovaccines that have superior mucoadhesion and mucus penetration capacity, designing nanovaccines with better targeting efficiency to M cells or antigen-presenting cells, and co-delivering adjuvants by using nanovaccines. The reported applications of mucosal nanovaccines were also briefly discussed, including prevention of infectious diseases, and treatment of tumors and autoimmune diseases. Future research progresses in mucosal nanovaccines may promote the clinical translation and application of mucosal vaccines.

Photothermal therapy has been intensively investigated for treating cancer in recent years. However, the long-term therapeutic outcome remains unsatisfying due to the frequently occurred metastasis and recurrence. To address this challenge, immunotherapy has been combined with photothermal therapy to activate anti-tumor immunity and relieve the immunosuppressive microenvironment within tumor sites. Here, we engineered silica-based core‒shell nanoparticles (JQ-1@PSNs-R), in which silica cores were coated with the photothermal agent polydopamine, and a bromodomain-containing protein 4 (BRD4) inhibitor JQ-1 was loaded in the polydopamine layer to combine photothermal and immune therapy for tumor elimination. Importantly, to improve the therapeutic effect, we increased the surface roughness of the nanoparticles by hydrofluoric acid (HF) etching during the fabrication process, and found that the internalization of JQ-1@PSNs-R was significantly improved, leading to a strengthened photothermal killing effect as well as the increased intracellular delivery of JQ-1. In the animal studies, the multifunctional nanoparticles with rough surfaces effectively eradicated melanoma via photothermal therapy, successfully activated tumor-specific immune responses against residual tumor cells, and further prevented tumor metastasis and recurrence. Our results indicated that JQ-1@PSNs-R could serve as an innovative and effective strategy for combined cancer therapy.

Objective:To evaluate our novel path based on anatomical division of the anterior lateral wall of calcaneus and the sustentaculum tali for precise sustentacular screw placement in the surgical treatment of calcaneal fractures of Sanders types Ⅱ and Ⅲ.Methods:The anterior lateral wall of the calcaneus was divided into the anterior-superior zone S 1, the anterior-inferior zone S 2, the posterior-superior zone S 3 and the posterior-inferior zone S 4 for demarcation of the screw insertion points by our method of Four Zones, and into the front, middle and rear sections by our method of Three Sections for demarcation of the screw target points. The specimens were scanned by CT and modeled by Mimics. On the 3D virtual model of the calcaneus, one screw was placed from each zone of the anterior lateral wall of the calcaneus to the sustentaculum tali body. The screw placement target for S 1 and S 2 was the medial intersection point P 1 of the front and middle sections of the sustentaculum tali, and that for S 3 and S 4 was the medial intersection point P 2 of the middle and rear sections of the sustentaculum tali. It was observed whether the screws were placed in the bone channel. A total of 72 patients were included who had been admitted to Department of Orthopaedics, Suqian Hospital Affiliated to Xuzhou Medical University for calcaneal fractures of Sanders types Ⅱ and Ⅲ from January 2017 to January 2021. They were divided into an anatomical division group and a 3D printing group according to their screw placement method for the sustentaculum tali. In the anatomical division group of 32 patients subjected to screw placement based on our anatomical division, there were 25 males and 7 females, aged from 24 to 60 years; in the 3D printing group of 40 patients subjected to screw placement assisted by 3D printing, there were 31 males and 9 females, aged from 25 to 58 years. The disparities between the parameters of sustentacular screw placement and the actual values were compared in the anatomical division group, and the total number of screws, screws on average, distribution of screws, and accuracy of screw placement were compared between the 2 groups. Results:All the screws which were virtually placed in the specimens of the calcaneus from S 1 and S 2 to P 1 and from S 3 and S 4 to P 2 passed through the bony channel, with no perforation into the tarsal sinus. There was no significant difference in the general date between the anatomical division group and the 3D printing group, showing they were comparable ( P > 0.05). In the anatomical division group, a total of 52 screws were placed to the sustentaculum tali with an average of (1.63 ± 0.48) screws per patient, and 2 screws were placed in 20 patients, yielding an accuracy rate of screw placement of 92.3% (48/52). There were no statistically significant differences between the parameters and the actual values of screw placement in the anatomical division group ( P > 0.05). In the 3D printing group, a total of 63 screws were placed to the sustentaculum tali with an average of (1.58 ± 0.49) screws per patient, and 2 screws were placed in 23 patients, yielding an accuracy rate of screw placement of 93.7% (59/63). There were no significant differences in the above comparisons between the anatomical division group and the 3D printing group ( P > 0.05). Conclusion:In the surgical treatment of calcaneal fractures of Sanders types Ⅱ and Ⅲ, the sustentacular screw placement based on our anatomical division of the anterior lateral wall of the calcaneus and the sustentaculum tali can lead to similar clinical accuracy as 3D printing-assisted screw placement does.

Objective:To compare the difference of low-level assisted ventilation and T-piece method on respiratory mechanics of patients with invasive mechanical ventilation during spontaneous breathing trial (SBT) within 3 days before extubation.Methods:A retrospective observational study was conducted. Twenty-five patients with difficulty in weaning or delayed weaning from invasive mechanical ventilation who were admitted to department of critical care medicine of the First Affiliated Hospital of Guangzhou Medical University from December 2018 to June 2020, and were in stable condition and entered the weaning stage after more than 72 hours of invasive mechanical ventilation were studied. A total of 119 cases of respiratory mechanical indexes were collected, which were divided into the low-level assisted ventilation group and the T-piece group according to the ventilator method and parameters used during the data collection. The different ventilation modes related respiratory mechanics indexes such as the esophageal pressure (Pes), the gastric pressure (Pga), the transdiaphragmatic pressure (Pdi), the maximum Pdi (Pdimax), Pdi/Pdimax ratio, the esophageal pressure-time product (PTPes), the gastric pressure-time product (PTPga), the transdiaphragmatic pressure-time product (PTPdi), the diaphragmatic electromyography (EMGdi), the maximum diaphragmatic electromyography (EMGdimax), PTPdi/PTPes ratio, Pes/Pdi ratio, the inspiratory time (Ti), the expiratory time (Te) and the total time respiratory cycle (Ttot) at the end of monitoring were recorded and compared between the two groups.Results:Compared with the T-piece group, Pes, PTPes, PTPdi/PTPes ratio, Pes/Pdi ratio and Te were higher in low-level assisted ventilation group [Pes (cmH 2O, 1 cmH 2O = 0.098 kPa): 2.84 (-1.80, 5.83) vs. -0.94 (-8.50, 2.06), PTPes (cmH 2O·s·min -1): 1.87 (-2.50, 5.93) vs. -0.95 (-9.71, 2.56), PTPdi/PTPes ratio: 0.07 (-1.74, 1.65) vs. -1.82 (-4.15, -1.25), Pes/Pdi ratio: 0.17 (-0.43, 0.64) vs. -0.47 (-0.65, -0.11), Te (s): 1.65 (1.36, 2.18) vs. 1.33 (1.05, 1.75), all P < 0.05], there were no significant differences in Pga, Pdi, Pdimax, Pdi/Pdimax ratio, PTPga, PTPdi, EMGdi, EMGdimax, Ti and Ttot between the T-piece group and the low-level assisted pressure ventilation group [Pga (cmH 2O): 6.96 (3.54,7.60) vs. 7.74 (4.37, 11.30), Pdi (cmH 2O): 9.24 (4.58, 17.31) vs. 6.18 (2.98, 11.96), Pdimax (cmH 2O): 47.20 (20.60, 52.30) vs. 29.95 (21.50, 47.20), Pdi/Pdimax ratio: 0.25 (0.01, 0.34) vs. 0.25 (0.12, 0.41), PTPga (cmH 2O·s·min -1): 7.20 (2.54, 9.97) vs. 7.97 (5.74, 13.07), PTPdi (cmH 2O·s·min -1): 12.15 (2.95, 19.86) vs. 6.87 (2.50, 12.63), EMGdi (μV): 0.05 (0.03, 0.07) vs. 0.04 (0.02, 0.06), EMGdimax (μV): 0.07 (0.05, 0.09) vs. 0.07 (0.04, 0.09), Ti (s): 1.20 (0.95, 1.33) vs. 1.07 (0.95, 1.33), Ttot (s): 2.59 (2.22, 3.09) vs. 2.77 (2.35, 3.24), all P > 0.05]. Conclusions:When mechanically ventilated patients undergo SBT, the use of T-piece method increases the work of breathing compared with low-level assisted ventilation method. Therefore, long-term use of T-piece should be avoided during SBT.

Objective:To understand the function of diaphragm and analyze the clinical factors affecting the function of diaphragm by measuring twitch tracheal pressure (TwPtr) in patients with mechanical ventilation and in the weaning phase.Methods:Patients with more than 48 hours of invasive mechanical ventilation admitted to the department of critical care medicine of the First Affiliated Hospital of Guangzhou Medical University from December 2015 to March 2017 were enrolled. After the patient entered the weaning stage, TwPtr of patients was monitored by two-way non repetitive automatic respiratory trigger device, the effects of duration of mechanical ventilation, severe pulmonary infection, sedative application and chronic obstructive pulmonary disease (COPD) on weaning were analyzed.Results:A total of 62 patients were included, of which 45 were male and 17 were female. The average age was (66.8±11.7) years old. Twenty-three cases had severe pneumonia. The absolute value of TwPtr in severe pneumonia group was lower than that in non-severe pneumonia group [cmH 2O (1 cmH 2O = 0.098 kPa): 10.40±5.81 vs. 14.35±5.22, P = 0.021]. However, there was no significant difference in the duration of mechanical ventilation between the severe pneumonia group and non-severe pneumonia group [days: 26 (17, 43) vs. 15 (11, 36), P = 0.091]. In 62 patients with mechanical ventilation, there was a negative correlation between TwPtr and duration of mechanical ventilation ( r = 0.414, P = 0.002), there was also a negative correlation between the duration of mechanical ventilation and TwPtr after the assessment of diaphragm function ( r = 0.277, P = 0.039). There was a linear relationship between TwPtr and sedatives ( r = 0.220, P = 0.040), but there was no correlation between TwPtr and COPD ( r = -0.178, P = 0.166). Conclusions:For patients in the weaning stage of mechanical ventilation, severe pulmonary infection is one of the factors that affect the diaphragm dysfunction. There is a certain correlation between the diaphragm dysfunction and the use of sedatives.