Immunomodulatory cancer therapy is witnessing the rise of viral immunotherapy. The oncolytic influenza A virus, although promising in preclinical investigations, remains to be implemented in clinical practice. Recent progress in genetic engineering, coupled with experiential insights, offers opportunities to enhance the therapeutic efficacy of the influenza A virus. This review explores the use of the influenza virus, its attenuated forms, and associated vaccines in cancer immunotherapy, highlighting their respective advantages and challenges. We further elucidate methods for engineering influenza viruses and innovative approaches to augment them with cytokines or immune checkpoint inhibitors, aiming to maximize their clinical impact. Our goal is to provide insights essential for refining influenza A virus-based viral tumor immunotherapies.
Humans ; Neoplasms/immunology* ; Immunotherapy/trends* ; Influenza A virus/immunology* ; Oncolytic Virotherapy/trends* ; Animals ; Cancer Vaccines/therapeutic use* ; Oncolytic Viruses ; Genetic Engineering ; Immune Checkpoint Inhibitors/therapeutic use*

Objective:To investigate the effects of gelatin methacrylate anhydride (GelMA) hydrogel loaded with small extracellular vesicles derived from human umbilical cord mesenchymal stem cells (hUCMSCs-sEVs) in the treatment of full-thickness skin defect wounds in mice.Methods:This study was an experimental study. hUCMSCs-sEVs were extracted by ultracentrifugation, their morphology was observed through transmission electron microscope, and the expression of CD9, CD63, tumor susceptibility gene 101 (TSG101), and calnexin was detected by Western blotting. The human umbilical vein endothelial cells (HUVECs), the 3 rd and 4 th passages of human epidermal keratinocytes (HEKs) and human dermal fibroblasts (HDFs) were all divided into blank control group (routinely cultured) and hUCMSC-sEV group (cultured with the cell supernatant containing hUCMSCs-sEVs). The cell scratch test was performed and the cell migration rates at 6, 12, and 24 h after scratching were calculated, the cell Transwell assay was performed and the number of migration cells at 12 h after culture was calculated, and the proportion of proliferating cells was detected by 5-acetylidene-2'-deoxyuridine and Hoechst staining at 24 h after culture, with sample numbers being all 3. The simple GelMA hydrogel and the GelMA hydrogel loaded with hUCMSCs-sEVs (hereinafter referred to as hUCMSC-sEV/GelMA hydrogel) were prepared. Then the micromorphology of 2 kinds of hydrogels was observed under scanning electron microscope, the distribution of hUCMSCs-sEVs was observed by laser scanning confocal microscope, and the cumulative release rates of hUCMSCs-sEVs at 0 (immediately), 2, 4, 6, 8, 10, and 12 d after soaking hUCMSC-sEV/GelMA hydrogel in phosphate buffer solution (PBS) were measured and calculated by protein colorimetric quantification ( n=3). Twenty-four 6-week-old male C57BL/6J mice were divided into PBS group, hUCMSC-sEV alone group, GelMA hydrogel alone group, and hUCMSC-sEV/GelMA hydrogel group according to the random number table, with 6 mice in each group, and after the full-thickness skin defect wounds on the back of mice in each group were produced, the wounds were performed with PBS injection, hUCMSC-sEV suspenson injection, simple GelMA coverage, and hUCMSC-sEV/GelMA hydrogel coverage, respectively. Wound healing was observed on post injury day (PID) 0 (immediately), 4, 8, and 12, and the wound healing rates on PID 4, 8, and 12 were calculated, and the wound tissue was collected on PID 12 for hematoxylin-eosin staining to observe the structure of new tissue, with sample numbers being both 6. Results:The extracted hUCMSCs-sEVs showed a cup-shaped structure and expressed CD9, CD63, and TSG101, but barely expressed calnexin. At 6, 12, and 24 h after scratching, the migration rates of HEKs (with t values of 25.94, 20.98, and 20.04, respectively), HDFs (with t values of 3.18, 5.68, and 4.28, respectively), and HUVECs (with t values of 4.32, 19.33, and 4.00, respectively) in hUCMSC-sEV group were significantly higher than those in blank control group ( P<0.05). At 12 h after culture, the numbers of migrated HEKs, HDFs, and HUVECs in hUCMSC-sEV group were 550 ±23, 235 ±9, and 856 ±35, respectively, which were significantly higher than 188 ±14, 97 ±6, and 370 ±32 in blank control group (with t values of 22.95, 23.13, and 17.84, respectively , P<0.05). At 24 h after culture, the proportions of proliferating cells of HEKs, HDFs, and HUVECs in hUCMSC-sEV group were significantly higher than those in blank control group (with t values of 22.00, 13.82, and 32.32, respectively, P<0.05). The inside of simple GelMA hydrogel showed a loose and porous sponge-like structure, and hUCMSCs-sEVs was not observed in it. The hUCMSC-sEV/GelMA hydrogel had the same sponge-like structure, and hUCMSCs-sEVs were uniformly distributed in clumps. The cumulative release rate curve of hUCMSCs-sEVs from hUCMSC-sEV/GelMA hydrogel tended to plateau at 2 d after soaking, and the cumulative release rate of hUCMSCs-sEVs was (59.2±1.8)% at 12 d after soaking. From PID 0 to 12, the wound areas of mice in the 4 groups gradually decreased. On PID 4, 8, and 12, the wound healing rates of mice in hUCMSC-sEV/GelMA hydrogel group were significantly higher than those in the other 3 groups ( P<0.05); the wound healing rates of mice in GelMA hydrogel alone group and hUCMSC-sEV alone group were significantly higher than those in PBS group ( P<0.05). On PID 8 and 12, the wound healing rates of mice in hUCMSC-sEV alone group were significantly higher than those in GelMA hydrogel alone group ( P<0.05). On PID 12, the wounds of mice in hUCMSC-sEV/GelMA hydrogel group showed the best wound epithelization, loose and orderly arrangement of dermal collagen, and the least number of inflammatory cells, while the dense arrangement of dermal collagen and varying degrees of inflammatory cell infiltration were observed in the wounds of mice in the other 3 groups. Conclusions:hUCMSCs-sEVs can promote the migration and proliferation of HEKs, HDFs, and HUVECs which are related to skin wound healing, and slowly release in GelMA hydrogel. The hUCMSC-sEV/GelMA hydrogel as a wound dressing can significantly improve the healing speed of full-thickness skin defect wounds in mice.

The final outcome of the retinal degenerative diseases is the massive loss of photoreceptors, resulting in irreversible visual impairment which lacks effective treatment at present.As a potential therapeutic approach, photoreceptor transplantation can be used to restore retinal function to a certain extent by replacing the lost photoreceptors and rebuilding the retinal circuits.However, the discovery of material exchange unveiled a number of problems in previous studies, including low cellular integration, insufficient outer segment and synapse formation, highlighting the challenges of clinical translation.To explore the possibility of increasing the functional integration of photoreceptors, this article reviewed a variety of strategies, including selection of the transplanted cells with optimal developmental stage to enhance the interaction with the host retina, disruption of the outer limiting membrane and alleviation of retinal remodeling to improve the migration and integration of the transplanted photoreceptors, regulation of immunity can be used to reduce microglial activation to create a better host microenvironment for transplantation, use of retinal sheets or biological scaffolds to improve photoreceptor organization, rational development and use of biomaterials to optimize the physiological microenvironment of the transplanted cells, adequate evaluation of surgical parameters to reduce the effect of surgery on the transplanted cells and the host retina.

Objective To explore the correlations between serum Mas-related G protein-coupled receptor X2 (MRGPRX2), interleukin (IL)-4, IL-5, IL-6, IL-13, IL-23 and IL-33 levels and chronic spontaneous urticaria (CSU). Methods The clinical characteristics and laboratory data from 55 patients with CSU and 21 healthy controls at Huashan Hospital, Fudan University from February 2021 to September 2023 were collected. The disease activity and severity of CSU patients were assessed. Serum level of MRGPRX2 was tested using enzyme-linked immunosorbent assay (ELISA), and levels of IL-4, IL-5, IL-6, IL-13, IL-23, and IL-33 were measured using Luminex multiplex assay in all subjects. Spearman correlation analysis was used to evaluate the correlations between biomarkers and other parameters in CSU patients, and logistic regression analysis was performed to identify factors influencing CSU. Results CSU patients exhibited significantly higher serum levels of MRGPRX2 (2.41[0, 11.51] ng/mL vs 0[0, 2.86] ng/mL, P＝0.015) and IL-23 (0.09[0.04, 0.56] pg/mL vs 0.05[0.03, 0.08] pg/mL, P＝0.033) than healthy controls. There was no difference in levels of other cytokines between the two groups. There was no difference in levels of MRGPRX2 and cytokines between severe and non-severe CSU patients. Correlation analysis showed that serum MRGPRX2 levels in CSU patients were positively correlated with IL-4 (r＝0.345, P＝0.010) and IL-6 (r＝0.395, P＝0.003) levels. Logistic regression analysis indicated that MRGPRX2≥0.055 ng/mL and IL-23≥0.135 pg/mL were independent risk factors for CSU (P＜0.05). Conclusions Serum levels of MRGPRX2 and IL-23 in CSU patients are elevated, which may be involved in the pathogenesis of CSU.

Shear thinning is an ideal feature of bioink because it can reduce the chance of blocking. For extrusion based biological printing, bioink will experience shear force when passing through the biological printer. The shear rate will increase with the increase of extrusion rate, and the apparent viscosity of shear-thinning bioink will decrease, which makes it easier to block, thus achieving the structural fidelity of 3D printing tissue. The manufacturing of complex functional structures in tissue trachea requires the precise placement and coagulation of bioink layer by layer, and the shear-thinning bioink may well meet this requirement. This review focuses on the importance of mechanical properties, classification and preparation methods of shear-thinning bioink, and lists its current application status in 3D printing tissue trachea to discuss the more possibilities and prospects of this biological material in tissue trachea.

Objective To investigate the value of Thromboelastogram(TEG)combined with conventional coagulation indexes and peripheral blood inflammation indexes in the diagnosis of threatened abortion.Methods A total of 33 patients with threatened abortion who were treated in the hospital from July 1,2022 to July 1,2023 were selected as the study group,and 40 healthy pregnant women in the first and second trimester of pregnan-cy were randomly selected as the control group.The blood routine,C-reactive protein(CRP),routine coagula-tion indexes and TEG were detected in the two groups,and the diagnostic value of TEG,routine coagulation indexes and peripheral blood inflammatory indexes in threatened abortion was analyzed.Results D-dimer(DD),white blood cell count(WBC),neutrophil count(NEU),neutrophil to lymphocyte ratio(NLR),CRP,coagulation Angle(α),maximum amplitude(MA),coagulation comprehensive index(CI)in the study group were higher than those in the control group(P＜0.05).The coagulation reaction time(R)and blood clot for-mation time(K)in the observation group were lower than those in the control group(P＜0.05).The correla-tion analysis between TEG and conventional coagulation indexes and inflammatory indexes showed that R was positively correlated with prothrombin time(PT)and activated partial thromboplastin time(APTT)(P＜0.05),K was negatively correlated with fibrinogen(FIB)(P＜0.05),and α was positively correlated with FIB(P＜0.05).MA and CI were positively correlated with PLT and FIB(P＜0.05).R and K were negatively cor-related with WBC,NEU and NLR(P＜0.05),α was positively correlated with CRP,WBC,NEU and NLR(P＜0.05),MA and CI were positively correlated with WBC,NEU and NLR(P＜0.05).Receiver operating characteristic curve analysis showed that the area under the curve of DD,CRP,α and the combination of the three were 0.775,0.748,0.922 and 0.959,respectively(P＜0.001).Conclusion TEG can be used to evaluate the coagulation function of patients with threatened abortion,and its combination with conventional coagula-tion indexes and peripheral blood inflammation indexes can provide an important basis for clinical diagnosis of threatened abortion,and has certain guiding significance for clinical diagnosis and treatment.

Medical homogenization in multi-campus hospital plays an essential role in leveraging the advantages of public hospitals, promoting the expansion of high-quality medical resources and balancing regional layout. The Second Affiliated Hospital Zhejiang University School of Medicine deeply used digital intelligence technology to build a new integrated mobile health service system consisting of internet hospital and 5G intelligent applications, which empowered medical efficiency in multi-campus hospital. This system broke the limitations of inconsistent medical resources, unbalanced discipline layout, and insufficient information connectivity in the construction of multi-campus hospitals, and achieved remarkable results in practice. It could provide reference for the multi-campus construction of other large public hospitals.

Objective:To observe the incidence and risk factors of microcystic macular edema (MME) in patients with idiopathic macular epiretinal membrane (IMEM) preoperatively and postoperatively.Methods:A retrospective case series study. From January 2017 to May 2021, 72 eyes of 72 patients with IMEM in Eye Hospital of Wenzhou Medical University at Hangzhou were included. There were 18 male and 54 female. Average age was 64.8±7.8 years. Eyes were all monocular. All patients received the examination of best corrected visual acuity (BCVA) by standard logarithmic visual acuity chart, which was represented logarithmic of minimum angle of resolution (logMAR). Optical coherence tomography was used to measure central macular thickness (CMT). MME was defined as small, vertically bounded cystic space located outside the fovea in the inner layer of the retina. According to the presence or absence of MME before surgery, the affected eyes were divided into two groups: non-MME group (35 eyes) and MME group (37 eyes). The difference of logMAR BCVA and CMT was statistically significant between Group A and B ( t=3.117, 2.589; P=0.003, 0.012). All patients with IMEM were treated with 23G three-channels pars plana vitrectomy (PPV) with epiretinal membrane and inner limiting membrane (ILM) peeling. The two groups were further divided into four groups according to whether there was MME in the postoperative follow-up time. The group A1 was without MME before and after surgery, group A2 was without MME before surgery and with MME after surgery. The MME group was subdivided into the group with MME before surgery and without MME after surgery (group B1) and the group with MME before and after surgery (group B2). The mean follow-up time was 8.8±7.7 months. The same equipment and methods were used to exam the patients during the follow-up. Paired t test was used to compare the changes of MME, BCVA and CMT before and after surgery. The differences of CMT and BCVA among groups before and after surgery were compared by independent sample t test and one-way ANOVA. Logistic regression was used to analyze the influencing factors of MME before and after surgery, and multiple linear regression was used to analyze the influencing factors of postoperative BCVA. Results:There were 35 eyes in the non-MME group, 18 eyes (51.43%, 18/35) in the A1 group and 17 eyes (48.57%, 17/35) in the A2 group, respectively. There were 37 eyes in MME group, 6 eyes (16.22%, 6/37) in group B1 and 31 eyes (83.78%, 31/37) in group B2, respectively. At last follow-up, the logMAR BCVA was 0.10±0.12, 0.25±0.17, 0.09±0.11, 0.30±0.26 in group A1, A2, B1, and B2, respectively. Compared with the logMAR BCVA before surgery, the differences were statistically significant ( t=3.779, 4.253, 7.869, 6.668; P<0.01). There was significant difference in logMAR BCVA among the four groups ( F=4.460, P<0.01). There was a significant difference in logMAR BCVA between group A1 and group A2 ( t=-2.930, P=0.006). There was no significant difference between group B1 and group B2 ( t=-1.921, P=0.063). The CMT of group A1, A2, B1 and B2 were 371.83±73.24, 431.24±83.13, 407.00±28.07 and 425.19±70.97 μm, respectively. Compared with those before operation, the differences were statistically significant ( t=5.197, 2.465, 3.055, 6.078; P<0.05). There was no significant difference in CMT among the four groups ( F=2.597, P=0.059). Logistic regression analysis showed that pre-operation MME was correlated with pre-operation IMEM stage ( β=1.494, P=0.004). New MME after surgery was correlated with age ( β=0.153, P=0.013). Multiple linear regression analysis showed that postoperative visual acuity was significantly correlated with CMT before surgery and MME after surgery ( β=0.001, 0.134; P=0.015, 0.019). Conclusions:PPV combined with epiretinal membrane and ILM peeling surgery for IMEM can improve visual acuity and decrease CMT. MME regress or regenerate after surgery. Age is an independent predictor of the risk of newly formed MME after surgery.

Focal cortical dysplasia (FCD) is one of the most common causes of drug-resistant epilepsy. Dysmorphic neurons are the major histopathological feature of type II FCD, but their role in seizure genesis in FCD is unclear. Here we performed whole-cell patch-clamp recording and morphological reconstruction of cortical principal neurons in postsurgical brain tissue from drug-resistant epilepsy patients. Quantitative analyses revealed distinct morphological and electrophysiological characteristics of the upper layer dysmorphic neurons in type II FCD, including an enlarged soma, aberrant dendritic arbors, increased current injection for rheobase action potential firing, and reduced action potential firing frequency. Intriguingly, the upper layer dysmorphic neurons received decreased glutamatergic and increased GABAergic synaptic inputs that were coupled with upregulation of the Na⁺-K⁺-Cl^- cotransporter. In addition, we found a depolarizing shift of the GABA reversal potential in the CamKII-cre::PTEN^flox/flox mouse model of drug-resistant epilepsy, suggesting that enhanced GABAergic inputs might depolarize dysmorphic neurons. Thus, imbalance of synaptic excitation and inhibition of dysmorphic neurons may contribute to seizure genesis in type II FCD.
Animals ; Drug Resistant Epilepsy/surgery* ; Epilepsy/pathology* ; Malformations of Cortical Development/pathology* ; Malformations of Cortical Development, Group I ; Mice ; Neurons/pathology* ; Seizures/pathology*

Objective:To evaluate the application of intraoperative optical coherence tomography (iOCT) in vitreoretinal surgery.Methods:An observational case series study was conducted.A total of 316 patients (316 eyes) who received vitreoretinal surgery and iOCT from January 2015 to December 2017 in Hangzhou Branch of Eye Hospital, Wenzhou Medical University were enrolled.The iOCT scanning time and result, the consistency between iOCT result and observation under the operating microscope as well as the influence of iOCT on surgical strategy were recorded.The postoperative adverse reactions and complications were observed.The study protocol was approved by an Ethics Committee of Eye Hospital, Wenzhou Medical University(No.2019-168-K-160). Written informed consent was obtained from all patients prior to any medical examination.Results:There were 85.8% (271/316) of patients who successfully completed iOCT scanning, with an average scanning time of (3.54±2.30)minutes.Among the 271 successful eyes, there were 51 with idiopathic macular hole (IMH), 95 with epiretinal membrane, 50 with dense vitreous hemorrhage (VH), 30 with macular lamellar hole, 12 with vitreous macular traction syndrome, 33 with myopic maculopathy.There were 45 eyes, accounting for 16.6%, the iOCT results of which were inconsistent with the observation of operator under the operating microscope.There were 27 eyes, accounting for 10.0%, the surgical strategy of which was changed.The iOCT results of IMH showed that intra-retinal bridge connection appeared in 2 eyes after ILM peeling, then air tamponade was used.High-reflection strips on the edge of the MH after ILM peeling, called the Hole-door phenomenon, were found in 15 eyes, accounting for 32.6%.In MH with a diameter >400 μm, the postoperative best corrected visual acuity and continuity of outer limiting membrane were better in eyes with Hole-door phenomenon than eyes without Hole-door phenomenon.There were 8 eyes, accounting for 8.4%, showing residual membrane, 3 eyes of which received extra ERM peeling.There were 56 eyes, accounting for 58.9%, showing the secondary changes after membrane peeling.For 50 eyes with dense VH, there were 17 eyes showing the normal macular structure, accounting for 34%, and 33 eyes showing the abnormal macular structure, accounting for 66.0%.In addition, the macular structures of 14 eyes, accounting for 28.0%, observed in iOCT image were inconsistent with the intraocular microscope finding, and 11 of them received extra ERM peeling.Conclusions:The application of iOCT in vitreoretinal surgery can guide the selection of reasonable surgical methods during operation, predict postoperative recovery as well as improve postoperative outcomes.