This study aims to investigate the potential of oncolytic nanoparticles encapsulating Coxsackievirus A21 (CVA21) full-genome mRNA (CVA21@ONP) to resurrect CVA21 and induce apoptosis in host cells, as well as the antitumor immune effects of CVA21@ONP in immunocompetent tumor-bearing BALB/c mice. We used lipid nanoparticles (LNPs) to encapsulate CVA21 full-genome mRNA, thus preparing CVA21@ONP. The killing efficacy of CVA21@ONP was determined by the plaque assay and cell counting kit-8 (CCK-8), and the apoptosis in HT29 and CT26-iRFP cells was evaluated by flow cytometry. Mice were administrated with CVA21@ONP at high and low doses intratumorally, and the growth of tumors expressing infra-red fluorescent protein (iRFP) was monitored. Additionally, the types and changes of immune cells in the spleen were analyzed by flow cytometry. The results demonstrated that CVA21@ONP successfully resurrected CVA21 in both HT29 and U87MG cells. The plaque assay revealed robust killing effects of CVA21@ONP against both human and murine cell lines, and flow cytometry results showed increased early and late apoptotic cells. Notably, intratumoral detection revealed significantly down-regulated expression of iRFP in both high- and low-dose CVA21@ONP groups. Flow cytometry results further indicated that CVA21@ONP treatment effectively reduced the levels of immunosuppressive cells, including myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs), in the spleen, while enhancing T cell-dependent antitumor immune responses. These findings suggest that CVA21@ONP can replicate and survive extensively both in vitro and in vivo, activating the immune system of mice administrated with CVA21@ONP to target cells at the tumor site, thereby remodeling the tumor immune microenvironment and accelerating the suppression or even complete regression of tumors. The oncolytic performance of CVA21@ONP has been verified through intratumoral injection administration in this study, aimed at further exploring its therapeutic potential and promoting the development of the field of tumor treatment.
Animals ; Nanoparticles/chemistry* ; Mice ; Mice, Inbred BALB C ; Humans ; Apoptosis ; Oncolytic Viruses/genetics* ; Oncolytic Virotherapy/methods* ; Cell Line, Tumor ; RNA, Messenger/genetics* ; HT29 Cells

Objective To investigate the expression levels of platelet activating factor and rhythm nuclear receptor Rev-erbα in patients after ischemic stroke on different onset times.Methods A total of 158 patients after ischemic stroke admitted from June 2022 to May 2023 were divided into four groups according to the onset time T1(0-6 o'clock),T2(6-12 o'clock),T3(12-18 o'clock),and T4(18-24 o'clock).Matched to the ischemic stroke groups,126 patients hospitalized at the same time with non-ischemic stroke were divided into four groups.General data and serum samples of the subjects were collected,and the expression levels of CD62P,CD40L and Rev-erbα were detected with enzyme-linked immunosorbent assay(ELISA).Results The expressions of platelet activating factor CD62P,tumor necrosis factor CD40L,and rhythm nuclear receptor Rev-erbα in T2 group after ischemic stroke were significantly higher than that in the other groups(P＜0.05),whilst no significant difference were observed in other three groups(P＞0.05).The expression of CD62P,CD40L and rev-erbα in ischemic stroke groups were significantly higher than that in control groups.Conclusion The expression levels of the rhythm nuclear receptor Rev-erbα,CD62P,and CD40L in ischemic stroke groups is higher than that in control groups,and the expression of these cytokines is more significant in T2 group(i.e.early in the morning)than other time slot.On the other hand,the expressions of CD62P and CD40L are consistent with Rev-erbα on the time slots,indicating a clear correlation.It is indicated Rev-erbα and platelet activation play a key role on attacking of ischemic stroke.

Objective To investigate the expression levels of platelet activating factor and rhythm nuclear receptor Rev-erbα in patients after ischemic stroke on different onset times.Methods A total of 158 patients after ischemic stroke admitted from June 2022 to May 2023 were divided into four groups according to the onset time T1(0-6 o'clock),T2(6-12 o'clock),T3(12-18 o'clock),and T4(18-24 o'clock).Matched to the ischemic stroke groups,126 patients hospitalized at the same time with non-ischemic stroke were divided into four groups.General data and serum samples of the subjects were collected,and the expression levels of CD62P,CD40L and Rev-erbα were detected with enzyme-linked immunosorbent assay(ELISA).Results The expressions of platelet activating factor CD62P,tumor necrosis factor CD40L,and rhythm nuclear receptor Rev-erbα in T2 group after ischemic stroke were significantly higher than that in the other groups(P＜0.05),whilst no significant difference were observed in other three groups(P＞0.05).The expression of CD62P,CD40L and rev-erbα in ischemic stroke groups were significantly higher than that in control groups.Conclusion The expression levels of the rhythm nuclear receptor Rev-erbα,CD62P,and CD40L in ischemic stroke groups is higher than that in control groups,and the expression of these cytokines is more significant in T2 group(i.e.early in the morning)than other time slot.On the other hand,the expressions of CD62P and CD40L are consistent with Rev-erbα on the time slots,indicating a clear correlation.It is indicated Rev-erbα and platelet activation play a key role on attacking of ischemic stroke.

Objective To explore the mechanism by which puerarin alleviates the cardiotoxicity induced by doxorubicin in myocardial cells. Methods Cells in the logarithmic growth phase were divided into normal control group,model group,low-(20 mmol·L-1),medium-(40 mmol·L-1) and high-(80 mmol·L-1) dose puerarin groups,and positive control group(captopril,1 mmol·L-1). Except for the normal control group,the other groups were co-incubated with 5 mmol·L-1 doxorubicin. Cell viability was assessed using CCK-8 and lactate dehydrogenase (LDH) assays. ROS levels were detected using a ROS probe. Autophagy flux was detected by transfection with HBAD-mcherry-EGFP-LC3 adenovirus. Western Blot was used to measure the protein expression levels of Beclin-1,LC3,p62,p-AMPKα,and AMPKα. Lysosomal function was assessed using a lysosomal probe. Immunofluorescence was used to detect the relative intensity and co-localization of ASMase and LAMP1. Molecular docking analysis was performed to predict the binding capacity of PUE with ASMase. Differential gene expression was analyzed by gene set enrichment analysis. Results Compared to the normal control group,the model group showed reduced cell viability (P<0.01),increased release levels of LDH and ROS (P<0.05,P<0.01),increased number of autophagosomes (P<0.01),and decreased number of autophagic lysosomes (P<0.05). Beclin-1 protein expression and LC3-II/LC3-I ratio decreased(P<0.01),but p62 protein expression increased(P<0.01). Fluorescence intensity of lysosome decreased(P<0.01),whereas fluorescence intensity of ASMase increased(P<0.01). Immunofluorescence co-localization of ASMase and LAMP1 increased (P<0.01),the ratio of p-AMPKα/AMPKα decreased(P<0.05). Compared to the model group,the high-dose puerarin group showed a rebound in cell viability (P<0.05). The medium-and high-dose puerarin groups showed a decreasing trend in LDH level (P<0.05),and all puerarin groups showed a decreasing trend in ROS level (P<0.01). The number of autophagosomes in high-dose puerarin group reduced (P<0.01). The number of autophagic lysosomes in all puerarin groups increased (P<0.05,P<0.01). The high-dose puerarin group showed increased expression of Beclin-1 (P<0.05) and LC3-II/LC3-I ratio,and decreased p62 expression (P<0.01). All puerarin groups showed increased lysosomal fluorescence intensity (P<0.05,P<0.01). The medium-and high-dose puerarin groups showed a decrease in ASMase fluorescence intensity(P<0.05),a reduction in the immunofluorescence co-localization of ASMase with LAMP1 (P<0.01),and an increase in the p-AMPKα/AMPKα ratio (P<0.01). Molecular docking analysis discovered puerarin showed a binding energy of-8.6 kcal·mol-1 with ASMase. Gene enrichment analysis indicated that the differentially expressed genes in the doxorubicin cardiotoxicity model were related to apoptosis,autophagy,and lysosomal function. Conclusion Puerarin can alleviate doxorubicin-induced cardiotoxicity in myocardial cells and protect myocardial cells by regulating autophagy through AMPK/ASMase,as well as restoring autophagic flux.

To summarize the clinical effect of using boosters for orbital autologous fat injection transplantation.

To summarize the clinical effect of using boosters for orbital autologous fat injection transplantation.

OBJECTIVE To evaluate the effect of “Cloud Pharmacy Service System” electronic management platform in medication therapy management （MTM） of chronic airway diseases. METHODS MTM module setting of “Cloud Pharmaceutical Service System” was introduced. Totally 371 patients with chronic airway disease admitted to our hospital from January 2021 to March 2022 were selected for MTM based on the “Cloud Pharmacy Service System”. The standardization of inhalation device mastery and compliance of patients before and after intervention were compared with self-made inhalation device evaluation scale and Morisky medication adherence scale-8 （MMAS-8）. The satisfaction of patients with pharmacist after intervention was investigated. RESULTS “Cloud Pharmaceutical Service System” is mainly divided into 4 modules， such as medication therapy review， pharmacist intervention， personal medication record and the medication-related action plan， other functions. Among 371 patients， there were 237 outpatients （142 cases of asthma， 95 cases of chronic obstructive pulmonary disease） and 134 inpatients （19 cases of asthma and 115 cases of chronic obstructive pulmonary disease）. The score of the patients using inhalation device increased from 74.76±24.71 before intervention to 99.45±2.12 after intervention （P＜0.05）. MMAS-8 score increased from 7.14± 1.15 before intervention to 7.88±0.44 after intervention （P＜0.05）. The degree of patients’ satisfaction with pharmacists reached 100% after intervention. CONCLUSIONS “Cloud Pharmacy Service System” is helpful to improve the effects of pharmaceutical service for patients with chronic airway disease by providing whole-process， online， visual and immediate MTM.

Objective:To analyze the failure causes and repair methods of conjoint fascial sheath (CFS) suspension in ptosis correction.Methods:From 2021 September to September 2022, 13 cases (5 males and 8 females, aged from 19 to 49 years with an average of 25 years) were collected, and there were 18 eyes of CFS suspension in the Department of Burn and Plastic Surgery Center, the 940th Hospital of the Joint Logistic Support Force of PLA with a failed ptosis surgeries, including levator palpebrae superioris muscle shortening, CFS suspension, and frontalis muscle suspension.Results:Among the 13 patients (18 eyes), the failure causes included a result of the unfamiliar anatomy of CFS, unstable suture fixation, dysplasia of CFS, and improper suture site, CFS suspension failed to correct ptosis. After following-up for 6 to 30 months after reconstructive surgery, 14 eyes were corrected satisfactorily, 2 eyes were relieved of ptosis, and 2 eyes had ptosis recured six months after restorative surgery and a frontalis muscle suspension was used in the recured cases with a satisfactory outcome.Conclusions:According to the patient′s eye condition, mastering the indications, be familiar with the local anatomy and accurate surgical procedures are the key to success.

ObjectiveTo determine the prevalence of ticks parasitic on domestic animals in Yiwu City of Zhejiang Province. MethodsWe monitored， identified and analyzed ticks parasitic on domestic animals in Yiwu in 2021. ResultsA total of 452 domestic animals were monitored， of which 306 were infected with ticks， suggesting a prevalence of 67.70%. A total of 2，603 ticks were captured， and the tick index was determined to be 5.76. The ticks were identified to belong to one family， four genera， and 10 species. Haemaphysalis longicornis was the dominant tick （59.16%）， followed by Rhipicephalus haemaphysaloides （26.32%） and Rhipicephalus microplus （6.45%）. Prevalence of ticks， tick index， and number of tick species were listed for domestic animals as follows： goats （74.37%， 7.45， 7 species）， yellow cattle （62.50%， 4.14，6 species）， and dogs （54.62%， 2.59， 8 species）. Moreover， goat was the main domestic animal infected with more than two tick species （25.99%）； H.longicornis （91.26%） was the species with most repated infections. The dominant species of ticks were H.longicornis （65.15%） and R.haemaphysaloides （29.42%） in goats， R.microplus （67.67%） and R.haemaphysaloides （20.69%） in yellow cattle， and H.longicornis （57.14%） and Haemaphysalis porcupines （21.10%） in dogs. However， seasonal fluctuation in tick index differed among goats， yellow cattle and dogs. The fluctuation in tick index in goats and dogs was consistent with that of H.longicornis， with a major peak in March and the minor peaks in September and October. The fluctuation in tick index in yellow cattle was consistent with that of R.microplus， with two peaks in April and August-September. In addition， larval ticks， nymphal ticks and adult ticks were caught in this study， among which nymphal ticks were most common. We found that single domestic animals were infected with ticks of the same species and different stages， which was more common in goats and those animals infected with H.longicornis. ConclusionTicks parasitic on domestic animals are common in Yiwu. Multiple species of ticks have been identified， with H.longicornis as the dominant species. It warrants further monitoring and countermeasures.

Objective:To explore the importance of managing retro-orbicularis oculi fat (ROOF) in double-fold eyelid operation for patients with swollen upper-eyelid.Methods:From January 2018 to December 2020, 76 cases of swollen upper-eyelid were treated with double-fold eyelid operation by PARK method. In operation, an oval shape ROOF was removed from the outside-to the inside, and then appropriate lateral intraorbital fat was also removed. If fleshy orbicularis oculi muscle and anterior tarsus fat affected appearance of double-fold eyelid, both of them might be removed properly.Results:All patients were followed up from seven days to two years, two cases presented with shallowed double-fold eyelid, 5 cases of lacrimal gland prolapse had good lacrimal gland reduction and no recurrence; 4 cases of ptosis had normal corneal exposure and natural appearance after correction; the swollen upper-eyelid was obviously improved in 74 cases, and restoration stage was short, the apperaence and radian of doule-fold eyelid were beautiful, local scar was slight, satisfactory operative effect was achieved.Conclusions:In double-fold eyelid operation by PARK method for patients with swollen upper-eyelid, an ideal effect would be achieved, if transconjunctival route and fleshy orbicularis oculi muscle are appropriately removed in turn according to the reasons to give rise to swollen upper-eyelid.