The rapid emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants that evade immunity elicited by vaccination has posed a global challenge to the control of the coronavirus disease 2019 (COVID-19) pandemic. Therefore, developing countermeasures that broadly protect against SARS-CoV-2 and related sarbecoviruses is essential. Herein, we have developed a lipid nanoparticle (LNP)-encapsulated mRNA (mRNA-LNP) encoding the full-length Spike (S) glycoprotein of SARS-CoV-2 (termed RG001), which confers complete protection in a non-human primate model. Intramuscular immunization of two doses of RG001 in Rhesus monkey elicited robust neutralizing antibodies and cellular response against SARS-CoV-2 variants, resulting in significantly protected SARS-CoV-2-infected animals from acute lung lesions and complete inhibition of viral replication in all animals immunized with low or high doses of RG001. More importantly, the third dose of RG001 vaccination elicited effective neutralizing antibodies against current epidemic XBB and JN.1 strains and similar cellular response against SARS-CoV-2 Omicron variants (BA.1, XBB.1.16, and JN.1) were observed in immunized mice. All these results together strongly support the great potential of RG001 in preventing the infection of SARS-CoV-2 variants of concern (VOCs).

Objective:To explore the effect of exercise on the memory of rats modeling vascular cognitive impairment (VCI) and also its effects on the hippocampal Sema3G/neuropilin-2 (Nrp2)/PlexinA4 signaling pathway.Methods:Eighteen male Sprague-Dawley rats were randomly divided into a sham-operated group, a model group, and an exercise group, each of 6. The model and exercise groups underwent VCI modeling via bilateral common carotid artery occlusion, while the sham-operated group received the same surgical procedure without vessel ligation or transection. Beginning forty-eight hours after the surgery, the exercise group carried out daily 30-minute treadmill training sessions, 5 days a week, for a total of 4 weeks, while the other two groups were placed on the same treadmill with it not in operation. After the intervention, cognitive functioning was assessed using the novel object recognition (NOR) test and a Y-maze test. Western blotting was employed to evaluate the expression of Sema3G, Nrp2, PlexinA4, and Rac1 in the hippocampus. Immunofluorescence staining was used to observe the distribution of Nrp2 and PlexinA4 in the hippocampus.Results:Compared with the model group, the exercise group exhibited significantly higher NOR indices during both the short-term and long-term memory testing phases after the intervention. Those rats also tended to have significantly longer total exploration times in the novel arm of the Y-maze test. The western blotting revealed that the expression levels of Sema3G, PlexinA4, and Rac1 in the hippocampus were significantly higher in the exercise group compared to the model group, on average. Immunofluorescence showed significantly increased PlexinA4 fluorescence intensity in the CA1, CA3, and dentate gyrus (DG) regions of the hippocampus, and significantly elevated Nrp2 fluorescence intensity in the CA3 region in the exercise group compared to the model group. The Pearson correlation coefficients for Nrp2/PlexinA4 co-localization in the CA1, CA3 and DG regions were significantly higher in the exercise group than in the model group.Conclusions:Exercise training significantly improves memory function in rats with VCI, and this effect may be associated with activation of the hippocampal Sema3G/Nrp2/PlexinA4 signaling pathway.

OBJECTIVES: To explore the mechanism through which moxibustion promotes endometrial repair in rats with in thin endometrium (TE). METHODS: Female SD rats were randomized into control group, 95% anhydrous ethanol-induced TE model group and moxibustion (at "Guan Yuan") group. High-throughput sequencing was used to identify the target genes of TE, and the targeting relationship between miR-223-3p and NLRP3 was verified using a dual luciferase assay. Histopathological of rat uterus was observed with HE staining, and expressions of miR-223-3p and NLRP3 were detected using RT-qPCR; serum levels of IL-1β and IL-18 of the rats were detected using ELISA, and protein expressions of NLRP3, ASC, caspase-1 and GSDMD in the uterus were detected with Western blotting. The pregnancies of the rats after treatment were counted. RESULTS: Enrichment analysis of the differential genes suggested up-regulated inflammatory response in TE, and dual luciferase assay verified targeted inhibition of NLRP3 expression by miR-223-3p. The rat models of TE had significantly decreased endometrial thickness and reduced endometrial glands and blood vessels with enhanced mRNA expression of NLRP3, increased serum levels of IL-1β and IL-18, up-regulated protein expressions of NLRP3, ASC, caspase-1 and GSDMD, lowered pregnancy rates on both the affected and unaffected sides and the overall number of pregnancies. Treatment of the rat models with mo-xibustion obviously increased the endometrial thickness and the density of glands and blood vessels, up-regulated miR-223-3p expression, lowered serum IL-1β and IL-18 levels and the protein expressions of NLRP3, ASC, caspase-1 and GSDMD, and significantly increased the number of pregnancies. CONCLUSIONS Moxibustion at "Guan Yuan" acupoint up-regulates the expression of miR-223-3p, which results in targeted inhibition of NLRP3 to suppress pyroptosis and promote endometrial repair in rat models of TE.
Animals ; Female ; MicroRNAs/genetics* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Endometrium/pathology* ; Rats, Sprague-Dawley ; Rats ; Moxibustion ; Pyroptosis ; Up-Regulation ; Interleukin-1beta/metabolism* ; Interleukin-18 ; Caspase 1/metabolism*

Objective:To explore the effect of exercise on the memory of rats modeling vascular cognitive impairment (VCI) and also its effects on the hippocampal Sema3G/neuropilin-2 (Nrp2)/PlexinA4 signaling pathway.Methods:Eighteen male Sprague-Dawley rats were randomly divided into a sham-operated group, a model group, and an exercise group, each of 6. The model and exercise groups underwent VCI modeling via bilateral common carotid artery occlusion, while the sham-operated group received the same surgical procedure without vessel ligation or transection. Beginning forty-eight hours after the surgery, the exercise group carried out daily 30-minute treadmill training sessions, 5 days a week, for a total of 4 weeks, while the other two groups were placed on the same treadmill with it not in operation. After the intervention, cognitive functioning was assessed using the novel object recognition (NOR) test and a Y-maze test. Western blotting was employed to evaluate the expression of Sema3G, Nrp2, PlexinA4, and Rac1 in the hippocampus. Immunofluorescence staining was used to observe the distribution of Nrp2 and PlexinA4 in the hippocampus.Results:Compared with the model group, the exercise group exhibited significantly higher NOR indices during both the short-term and long-term memory testing phases after the intervention. Those rats also tended to have significantly longer total exploration times in the novel arm of the Y-maze test. The western blotting revealed that the expression levels of Sema3G, PlexinA4, and Rac1 in the hippocampus were significantly higher in the exercise group compared to the model group, on average. Immunofluorescence showed significantly increased PlexinA4 fluorescence intensity in the CA1, CA3, and dentate gyrus (DG) regions of the hippocampus, and significantly elevated Nrp2 fluorescence intensity in the CA3 region in the exercise group compared to the model group. The Pearson correlation coefficients for Nrp2/PlexinA4 co-localization in the CA1, CA3 and DG regions were significantly higher in the exercise group than in the model group.Conclusions:Exercise training significantly improves memory function in rats with VCI, and this effect may be associated with activation of the hippocampal Sema3G/Nrp2/PlexinA4 signaling pathway.

Objective:To evaluate the wound healing of recipient and donor sites following keratinized mucosa augmentation(KMA)around implants in reconstructed jaw areas and to compare these outcomes with gingival grafts in native jawbone,so as to provide clinical guidance for postoperative maintenance,and to investigate the impact of clinical experience on the evaluation of KMA postoperative healing through subgroup comparisons.Methods:This study included patients who underwent resection of maxil-lofacial tumors,fibular or iliac flap reconstruction,and implant placement at Peking University Dental Hospital from October 2020 to April 2023.Three months post-implant placement,the patients were re-ferred for KMA procedures.Clinical photographs of the reconstructed area were taken preoperatively,im-mediately postoperatively,and 3 weeks and 3 months post-surgery.Additionally,photographs of the pala-tal donor site were obtained preoperatively and 3 weeks later.Wound healing was assessed by four junior and three senior clinicians utilizing the early healing index(EHI),early wound healing score(EHS),and pink esthetic score(PES).And senior clinicians evaluated the healing effect compared with gingival transplantation on natural jawbone using a 10-point scale.Results:A total of 26 patients with jawbone reconstruction were included,with an average age of(34.2±10.2)years,11 males(42.3％)and 15 females(57.7％).Among them,13 cases(50.0％)underwent fibula flap reconstruction,and 13 cases(50.0％)underwent iliac flap reconstruction.The average number of implants per patient was 3.2±0.7.In the recipient area,3 weeks postoperatively,the EHS was 7.0(4.0,9.0),with sub-item scores as follows:Clinical signs of re-epithelialization(CSR)6.0(3.0,6.0),clinical signs of haemostasis(CSH)1.5(1.0,2.0),and clinical signs of inflammation(CSI)1.0(0.0,1.0),indicating that the average appearance of the wound in the recipient area was characterized by generally well-approximated wound edges with minimal fibrin lines and mild erythema and swelling.The EHI for the recipient area was 2.0(1.5,2.5),suggesting that the incision was mostly closed with some fibrin lines 3 weeks post-operatively.The long-term healing evaluation system,PES,was 2.5(2.0,3.0),with sub-scores for color[1.0(1.0,1.5)]and texture[1.5(1.0,2.0)],which were slightly different from the reference values.In the palatal donor area,3 weeks postoperatively,the EHI score was lower at 1.3(1.0,2.5),while the EHS score was higher at 8.5(6.0,10.0),indicating better soft tissue healing in the donor area compared with the recipient area.Among the clinicians with different levels of experience,the assessment of wound healing revealed that except for the CSI sub-item,where the junior group scored higher than the senior group,all other sub-items showed significantly higher scores in the senior group compared with the junior group.In the EHS evaluation system,the CSH sub-item demonstrated no significant differences between the groups with varying levels of experience.Experienced clinicians'evaluation outcomes of healing effect compared with gum graft on natural alveolar bone was 8.5(7.5,9.5),showing high consistency[intraclass correlation coefficient(ICC):0.892;95％confidence interval(CI):0.791-0.949],suggesting slightly suboptimal healing results after KMA surgery.Conclusion:The healing process following KMA in the context of jawbone reconstruction is relatively protracted,em-phasizing the necessity for comprehensive postoperative management.Moreover,clinician experience plays a significant role in the assessment of wound healing outcomes for KMA in maxillofacial reconstruction.

Objective:To analyze the histopathological characteristics of peri-implant soft tissue in re-constructed jaws and the changes after keratinized mucosa augmentation(KM A)with free gingival graft(FGG).Methods:Twenty patients were enrolled in this study.Five patients of them,who were perio-dontal and systemic healthy and referred for crown lengthening before restoration with healthy keratinized gingiva collected were enrolled as healthy controls.15 patients of them were with fibula or iliac bone flaps jaw reconstruction(10 with fibula flap and 5 with iliac flap),who were referred to FGG and implant exposures before restoration.Soft tissue was collected before FGG in reconstructed jaws,and in 5 patients(3 with fibula flap and 2 with iliac flap)8 weeks after FGG if a second surgery was conducted.Histologi-cal analysis with hematoxylin-eosin stain and immunological analysis to interlukin-1(IL-1),interlukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were performed.Results:Thickness from the bottom of stratum basale to the top of stratum granulosum and thickness of keratinized layer in reconstructed jaws were significantly lower compared with that of natural healthy keratinized gingiva[0.27(0.20,0.30)mmvs.0.36(0.35,0.47)mm,P＜0.05;16.49(14.90,23.37)μm vs.26.37(24.12,31.53)μm,P＜0.05].In the reconstructed area,thickness from the bottom of stratum basale to the top of stra-tum granulosum increased after KMA with FGG[0.19(0.16,0.25)mm vs.0.38(0.25,0.39)mm,P=0.059]and the thickness of keratinized layer significantly increased after KMA with FGG[16.42(14.16,22.35)μm vs.28.57(27.16,29.14)μm,P＜0.05],which was similar to that in the con-trol group.Furthermore,the number of positive cells of IL-1,IL-6 and TNF-α significantly increased after KMA[0.67(0.17,8.93)vs.11.00(9.16,18.00);13.00(8.50,14.14)vs.21.89(15.00,28.12);0.22(0.04,0.63)vs.2.83(1.68,5.00),respectively,P＜0.05]as well as the average optical density value[0.15(0.14,0.17)vs.0.18(0.17,0.21);0.28(0.26,0.33)vs.0.36(0.33,0.37);0.23(0.22,0.29)vs.0.30(0.28,0.42),respectively,P＜0.05],which was similar to that in the healthy keratinized gingiva.Conclusion:The lack of rete pegs and inflammato-ry factors were common in soft tissue with jaw reconstruction.FGG can improve the quality of the epithe-lium and may improve the stability of the mucosa around implants.

Objective To compare the reparative effects of moxibustion at"Guanyuan"and"Shenshu"points in rats with a thin endometrium.Methods Thirty-two female Sprague-Dawley rats were divided randomly into control(CON),model(MOD),Guanyuan(GY),and Shenshu(SS)groups.A thin endometrium model was established using 95％anhydrous alcohol.The Guanyuan and Shenshu groups underwent moxibustion at the Guanyuan and Shenshu points,respectively,and the other two groups of rats were fixed in the same way.Temperature changes at the acupoint area were measured at different time points.The pathological morphology of the uterus was observed by hematoxylin and eosin(HE)staining,and serum levels of inflammatory factors and hormone levels were detected by enzyme-linked immunosorbent assay(ELISA).Expression levels of proliferation and endometrial receptivity factors were detected by immunohistochemistry(IHC),and angiogenesis factors were detected by Western blot.Results The temperatures immediately after moxibustion and after 5 min were higher at the Shenshu compared with the Guanyuan acupoint(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were decreased in the MOD group compared with the CON group(P＜0.01),serum interleukin 1 β(IL-1 β)was increased(P＜0.01)and PROG was decreased(P＜0.01),and Ki67,proliferating cell nuclear antigen(PCNA),leukemia inhibitory factor(LIF),integrin β3(ITG-β3),angiopoietin-1(ANG-1),and CD34 protein in uterine tissue were all decreased(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were increased in the GY and SS groups compared with the MOD group(P＜0.01),while serum IL-1 β was decreased(P＜0.05),E2 and PROG were increased(P＜0.05),and serum interleukin 10(IL-10)was increased in the SS group(P＜0.05).Ki67,PCNA,LIF,and ITG-β3 were all increased in both treatment groups compared with the MOD group(P＜0.01).PCNA was higher in the GY group compared with the SS group(P＜0.05),while LIF and ITG-β3 were higher in the SS group compared with the GY group(P＜0.01,P＜0.05,respectively).CD34 and VEGFA levels were increased in the two treatment groups compared with the model group(P＜0.01).ANG-1 was higher in the GY group than that in the SS group(P＜0.01),while CD34 was higher in the SS group than that in the GY group(P＜0.05).Conclusions Moxibustion at the Guanyuan and Shenshu points could help to repair the pathological thin endometrium in rats.The GY group may be superior to the SS group for reducing the levels of serum IL-1 β,increasing E2,and promoting the expression of proliferation factors,while the SS group may be superior to the GY group for increasing the levels of PROG and promoting the expression of endometrium receptor-related factors.Both acupoints can promote the expression of angiogenesis factors,potentially acting via different pathways.

Objective To compare the reparative effects of moxibustion at"Guanyuan"and"Shenshu"points in rats with a thin endometrium.Methods Thirty-two female Sprague-Dawley rats were divided randomly into control(CON),model(MOD),Guanyuan(GY),and Shenshu(SS)groups.A thin endometrium model was established using 95％anhydrous alcohol.The Guanyuan and Shenshu groups underwent moxibustion at the Guanyuan and Shenshu points,respectively,and the other two groups of rats were fixed in the same way.Temperature changes at the acupoint area were measured at different time points.The pathological morphology of the uterus was observed by hematoxylin and eosin(HE)staining,and serum levels of inflammatory factors and hormone levels were detected by enzyme-linked immunosorbent assay(ELISA).Expression levels of proliferation and endometrial receptivity factors were detected by immunohistochemistry(IHC),and angiogenesis factors were detected by Western blot.Results The temperatures immediately after moxibustion and after 5 min were higher at the Shenshu compared with the Guanyuan acupoint(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were decreased in the MOD group compared with the CON group(P＜0.01),serum interleukin 1 β(IL-1 β)was increased(P＜0.01)and PROG was decreased(P＜0.01),and Ki67,proliferating cell nuclear antigen(PCNA),leukemia inhibitory factor(LIF),integrin β3(ITG-β3),angiopoietin-1(ANG-1),and CD34 protein in uterine tissue were all decreased(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were increased in the GY and SS groups compared with the MOD group(P＜0.01),while serum IL-1 β was decreased(P＜0.05),E2 and PROG were increased(P＜0.05),and serum interleukin 10(IL-10)was increased in the SS group(P＜0.05).Ki67,PCNA,LIF,and ITG-β3 were all increased in both treatment groups compared with the MOD group(P＜0.01).PCNA was higher in the GY group compared with the SS group(P＜0.05),while LIF and ITG-β3 were higher in the SS group compared with the GY group(P＜0.01,P＜0.05,respectively).CD34 and VEGFA levels were increased in the two treatment groups compared with the model group(P＜0.01).ANG-1 was higher in the GY group than that in the SS group(P＜0.01),while CD34 was higher in the SS group than that in the GY group(P＜0.05).Conclusions Moxibustion at the Guanyuan and Shenshu points could help to repair the pathological thin endometrium in rats.The GY group may be superior to the SS group for reducing the levels of serum IL-1 β,increasing E2,and promoting the expression of proliferation factors,while the SS group may be superior to the GY group for increasing the levels of PROG and promoting the expression of endometrium receptor-related factors.Both acupoints can promote the expression of angiogenesis factors,potentially acting via different pathways.

Objective:A mouse model of pancreatitis induced by coxsackievirus B3 (CVB3) was established. The pathological change of pancreas and the infiltration of Th17/Treg cells were observed.Methods:The BALB/c mice were inoculated intraperitoneally with CVB3 to induce acute viral pancreatitis model. Then the pathological changes of pancreas were observed by HE staining; the viral RNA load and relative expression of cytokines (IFN-γ, IL-6 and IL-17) mRNA were detected by q-PCR; the proportion of infiltrated CD45 + CD3 + T cells, CD4 + and CD8 + T cells, Th17 and Treg cells in the pancreas was determined by flow cytometry. Results:Three days after CVB3 infection, the viral RNA load in pancreas was the highest (0.96±0.18) and gradually decreased with prolongation of infection. Compared with the 3 dpi group, the viral RNA load in pancreas was decreased (0.96±0.18 vs. 0.62±0.14) at 7 dpi, but there was no statistically significant difference. In addition, the infiltration of immune cell in pancreas increased significantly after 7dpi and the pathological score >2. The percent of infiltrated Th17 cells (1.05±0.21 vs. 22.13±5.79) and Treg cells (3.11±0.78 vs. 8.25±1.30) among CD4 + T cells significantly increased after infection (P<0.05), and the Th17/Treg also increased (P<0.01). Compared with the control group, the relative mRNA expression of IFN-γ (1.05±0.23 vs. 672.6±47.67), IL-6 (1.00±0.38 vs. 68.28±4.57), and IL-17 (1.01±0.11 vs. 54.15±7.94) in pancreas increased at 7 days after CVB3 infection ( P<0.01). Conclusions:The infiltration of Th17/Treg cells and the expression of related cytokines related cytokines IL-6 and IL-17 mRNA were upregulated in pancreas, which promoted the process of CVB3-induced pancreatitis.