IntroductionRotavirus remains a major cause of mortality among young children worldwide, underscoring the urgent need for new vaccines. The present study evaluated the repeated-dose toxicity and immunogenicity of a bivalent rotavirus subunit candidate vaccine PP-P[6]/P[8]-VP8 Mix (containing equal amounts of P[6]-VP8 and P[8]-VP8 chimeric antigens) based on norovirus P particles in cynomolgus monkeys.MethodsThe monkeys (5 per sex per group) were randomly assigned to a blank control group, an adjuvant control group, and low- and high-dose vaccine groups (60 μg/monkey and 120 μg/monkey). The animals received four intramuscular injections at 4-week intervals, followed by a 6-week recovery period. General behavior, body temperature, body weight, food intake, hematology, serum biochemistry, ophthalmic examination, electrocardiogram, urine analysis, immunogenicity parameters, and pathological examination of the animals were detected.ResultsNo toxicologically significant abnormalities were observed throughout the study. Histopathological examination showed that there were deposition foci of aluminum hydroxide adjuvant and mild subacute inflammatory reactions at the injection site in the vaccine group. Lymphoid follicle hyperplasia and lymph sinus histiocytosis were observed in the draining lymph nodes, which are typical local immune response manifestations of aluminum adjuvant vaccines. No other adverse pathological changes were found.ConclusionsOur regulatory toxicology study has demonstrated the safety and immunogenicity of PP-P[6]/P[8] - VP8 Mix under the experimental conditions, supporting its entry into clinical research.

Ethnopharmacological relevance Suhexiang pills (SHXP), a classic “orifice-opening formula” in traditional Chinese medicine, comprise 15 herbal ingredients and possess aromatic properties that open orifices, regulate qi, and alleviate pain. It has been approved by China's National Medical Products Administration for the treatment of phlegm-induced coma caused by obstruction of the heart orifice and hemiplegia following stroke. Aim of the study The present study aimed to evaluate the potential therapeutic effects of SHXP on cerebral ischemia-reperfusion injury in rats using a transient middle cerebral artery occlusion (tMCAO) model. Additionally, adult Sprague-Dawley (SD) rats were intragastrically administered SHXP for 91 days to assess potential toxicities. Materials and methods In a tMCAO model, the neuroprotective effects of SHXP were assessed. Male rats received SHXP (320, 640, and 1280 mg/kg/day) starting 7 days before ischemia and continuing until 48 h post-ischemia. Infarct volume was then measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Concurrently, a 91-day repeated-dose toxicity experiment of SHXP was conducted in rats. Rats were orally administered SHXP (0.6, 1.5, and 4.5 g/kg/day), and the assessment included in vivo observations (survival, body weight, food consumption) as well as hematological, biochemical, urinalysis, and histopathological analyses. Results The results indicated that SHXP exerted therapeutic efficacy against ischemic stroke. Additionally, no significant systemic toxicity was observed after 91 days of repeated SHXP administration. The data established the no-observed-adverse-effect level (NOAEL) of SHXP at 4.5 g/kg. Conclusions This study demonstrates that SHXP is an effective therapeutic agent for ischemic stroke with a favorable safety profile for long-term use.

Doxorubicin (DOX)-induced cardiotoxicity has become a major concern and is considered a limitation for the use of DOX in oncology treatment. Ginsenoside Rh2 (Rh2) is a ginseng extract with anti-inflammatory, antioxidant and cell cycle regulating activities. The aim of this study was to investigate the mechanism of cardioprotective effects of Rh2 in DOX-induced cardiotoxicity. This study utilized network pharmacology to search for potential targets and pathways of Rh2 against doxorubicin-induced heart failure. The mechanism of Rh2 protection of myocardial tissue was further examined using a doxorubicin-formed rat model of heart failure. Network pharmacology predicted 128 potential targets for Rh2 treating to heart failure. Autophagy and apoptosis pathways play critical roles in Rh2 treatment of heart failure accessed by GO and KEGG enrichment analysis. Animal experiment results showed that Rh2 attenuated DOX-induced cardiotoxicity, normalized the morphology of cardiac tissue and reduced cardiomyocyte autophagy as well as apoptosis by up-regulation of the PI3K-AKT-mTOR signaling pathway to antagonize the effect of DOX on cardiomyocyte damage. These results suggest that Rh2 was able to inhibit DOX-activated autophagy signaling and apoptotic pathways in myocardial tissues and reduced cardiomyocyte apoptosis. It has potential effects to protect myocardial tissue as well as antagonize DOX-induced cardiotoxicity.

BackgroundStem cell therapy shows promise for treating skin diseases and enhancing medical aesthetics. However, safety data for subcutaneous injection of stem cells remain limited. In this study, we evaluated the toxicity of human umbilical cord mesenchymal stem cells (hUC-MSCs) in NOD. Cg-PrkdcscidIL2rgtm1Sug/JicCrl (NOG) mice.MethodsMice received subcutaneous hUC-MSC injections at doses of 2.5 × 107 and 2.0 × 108 cells/kg on days 1, 8, 12, 16, and 20, followed by withdrawal and observation for 6 weeks. Toxicity was assessed through clinical observation, behavioral analysis, pathology, organ weight measurements, and histopathology. hUC-MSC distribution was determined via validated quantitative (q)PCR and colonization was assessed using immunohistochemistry.ResultsNo abnormal effects on clinical responses, body weight, or food intake were observed following five repeated hUC-MSCs administrations, except for masses at the administration site in the high-dose group. Mouse activity levels increased in both dose groups 6 h post-final injection. Foamy cells were observed under the pleural membrane in high-dose mice. hUC-MSCs primarily colonized and were distributed within skin tissues 24 h after the last administration.ConclusionThe no-observed-adverse-effect level for subcutaneous hUC-MSC administration in NOG mice over 3 weeks was 2.5 × 107 cells/kg. Our results will help in advancing the clinical use of hUC-MSCs, particularly for treating conditions such as atopic dermatitis.

Human umbilical cord mesenchymal stem cells (hUC-MSCs) are proposed for the treatment of acute lung injury and atopic dermatitis. To advance hUC-MSC entry into clinical trials, the effects of hUC-MSCs on the general toxicity, immune perturbation and toxicokinetic study of hUC-MSCs in cynomolgus monkeys were assessed. hUC-MSCs were administered to cynomolgus monkeys by intravenous infusion of 3.0 × 106 or 3.0 × 107cells/kg or by subcutaneous injection of 3.0 × 107cells/kg twice a week for 3 weeks followed by withdrawal and observation for 6 weeks. Toxicity was assessed by clinical observation, clinical pathology, ophthalmology, immunotoxicology and histopathology. Moreover, toxicokinetic study was performed using a validated qPCR method after the first and last dose. After 3rd or 4th dosing, one or three the monkeys in the intravenous high-dose group exhibited transient coma, which was eliminated by slow-speed infusion after 5th or 6th dosing. In all dose groups, hUC-MSCs significantly increased NEUT levels and decreased LYMPH and CD3+ levels, which are related to the immunosuppressive effect of hUC-MSCs. Subcutaneous nodules and granulomatous foci were found at the site of administration in all monkeys in the subcutaneous injection group. Other than above abnormalities, no obvious systemic toxicity was observed in any group. The hUC-MSCs was detectable in blood only within 1 h after intravenous and subcutaneous administration. The present study declared the preliminary safety of hUC-MSCs, but close monitoring of hUC-MSCs for adverse effects, such as coma induced by intravenous infusion, is warranted in future clinical trials.

Peach gum is a homologous substance of medicine and food, mainly composed of polysaccharides. This study aimed to investigate the effectiveness of Peach gum polysaccharides (PGP) in treatment of ulcerative colitis (UC) and explore its mechanisms. Firstly, we found PGP could decrease the disease activity index, increase length of shortened colon and improve colonic tissue lesions of UC observably in vivo. Further, we explored its mechanisms of anti-UC from aspects of immune regulation and intestinal barrier. On one hand, we found PGP could signif-icantly alleviate inflammation in the colon, but promote the secretion of inflammatory mediators such as IL-6 and TNF-& alpha; by RAW264.7 cells in vitro. It is suggested that PGP does not have direct anti-inflammatory effect. On other hand, we found PGP could increase the ki67 protein to promote the proliferation of colonic epithelial cells in vivo and protect the monolayer barrier function formed by Caco-2 from LPS damage via PI3K/AKT pathway in vitro. In conclusion, PGP could promote the proliferation of colonic epithelial cells via PI3K/AKT pathway to treat UC.

Although the new coronavirus disease 2019 (COVID-19) outbreak occurred in late 2019, it is still endemic worldwide, and has become a global public health problem. Vaccination against SARS-CoV-2 is considered to be the most effective intervention to prevent the spread of COVID-19. ZF2001 is a recombinant protein vaccine based on SARS-CoV-2 receptor-binding domain (RBD) subunit which contains aluminum adjuvant. In order to advance our research on ZF2001 into clinical trial, we investigated the general toxicity and immunogenicity of ZF2001 in cynomolgus monkeys and assessed the possible target organs for vaccine-induced toxicity. In the present research, we observed no significant systemic toxicities and abnormal cardiovascular and respiratory events following four times injections of intramuscular ZF2001 in cynomolgus monkeys. Histological examination revealed recoverable inflammatory changes in quadricep muscle and adjacent lymph node at the vaccine injection site. As expected, the vaccine can produce a strongly specific binding antibody and neutralizing antibodies in cynomolgus monkeys after inoculation. Taken together, our regulatory toxicology research proves the safety and immunogenicity of the ZF2001 vaccine, supporting its entry into large scale clinical trials.

BACKGROUND:CHMP1A, a member of the ESCRT-III complex family, has been indicated as a brand-new inhibitor gene of tumors. Our previous research has revealed that CHMP1A plays a vital role in the development and progression of renal cell carcinoma (RCC).OBJECTIVE:To investigate the potential target pathway of the regulation of the tumor cell growth by CHMP1A.METHODS:The effect of CHMP1A on mTOR pathway was elucidated by western blotting. The effect of CHMP1A on the expression of p53 was evaluated, and A498 cell growth was assessed by colony formation and MTT assays. The expression of p53 was knocked down by shRNA-p53, and the effect of CHMP1A on mTOR after knockdown of p53 was evaluated. The effect of CHMP1A on apoptosis and its relationship with MDM2 pathway were detected by western blotting and FCM. Finally, the relationship between the regulation of p53 by CHMP1A and the PI3K/mTOR pathway was detected.RESULTS:This study showed that the mTOR pathway was suppressed significantly in CHMP1A-overexpressing A498 and 786-0 cells; moreover, the enhanced expression of p53 and the reduced proliferation were shown in CHMP1A-overexpressing A498 cells. Furthermore, CHMP1A was able to regulate the PI3K/PTEN/mTOR and MDM2/p53 pathways in order to suppress RCC. In addition, CHMP1A regulated Bax and Bcl-2 via MDM2/p53 to induce the apoptosis of tumor cells and upregulated the expression of p53 via the PI3K/mTOR pathway.CONCLUSIONS:The results convey that CHMP1A-related suppression of RCC is closely related to the PI3K/mTOR/p53 pathway.

Objective: To evaluate the anti-inflammatory activity of Crotalaria ferruginea extract (CFE) and its mechanism. Methods: An intratracheal lipopolysaccharide (LPS) instillation-induced acute lung injury (ALI) model was used to study the anti-inflammatory activity of CFE in vivo. The LPS-induced shock model was used to analyze the effect of CFE on survival. LPS-stimulated RAW264.7 cell model was used to investigate the anti-inflammatory activity of CFE in vitro and the effects on mitogen-activated protein kinase (MAPK) or nuclear factor-κB (NF-κB) signaling pathways. Results: CFE administration decreased the number of inflammatory cells, reduced the levels of tumor necrosis factor-α (TNF-a), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), and interferon-γ, and diminished protein content in the bronchoalveolar lavage fluid of mice. CFE also reduced lung wet-to-dry weight ratio, myeloperoxidase, and lung tissue pathological injury. CFE pre-administration improved the survival rate of mice challenged with a lethal dose of LPS. CFE reduced LPS-activated RAW264.7 cells to produce nitric oxide, TNF-α, MCP-1, and IL-6. Furthermore, CFE inhibited nuclear translocation and phosphorylation of NF-κB P65, extracellular signal-regulated kinase, c-Jun N-terminal kinases, and P38 MAPKs. Conclusions: CFE exhibits potent anti-inflammatory activity in LPS-induced ALI mice, LPS-shock mice, and RAW264.7 cells, and its mechanism may be associated with the inhibition of NF-κB and MAPK signaling pathways. Crotalaria ferruginea may be a useful therapeutic drug for the treatment of ALI and other respiratory inflammations.

Naming is a commonly impaired language domain in various types of aphasia. Emerging evidence supports the cortico-subcortical circuitry subserving naming processing, although neurovascular regulation of the non-dominant thalamic and basal ganglia subregions underlying post-stroke naming difficulty remains unclear. Data from 25 subacute stroke patients and 26 age-, sex-, and education-matched healthy volunteers were analyzed. Region-of-interest-wise functional connectivity (FC) was calculated to measure the strength of cortico-subcortical connections. Cerebral blood flow (CBF) was determined to reflect perfusion levels. Correlation and mediation analyses were performed to identify the relationship between cortico-subcortical connectivity, regional cerebral perfusion, and naming performance. We observed increased right-hemispheric subcortical connectivity in patients. FC between the right posterior superior temporal sulcus (pSTS) and lateral/medial prefrontal thalamus (lPFtha/mPFtha) exhibited significantly negative correlations with total naming score. Trend-level increased CBF in subcortical nuclei, including that in the right lPFtha, and significant negative correlations between naming and regional perfusion of the right lPFtha were observed. The relationship between CBF in the right lPFtha and naming was fully mediated by the lPFtha-pSTS connectivity in the non-dominant hemisphere. Our findings suggest that perfusion changes in the right thalamic subregions affect naming performance through thalamo-cortical circuits in post-stroke aphasia. This study highlights the neurovascular pathophysiology of the non-dominant hemisphere and demonstrates thalamic involvement in naming after stroke.

目的 探究天台乌药对TNBS诱导的溃疡性结肠炎模型大鼠的抗炎作用.方法 36只雄性SD大鼠随机分为正常对照组、模型对照组、阳性对照组、乌药醇提物低、中、高剂量组.采用2,4,6-三硝基苯磺酸(TNBS)50％乙醇溶液灌肠建立UC大鼠模型.造模同时开始灌胃给予天台乌药醇提物干预9 d,实验期间通过观测大鼠体重、便血等疾病症状表现及结肠组织病变程度等指标评价乌药给药后的药效作用;通过检测大鼠结肠组织髓过氧化物酶(MPO)活性、血清中IL-6、TNF-α含量及外周血中Treg等淋巴细胞亚群比例等指标初步探讨其可能机制.结果 与模型对照组比较,天台乌药各剂量组大鼠疾病症状及结肠组织病变均有所减轻;外周血中Treg占辅助性T细胞比例均有所升高;结直肠重量及单位长度重量显著降低(P<0.05).天台乌药中剂量组结直肠长度显著增加(P<0.05).天台乌药各剂量组大鼠血清中IL-6含量均显著降低(P<0.01);天台乌药低、中剂量组大鼠血清中TNF-α 含量及结肠组织中MPO活性均显著降低(P<0.05).结论 天台乌药能够改善TNBS诱导的UC模型大鼠的疾病症状及组织病理学改变,具有良好的抗UC作用.乌药的抗UC作用可能与其降低IL-6等炎性细胞因子含量有关,具体机制仍有待进一步研究.