Smart hydrothermally responsive microneedle for topical tumor treatment.

Locoregional therapy has attracted increasing attention for subcutaneous tumors owing to its merits of minimally invasive operation and negligible systematic toxicity. However, to accelerate clinical translation, further promotions in deep-seated penetration, temporal-spatial controllability, personalized adaptability, as well as easy operation are still urgently needed. This work proposed a self-heating-cooking-package-inspired hydrothermally responsive multi-round acturable microneedle (HRMAM) system, which loaded docetaxel (DTX) in the polycaprolactone (PCL), to serve as deeply penetrable, hydrothermal-chemotherapy synergetic, on-demand and self-service anti-tumor treatment. The optimized hydrothermally responsive formulation served as base components for water-based self-heating responsive drug release with synergistic anti-tumor thermal therapy, and simultaneously in combination with well-designed grooved-structure needle tips for directional deep delivery and enhanced transfer efficiency. To satisfy spatial precision and flexible controllability, this engineering-based detachable HRMAM system was divided into three relatively independent segments, which were fitted perfectly with an original-designed applicator for ensuring easy operation in a smart self-service manner. Impressively, the HRMAM system achieved encouraging tumor growth inhibition of 75.11% and 72.29% in animal model of melanomas and breast carcinoma, respectively, much higher than those of other groups receiving traditional treatment, without obvious side effects. It was anticipated that the HRMAM system would manifest great promise to combat unreachable and deep-seated subcutaneous tumors, bellowing clinical values upon locoregional therapy products with high efficiency, low toxicity, flexible controllability, temporal-spatial precision, easy operation, as well as patient's painless, comfort and compliance.

Microneedle Device Delivering Aggregation-Induced Emission Photosensitizers for Enhanced Metronomic Photodynamic Therapy of Cancer.

Metronomic photodynamic therapy (mPDT), which induces cancer cell death by prolonged intermittent continuous irradiation at lower light power, has profoundly promising applications. However, the photobleaching sensitivity of the photosensitizer (PS) and the difficulty of delivery pose barriers to the clinical application of mPDT. Here, we constructed a microneedle-based device (Microneedles@AIE PSs) that combined with aggregation-induced emission (AIE) PSs to achieve enhanced mPDT for cancer. Due to the strong anti-photobleaching property of the AIE PS, it can maintain superior photosensitivity even after long-time light exposure. The delivery of the AIE PS to the tumor through a microneedle device allows for greater uniformity and depth. This Microneedles@AIE PSs-based mPDT (M-mPDT) offers better treatment outcomes and easier access, and combining M-mPDT with surgery or immunotherapy can also significantly improve the effectiveness of these clinical therapies. In conclusion, M-mPDT offers a promising strategy for the clinical application of PDT due to its better efficacy and convenience.

Exploring the mechanism of Xiaoqinglong decoction in the treatment of infantile asthma based on network pharmacology and molecular docking.

To explore the mechanism of Xiaoqinglong decoction (XQLD) in the treatment of infantile asthma (IA) based on network pharmacology and molecular docking. The active ingredients of fdrugs in XQLD were retrieved from Traditional Chinese Medicine Systems Pharmacology database and then the targets of drug ingredients were screened. The disease targets of IA were obtained from OMIM and Gencards databases, and the intersection targets of XQLD in the treatment of IA were obtained by Venny 2.1 mapping of ingredient targets and disease targets. Cytoscape software was used to construct active ingredient-intersection target network. The potential targets of XQLD in the treatment of IA were analyzed by protein-protein interaction network using STRING platform, and the Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were obtained by R Studio software. AutoDock was used to perform molecular docking for verification. In this study, 150 active ingredients of XQLD were obtained, including quercetin, kaempferol, ß-sitosterol, luteolin, stigmasterol, and so on. And 92 intersection targets of drugs and diseases were obtained, including interleukin 6 (IL6), cystatin 3, estrogen receptor 1, hypoxia inducible factor 1A, HSP90AA1, epidermal growth factor receptor and so on. There were 127 items of Gene Ontology enrichment analysis and 125 Kyoto Encyclopedia of Genes and Genomes enrichment results, showing that apoptosis, IL-17 signaling pathway, tumor necrosis factor signaling pathway, P13K-Akt signaling pathway and other pathways may play a key role in the treatment of IA by XQLD. The results of molecular docking showed that the key active ingredients including quercetin, kaempferol, ß-sitosterol, luteolin, stigmasterol, and the core targets including IL6, cystatin 3, estrogen receptor 1, hypoxia inducible factor 1A, HSP90AA1, and epidermal growth factor receptor had good binding activity. Through network pharmacology and molecular docking, the potential targets and modern biological mechanisms of XQLD in the treatment of IA were preliminarily revealed in the study, which will provide reference for subsequent animal experiments and clinical trials.

Development and validation of a nomogram for risk prediction of nephrolithiasis recurrence in patients with primary hyperparathyroidism.

Background: Nephrolithiasis is a common complication of primary hyperparathyroidism (PHPT), and the recurrence of nephrolithiasis in patients with PHPT is also an urgent concern. What is worse, there is a scarcity of recommended evaluation to predict the risk of nephrolithiasis recurrence in patients with PHPT. This study was aimed to develop and validate a nomogram to facilitate risk assessment in patients with PHPT. Methods: A total of 197 patients with PHPT were retrospectively included in this study from September 2016 to August 2021. Patients' demographic data, blood test parameters, urinalysis, stone parameters, and surgical intervention were collected. Extracted variables were submitted to a least absolute shrinkage and selection operator (LASSO) regression model. A nomogram was built and validated according to the area under the curve (AUC) value, calibration curve, and decision curve analysis. Results: According to the LASSO regression and logistic regression analyses, five predictors were derived from 22 variables: creatinine, uric acid, bilateral stone, multiplicity, and surgery. The AUC and concordance index of the training cohort and validation cohort were 0.829 and 0.856, and 0.827 and 0.877, respectively. The calibration curve analysis and the decision curve analysis showed that the nomogram had an adequate prediction accuracy. Conclusion: We built a useful nomogram model to predict the risk of nephrolithiasis recurrence in patients with PHPT. This would assist clinicians to provide appropriate advices and managements for these patients.

Prognostic value of SOX2, Cyclin D1, P53, and ki-67 in patients with esophageal squamous cell carcinoma.

In this study, we evaluated SOX2, Cyclin D1, p53, and ki-67 expression immunohistochemically in 117 samples of surgically resected esophageal squamous cell carcinoma (ESCC) and matched normal tumor adjacent tissues and correlated the expression with clinicopathological finding and patient survival. Lymph node metastasis was observed in 36.8% of patients, and organ metastasis was observed in 17.9%. We detected high expression of SOX2, Cyclin D1, p53, and ki-67 in 46.1%, 70.1%, 54.7%, and 32.5% of ESCC tissues, respectively. SOX2 is localized in the tumor cell nuclei, and its expression was significantly associated with N stage (p=0.034) and differentiation (p=0.003) and ki-67 expression (p=0.001), whereas increased Cyclin D1 expression was correlated with high p53 (p=0.015). With regard to survival, we found that ESCC patients with high SOX2 expression had significantly better survival time than those with low SOX2 expression (p=0.021). A multivariate Cox analysis revealed that therapy and high p53 expression and venous invasion were independent predictors of unfavorable prognosis in overall survival (p=0.039, p=0.004, and p=0.023, respectively). Furthermore, higher T stage, clinical stage (pTNM), venous invasion, and high p53 expression were independent predictors of a worse progression-free survival. Notably, co-overexpression of p53 and Cyclin D1 was significantly correlated with poor overall survival and progression-free survival (p=0.029 and p=0.0227, respectively). Therefore, SOX2 might be considered as a potential prognostic indicator and a potential target for therapeutic targets in ESCC. p53 staining and combined p53 and Cyclin D1 expression had significantly unfavorable prognostic value for patients with ESCC. These findings provide more insight into ESCC; thus, further investigations into molecular mechanisms of drug resistance are essential.

Altered expression of the mismatch repair genes in DF-1 cells infected with the avian leukosis virus subgroup A.

The absence or deficiency of DNA mismatch repair (MMR) activity results in microsatellite instability (MSI) in cancer. The avian leukosis virus (ALV) causes neoplastic disease in chickens. In this study, the status of MMR, MSI, the cell cycle and apoptosis were detected in DF-1 cells after avian leukosis virus subgroup A infection. Flow cytometry analysis results indicated that there was no significant difference in cell apoptosis between the control and infected groups. The percentage of cells in S and G2 phases were increased in the infected group. MSI and mutation of MSH2 and MLH1 gene exons were absent in DF-1 cells after infection. Levels of MSH2 and MLH1 mRNA were dramatically increased in DF-1 cells after infection. These results demonstrated that ALV RAV-1 infection may promote the expression of MSH2 and MLH1 genes rather than resulting in gene mutations. Mismatch repair functions were normal and may be have relationships with the arrest of S phase and G2 phase.

Heme oxygenase-1 enhances autophagy in podocytes as a protective mechanism against high glucose-induced apoptosis.

Injury and loss of podocytes play vital roles in diabetic nephropathy progression. Emerging evidence suggests autophagy, which is induced by multiple stressors including hyperglycemia, plays a protective role. Meanwhile, heme oxygenase-1 (HO-1) possesses powerful anti-apoptotic properties. Therefore, we investigated the impact of autophagy on podocyte apoptosis under diabetic conditions and its association with HO-1. Mouse podocytes were cultured in vitro; apoptosis was detected by flow cytometry. Transmission electron microscopy and biochemical autophagic flux assays were used to measure the autophagy markers microtubule-associated protein 1 light chain 3-II (LC3-II) and beclin-1. LC3-II and beclin-1 expression peaked 12-24h after exposing podocytes to high glucose. Inhibition of autophagy with 3-methyladenine or Beclin-1 siRNAs or Atg 5 siRNAs sensitized cells to apoptosis, suggesting autophagy is a survival mechanism. HO-1 inactivation inhibited autophagy, which aggravated podocyte injury in vitro. Hemin-induced autophagy also protected podocytes from hyperglycemia in vitro and was abrogated by HO-1 siRNA. Adenosine monophosphate-activated protein kinase phosphorylation was higher in hemin-treated and lower in HO-1 siRNA-treated podocytes. Suppression of AMPK activity reversed HO-1-mediated Beclin-1 upregulation and autophagy, indicating HO-1-mediated autophagy is AMPK dependent. These findings suggest HO-1 induction and regulation of autophagy are potential therapeutic targets for diabetic nephropathy.

Microsatellite instability in chicken lymphoma induced by gallid herpesvirus 2.

Microsatellite instability (MSI) has been found in a range of human tumors, and little is known of the links between MSI and herpesvirus. In order to investigate the relationship between MSI and Gallid herpesvirus 2 (GaHV-2)-induced lymphoma, fifteen Marek's disease (MD) lymphomas were analyzed through using 46 microsatellite markers, which were amplified by PCR from DNA specimens of lymphoma and normal muscular tissues from the same chicken. PCR products were evaluated by denaturing polyacrylamide gel electrophoresis for MSI analysis. MSI was proved in all lymphomas, at least in one locus. Thirty of the 46 microsatellite markers had microsatellite alterations. These results suggested that GaHV-2-induced lymphoma in chickens is related to MSI, and this is the first report to demonstrate that MSI is associated with the GaHV-2 induced lymphoma in chicken.

Analysis of koi herpesvirus latency in wild common carp and ornamental koi in Oregon, USA.

Koi herpesvirus (KHV) infection is associated with high mortalities in both common carp (Cyprinus carpio carpio) and koi carp (Cyprinus carpio koi) worldwide. Although acute infection has been reported in both domestic and wild common carp, the status of KHV latent infection is largely unknown in wild common carp. To investigate whether KHV latency is present in wild common carp, the distribution of KHV latent infection was investigated in two geographically distinct populations of wild common carp in Oregon, as well as in koi from an Oregon-based commercial supplier. Latent KHV infection was demonstrated in white blood cells from each of these populations. Although KHV isolated from acute infections has two distinct genetic groups, Asian and European, KHV detected in wild carp has not been genetically characterized. DNA sequences from ORF 25 to 26 that are unique between Asian and European were investigated in this study. KHV from captive koi and some wild common carp were found to have ORF-25-26 sequences similar to KHV-J (Asian), while the majority of KHV DNA detected in wild common carp has similarity to KHV-U/-I (European). In addition, DNA sequences from IL-10, and TNFR were sequenced and compared with no differences found, which suggests immune suppressor genes of KHV are conserved between KHV in wild common carp and koi, and is consistent with KHV-U, -I, -J.

[Impact of techniques for gastrointestinal tract reconstruction following gastrectomy on pancreatic ß-cell function in patients with type 2 diabetes mellitus].

OBJECTIVE: To evaluate the impact of different techniques for gastrointestinal tract reconstruction on postoperative pancreatic ß-cell function in patients with type 2 diabetes mellitus (T2DM). METHODS: Twenty-three patients with gastric cancer and T2DM were studied. Techniques for reconstruction included Billroth I (n=13) and bypass procedures(Billroth II n=4 and Roux-en-Y anastomosis n=6). Pancreatic ß-cell function was evaluated by oral glucose tolerance test (OGTT). Serum insulin was measured by electrochemiluminescence immunoassay and blood glucose by glucose oxidase method. HOMA-IR and HOMA-ß were assessed. RESULTS: T2DM remission rate was 90% (9/10) in the bypass group, and 23% (3/13) in Billroth I group (P<0.01). Glycosylated hemoglobin A1c and glycated hemoglobin HbA1 were improved significantly in patients after bypass procedures(P<0.05), but the difference in Billroth I group was not statistically significant (P>0.05). OGTT showed that fasting and post-glucose load plasma glucose at each time point were significantly lower in the bypass group compared to the Billroth I group. At 30 minutes and 60 minutes after glucose load, insulin levels and insulin release index were significantly higher in the bypass group compared to Billroth I( group, as were levels of HOMA-ß and ΔI30/ΔG30 in the bypass group(P<0.05). CONCLUSION: Gastrointestinal bypass following gastrectomy may induce resolution of T2DM and improve ß-cells function.

Immunogenicity and protective efficacy of a replication-defective infectious bronchitis virus vaccine using an adenovirus vector and administered in ovo.

In ovo vaccination remains an attractive option for a cost effective, uniform and mass application of vaccines for commercial poultry. However, the vaccines which can be delivered safely by this method are limited and there is no currently licensed embryo-safe vaccine against infectious bronchitis virus (IBV). In this study, a recombinant adenovirus expressing the S1 gene of nephropathogenic IBV (rAd-S1) was constructed and the immune responses and protective efficacy against homologous challenge were evaluated after in ovo vaccination. The results showed that the rAd-S1 led to dramatic augmentation of humoral and cellular responses in birds vaccinated in ovo followed by an intramuscular inoculation. Both IFN-gamma and IL-4 in chicken's lymphocytes were produced by this strategy. Following challenge with IBV, the chickens vaccinated with recombinant adenovirus showed fewer nephropathic lesions and less severe clinical signs as compared to those receiving wild-type adenovirus or PBS. The construction of non-replicating human adenovirus vector encoding S1 gene of IBV and its in ovo delivery demonstrated the potential of an alternative vaccination strategy against IBV.

Resveratrol attenuates renal hypertrophy in early-stage diabetes by activating AMPK.

BACKGROUND: Recent studies suggest the involvement of the adenosine monophosphate-activated serine/threonine protein kinase (AMPK) pathway in the pathogenesis of diabetic nephropathy (DN). Resveratrol, an agent that activates AMPK, may have the potential to protect against the development of DN. This study was designed to investigate the therapeutic effects of resveratrol on renal hypertrophy in early-stage diabetes and the underlying mechanisms. METHOD: Molecular and structural changes involved in the pathogenesis of DN were tested in a rat model of early-stage diabetes. Renal mesangial cells (RMCs) were cultured in media containing different concentrations of glucose with or without resveratrol. Cellular DNA synthesis was assayed by measuring (3)H-thymidine incorporation. The phosphorylation status of AMPK, eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1), and phospho- ribosomal protein S6 (S6) was analyzed by Western blot. RESULTS: Resveratrol reduced plasma creatinine and urinary albumin excretion and attenuated renal hypertrophy without affecting blood glucose levels. Moreover, resveratrol activated AMPK and inhibited phosphorylation of 4E-BP1 and S6 in diabetic rat kidneys. In vitro, resveratrol blocked high glucose-induced dephosphorylation of AMPK and phosphorylation of 4E-BP1 and S6 and strongly inhibited both the DNA synthesis and proliferation of RMCs. CONCLUSION: These findings suggest the possibility that resveratrol exerts antiproliferative, antihypertrophic effects by activating AMPK and reducing 4E-BP1 and S6 phosphorylation, thus suppressing the development and progression of DN.

Palliative gastrectomy and chemotherapy for stage IV gastric cancer.

PURPOSE: To investigate the value of palliative gastrectomy and chemotherapy in a large series of patients with stage IV gastric cancer. METHODS: A total of 389 patients were identified in survival analysis. Among which, 183 cases received palliative gastrectomy (PG) and 206 cases received unresectable operation, 184 cases received palliative chemotherapy (PC) and 205 cases did not receive chemotherapy. The survival advantages of patients, based on treatments modality, were also analyzed in patients with liver metastasis, peritoneal dissemination and lymph node metastasis. RESULTS: The 1-year, 3-year, 5-year survival rate of those patients who were treated with PG + PC were 85.7% (96/112), 32.1% (36/112), and 8.9% (10/112), which were far better than those who were not. For those patients with liver metastasis, peritoneal dissemination, and/or N3 lymph node metastasis, survival advantages were also present if treated with this multimodality approach. CONCLUSION: The survival time and palliative duration were significantly longer in patients after PG than after non-resection operations. Postoperative chemotherapy prolonged the survival time of patients after palliative surgery. PG combined with adjuvant chemotherapy may improve survival in patients with stage IV gastric cancer, even with liver metastasis, peritoneal dissemination, and lymph node metastasis.

Construction and immunogenicity of recombinant adenovirus expressing the capsid protein of porcine circovirus 2 (PCV2) in mice.

Porcine circovirus 2 (PCV2) has been implicated as the etiological agent of some diseases, mainly postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS). The capsid (Cap) protein encoded by the PCV2 ORF2 gene may be an excellent candidate for vaccination. In this study, the Cap protein gene was amplified by PCR, and cloned into the transfer vector pShuttle-CMV. After co-transformation of PmeI-linearized recombinant plasmid pShuttle-CMV-ORF2 and the bone vector pAdEasy-1 into Escherichia coli bacteria strain BJ5183, recombinant plasmid containing Cap protein gene (pAd-ORF2) was obtained and identified with PCR. Upon transfection of PacI-linearized plasmid pAd-ORF2 in 293 cell line, a recombinant adenovirus was obtained and named as rAd-Cap with viral titer of 10(13.0) TCID(50)/ml. The expression of the Cap protein in the 293 cells infected with rAd-Cap was confirmed with specific antibody to PCV2 by Western blotting and IPMA. Mice were inoculated with 10(8), 10(10) and 10(12) TCID(50)/mouse of rAd-Cap and boosted 2 weeks later, and they could generate antibody against PCV2 detected by indirect ELISA, Western blot and neutralizing activity assay. It indicated that the rAd-Cap was able to express the capsid of PCV2 and could elicit immune responses against the PCV2 in mice. The recombinant adenovirus might be an attractive candidate vaccine for preventing the disease associated with PCV2 infection.

Humoral immune response induced by oral administration of S. typhimurium containing a DNA vaccine against porcine reproductive and respiratory syndrome virus.

The ORFs-encoded major envelope glycoprotein (GP5) of porcine reproductive and respiratory syndrome virus (PRRSV) is one of the major structural proteins of this virus. In this report, we described the induction of a PRRSV GP5-specific immune response by oral vaccination of mice with eukaryotic expression vectors containing the GP5 gene of PRRSV (pcDNA3-GP5), delived by attenuated Salmonella typhimurium aroA. It demonstrated that oral administration of the transformants resulted in expression of the GP5 transcript in the intestinal epithelium. The level of serum neutralization antibodies to PRRSV was not significantly different between the mice immunized with the transformants and the naked plasmid DNA. But the neutralizing antibody titres in sera of the mice immunized with SL7207/pcDNA3-GP5' and pcDNA3-GP5' (resides 2-25 deletion mutant of GP5) were significantly lower than those immunized with the complete GP5 gene. These results show that oral inoculation of the transformants can induce humoral immune response to PRRSV. The signal peptide of the GP5 protein of PRRSV is associated with the neutralizing epitope of the protein. The attenuated S. typhimurium may be used as a delivery system for oral DNA vaccines containing PRRSV GP5 glycoprotein.

Efficacy of DNA vaccines against infectious bursal disease virus in chickens enhanced by coadministration with CpG oligodeoxynucleotide.

The objective of the present study was to investigate the feasibility of a DNA vaccine and CpG oligodeoxynucleotide (ODN) to protect chickens against infectious bursal disease virus (IBDV) infection. Two plasmids DNA carrying VP2 genes of the very virulent (vv) strain of IBDV were constructed with reverse transcription-polymerase chain reaction and designated as pcDNA3.1-VP2 and pCI-VP2. The VP2 protein expressed in COS-7 cells transfected with the plasmid was confirmed by indirect immunofluorescence assay. Seven-day-old chickens were intramuscularly injected with the plasmids alone or plus commercial attenuated infectious bursal disease (IBD) vaccine or synthetic CpG ODN twice at weekly intervals. Chickens at 5 wk old were orally inoculated with vvIBDV strain 99J1 and observed for 7 days after challenge. Immunization with plasmid plus commercial attenuated IBD vaccine or CpG ODN conferred protection for 70%-80% of chickens, as evidenced by the absence of dinical signs, mortality, and atrophy in the cloacal bursa. About 25%-45% of chickens vaccinated with commercial attenuated IBD vaccine or pcDNA3.1-VP2 or pCI-VP2 plasmid alone had dinical signs and died after challenge. Furthermore, there were significantly different histopathologic lesion scores in the clocal bursae between the pcDNA3.1-VP2 or pCI-VP2 plus CpG or live vaccine and pcDNA3.1-VP2, pCI-VP2, or live vaccine vaccinated group. Enzyme-linked immunosorbent assay antibody titers in chickens vaccinated the constructs DNA plus live vaccine or CpG ODN were significantly higher than in those inoculated with the constructs or the live vaccine alone. These results suggest that coadministration of the constructed plasmid pcDNA3.1-VP2 or pCI-VP2 with CpG ODN or commercial attenuated IBD vaccine could protect chickens efficiently from direct vvIBDV challenge.