Podocyte-specific Nup160 knockout mice develop nephrotic syndrome and glomerulosclerosis.

More than 60 monogenic genes mutated in steroid-resistant nephrotic syndrome (SRNS) have been identified. Our previous study found that mutations in nucleoporin 160 kD (NUP160) are implicated in SRNS. The NUP160 gene encodes a component of the nuclear pore complex. Recently, two siblings with homozygous NUP160 mutations presented with SRNS and a nervous system disorder. However, replication of nephrotic syndrome (NS)-associated phenotypes in a mammalian model following loss of Nup160 is needed to prove that NUP160 mutations cause SRNS. Here, we generated a podocyte-specific Nup160 knockout (Nup160podKO) mouse model using CRISPR/Cas9 and Cre/loxP technologies. We investigated NS-associated phenotypes in these Nup160podKO mice. We verified efficient abrogation of Nup160 in Nup160podKO mice at both the DNA and protein levels. We showed that Nup160podKO mice develop typical signs of NS. Nup160podKO mice exhibited progression of proteinuria to average albumin/creatinine ratio (ACR) levels of 15.06 ± 2.71 mg/mg at 26 weeks, and had lower serum albumin levels of 13.13 ± 1.34 g/l at 30 weeks. Littermate control mice had urinary ACR mean values of 0.03 mg/mg and serum albumin values of 22.89 ± 0.34 g/l at the corresponding ages. Further, Nup160podKO mice exhibited glomerulosclerosis compared with littermate control mice. Podocyte-specific Nup160 knockout in mice led to NS and glomerulosclerosis. Thus, our findings strongly support that mutations in NUP160 cause SRNS. The newly generated Nup160podKO mice are a reliable mammalian model for future study of the pathogenesis of NUP160-associated SRNS.

Successful ablation of a right epicardial accessory pathway via the right ventricular diverticulum in a patient with Wolff-Parkinson-White syndrome.

INTRODUCTION: We describe one rare case of successful ablation of a right epicardial accessory pathway (AP) via the right ventricular diverticulum in a patient with Wolff-Parkinson-White syndrome. METHODS: A 42-year-old woman was referred to the hospital for a catheter ablation of Wolf-Parkinson-White syndrome. The earliest activation was shown to be present in the region of the tricuspid annulus. However, ablation had no effect on the AP. RESULTS: We decided to do a selected angiography, in which a big diverticulum near the right tricuspid annulus was shown to be present. Ablation in this region successfully repressed the AP without any recurrences within a follow-up period of 12 months. CONCLUSION: The ventricular diverticulum-mediated AP is a novel variant of pre-excitation. It can serve as an anatomical substrate of supraventricular tachycardia, and can be ablated endocardially using an irrigation tip catheter within the diverticulum.

The characteristics of a 12-lead electrocardiogram measuring premature ventricular contractions originating from the tricuspid annulus using the clock position method.

OBJECTIVE: This study aimed to analyze the conventional surface electrocardiogram (ECG) characteristics of premature ventricular contractions (PVCs) originating from the tricuspid annulus and to investigate the efficacy of locating their origins according to ECG results. METHODS: Eight patients who underwent radiofrequency ablation in the First Hospital of Shanxi Medical University (China) were included in the study. Pace mapping (PM) was used to analyze the characteristics of the PVCs originating from the tricuspid annulus recorded via 12-lead body surface ECGs. RESULTS: An R-wave was found in leads I, V5 , and V6 . The QRS wave was narrower when the PVCs originated from the septum and shifted in lead V3 (R-wave amplitude/S-wave amplitude in the precordial lead-1). The QRS wave was broadest when the PVCs originated from the 7 to 9 o'clock position. The augmented vector left lead showed RS, QS, or RSR-type waves with a low amplitude when the PVCs originated from the upper part of the annulus. When the PVCs originated from the lower part of the annulus, the augmented vector right lead reflected multidirectional and QS-type waves. CONCLUSION: The ECG-lead characteristics related to the origin of PVCs in the tricuspid annulus indicate some level of significance and can be used to formulate a specific diagnosis.

Comparison of the lesion formation and safety in ex vivo porcine heart study: Using ThermoCool SmartTouch and ThermoCool SmartTouch-SF catheters.

BACKGROUND: The study was performed to compare the efficacy and safety during radiofrequency ablation (RFA) using ThermoCool SmartTouch (ST) and ThermoCool SmartTouch-SF (STSF) catheters in the porcine heart. METHODS AND RESULTS: RFA was performed on the porcine myocardium by using two irrigated ablation catheters. Three groups were divided based on the different contact forces (CFs): low contact force (LCF) (1-3 g), medium contact force (MCF) (5-10 g), and high contact force (HCF) (15-20 g). In each group, RFA was delivered at four power settings of 30, 40, 50, 60 W. At each power, RFA was applied to reach the target ablation index (AI) of 350, 450, and 500. Altogether, 360 RF lesions were created by using 72 ablation conditions. AI value was positively correlated with lesion size using ST and STSF catheters. At a fixed power, lesion dimensions significantly smaller in the LCF group, whereas did not differ between MCF and HCF groups. Furthermore, at a fixed CF, lesion dimensions increased with power set at 40 W compared with 30 W but decreased with high-power RF energy (50 and 60 W). Although the average lesion surface diameter and the maximum diameter was increased using the STSF catheter, there were no significant differences in LV between the two catheters. The steam pop provoked more frequently using ST catheter and showed a negative correlation with CF and positive correlation with high-power energy. CONCLUSION: The STSF catheter is safer and equally effective in lesion formation compared with the ST catheter. LV was increased along with the early increase of CF and power, whereas a further increase of CF and power significantly reduces the lesion size.

Gastric intestinal metaplasia assessment between linked color imaging based on endoscopy and pathology.

OBJECTIVE: Cumulative evidence suggests that linked color imaging (LCI) can be used to identify gastric intestinal metaplasia (GIM). We aimed to develop endoscopic grading for GIM (EGGIM) with LCI. METHODS: Two hundred and seventy-seven patients who underwent high-resolution white-light gastroscopy followed by LCI for EGGIM estimation were included. LCI was performed for the entire mucosa, and images of five areas each were recorded from the lesser and greater curvatures of the antrum and corpus, and for the incisura. For each area, scores of 0 (no GIM), 1 (focal GIM, ≤30% of the area), and 2 (extensive GIM, >30% of the area) were attributed for 10 points. If GIM was suspected based on endoscopy findings, targeted biopsies were performed; if GIM was not evident, random biopsies were performed according to the Sydney system to estimate the operative link on GIM (OLGIM). RESULTS: GIM was staged as OLGIM 0, I, II, III, and IV in 136, 70, 37, 28, and 6 patients, respectively. For OLGIM III/IV diagnosis, the area under the receiver operating curve was 0.949 (95% CI 0.916-0.972). EGGIM of 4, with sensitivity and specificity of 94.12% (95% CI 80.3%-99.3%) and 86.42% (95% CI 81.5%-90.5%), respectively, was determined the best cut-off value for identifying OLGIM III/IV patients. CONCLUSIONS: Our findings demonstrated the ability of EGGIM for diagnosing the extent of intestinal metaplasia and showed that EGGIM is related to OLGIM staging. EGGIM of 4 was the best cut-off value for identifying OLGIM III/IV patients.

Renal injury following long-term exposure to carbon disulfide: analysis of a case series.

BACKGROUND: To investigate the clinicopathological characteristics of renal damage caused by long-term exposure to carbon disulfide (CS2) in nine patients. METHODS: All the patients underwent ultrasound-guided renal biopsy. All specimens were examined by light microscopy and immunohistochemistry (IHC). Samples form one patient were further analyzed using transmission electron microscopy. RESULTS: Similar pathological changes were observed in all patients, but the degrees of lesions were different. All cases had moderate to severe nodular mesangial hyperplasia; among these, type "Kimme1stie1-Wi1son" (K-W nodule for short) was observed in four cases, type "K - W nodule" refer to nodular hyperplasia of mesangial membrane like letter K or W. four cases had proliferative extracapillary glomerulonephritis (GN), while there were no concomitant changes in one patient. Besides, six cases had diffuse basement membrane thickening, focal segmental sclerosis or bulbar sclerosis; two cases had diffuse glomerular sclerosis, and one case had focal segmental capillary hyperplasia. Moreover, all patients had renal tubular atrophy/interstitial fibrosis with less to moderate chronic inflammatory cell infiltration, as well as renal arteriosclerosis. IHC showed that the depositions of IgA, IgM, C3d, C4d, C1q and Fib were not specific; while IgG, type III collagen, Fibronectin, Amyloid A, Igκ, Igλ, HBsAg and HBcAg were all negative. CONCLUSION: Diffuse nodular mesangial hyperplasia/sclerosing glomerular nephropathy is characterized by nodular mesangial hyperplasia with type "K-W nodules" formation, which we speculate is a special pathological manifestation of renal damage caused by carbon disulfide (CS2).

Global mapping of binding sites for phic31 integrase in transgenic maden-darby bovine kidney cells using ChIP-seq.

BACKGROUND: ΦC31 integrase, a site-specific recombinase, can efficiently target attB-bearing transgenes to endogenous pseudo attP sites within mammalian genomes. The sequence features of endogenous binding sites will help us to fully understand the site-specific recognition function by ΦC31 integrase. The present study was aimed to uncover the global map of ΦC31 integrase binding sites in bovine cells and analysis the features of these binding sites by comprehensive bioinformatics methods. RESULTS: In this study, we constructed a ChIP-seq method that can be used to uncover the global binding sites by phiC31 integrase. 6740 potential ΦC31 integrase binding sites were identified. A sequence motif was found that contains inverted repeats and has similarities to wild-type attP site. Using REPEATMASKER, we identified a total of 20,183 repeat-regions distributed in 50 repeat types for the 6740 binding sites. These sites enriched in "regulation of GTPase activity" of in the GO category of biological process and KEGG pathway of signal transmembrane transporter activity. CONCLUSION: This study is the first time to uncover the global map of binding sites for ΦC31 integrase using ChIP-sequencing method and analysis the features of these binding sites. This method will help us to fully understand the mechanism of the site-specific integration function by phiC31 integrase and will potentially boost its genetic manipulations in both gene therapy and generation of transgenic animals.

Identification of New Epididymal Luminal Fluid Proteins Involved in Sperm Maturation in Infertile Rats Treated by Dutasteride Using iTRAQ.

BACKGROUND: Spermatozoa become mature and acquire fertilizing capacity during their passage through the epididymal lumen. In this study, we identified new epididymal luminal fluid proteins involved in sperm maturation in infertile rats by dutasteride, a dual 5α-reductase inhibitor, in order to provide potential epididymal targets for new contraceptives and infertility treatment. METHODS: Male rats were treated with dutasteride for 28 consecutive days. We observed the protein expression profiles in the epididymal luminal fluids in infertile and normal rats using isobaric tags for relative and absolute quantitation (iTRAQ) technique. The confidence of proteome data was validated by enzyme-linked immunosorbent assays. RESULTS: 1045 proteins were tested, and 23 of them presented different expression profiling in the infertile and normal rats. The seven proteins were down-regulated, and 16 proteins were up-regulated. Among the seven proteins which were significantly down-regulated by dutasteride in the epididymal luminal fluids, there were three ß-defensins (Defb2, Defb18 and Defb39), which maybe the key proteins involved in epididymal sperm maturation and male fertility. CONCLUSIONS: We report for the first time that dutasteride influences the protein expression profiling in the epididymal luminal fluids of rats, and this result provides some new epididymal targets for male contraception and infertility therapy.

[Inhibitory effect of dutasteride on the expressions of epididymal Claudin1 and ß-catenin in male rats].

OBJECTIVE: To explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and ß-catenin in rats. METHODS: Sixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and ß-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated. RESULTS: Dutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and ß-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05). CONCLUSION: Dutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and ß-catenin expressions, and damaging epididymal epithelial cell junctions.

C3d deposition in the media of renal arterioles is a useful marker for arteriolosclerosis in IgA nephropathy.

C3d deposition in peritubular capillaries has been demonstrated to indicate antibody-mediated alloresponse during renal transplantation. C3d deposition in renal arterioles in IgA nephropathy (IgAN), however, is poorly documented. Especially, its significance to the pathology of primary glomerulonephritis remains unclear. This retrospective study included 340 patients with IgAN who underwent renal biopsy at our center. C3d strongly positive deposition in arterioles was observed in 123 (36.2%) of the 340 cases, and weakly positive deposition of C3d was observed in 217 cases (63.8%). In the weakly positive group, C3d mainly deposited in the intima of arterioles. In the strongly positive group, C3d deposited in the intima and the media of arterioles, presenting as the medial thickening and sclerosis of varying severities. The prognosis was worse in the C3d strongly positive group than in the weakly positive group during a 2-year follow-up (P = .027). The predictive value of C3d deposition in the media of arterioles in patients with IgAN may be a useful marker for arteriolosclerosis indicating unfavorable clinical outcomes.

Identification of common mechanisms and biomarkers of atrial fibrillation and heart failure based on machine learning.

AIMS: Atrial fibrillation (AF) is the most common arrhythmia. Heart failure (HF) is a disease caused by heart dysfunction. The prevalence of AF and HF were progressively increasing over time. The co-existence of AF and HF presents a significant therapeutic challenge. In order to provide new ideas for the diagnosis of AF and HF, it is necessary to carry out biomarker related studies. METHODS AND RESULTS: The training set and validation set data of AF and HF patient samples were downloaded from the GEO database, 'limma' was used to compare the differences in gene expression levels between the disease group and the normal group to screen for differentially expressed genes (DEGs). Weighted correlation network analysis (WGCNA) identified the modules with the highest positive correlation with AF and HF. Functional enrichment and PPI network construction of key genes were carried out. Biomarkers were screened by machine learning. The infiltration of immune cells in AF and HF groups was evaluated by R-packet 'CIBERSORT'. The miRNA network was constructed and potential therapeutic agents for biomarker genes were predicted through the drugbank database. Through WGCNA analysis, it was found that the modules most positively correlated with AF and HF were MEturquoise (r = 0.21, P value = 0.09) and MEbrown (r = 0.62, P value = 8e-12), respectively. We screened 25 genes that were highly correlated with both AF and HF. Lasso regression analysis results showed 7 and 20 core genes in AF and HF groups, respectively. The top 20 important genes in AF and HF groups were obtained as core genes by RF model analysis. Four biomarkers were obtained after the intersection of core genes in four groups, namely, GLUL, NCF2, S100A12, and SRGN. The diagnostic efficacy of four genes in AF validation sets was good (AUC: GLUL 0.76, NCF2 0.64, S100A12 0.68, and SRGN 0.76), as well as in the HF validation set (AUC: GLUL 0.76, NCF2 0.84, S100A12 0.92, and SRGN 0.68). The highest correlation with neutrophils was observed for GLUL, NCF2, and S100A12, while SRGN exhibited the strongest correlation with T cells CD4 memory resting in the AF group. GLUL, NCF2, S100A12, and SRGN were most associated with neutrophils in the HF group. A total of 101 miRNAs were predicted by four genes, and GLUL, NCF2, and S100A12 predicted a total of 10 potential therapeutic agents. CONCLUSIONS: We identified four biological markers that are highly correlated with AF and HF, namely, GLUL, NCF2, S100A12, and SRGN. Our findings provide theoretical basis for the clinical diagnosis and treatment of AF and HF.

Pulmonary Epstein-Barr virus-associated smooth muscle tumor after kidney transplantation: two case reports with review of differential diagnosis.

Pulmonary nodules are a common complication in solid organ transplant recipients, and may have various underlying causes, with Epstein-Barr virus-associated smooth muscle tumor (EBV-SMT) being one of them. Given the rarity of this entity, we describe the diagnosis and therapeutic interventions for post-transplant EBV-SMT in two individuals. Both cases involved female patients who were diagnosed with multiple pulmonary nodules 60 months and 116 months, respectively, after receiving living-related kidney transplantation. Pathological examination revealed a spindle cell tumor, with immunophenotype and EBV in situ hybridization supporting the diagnosis of EBV-SMT. After diagnosis, these two patients underwent intervention by decreasing their intake of immunosuppressants. As of the latest follow-up, the patients' lesion size remained stable, and their overall condition was favorable. We also reviewed literature about the morphological and molecular pathological features of EBV-SMT and highlighted the diagnosis and differential diagnosis of pulmonary spindle cell lesions especially in the setting of immunosuppression.

Sodium butyrate attenuate hyperglycemia-induced inflammatory response and renal injury in diabetic mice.

The activation of the monocyte-macrophage system and the damage to the renal and pancreatic tissue are common complications in patients with diabetes induced by hyper-glycemia. This study aimed to evaluate the effect and mechanism of butyrate (NaB), a metabolite of intestinal flora, on inhibiting the inflammatory response of human monocyte-macrophages (THP-1 cells) induced by high glucose and the damage of pancreatic and renal tissue in diabetic mice. The results showed that high concentration glucose significantly up-regulated the expressions of IL-1ß, TNF-α, and NLRP3 in THP-1 cells and mouse spleen, and that NaB could inhibit the overexpression of those genes. The abundance of Beclin-1, LC3B and reactive oxygen species (ROS) in THP-1 cells is increased due to the high glucose concentration, and NaB can inhibit the genes responsible for upregulating the expression. In diabetic mice, vacuolar degeneration of renal tubules was observed. Then we observed that some of the epithelial cells of the renal tubules were exfoliated and some formed tubules. NaB could alleviate these pathological lesions, but NaB cannot alleviate pancreatic injury. Our results indicated that NaB could be used for the prevention and adjuvant treatment of diabetic kidney injury.

Expert Consensus on Pathological Diagnosis of Intrahepatic Cholangiocarcinoma (2022 version).

Intrahepatic cholangiocarcinoma (iCCA) can originate from the large bile duct group (segment bile ducts and area bile ducts), small bile duct group (septal bile ducts and interlobular bile ducts), and terminal bile duct group (bile ductules and canals of Hering) of the intrahepatic biliary tree, which can be histopathological corresponding to large duct type iCCA, small duct type iCCA and iCCA with ductal plate malformation pattern, and cholangiolocarcinoma, respectively. The challenge in pathological diagnosis of above subtypes of iCCA falls in the distinction of cellular morphologies, tissue structures, growth patterns, invasive behaviors, immunophenotypes, molecular mutations, and surgical prognoses. For these reasons, this expert consensus provides nine recommendations as a reference for standardizing and refining the diagnosis of pathological subtypes of iCCA, mainly based on the 5th edition of the World Health Organization Classification of Tumours of the Digestive System.

Establishment of an improved high-efficiency thermal asymmetric interlaced PCR for identification of genomic integration sites mediated by phiC31 integrase.

Streptomyces phage phiC31 integrase is widely used to mediate the integration of exogenous genes into host genomes for gene therapy and genomic modification, as it autonomously performs efficient, unidirectional, site-specific integration into pseudo attP sites of the host genome. Although pseudo attP sites are rarely found within exons, it is necessary to map their precise locations to avoid the risk of insertion mutagenesis. High-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) is a technique that has been developed to recover genomic sequences that flank insertion tags. We have found, however, that this technique is poorly efficient, as it amplifies many non-specific targets and frequently does not generate sufficient product for downstream analysis. Therefore, we have modified the hiTAIL-PCR procedure and re-designed the random primers. As a result, both the amount and specificity of the reaction product were enhanced for each integration site. Restriction analysis of known sequences within the integrated vector, which co-amplified with the flanking genomic sequences, validated 90% of these bands for sequencing. In contrast, only 30% of the bands produced by previous hiTAIL-PCR could be validated. Compared with the original hiTAIL-PCR, our improved hiTAIL-PCR procedure identified phiC31 integration sites more accurately and efficiently.

Effects of Nutritious Meal Combined with Online Publicity and Education on Postoperative Nutrition and Psychological State in Patients with Low Rectal Cancer After Colostomy.

Objective: To investigate the effects of nutritious meal combined with online publicity and education on postoperative nutrition and psychological state in patients with low rectal cancer after colostomy. Methods: The clinic data of 88 patients with low rectal cancer who received the colostomy in our hospital (August 2020-August 2021) were retrospectively reviewed. Among them, 44 patients received nutritious meal combined with online publicity and education and they made up the study group, and the others were given conventional care and they made up the reference group. The nutrition indicators, scores of the World Health Organization Quality of Life (WHOQOL)-BREF, and other materials of the patients in the two groups were compared. Results: After intervention, the various nutrition indicators, immune indexes, and WHOQOL-BREF score of the study group were all prominently higher than those of the reference group (P < 0.001). Compared with the reference group, the study group after intervention achieved markedly lower self-rating anxiety scale (SAS) score and self-rating depression scale (SDS) score (P < 0.001) and obviously lower total incidence of complications (P < 0.05). Conclusion: Combining nutritious meal with online publicity and education can effectively improve the postoperative nutrition and immune function of the patients with low rectal cancer after colostomy, and this intervention contributes to releasing the patients' adverse emotions. Further study helps to provide these patients with favorable solutions.

Effect of Baseline Impedance in Radiofrequency Delivery on Lesion Characteristics and the Relationship Between Impedance and Steam Pops.

Objective: To explore the effects of baseline impedance (R) and power (P) on radiofrequency ablation (RFA) lesion characteristics and their correlation with steam pops using ThermoCool SmartTouch-SF (STSF) catheters in the porcine heart. Method: A porcine left ventricle was submerged in 37°C saline ex vivo, and the experiment was performed with various P (P = 30, 40, 50, and 60 W) and multiple R loads (R = 80-100, 100-140, 140-180, and 180-220 Ω) to reach the target ablation index (AI; AI = 350, 450, and 500) or reach the target ablation time using a fixed contact force (CF; CF = 10-15 g) and the same saline irrigation (30 W/8 ml/min or 40-60 W/15 ml/min), repeated five times under each condition. Results: The surface diameter, maximum diameter, depth, and volume of the lesions were strongly correlated with the AI (P = 40 W, R = 100-140 Ω, CF = 10-15 g) (r = 0.5412; r = 0.7889; r = 0.9366; and r = 0.913, respectively; all p < 0.05). As the value of R increased, the maximum diameter, depth, and volume of the lesions significantly increased (AI = 350, P = 30 W). Moreover, the higher the baseline value of R, the greater the absolute value of the R decrease (r = 0.9035, p < 0.05, Y = 0.2759 × X - 18.33). Under high power and high impedance, the occurrence rate of steam pops was high (P = 60 W, R = 180-220 Ω, AI when a steam pop occurred: 480 ± 26.5, ablation time: 11.29 ± 1.04 s). Conclusion: Radiofrequency catheter ablation (RFCA) in power-controlled mode resulted in various lesion characteristics that were related to diverse baseline Rs. In addition, the incidence of steam pops was strongly correlated with high baseline R and high P.

Targeted delivery of exosomal miR-484 reprograms tumor vasculature for chemotherapy sensitization.

The vascular dysfunction of ovarian cancer (OC) contributes to the chemotherapeutic resistance. In this study, we aimed to explore whether exosome-mediated angiogenesis blocking could improve the chemotherapy sensitivity via vascular normalization. Exosomes were armed with RGD on the surface by fusing Lamp2b. Candidate miRNAs related to tumor angiogenesis was detected by qRT-PCR. RGD-modified exosomes were loaded with miRNAs via electroporation. The therapeutic effects of the exosomes on angiogenesis, vascular normalization, and chemotherapy sensitivity were systemically analyzed in the xenograft model. RGD-modified exosomes were relatively enriched in the tumor mass, both the tumor cell and the endothelial cells. Among the miRNA candidates, miR-484 was found down-regulated in both the cancer cells and the angiogenic endothelial cells. In vivo xenograft model experiment revealed that injection of RGD-modified exosomes loaded with miR-484 induced vessel normalization and in turn sensitized the cancer cells to chemotherapy induced apoptosis. Mechanistically, miR-484 simultaneously inhibited the expression of VEGF-A from the cancer cells and the corresponding receptors in the endothelial cells. Targeted delivery of miR-484 via RGD-modified exosomes improves the vascular normalization, sensitizes the cancer to chemotherapy, and prolongs the survival time of tumor-bearing mice after chemotherapy, opening an avenue for the clinical management of chemotherapy resistance.

[An experimental study on the safety of intratracheal drug administration].

OBJECTIVE: To explore whether injury and repair occur in the trachea and the lung after intra-tracheal administration of different drugs. METHODS: Wistar rats were randomly divided into 5 groups, a normal group, a blank control (BC) group, a normal saline (NS) group, a lidocaine (LD) group and an amikacin (AK) group. For the latter 3 groups, normal saline, lidocaine and amikacin were injected into trachea by needle puncture. Scanning electron microscope was used to observe the ultra-structural changes of the epithelium, and the percentage of the area of damage (PAD) in tracheal mucosa was calculated. Moreover, pathological changes of the mucous membrane of bronchioles and alveolar epithelial cells were also examined, and the degree of lung pathology was semi-quantified. RESULTS: Two hours after the injection of the 3 drugs, derangement and edema of the cilia were evident by scanning electron microscopy. The PAD of the NS group, the LD group and the AK group were (94.2 ± 3.2)%, (93.1 ± 3.0)% and (95.5 ± 1.8)%, respectively; all being significantly higher than that of the BC group (1.3 ± 0.3)%. For the NS group and the LD group, the PAD decreased significantly after 24 h, which were (73.7 ± 7.8)% and (81.0 ± 4.6)% respectively, and returned to normal at 48 h and 96 h. While for the AK group, the damage began to improve at 72 h [PAD (62.1 ± 5.2)%], and recovered at 96 h. Airway epithelial derangement and cell edema in the alveoli and the bronchioles also occurred 2 h after drug injection, and inflammatory cell infiltration became evident at 24 h. At this time, the score of pathology was 1.80 ± 0.84, 2.60 ± 0.55 and 2.80 ± 0.45 for the NS group, the LD group and the AK group, respectively; all being higher than that of the BC group (0). These pathological changes recovered totally after 72 h for the NS and the LD groups, and 96 h for the AK group. CONCLUSIONS: Intra-tracheal administration of normal saline, lidocaine and amikacin in rats led to reversible airway mucosal and lung tissue damages.

WDR74 induces nuclear ß-catenin accumulation and activates Wnt-responsive genes to promote lung cancer growth and metastasis.

Lung cancer has been notorious for its lack of advance in clinical therapy, urging for effective therapeutic targets. WD repeat-containing protein 74 (WDR74) has previously been implicated in tumorigenesis, but its mechanistic functions remain not well understood. Herein, WDR74 expression was observed to be increased upon lung cancer progression from healthy normal tissues to the primary cancer and further to the metastatic cancer. Through gain- and loss-of-function approaches, we found that WDR74 regulated lung cancer cell proliferation, cell cycle progression, chemoresistance and cell aggressiveness in vitro. Moreover, a xenograft mouse model disclosed that WDR74 knockout inhibited lung cancer growth and metastasis, whereas WDR74 overexpression reciprocally enhanced these characteristics. Mechanistically, WDR74 promoted nuclear ß-catenin accumulation and drove downstream Wnt-responsive genes, thus revealing that WDR74 activated the Wnt/ß-catenin signaling pathway. Collectively, WDR74 inducing nuclear ß-catenin accumulation and driving the downstream Wnt-responsive genes expression facilitates lung cancer growth and metastasis. WDR74 can serve as a candidate target for the prevention and treatment of lung cancer in clinic.