Single-cell analysis of Crohn's disease: Unveiling heterogeneity and evaluating ustekinumab outcomes.

BACKGROUND: The present study aimed to dissect the cellular complexity of Crohn's disease (CD) using single-cell RNA sequencing, focusing on identifying key cell populations and their transcriptional profiles in inflamed tissue. METHODS: We applied scRNA-sequencing to compare the cellular composition of CD patients with healthy controls, utilizing Seurat for clustering and annotation. Differential gene expression analysis and protein-protein interaction networks were constructed to identify crucial genes and pathways. RESULTS: Our study identified eight distinct cell types in CD, highlighting crucial fibroblast and T cell interactions. The analysis revealed key cellular communications and identified significant genes and pathways involved in the disease's pathology. The role of fibroblasts was underscored by elevated expression in diseased samples, offering insights into disease mechanisms and potential therapeutic targets, including responses to ustekinumab treatment, thus enriching our understanding of CD at a molecular level. CONCLUSIONS: Our findings highlight the complex cellular and molecular interplay in CD, suggesting new biomarkers and therapeutic targets, offering insights into disease mechanisms and treatment implications.

[Retracted] TRIM16 suppresses the progression of prostate tumors by inhibiting the Snail signaling pathway.

Following the publication of this paper, it was drawn to the Editor's attention by concerned readers that the western blotting data shown in Figs. 4C and 7B and D, the scratch­wound assay images shown in Figs. 5A and 6A, and certain of the cell migration and invasion assay data shown in Figs. 5B and 6B were strikingly similar to data that had previously appeared in different form in other articles by different authors. Owing to the fact that the contentious data in the above article had already been published elsewhere, or were already under consideration for publication, prior to its submission to International Journal of Molecular Medicine, the Editor has decided that this paper should be retracted from the Journal. After having been in contact with the authors, they accepted the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. [International Journal of Molecular Medicine 38: 1734­1742, 2016; DOI: 10.3892/ijmm.2016.2774].

[Research Progress of FAM60A in the Regulation of Cellular Function].

FAM60A,a cell cycle protein,is a subunit of the SIN3 transcription regulator family member A/histone deacetylase(SIN3-HDAC)complex and plays an important role in cell cycle regulation,cell morphology change,cell proliferation,differentiation and migration,early embryogenesis and so on.Studies in recent years have shown that FAM60A plays a role in the occurrence and development of tumors including human osteosarcoma,esophageal cancer,gastric cancer,lung cancer and liver cancer,providing a new research direction for tumor diagnosis and treatment.Based on the research results in recent years at home and abroad,this paper discussed the effects of FAM60A on cellular functions.

[Effect of particulate matter 2.5 at urban centre of Hangzhou on lung impairment in rats].

OBJECTIVE: To explore the effect of ambient particle matter 2.5 (PM2.5) collected in the urban center of Hangzhou on the lung injury of rats and on the activating of endoplasmic reticulum pathway. METHODS: PM2.5 samples were collected on quartz fiber filters using a PM2.5 high-volume air sampler in the urban area of Hangzhou. The collected PM2.5 particles were extracted in ultrapure water and concentrated by vacuum freeze-drying. Twenty-four male Sprague-Dawly (SD) rats were randomly divided into 3 groups:saline control group, low dose PM2.5 exposure group (5 mg/kg BW) and high dose PM2.5 exposure groups (25 mg/kg BW). Each group received intratracheal instillation of PM2.5, once a week for 4 weeks. Twenty-four hours after the last exposure, the rats were narcotized and sacrificed, left lung was isolated and fixed with 4% paraformaldehyde for histopathological detection. The bronchoalveolar lavage fluid (BALF) was collected from the right lung. The total antioxidant capacity (T-AOC) level, the activities of superoxide dismutase (SOD) and lactic dehydrogenase (LDH) in BALF were detected by chemical colorimetry. The level of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß) and interleukin-6 (IL-6) cytokines in BALF was measured by enzyme linked immunosorbent assay (ELISA). And the protein expressions of glucose-regulated protein 78 (GRP78), phosphorylated protein kinase receptor-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic translation initiation factor (p-eIF2α), transcription factors C/EBP homologue protein (CHOP), inositol-requiring enzyme 1α (IRE1α) and X-box binding protein 1 (XBP1) in lung tissue were determined by Western blotting. RESULTS: Compared with control group, rats in both low dose (5 mg/kg) and high dose (25 mg/kg) PM2.5-treated groups showed obviously dose-dependent pulmonary toxicity including thickening of alveolar walls, narrowing of alveolar space, interstitial hyperplasia and inflammatory cell infiltration. Compared with control group, T-AOC level and the SOD activity in BALF in both PM2.5-treated groups were decreased dose-dependently (P<0.05), whereas the LDH activity in BALF were increased in a dose-dependent manner (P<0.05). Exposure to PM2.5 resulted in a increasing of the release of proinflammatory cytokines including TNF-α, IL-1ß and IL-6 in rat lung in a dose-dependent manner (P<0.05). The levels of GRP78, p-PERK, p-eIF2α, CHOP, IRE1α and spliced XBP1 (XBP1-S) were significantly up-regulated, whereas the level of unspliced XBP1 (XBP1-U) was down-regulated in the rat lung tissue of high-dose PM2.5 treated group. CONCLUSIONS: The PM2.5 in the urban area of Hangzhou can significantly cause lung inflammatory injury in rats. Both oxidative stress and activation of ER stress pathways may be related to such PM2.5 inhalation-induced lung inflammatory injury.

[Study on the Relationship Between Normalization of Tumor Microvessels and CA9 for Rh-Endostatin to Inhibit Lewis Lung Cancer].

OBJECTIVES: To explore the relationship between normalization of tumor microvessels and CA9 for rh-Endostatin to inhibit Lewis lung cancer (LLC) and the expression level of CA9 in LLC. METHODS: Lewis cells of logarithmic growth phase were collected and made into 1×106 mL-1 cell suspensions were prepared. The transplanted tumor model of LLC was established on C57/BL6 mice by injected 0.2 mL cell suspensions/mice into 40 C57/BL6 mice. 40 LLC mice were randomly divided into control group and rh-ES group (20 mice per group). Control group experienced treatment of intraperitoneal injection (ip) for 0.2 mL NS/d, while rh-ES group was treated for 5 mg rh-ES/(kg·d) from the first to the ninth day. The samples of 5 mice were obtained from day 2, day 4, day 6 and day 9 after treatment in control group or rh-ES group, respectively. CA9 was tested by IHC in LLC and paracancerous tissues and estimated by RT-PCR and ELISA in the each time point of both rh-ES group and control group,respectively. RESULTS: The transplanted tumor model of LLC on C57/BL6 mice was established successfully. The expression of CA9 decreased on day 4 and day 6 in rh-ES group estimated by RT-PCR and ELISA, which indicated some great significance when compared with day 2, day 9 in rh-ES group and day 4, day 6 in control group (P<0.05), and the expression of CA9 in day 2, day 4, day 6, day 9 tested by IHC was higher in LLC than in paracancerous tissues in control group (P<0.05). CONCLUSIONS: The expression of CA9 was higher in LLC. Rh-ES could have positive effect on LLC model of C57/BL6 mice, in day 4-6 (a brief normalized time course) decreased the expression of CA9 and reversed the tumor hypoxia.

Puerarin Prevents LPS-Induced Osteoclast Formation and Bone Loss via Inhibition of Akt Activation.

Osteolysis induced by chronic Gram-negative bacterial infection underlies many bone diseases such as osteomyelitis, septic arthritis, and periodontitis. Drugs that inhibit lipopolysaccharide (LPS)-induced osteolysis are critically needed for the prevention of bone destruction in infective bone diseases. In this study, we assessed the effect of puerarin, a natural isoflavone isolated from Pueraria lobata OHWI root, on LPS-induced osteoclastogenesis and bone loss. Our in vitro study showed that puerarin significantly inhibited LPS-induced osteoclast differentiation from osteoclast precursor RAW264.7 cells. The inhibition occurred through suppressing the production of osteoclast activating factor tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and prostaglandin E2 (PGE2), which led to down-regulating mRNA expression of osteoclastogenic genes including tartrate-resistant acid phosphatase (TRAP), cathepsin K and matrix metalloprotein 9 (MMP-9). Furthermore, LPS triggered activation of Akt in osteoclast precursor RAW264.7 cells, which was inhibited by puerarin treatment. In vivo, puerarin attenuated LPS-induced bone loss in a murine calvarial osteolysis model. Collectively, puerarin prevents LPS-induced osteoclast formation, function and bone loss, where the inhibition of Akt activation plays an important role. These findings provide evidences that puerarin might be beneficial as a promising candidate drug for the prevention and treatment of bacteria-induced bone destruction disease, and give new insights for understanding its possible mechanism.

TRIM16 suppresses the progression of prostate tumors by inhibiting the Snail signaling pathway.

Prostate carcinoma is a devastating disease which is characterized by insidious early symptoms, rapid progression and a poor prognosis. Tripartite motif-containing protein 16 (TRIM16) was identified as an estrogen- and antiestrogen-regulated gene in epithelial cells stably expressing estrogen receptors. The protein encoded by this gene contains two B-box domains and a coiled-coiled region that are characteristic of the B-box zinc finger protein family. Proteins belonging to this family have been reported to be involved in a variety of biological processes including cell growth, differentiation and pathogenesis. TRIM16 expression has been detected in most tissues. However, the funtions of this gene remain to be elucidated. In the present study, immunohistochemical staining revealed that the expression of TRIM16 was decreased in prostate adenocarcinoma compared with that in normal prostate tissues. The patients with high TRIM16-expressing tumors had a significantly greater survival than those with low TRIM16-expressing tumors. Western blot analysis showed that TRIM16 was downregulated in distant metastatic cancer tissues compared with that in non-distant metastatic cancer tissues. The overexpression of TRIM16 inhibited the migration and invasion of prostate cancer cells as well as inhibiting the epithelial-to-mesenchymal transition process, whereas TRIM16 depletion enhanced these processes. Moreover, TRIM16 inhibited the Snail signaling pathway. The silencing of Snail by small interfering RNA was performed in order to determine the role of Snail in the TRIM16-mediated tumor phenotype. Taken together, these findings suggest that TRIM16 may be an important molecular target which may aid in the design of novel therapeutic agents for prostate cancer.

Relationship between genotypes and clinical manifestation, pathology, and cccDNA in Chinese children with hepatitis B virus-associated glomerulonephritis.

BACKGROUND: Hepatitis B virus-associated glomerulonephritis (HBV-GN) is one of the extrahepatic manifestations after HBV infection, which would cause great clinical harm to people. The present study was undertaken to investigate the HBV-GN genotypes and its clinical relevance in Chinese children. METHODS: A total of 41 HBV-infected children diagnosed with HBV-GN were enrolled in the study. All patients underwent liver and kidney biopsy. The genotypes and cccDNA were detected in their serum samples to analyze the relationship between HBV genotypes and clinical characteristics, cccDNA, and pathology. RESULTS: Among the 41 children with HBV-GN, 29 (70.7%) had genotype C, 10 (24.4%) had genotype B, 2 (4.9%) had genotype B/C, and none of them had genotype non-B/C. Most children had genotypes B or C; moreover, the genotype C was the most frequent one. The incidence of hematuria and albuminuria, reduction in complement C3, increase in serum alanine aminotransferase levels and renal insufficiency in the children with genotype C were significantly higher than those in the children with genotype B (P<0.05); however, there was no statistically significant difference in hypertension and hepatomegaly (P>0.05). The frequency of HBV cccDNA positive in the genotype C group was significantly higher than that in the genotype B group (72.4% vs. 30.0%, P<0.05). No difference was observed in hepatic inflammation grades and stages of fibrosis between the two groups (P>0.05). CONCLUSIONS: Genotype C was the most frequent genotype in the described group of patients with HBV-GN, and the liver and kidney damage indicators were more likely to occur in patients with genotype C.

[Clinical and pathological differences between children with various genotypes of hepatitis B virus-associated glomerulonephritis].

OBJECTIVE: To compare the clinical and pathological features between children with various genotypes of hepatitis B virus-associated glomerulonephritis (HBV-GN). METHODS: Forty-one children with HBV-GN concurrently undergoing liver and renal biopsy were randomly selected. Serum specimens were collected for genotyping and hepatitis B virus (HBV) cccDNA assay. The clinical, pathological, and HBV cccDNA differences between HBV-GN children of various genotypes were analyzed. RESULTS: Among the 41 HBV-GN children, 29 (71%) were genotype C, 10 (24%) were genotype B, and 2 (5%) were genotype B/C. The incidence rates of hematuria, albuminuria, complement 3 decrease, alanine transaminase increase, and renal insufficiency in the genotype C group were significantly higher than those in the genotype B group (P<0.05). Similarly, the HBV cccDNA positive rate was significantly higher in the genotype C group than that in the genotype B group. No difference was observed in the distribution of pathological types of renal tissues betwee the two geonotype groups. There were no significant differences in the degrees of hepatic inflammation and fibrosis between the two groups. CONCLUSIONS: Mainly genotypes C and B occur in children with HBV-GN and the former genotype is dominant. The clinical symptoms of patients with genotype C are more serious than those with genotype B. However, there is no difference in the pathological features between them.

ABCB1 polymorphisms correlate with susceptibility to adult acute leukemia and response to high-dose methotrexate.

The aim of this study is to investigate the association of ABCB1 polymorphisms with susceptibility to adult acute leukemia, and the influence of ABCB1 polymorphisms on the efficacy of high-dose methotrexate (HDMTX). ABCB1 polymorphisms in 178 acute leukemia patients (case group) and 150 healthy subjects (control group) were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. All patients received HDMTX therapy. Correlation analysis was performed to explore the associations of ABCB1 polymorphisms with MTX concentration and efficacy of MTX therapy. All statistical analyses were conducted with SPSS 19.0 software. The frequency of TT genotype and T allele on ABCB1 3435C > T in case group were significantly higher than the control group (P < 0.05), while no statistical difference between the two groups was observed in genotypic distribution and allele frequencies of ABCB1 2677G > T/A (P > 0.05). Furthermore, 24-h MTX concentration of patients carrying TT and TA genotypes on 2677G > T/A was higher than carriers with other genotypes (P < 0.05), and 24-h MTX concentration of patients with TT and CT genotypes on 3435C > T was also apparently higher than carriers with CC genotype (P < 0.05). In addition, ABCB1 polymorphisms were connected with increased risk of liver dysfunction and infection (P < 0.05). Complete remission (CR) rate in patients carrying GG on 2677G > T/A was markedly lower than carriers with non-GG genotype (P < 0.05). ABCB1 3435C > T polymorphisms may be associated with susceptibility to acute leukemia, and ABCB1 polymorphisms might be a sensitive indicator for predicting efficacy of MTX therapy in the treatment of acute leukemia.

[Relationship between B/C genotype of hepatitis B virus and hepatitis B virus related-nephritis in children].

OBJECTIVE: To investigate the relationship between genotype of hepatitis B virus and hepatitis B virus related-glomerular nephritis in (HBV-GN) children. METHOD: Totally 176 HBV-DNA positive children with chronic hepatitis B were randomly collected. Among the 176 patients, 92 were HBV carriers, 84 were cases with chronic hepatitis. The genotypes of their serum HBV, liver function, and HBV-DNA load were detected. When children showed nephrotic syndrome, renal biopsy was performed. RESULT: Of the serum samples of 176 cases, 85 (48.3%) were genotype C, 72 (40.9%) were genotype B, 13 (7.4%) were genotype B/C, and 6 (3.4%) were non-B/C genotype which were excluded. Among the analyzed 157 cases, the ratio of HBV-GN in the HBeAg positive group (78.3%) was significantly higher than that in the negative group (21.7%) (χ(2) = 18.301, P < 0.001). And, the ratio of HBV-GN in the genotype C group (73.9%) was significantly higher than that in the genotype B group (26.1%) (P < 0.039). The ratio of hematuria or proteinuria in the genotype C group (20%, 18.8%) was significantly higher than that in the genotype B group (8.3%, 5.6%) (P < 0.039; P value = 0.013); and the alteration of ALT or C3 in the genotype C group (10.2%, 15.3%) was more frequent than those in the genotype B group (2.8%, 2.8%) (P = 0.005; P = 0.008). There were no significant differences in kidney dysfunction or hepatomegaly. Further, the ratio of HBV-GN was more significantly frequent in HBV-DNA highly loading group (79.2%) than which in HBV-DNA lowly loading group (20.8%) (P = 0.000). Finally, in HBV-GN group, genotype C cases (88.2%) more frequently had high HBV-DNA load condition than genotype B cases (11.8%) (P = 0.021). CONCLUSION: Children with HBV infection in Gansu province showed mainly genotypes C or B, while genotype C seemingly predominant. Patients with genotype C more frequently showed proteinuria or hematuria. The high HBV-DNA load may be related with HBV-GN. It is a potential reason in the mechanism of HBV-GN that patients with genotype C had more possibility to have HBV-DNA high load. Analysis of HBV genotype for HBV patients maybe helpful in diagnosis and treatment.

Multiple primary malignant tumors of upper gastrointestinal tract: a novel role of 18F-FDG PET/CT.

AIM: To evaluate the capacity of (18)F-fluorodeoxyglucose positron emission tomography/computed tomography ((18)F-FDG PET/CT) for detecting multiple primary cancer of upper gastrointestinal (UGI) tract. METHODS: Fifteen patients (12 without cancer histories and 3 with histories of upper GI tract cancer) were investigated due to the suspicion of primary cancer of UGI tract on X-ray barium meal and CT scan. Subsequent whole body (18)F-FDG PET/CT scan was carried out for initial staging or restaging. All the patients were finally confirmed by endoscopic biopsy or surgery. The detection rate of multiple primary malignant cancers was calculated based on (18)F-FDG PET/CT and endoscopic examinations. RESULTS: (18)F-FDG PET/CT scan was positive in 32 suspicious lesions, 30/32 were true positive primary lesions, and 2/32 were false positive. In 15 suspicious lesions with negative (18)F-FDG PET/CT scan, 12/15 were true negative and 3/15 were false negative. Among the 15 patients, 12 patients had 29 primary synchronous tumors confirmed by pathology, including 8 cases of esophageal cancers accompanied with gastric cancer and 4 of hypopharynx cancers with esophageal cancer. The other 3 patients had 4 new primary metachronous tumors, which were multiple primary esophageal cancers. PET/CT imaging detected local lymph node metastases in 11 patients. Both local lymph node metastases and distant metastases were detected in 4 patients. On a per-primary lesion basis, the sensitivity, specificity, accuracy, negative predictive value and positive predictive value of (18)F-FDG PET/CT for detecting multiple primary cancer of UGI tract were 90.9%, 85.7%, 89.4%, 80% and 93.7%, respectively. CONCLUSION: The whole body (18)F-FDG PET/CT may play an important role in evaluating the multiple primary malignant tumors of UGI tract cancer.

Effects of astragaloside IV on pathogenesis of metabolic syndrome in vitro.

AIM: To investigate the diverse pharmacological actions of astragaloside IV from the perspective of metabolic syndrome, and to investigate the effect of the drug on the pathogenesis of metabolic syndrome. METHODS: Adipogenesis was used as an indicator of the effect of astragaloside IV on preadipocyte differentiation, and was measured by using an oil red O assay. Glucose uptake was determined by measuring the transport of [2-(3)H]-deoxyglucose into the cells. The concentrations of peroxisome proliferator-activated receptor-gamma (PPARgamma) and aP2 mRNA were determined by using reverse transcription-polymerase chain reaction. Apoptosis and viability loss of endothelial cells were detected by using flow cytometry and the WST-1 assay, respectively. Intracellular free Ca2+ was labeled with Fluo-3 AM and measured by using a laser scanning confocal microscope. RESULTS: Astragaloside IV can significantly potentiate insulin-induced preadipocyte differentiation at concentrations of 3, 10, and 30 microg/mL, improve high glucose-induced insulin resistance in adipocytes at a concentration of 30 microg/mL, and prevent tumor necrosis factor (TNF)-alpha-induced apoptosis and viability loss at concentrations of 10 and 30 microg/mL, and 30 microg/mL, respectively, in endothelial cells. Furthermore, we found that these effects were partly due to the promotion of PPARgamma expression and to the inhibition of abnormal TNF-alpha-induced intracellular free Ca(2+) accumulation in endothelial cells. CONCLUSION: The diverse pharmacological actions of astragaloside IV can all be linked to metabolic syndrome pathogenesis. Our study provides a new insight into the mechanism by which astragaloside IV exerts its effect.

The study of anti-metabolic syndrome effect of puerarin in vitro.

Puerarin is an isoflavone extracted from Chinese plant, Pueraria lobata (Wild.) Ohwi. It has been reported to have comprehensive pharmacological action in treatment of diabetes and cardiovascular diseases. The purpose of this study was to link the scattered effects of puerarin and to find the common mechanisms underlying. We investigated the effect of puerarin on the pivotal common pathogenic factors of metabolic syndrome, which includes obesity, Type II diabetes and cardiovascular diseases. Recently, a large body of evidence indicates that there is a complicated interplay among insulin resistance, adipocytes and endothelial dysfunction that links the abnormalities of metabolic syndrome. Results of present study showed that puerarin could potentiate insulin-induced preadipocyte differentiation, promote glucose-uptake of adipocytes that have been induced insulin resistance by high glucose, and prevent TNF-a-induced apoptosis and viability loss of endothelial cells. Furthermore, we found that these effects are probably due to promote PPARgamma expression and partly through inhibiting abnormal TNF-a-induced intracellular-free Ca(2+) accumulation of endothelial cells. Overall, our synthetical study links the comprehensive pharmacological actions of puerarin to the recognized common pathogenesis of metabolic syndrome, and provides a new insight into the mechanism of puerarin effect.

In situ immobilization of proteins and RGD peptide on polyurethane surfaces via poly(ethylene oxide) coupling polymers for human endothelial cell growth.

A "CBABC"-type pentablock coupling polymer, mesylMPEO, was designed and synthesized to promote human endothelial cell growth on the surfaces of polyurethane biomaterials. The polymer was composed of a central 4,4'-methylenediphenyl diisocyanate (MDI) coupling unit and poly(ethylene oxide) (PEO) spacer arms with methanesulfonyl (mesyl) end groups pendent on both ends. As the presurface modifying additive (pre-SMA), the mesylMPEO was noncovalently introduced onto the poly(ether urethane) (PEU) surfaces by dip coating, upon which the protein/peptide factors (gelatin, albumin, and arginine-glycine-aspartic acid tripeptide [RGD]) were covalently immobilized in situ by cleavage of the original mesyl end groups. The pre-SMA synthesis and PEU surface modification were characterized using nuclear magnetic resonance spectroscopy ((1)H NMR), attenuated total reflection infrared spectroscopy (ATR-FTIR), and X-ray photoelectron spectroscopy (XPS). Human umbilical vein endothelial cells (HUVEC) were harvested manually by collagenase digestion and seeded on the modified PEU surfaces. Cell adhesion ratios (CAR) and cell proliferation ratios (CPR) were measured using flow cytometry, and the individual cell viability (ICV) was determined by MTT assay. The cell morphologies were investigated by optical inverted microscopy (OIM) and scanning electrical microscopy (SEM). The gelatin- and RGD-modified surfaces were HUVEC-compatible and promoted HUVEC growth. The albumin-modified surfaces were compatible but inhibited cell adhesion. The results also indicated that, for HUVEC in vitro cultivation, the cell adhesion stage was of particular importance and had a significant impact on the cell responses to the modified surfaces.