Endoscopic ultrasound-based application system for predicting endoscopic resection-related outcomes and diagnosing subepithelial lesions: Multicenter prospective study.

OBJECTIVES: Subepithelial lesions (SELs) are associated with various endoscopic resection (ER) outcomes and diagnostic challenges. We aimed to establish a tool for predicting ER-related outcomes and diagnosing SELs and to investigate the predictive value of endoscopic ultrasound (EUS). METHODS: Phase 1 (system development) was performed in a retrospective cohort (n = 837) who underwent EUS before ER for SELs at eight hospitals. Prediction models for five key outcomes were developed using logistic regression. Models with satisfactory internal validation performance were included in a mobile application system, SEL endoscopic resection predictor (SELERP). In Phase 2, the models were externally validated in a prospective cohort of 200 patients. RESULTS: An SELERP was developed using EUS characteristics, which included 10 models for five key outcomes: post-ER ulcer management, short procedure time, long hospital stay, high medication costs, and diagnosis of SELs. In Phase 1, 10 models were derived and validated (C-statistics, 0.67-0.99; calibration-in-the-large, -0.14-0.10; calibration slopes, 0.92-1.08). In Phase 2, the derived risk prediction models showed convincing discrimination (C-statistics, 0.64-0.73) and calibration (calibration-in-the-large, -0.02-0.05; calibration slopes, 1.01-1.09) in the prospective cohort. The sensitivities and specificities of the five diagnostic models were 68.3-95.7% and 64.1-83.3%, respectively. CONCLUSION: We developed and prospectively validated an application system for the prediction of ER outcomes and diagnosis of SELs, which could aid clinical decision-making and facilitate patient-physician consultation. EUS features significantly contributed to the prediction. TRIAL REGISTRATION: Chinese Clinical Trial Registry, http://www.chictr.org.cn (ChiCTR2000040118).

Influence of Sedation on the Detection Rate of Early Cancer and Precancerous Lesions During Diagnostic Upper Gastrointestinal Endoscopies: A Multicenter Retrospective Study.

INTRODUCTION: The influence of sedation on the endoscopic detection rate of upper gastrointestinal (UGI) early cancer (EC) and precancerous lesions, including high-grade intraepithelial neoplasia (HGIN) and low-grade intraepithelial neoplasia, has not been assessed. The aim of this research is to assess whether the use of sedation can help improve the detection rate of UGI EC and precancerous lesions. The second objective is to evaluate its potential influencing factors. METHODS: The study includes 432,202 patients from a multicenter database from January 2012 to July 2019. Information on endoscopic findings and histology biopsies was obtained from endoscopy quality-control system. Associations of sedation with the detection rate of EC and precancerous lesions were assessed. RESULTS: The sedation group has a higher detection rate of UGI EC and HGIN compared with the no-sedation group, whereas the detection rate of low-grade intraepithelial neoplasia was similar between the 2 groups. There were more cases examined by using staining, image enhancement, or magnifying techniques in the sedation group (P < 0.001). And, the mean observation time was also longer in the sedation group (P < 0.001). The type 0-IIb esophageal HGIN and EC cases were significantly increased in the sedation group. No significant difference was detected on lesion subtypes for gastric HGIN and EC according to the Paris classification. More gastric HGIN and EC were detected at gastric body in the sedation group (P = 0.001). DISCUSSION: Sedation may improve the endoscopic detection rate of EC and HGIN in the UGI tract probably through enhancing the use of accessary endoscopic techniques, prolonging observation time, and taking more biopsies in different locations (see Visual Abstract, Supplementary Digital Content 2, http://links.lww.com/AJG/B926).

Antibiotics prescription for targeted therapy of pediatric invasive pneumococcal diseases in China: a multicenter retrospective study.

BACKGROUND: Streptococcus pneumoniae (S. pneumoniae) is a major cause of bacterial meningitis, septicemia and pneumonia in children. Inappropriate choice of antibiotic can have important adverse consequences for both the individual and the community. Here, we focused on penicillin/cefotaxime non-susceptibility of S. pneumoniae and evaluated appropriateness of targeted antibiotic therapy for children with IPD (invasive pneumococcal diseases) in China. METHODS: A multicenter retrospective study was conducted in 14 hospitals from 13 provinces in China. Antibiotics prescription, clinical features and resistance patterns of IPD cases from January 2012 to December 2017 were collected. Appropriateness of targeted antibiotics therapy was assessed. RESULTS: 806 IPD cases were collected. The non-susceptibility rates of S. pneumoniae to penicillin and cefotaxime were 40.9% and 20.7% respectively in 492 non-meningitis cases, whereas those were 73.2% and 43.0% respectively in 314 meningitis cases. Carbapenems were used in 21.3% of non-meningitis cases and 42.0% of meningitis cases for targeted therapy. For 390 non-meningitis cases with isolates susceptible to cefotaxime, vancomycin and linezolid were used in 17.9% and 8.7% of cases respectively for targeted therapy. For 179 meningitis cases with isolates susceptible to cefotaxime, vancomycin and linezolid were prescribed in 55.3% and 15.6% of cases respectively. Overall, inappropriate targeted therapies were identified in 361 (44.8%) of 806 IPD cases, including 232 (28.8%) cases with inappropriate use of carbapenems, 169 (21.0%) cases with inappropriate use of vancomycin and 62 (7.7%) cases with inappropriate use of linezolid. CONCLUSIONS: Antibiotic regimens for IPD definite therapy were often excessive with extensive prescription of carbapenems, vancomycin or linezolid in China. Antimicrobial stewardship programs should be implemented to improve antimicrobial use.

Down-regulation of MAP2K1 by miR-539 inhibits hepatocarcinoma progression.

Liver cancer has been considered as one of the major leading causes of cancer-related mortality. The incidence of liver cancer tends to increase in less developed regions. Increasing evidences have demonstrated that mircoRNAs (miRNAs) play crucial roles in the modulation of tumor growth and progression. Whereas, the functional role of miR-539 in hepatocellular carcinoma (HCC) is not well established. In our present study, we sought to explore biological role of miR-539 in HCC progression. qRT-PCR was utilized to evaluate the expression level of miR-539. Immunoblotting analysis, qRT-PCR and luciferase reporter assays were used for the identification of the potential target of miR-539. Proliferation, migration and invasion assays and flow cytometric were performed to assess the biological functional role of miR-539. The molecular signaling pathways related to the integration of miR-539 were also evaluated. MiR-539 was reduced in human HCC. Mitogen-activated protein kinase 1, also known as MAP2K1 was verified as the target of miR-539. Overexpression of miR-539 inhibited migration, invasion and cell proliferation, while apoptosis rate was increased. Knockdown or overexpression of MAP2K1 in HCC cell transfected with ag-miR-539 or in-miR-539 indicated that miR-539 suppresses the progression of HCC by directly targeting and regulating MAP2K1. Our results reveal that miR-539 might be a tumor suppressor in HCC, supporting a potential target for advanced therapeutic strategy for this disease.

Red cell distribution width as a potential index to assess the severity of hepatitis B virus-related liver diseases.

AIM: To investigate the association between red cell distribution width (RDW) and the severity of hepatitis B virus (HBV)-related liver diseases. METHODS: Sixty-nine patients with chronic hepatitis B (CHB) and 61 patients with HBV-related liver cirrhosis were enrolled in the present study. Forty-one healthy individuals were included as controls. Hematological parameters, hepatitis B e-antigen (HBeAg) status, HBV DNA levels and liver biochemistry were analyzed. Child-Pugh scores and Model for End-Stage Liver Disease (MELD) scores of the patients with HBV-related liver cirrhosis were calculated. RESULTS: The RDW was significantly higher in patients with HBV-related liver cirrhosis as compared with CHB patients and healthy controls. RDW was slightly higher in CHB patients as compared with healthy controls. An increasing correlation of RDW with Child-Pugh grades was found. RDW was positively correlated with Child-Pugh scores and MELD scores. In patients with HBV-related liver cirrhosis, RDW was also positively correlated with total bilirubin and negatively correlated with hemoglobin and serum albumin concentration. However, no significant difference was found between HBeAg positive and negative patients and no significant correlation between RDW and HBV DNA levels was found. CONCLUSION: The RDW was elevated in CHB patients and patients with HBV-related liver cirrhosis and was positively correlated with the severity of HBV-related liver cirrhosis. RDW is a potential index to assess the severity of HBV-related liver diseases.

Prognostic analysis of mutated genes and insight into effects of DNA damage and repair on mutational strand asymmetries in gastric cancer.

Gastric cancer (GACA) is a complex and multifaceted disease influenced by a variety of environmental and genetic factors. Somatic mutations play a major role in its development, and their characteristics, including the asymmetry between two DNA strands, are of great interest and appear as a signal of information and guidance, revealing mechanisms of DNA damage and repair. Here, we analyzed the impact of High-frequency mutated genes on patient prognosis and found that the effect of expression levels of tumor protein p53 (TP53) and lysine methyltransferase 2C (KMT2C) genes remained high throughout the development of GACA, with similar expression patterns. After investigating mutation asymmetry across mutagenic processes, we found that transcriptional asymmetry was dominated by T > G mutations under the influence of transcription couples repair and damage. The apolipoprotein B mRNA editing enzyme catalytic polypeptide like (APOBEC) enzyme that induces mutations during DNA replication has been identified here and we identified a replicative asymmetry, which was dominated by C > A mutations in left-replicating. Strand bias in different mutation classes at transcription factor binding sites and enhancer regions were also confirmed, which implies the important role of non-coding regulatory elements in the occurrence of mutations. This work systematically describes mutational strand asymmetries in specific genomic regions, shedding light on the DNA damage and repair mechanisms underlying somatic mutations in cohorts of GACA patients with gastric cancer.

Intestinal tuberculosis with small bowel stricture and hemorrhage as the predominant manifestation: Three case reports.

BACKGROUND: Intestinal tuberculosis is a chronic disease caused by Mycobacterium tuberculosis that mainly affects the ileum and cecum. Small bowel tuberculosis, characterized by predominant involvement of the small intestine, is an extremely rare condition with highly atypical clinical presentations, making diagnosis even more challenging. CASE SUMMARY: We report three cases of small intestinal tuberculosis, two of the patients presented primarily with abdominal pain, and one presented with gastrointestinal bleeding. All patients underwent blood tests and imaging examinations. Small bowel endoscopy (SBE) revealed that the main lesions in these patients were intestinal stenosis or gastrointestinal bleeding caused by small intestinal ulcers. One patient ultimately underwent surgical treatment. Following a complex diagnostic process and comprehensive analysis, all patients were confirmed to have small intestinal tuberculosis and received standard antituberculosis treatment, leading to an improvement in their condition. CONCLUSION: Patients with SBTs present with nonspecific symptoms such as abdominal pain, weight loss, and occasional gastrointestinal bleeding. Accurate diagnosis requires a thorough evaluation of clinical symptoms and various tests to avoid misdiagnosis and complications.

LncRNA HOXA10-AS promotes the progression of esophageal carcinoma by regulating the expression of HOXA10.

Esophageal cancer (EC) remains a primary cause of cancer-associated fatality worldwide and is characterized by poor prognosis. HOXA10-AS is reported to be relevant with the development of different human cancers. However, its role and regulatory mechanism in EC are still obscure. Our study targeted at investigating the functional and mechanical roles of HOXA10-AS in EC. We confirmed by RT-qPCR that HOXA10-AS presented a remarkably high expression in EC cells. Functional experiments demonstrated that knocking down HOXA10-AS weakened proliferation, invasion and migration in vitro and impeded tumorigenesis in vivo. Further, we found that HOXA10-AS positively regulated its neighbor gene HOXA10 and influenced EC cell biological activities depending on HOXA10. Mechanistically, we showed that HOXA10-AS combined with FMR1 to target and stabilize HOXA10 mRNA. Moreover, HOXA10 served as a transcriptional factor to stimulate the transcription of its target gene CHDH. Finally, rescue assays confirmed that HOXA10 influenced EC cell growth through modulating CHDH. In conclusion, our study first determines the function of HOXA10-AS in EC and demonstrates its mechanism relating to HOXA10/CHDH, suggesting HOXA10-AS as a potential novel target for EC treatment. [Figure: see text].

Stomatin-knockdown effectively attenuates sepsis-induced oxidative stress and inflammation of alveolar epithelial cells by regulating CD36.

Sepsis-induced acute lung injury is a type of lung disease with a high fatality rate that is characterized by acute inflammation. In the present study, the underlying role and potential mechanism of the stomatin (STOM) protein were investigated in lipopolysaccharide (LPS)-induced oxidative stress and inflammation in a mouse lung epithelial cell line, MLE-12. The expression levels of STOM and CD36 were measured using reverse transcription-quantitative PCR and western blotting. Subsequently, the expression levels of STOM and CD36 in LPS-treated MLE-12 cells were knocked down or overexpressed, respectively, via transfection with a small interfering RNA-STOM or a CD36-overexpression vector. An RNA immunoprecipitation (RIP) assay was used to determine the interaction between STOM and CD36, while Cell Counting Kit-8 assay and ELISA were performed to detect cell viability and oxidative stress, respectively. Moreover, western blotting and ELISA kits were used to detect the expression levels of associated inflammatory factors. The results of the present study demonstrated that STOM expression was upregulated in MLE-12 cells treated with LPS compared with the untreated control group. According to the Search Tool for the Retrieval of Interacting Genes/Proteins database, it was predicted that STOM and CD36 had the ability to interact with each other. The predicted binding between STOM and CD36 was verified using a RIP assay. The results demonstrated that STOM positively regulated the expression of CD36. Moreover, in LPS-treated MLE-12 cells, STOM-knockdown reversed the inhibitory effects of LPS on cell viability, and the promoting effects of LPS on oxidative stress and inflammation. These aforementioned changes were alleviated by the overexpression of CD36. To conclude, the results of the present study revealed that STOM may interact with CD36 to affect the levels of oxidative stress and inflammation in LPS-treated MLE-12 cells.

Ionic liquid-coated Fe3O4 magnetic nanoparticles as an adsorbent of mixed hemimicelles solid-phase extraction for preconcentration of polycyclic aromatic hydrocarbons in environmental samples.

In this paper, ionic liquid (IL)-coated magnetic Fe(3)O(4) nanoparticles (NPs) as an adsorbent of mixed hemimicelles solid-phase extraction (SPE) was investigated for the preconcentration of polycyclic aromatic hydrocarbons (PAHs) from environmental samples. Due to the high surface area and excellent adsorption capacity of the Fe(3)O(4) NPs after modification with ILs, satisfactory extraction recoveries can be achieved with only 80 mg Fe(3)O(4) NPs, 50 mg IL, 300 mL solution at pH = 10 and 10 min for equilibration. A comprehensive study of the adsorption conditions such as the amount of Fe(3)O(4) NPs and ILs, the solution pH, ionic strength, standing time, breakthrough volume, and desorption solvents was presented. The extraction ability of different coating agents, such as 1-hexadecyl-3-methylimidazolium bromide (C(16)mimBr), 1-decyl-3-methylimidazolium bromide (C(10)mimBr) and cationic surfactant cetyltrimethylammonium bromide (CTAB) was also compared. Under the optimized conditions, the recoveries for the water samples analysis were between 76 and 105% with relative standard deviations (RSDs) ranging from 3.9 to 6.9%, and the recoveries for soil samples were between 73 and 104% with RSDs ranging from 1.0 to 6.3%. In this method, only a small amount of C(16)mimBr (50 mg) and Fe(3)O(4) NPs (80 mg) was needed to obtain satisfactory recoveries.

Amino acid ionic liquids as chiral ligands in ligand-exchange chiral separations.

Recently, amino acid ionic liquids (AAILs) have attracted much research interest. In this paper, we present the first application of AAILs in chiral separation based on the chiral ligand exchange principle. By using 1-alkyl-3-methylimidazolium L-proline (L-Pro) as a chiral ligand coordinated with copper(II), four pairs of underivatized amino acid enantiomers-dl-phenylalanine (dl-Phe), dl-histidine (dl-His), dl-tryptophane (dl-Trp), and dl-tyrosine (dl-Tyr)-were successfully separated in two major chiral separation techniques, HPLC and capillary electrophoresis (CE), with higher enantioselectivity than conventionally used amino acid ligands (resolution (R(s))=3.26-10.81 for HPLC; R(s)=1.34-4.27 for CE). Interestingly, increasing the alkyl chain length of the AAIL cation remarkably enhanced the enantioselectivity. It was inferred that the alkylmethylimidazolium cations and L-Pro form ion pairs on the surface of the stationary phase or on the inner surface of the capillary. The ternary copper complexes with L-Pro are consequently attached to the support surface, thus inducing an ion-exchange type of retention for the dl-enantiomers. Therefore, the AAIL cation plays an essential role in the separation. This work demonstrates that AAILs are good alternatives to conventional amino acid ligands for ligand-exchange-based chiral separation. It also reveals the tremendous application potential of this new type of task-specific ILs.

Ionic liquid surfactant-mediated ultrasonic-assisted extraction coupled to HPLC: application to analysis of tanshinones in Salvia miltiorrhiza bunge.

Ionic liquid surfactants are a class of ionic liquids (ILs), which can form micelles in the aqueous solution. In this paper, we demonstrate a novel extracting system based on the use of IL surfactants in ultrasonic-assisted extraction followed by HPLC analysis. No organic solvents were used in the extraction, making this method environmentally friendly and more attractive than the conventional organic solvent-based extraction methods. As an example, this method was applied to determine tanshinones in Chinese herbal medicine Salvia miltiorrhiza bunge. The effect of the carbon chain length of the IL cation, as well as other influencing factors on ultrasonic-assisted extraction, was investigated in detail. Under the optimized conditions, satisfactory extraction efficiency was achieved with the recoveries ranging from 87.5 to 107.6%, and the RSDs were lower than 6%. This work shows a promising prospect of the IL surfactants in the extraction of active ingredients from herbs.

Curcumin protects the pancreas from acute pancreatitis via the mitogen­activated protein kinase signaling pathway.

Curcumin has been demonstrated to reduce markers of inflammation during acute pancreatitis (AP). However, the underlying mechanisms of the protective effects of curcumin are unknown. In the present study the effects of curcumin in an AP animal model and cell models was examined and the underlying mechanisms were investigated. An AP animal model was established by injection of 5% sodium taurocholate into the biliopancreatic duct of rats, and the cell model was established by treatment with 0.5 nM cerulein with an optimal concentration of lipopolysaccharide in AR42J rat pancreatic cancer cells. Amylase activity and arterial blood gas composition were assessed by automatic biochemical and blood gas analyzers. Pathological alteration of the pancreas was determined by hematoxylin and eosin staining. Interleukin (IL­6), tumor necrosis factor (TNF)­α and C­reactive protein (CRP) levels were measured by ELISA. Cell viability was determined by Cell Counting Kit­8 and protein expression levels were assessed by western blotting. Curcumin reduced the ascites volume after 12 and 24 h, the weight of pancreas after 12, 24 and 36 h of surgery, but also attenuated injury to the pancreas. Serum expression levels of TNF­α and CRP were reduced by curcumin. In addition, curcumin decreased the cell viability, amylase activity and the phosphorylation of p38 in AR42J cells, but did not affect the intracellular levels of IL­6 and TNF­α. Curcumin may lower the severity and inflammatory response via the mitogen­activated protein kinase­signaling pathway, to some extent. However, future studies are required to fully understand the protective effects of curcumin on AP.

Quantification of amino acid ionic liquids using liquid chromatography-mass spectrometry.

Amino acid ionic liquids (AAILs) containing imidazolium cations and amino acid (AA) anions, were synthesized and applied as task-specific ionic liquids. A sensitive and fast liquid chromatography-mass spectrometry (LC-MS) method was established for the quantitative analysis of 20 AAILs. Using ion pairing-reversed phase liquid chromatography technique, heptafluorobutyric acid was used as ion-pairing reagent to increase the retention of AAILs. Based on the zwitterionity of amino acid, this method was proposed to determine both the cation and the anion of AAILs simultaneously. The limit of detection of this method is down to 1-15ng/mL and the analysis time is less than 15min. According to the analytical data of seven selected AAILs, we found that the content of amino acid anion is always lower than that of butyl methyl imidazolium cation in AAILs. Moreover, the molar ratio of imidazolium cation to amino acid anion is dependent on the chemical property of the amino acid. These results supplied useful information on the interaction of imidazolium cation with acidic, basic, neutral and non-polar amino acids in AAILs.

Epidermal growth factor receptor-targeted immune magnetic liposomes capture circulating colorectal tumor cells efficiently.

AIM: To compare the capacity of newly developed epidermal growth factor receptor (EGFR)-targeted immune magnetic liposomes (EILs) vs epithelial cell adhesion molecule (EpCAM) immunomagnetic beads to capture colorectal circulating tumor cells (CTCs). METHODS: EILs were prepared using a two-step method, and the magnetic and surface characteristics were confirmed. The efficiency of capturing colorectal CTCs as well as the specificity were compared between EILs and EpCAM magnetic beads. RESULTS: The obtained EILs had a lipid nanoparticle structure similar to cell membrane. Improved binding with cancer cells was seen in EILs compared with the method of coupling nano/microspheres with antibody. The binding increased as the contact time extended. Compared with EpCAM immunomagnetic beads, EILs captured more CTCs in peripheral blood from colorectal cancer patients. The captured cells showed consistency with clinical diagnosis and pathology. Mutation analysis showed same results between captured CTCs and cancer tissues. CONCLUSION: EGFR antibody-coated magnetic liposomes show high efficiency and specificity in capturing colorectal CTCs.

Collagen-binding vascular endothelial growth factor attenuates CCl4-induced liver fibrosis in mice.

Vascular endothelial growth factor (VEGF) serves an important role in promoting angiogenesis and tissue regeneration. However, the lack of an effective delivery system that can target this growth factor to the injured site reduces its therapeutic efficacy. Therefore, in the current study, collagen­binding VEGF was constructed by fusing a collagen­binding domain (CBD) to the N­terminal of native VEGF. The CBD­VEGF can specifically bind to collagen which is the major component of the extracellular matrix in fibrotic liver. The anti­fibrotic effects of this novel material were investigated by the carbon tetrachloride (CCl4)­induced liver fibrotic mouse model. Mice were injected with CCl4 intraperitoneally to induce liver fibrosis. CBD­VEGF was injected directly into the liver tissue of mice. The liver tissues were stained with hematoxylin and eosin for general observation or with Masson's trichrome staining for detection of collagen deposition. The hepatic stellate cell activation, blood vessel formation and hepatocyte proliferation were measured by immunohistochemical staining for α­smooth muscle actin, CD31 and Ki67 in the liver tissue. The fluorescent TUNEL assay was performed to evaluate the hepatocyte apoptosis. The present study identified that the CBD­VEGF injection could significantly promote vascularization of the liver tissue of fibrotic mice and attenuate liver fibrosis. Furthermore, hepatocyte apoptosis and hepatic stellate cell activation were attenuated by CBD­VEGF treatment. CBD­VEGF treatment could additionally promote hepatocyte regeneration in the liver tissue of fibrotic mice. Thus, it was suggested that CBD­VEGF may be used as a novel therapeutic intervention for liver fibrosis.

Association between cytotoxic T-lymphocyte-associated antigen 4 +49A/G polymorphism and persistent hepatitis B virus infection in the Asian population: evidence from the current studies.

AIMS: A number of studies assessed the association between cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) +49A/G polymorphism and persistent hepatitis B virus (HBV) infection risk. However, the results are quite contradictory. In order to obtain a more precise estimation of the relationship, a meta-analysis was performed. METHODS: Pubmed, ISI Web of Knowledge, HuGE Navigator, Wanfang Database, and China National Knowledge Infrastructure were searched. Crude odds ratios (ORs) with 95% confidence intervals (95% CIs) were calculated to estimate the strength of this association. RESULTS: Six publications were included in the final meta-analysis with 1075 cases and 1321 controls for CTLA-4 +49A/G polymorphism according to our inclusion and exclusion criteria. We observed that the CTLA-4 +49A/G polymorphism was significantly correlated with chronic HBV infection risk (the homozygote codominant model: OR 1.67, 95% CI 1.30-2.15; the dominant model: OR 1.34, 95% CI 1.01-1.78; the recessive model: OR 1.56, 95% CI 1.24-1.96; the allele contrast model: OR 1.32, 95% CI 1.10-1.59). No publication bias was observed in this study according to Begg's funnel plot and Egger's test. CONCLUSION: CTLA-4 +49A/G polymorphism is assumed to confer a higher risk for persistent HBV infection in the Asian population.

Immunomodulatory effects of polysaccharopeptide in immunosuppressed mice induced by cyclophosphamide.

Polysaccharopeptide (PSP) is well known for its immunoregulatory effects. In the present study, the effect of PSP on white blood cell (WBC) count, T lymphocyte subsets, B lymphocytes, Th1/Th2 balance and negative immune regulators was investigated using an immunosuppressed mouse model. The results demonstrated that the WBC count and the absolute number of CD3+CD4+ T cells, CD3+CD8+ T cells and CD3­CD19+ B cells in the peripheral blood were increased in PSP­treated groups as compared with the cyclophosphamide (Cy) group. In addition, PSP reduced interleukin (IL)­4 and GATA binding protein 3 (GATA­3) mRNA relative expression levels and elevated the ratios of IL­2/IL­4 and the transcription factors, T­box­containing protein/GATA­3. The relative mRNA expression levels of the forkhead/winged­helix transcription factor box protein 3 (Foxp3), programmed death­1 (PD­1) and IL­10 were also downregulated by PSP. These observations indicate that the immunoregulatory effects of PSP are associated with restoration of WBC number, improving the absolute number of T lymphocyte subsets and B lymphocytes, inducing the Th1/Th2 response and downregulating the negative immune regulators, Foxp3, PD­1 and IL­10.

Observations on the survival and neovascularization of fat grafts interchanged between C57BL/6-gfp and C57BL/6 mice.

BACKGROUND: Autologous fat transplantation has become a prevalent option for soft-tissue augmentation throughout the body. However, there is still much controversy over whether the fat grafts have survived or have been replaced in the recipient sites and over how the vessels grow. METHODS: After C57BL/6-gfp mice and C57BL/6 mice were paired randomly, the inguinal fat was excised and cut into pieces with scissors, and the adipose granules, approximately 0.2 ml (0.195 g), were transplanted subcutaneously with syringes to the dorsa of the paired mice. Samples were obtained at different time intervals: 3 days, 7 days, 2 weeks, 4 weeks, 2 months, 3 months, and 4 months after transplantation. Each fat sample was weighed to evaluate the graft volume. Histology, origin, and densities of neovascularization were examined by immunohistochemical staining. RESULTS: At 4 months, there was no significant difference in either graft survival or histologic evaluation. Histologic evaluation manifested the normal physiologic process of inflammation, neovascularization, remodeling, and maturity at different time intervals. At the endpoint, the immunohistochemical staining of CD34 showed that the difference in capillary density of the fat graft-31.3 ± 3.9 capillaries/mm on the dorsa of the C57BL/6-gfp mice and 29.6 ± 3.2 capillaries/mm on the dorsa of the C57BL/6 mice-was not statistically significant. The α-smooth muscle actin staining indicated that there were neovascularized vessels in both C57BL/6-gfp and C57BL/6 fat grafts. CONCLUSIONS: Fat grafts can survive and neovascularized vessels can grow from the recipient sites. Fat transplantation is feasible and will be applied more widely if fat graft survival is improved.