Value of ctDNA methylation biomarkers in diagnosis of colorectal tumors.

Tweetable abstract DNA methylation alterations have been identified as promising biological markers for early-stage colorectal cancer detection. Here, the authors highlight some recent advances in DNA methylation and its role in the early diagnosis and overall disease course management of colorectal tumors. New insights into DNA methylation biomarkers for colorectal cancer early diagnosis and management are discussed.

The Protective Role of TLR4 in Intestinal Epithelial Cells through the Regulation of the Gut Microbiota in DSS-Induced Colitis in Mice.

BACKGROUND: The cause of ulcerative colitis (UC) is not yet fully understood. Previous research has pointed towards a potential role for mutations in nucleotide-binding oligomerization domain-containing protein 2 (NOD2) in promoting the onset and progression of inflammatory bowel disease (IBD) by altering the microbiota of the gut. However, the relationship between toll-like receptor 4 (TLR4) and gut microbiota in IBD is not well understood. To shed light on this, the interaction between TLR4 and gut microbiota was studied using a mouse model of IBD. METHODS: To examine the function of TLR4 signaling in intestinal injury repair, researchers developed Dextran Sulfate Sodium Salt (DSS)-induced colitis and injury models in both wild-type (WT) mice and TLR4 knockout (TLR4-KO) mice. To assess changes in the gut microbiota, 16S rRNA sequencing was conducted on fecal samples from both the TLR4-KO and WT enteritis mouse models. RESULTS: The data obtained depicted a protective function of TLR4 against DSS-induced colitis. The gut microbiota composition was found to vary considerably between the WT and TLR4-KO mice groups as indicated by ß-diversity analysis and operational taxonomic units (OTUs) cluster. Statistical analysis of microbial multivariate variables depicted an elevated abundance of Escherichia coli/Shigella, Gammaproteobacteria, Tenerlcutes, Deferribacteres, Enterobacteria, Rikenellaceae, and Proteobacteria in the gut microbiota of TLR4-KO mice, whereas there was a considerable reduction in Bacteroidetes at five different levels of the phylogenetic hierarchy including phylum, class, order, family, and genus in comparison with the WT control. CONCLUSIONS: TLR4 may protect intestinal epithelial cells from damage in response to DSS-induced injury by controlling the microbiota in the gut.

A pixelated liquid perovskite array for high-sensitivity and high-resolution X-ray imaging scintillation screens.

Scintillators with high spatial resolution at a low radiation dose rate are desirable for X-ray medical imaging. A low radiation dose rate can be achieved using a sufficiently thick scintillator layer to absorb the incident X-ray energy completely, however, often at the expense of low spatial resolution due to the issue of optical crosstalk of scintillation light. Therefore, to achieve high sensitivity combined with high-resolution imaging, a thick scintillator with perfect light guiding properties is in high demand. Herein, a new strategy is developed to address this issue by embedding liquid scintillators into lead-containing fiber-optical plates (FOPs, n = 1.5) via the siphon effect. The liquid scintillator is composed of perovskite quantum dots (QDs)/2,5-diphenyloxazole (PPO) and the non-polar high-refractive index (n = 1.66) solvent α-bremnaphthalene. Benefiting from the pixelated and thickness-adjustable scintillators, the proposed CsPbBr3 QDs/PPO liquid scintillator-based X-ray detector achieves a detection limit of 79.1 µGy s-1 and a spatial resolution of 4.6 lp mm-1. In addition, it displays excellent tolerance against radiation (>34 h) and shows outstanding stability under ambient conditions (>160 h). This strategy could also be applied to other liquid scintillators (such as CsPbCl3 QDs and Mn:CsPbCl3 QDs). The combination of high sensitivity, high spatial resolution and stability, easy fabrication and maintenance, and a reusable substrate matrix makes these liquid scintillators a promising candidate for practical X-ray medical imaging applications.

Discovery of extracellular vesicle-delivered miR-185-5p in the plasma of patients as an indicator for advanced adenoma and colorectal cancer.

BACKGROUND: We aimed to evaluate whether extracellular vesicles (EV)-derived microRNAs (miRNAs) can be used as biomarkers for advanced adenoma (AA) and colorectal cancer (CRC). METHODS: We detected the changes in the plasma EV-delivered miRNA profiles in healthy donor (HD), AA patient, and I-II stage CRC patient groups using miRNA deep sequencing assay. We performed the TaqMan miRNA assay using 173 plasma samples (two independent cohorts) from HDs, AA patients, and CRC patients to identify the candidate miRNA(s). The accuracy of candidate miRNA(s) in diagnosing AA and CRC was determined using the area under the receiver-operating characteristic curve (AUC) values. Logistic regression analysis was performed to evaluate the association of candidate miRNA(s) as an independent factor for the diagnosis of AA and CRC. The role of candidate miRNA(s) in the malignant progression of CRC was explored using functional assays. RESULTS: We screened and identified four prospective EV-delivered miRNAs, including miR-185-5p, which were significantly upregulated or downregulated in AA vs. HD and CRC vs. AA groups. In two independent cohorts, miR-185-5p was the best potential biomarker with the AUCs of 0.737 (Cohort I) and 0.720 (Cohort II) for AA vs. HD diagnosis, 0.887 (Cohort I) and 0.803 (Cohort II) for CRC vs. HD diagnosis, and 0.700 (Cohort I) and 0.631 (Cohort II) for CRC vs. AA diagnosis. Finally, we demonstrated that the upregulated expression of miR-185-5p promoted the malignant progression of CRC. CONCLUSION: EV-delivered miR-185-5p in the plasma of patients is a promising diagnostic biomarker for colorectal AA and CRC. Trial registration The study protocol was approved by the Ethics Committee of Changzheng Hospital, Naval Medical University, China (Ethics No. 2022SL005, Registration No. of China Clinical Trial Registration Center: ChiCTR220061592).

A retrospective case-series of influence of chronic hepatitis B on synchronous liver metastasis of colorectal cancer.

Main point: Our retrospective analysis of a large number of cases found in patients with primary colorectal cancer (CRC) carrying positive HBsAg inhibited the occurrence of synchronous liver metastases (SLM). However, liver cirrhosis caused by non-HBV factors promoted the occurrence of SLM. Objectives: This study aimed to investigate the effect of HBV on the occurrence of synchronous liver metastases (SLM) of colorectal cancer (CRC). Methods: Univariate and multivariate analyses were used to analyze the influence of clinical parameters on the occurrence of SLM. Results: A total of 6, 020 patients with primary CRC were included in our study, of which 449 patients carrying HBsAg(+) accounted for 7.46%. 44 cases of SLM occurred in the HBsAg(+) group, accounting for 9.80%, which was much lower than 13.6% (758/5571) in the HBsAg(-) group (X=5.214, P=0.022). Among CRC patients with HBsAg(-), the incidence of SLM was 24.9% and 14.9% in the group with high APRI and FIB-4 levels, respectively, which were significantly higher than that in the compared groups (12.3% and 12.5%, all P<0.05). Compared with the control group, female patients, late-onset patients, and HBV-infective patients had lower risks of SLM (HR=0.737, 95%CI: 0.614-0.883, P<0.001; HR=0.752, 95%CI: 0.603-0.943, P=0.013; HR=0.682, 95%CI: 0.473-0.961, P=0.034). Conclusions: The carriage of HBsAg(+) status inhibited the occurrence of SLM from CRC. HBV-causing liver cirrhosis did not further influence the occurrence of SLM, whereas non-HBV-factor cirrhosis promoted the occurrence of SLM. Nevertheless, this still required prospective data validation.

Interaction between the intestinal flora and the severity of diversion colitis after low anterior resection of rectal cancer.

Background: Diversion colitis (DC) is nonspecific inflammation of the distal intestinal mucosa following disruption of colonic continuity with colonic dysfunction. The colonscopic score is a good tool for differentiating the severity of patients with DC. At present, no studies have analyzed the pathogenesis of DC from the perspective of the diversity and and differences of intestinal flora. Methods: Retrospective study: Clinical information were collected from patients with low rectal cancer admitted to the Department of Anorectal Surgery, Changzheng Hospital, from April 2017 to April 2019. These patients underwent laparoscopic low anterior resection (LAR) combined with terminal ileum enterostomy (dual-chamber). We used chi-square test to comparethe clinical baseline information, clinical symptoms, and colonscopic characteristics between different severity of DC. Propsective oberservational study: We recruited 40 patients with laparoscopic anterior low resection combined with terminal ileum enterostomy and they were further classified into mild group and severe group according to the scores of colonscopic examinations for DC. 16s-rDNA sequencing was carried out to analyze the diversity and and differences of intestinal flora in the intestinal lavage fluid of the two groups. Results: In retrospective study, we found that age, BMI, history of diabetes, and symptoms associated with the stoma state were the independent risk factors that affect DC severity (P<0.05). Meanwhile, age, BMI, history of diabetes and colonscopic score were found to be independent risk factors affecting the severity of diarrhea after ileostomy closure surgery(P<0.05), which was consistent with our results of differentiating the severity of DC under endoscopy; In propsective oberservational study, 40 patients with low rectal cancer recruited by sample size calculation, 23 were in the mild group and 17 in the severe group. The results of 16s-rDNA sequencing showed that intestinal flora with high enrichment values primarily consisted of Bifidobacteriales and Prevotella in mild group, whereas that in the severe group consisted of Providencia and Dorea. The functional predictions on such two types of intestinal flora were mainly focused on lipid synthesis, glycan synthesis, metabolism, and amino acid metabolism pathways. Conclusion: After ileostomy closure surgery, a series of severe clinical symptoms might appear in DC patients. There are significant differences in local and systemic inflammatory responses, composition of intestinal flora between DC patients with different colonscopic scores, which provide a basis for the clinical interventional treatment for DC in patients with permanent stoma.

Constructing and Validating a Prognosis Predictive Nomogram for Cancer-Specific Survival in Rectal Cancer Patients Receiving Preoperative Radiotherapy.

Background A predictive tool is required to identify the cancer-specific survival in rectal cancer (RC) patients who have opted to receive preoperative radiotherapy.Methods A database containing the data on RC patients' records of Surveillance, Epidemiology, and End Results (SEER) receiving surgery during 2000-2014 was selected. All patients received neoadjuvant radiotherapy (NR). The correlation of clinicopathological parameters was analyzed using the Chi-square test and the survival risk factors were analyzed using the Cox proportional hazards analysis (univariate and multivariate). Finally, the nomogram was developed and validated to visually represent an accurate prediction of the probability of 3- and 5-year cancer-specific survival (CSS) based on the screened variables of the cohort.Results 11,499 rectal cancer patients were included in our cohort. Patients' records were randomly allocated to either the development or validation cohorts based on an equal ratio (1:1). Performing the multivariate Cox regression analysis incorporating these variables in the development cohort determined 11 independent prognostic factors. Statistically significant differences were recorded among subgroups using log-rank tests, which confirmed the appropriateness and acceptability of factor stratifications. Then, the nomogram was constructed and its concordance index (C-index) values in the development cohort (0.720) and validation cohort (0.717) were evaluated to be higher (P<0.05) than those of the AJCC stage (0.631 and 0.633 respectively). Also, the 3-year AUC values of this nomogram were higher than those of the AJCC stage in both the development cohort (0.746 vs. 0.631) and the validation cohort (0.745 vs. 0.640). Using DCA curves, the predictive potential of the currently developed nomogram outperformed the conventional AJCC staging system.Conclusion The nomogram model might be a more reliable tool to predict prognosis accurately in rectal cancer patients receiving preoperative radiotherapy.

White light interference demodulation of optical fiber Fabry-Perot micro-pressure sensors based on the Karhunen-Loeve transform and singular value decomposition.

In this paper, the Karhunen-Loeve transform (KLT) and wavelength domain interferometric spectral singular value decomposition (SVD) are used for the first time to demodulate the pressure of an optical fiber Fabry-Perot (F-P) micro-pressure sensor, and the feasibility of the proposed method is demonstrated experimentally. The eigenvalue decomposition of the dominant frequency part of the beam-domain interferometric spectrum after the fast Fourier transform (FFT) is performed using KLT, and the singular value decomposition of the wavelength domain interferometric spectrum is additionally performed using SVD. Both methods use high-order eigenvalues as a new metric and then derive the relation between the new metric and the reference pressure. The two demodulation methods are experimentally compared, and we used an optical fiber F-P pressure sensor with unknown structure and material for pressure measurements. Even though the interferometric spectral signal is acquired using a coarse spectrometer (2.5 nm wavelength resolution), one can still achieve high demodulation accuracy with both algorithms. However, the SVD demodulation accuracy decreases significantly after reducing the spectral data points in the wavelength domain from 1566 to 783. KLT still has high demodulation accuracy and linearity after spectral data points are reduced from 1024 to 256 in the wavenumber domain. The satisfactory linearity of the measured pressure versus reference pressure and low reading errors validate the feasibility of the proposed demodulation algorithm.

Significant Near-Field Enhancement over Large Volumes around Metal Nanorods via Strong Coupling of Surface Lattice Resonances and Fabry-Pérot Resonance.

Metal nanoparticles supporting plasmons are widely used to enhance electromagnetic fields, resulting in strong light-matter interactions at the nanoscale in a diverse range of applications. Recently, it has been shown that when metal nanorods are periodically arranged with proper lattice periods, surface lattice resonances (SLRs) can be excited and near fields can be greatly enhanced over extended volumes. In this work, we report significant near field enhancement over even larger volumes by placing the metal nanorod array within a Fabry-Pérot (F-P) microcavity. Simulation results show that by taking advantage of strong coupling between the SLR and the photonic F-P resonances, the electric field intensity of the bonding split mode can be enhanced by up to 1935 times, which is about three times of the enhancement of the SLR, and the greatly enhanced field can extend over most of the F-P microcavity. We further show that the F-P resonances of both odd and even orders can strongly couple to the SLR by varying the nanorods position from the middle of the microcavity. We expect that the proposed plasmonic-photonic coupling system will find promising applications in nanolasers, nonlinear optics and sensing.

A Highly Birefringent Photonic Crystal Fiber for Terahertz Spectroscopic Chemical Sensing.

A photonic crystal fiber (PCF) with high relative sensitivity was designed and investigated for the detection of chemical analytes in the terahertz (THz) regime. To ease the complexity, an extremely simple cladding employing four struts is adopted, which forms a rectangular shaped core area for filling with analytes. Results of enormous simulations indicate that a minimum 87.8% relative chemical sensitivity with low confinement and effective material absorption losses can be obtained for any kind of analyte, e.g., HCN (1.26), water (1.33), ethanol (1.35), KCN (1.41), or cocaine (1.50), whose refractive index falls in the range of 1.2 to 1.5. Besides, the PCF can also achieve high birefringence (∼0.01), low and flat dispersion, a large effective modal area, and a large numerical aperture within the investigated frequency range from 0.5 to 1.5 THz. We believe that the proposed PCF can be applied to chemical sensing of liquid and THz systems requiring wide-band polarization-maintaining transmission and low attenuation.

Progress in the diagnosis and treatment of COVID-19 and the role of surgeons in the front line of the pandemic.

The current struggle to control and contain COVID-19 is critical and surgeons are on the front line in the fight against this virus. Surgeons, and other medical workers in the field of surgery, have a solid foundation and experience in medical treatment and intensive care, and an understanding of the support of respiratory, circulatory, digestive, and other systemic organs. Furthermore, the operative standards of aseptic technique in their daily work enable surgeons to adapt to the working environment in infected areas. As surgeons in the anti-pandemic front line in China, we describe our experience with the diagnosis and treatment of COVID-19 in this country and how the work of surgeons is unfolding during the pandemic.

Toll-like receptor 4 regulates spontaneous intestinal tumorigenesis by up-regulating IL-6 and GM-CSF.

Inflammation is as an important component of intestinal tumorigenesis. The activation of Toll-like receptor 4 (TLR4) signalling promotes inflammation in colitis of mice, but the role of TLR4 in intestinal tumorigenesis is not yet clear. About 80%-90% of colorectal tumours contain inactivating mutations in the adenomatous polyposis coli (Apc) tumour suppressor, and intestinal adenoma carcinogenesis in familial adenomatous polyposis (FAP) is also closely related to the germline mutations in Apc. The ApcMin/+ (multiple intestinal neoplasia) model mouse is a well-utilized model of FAP, an inherited form of intestinal cancer. In this study, ApcMin/+ intestinal adenoma mice were generated on TLR4-sufficient and TLR4-deficient backgrounds to investigate the carcinogenic effect of TLR4 in mouse gut by comparing mice survival, peripheral blood cells, bone marrow haematopoietic precursor cells and numbers of polyps in the guts of ApcMin/+ WT and ApcMin/+ TLR4-/- mice. The results revealed that TLR4 had a critical role in promoting spontaneous intestinal tumorigenesis. Significant differential genes were screened out by the high-throughput RNA-Seq method. After combining these results with KEGG enrichment data, it was determined that TLR4 might promote intestinal tumorigenesis by activating cytokine-cytokine receptor interaction and pathways in cancer signalling pathways. After a series of validation experiments for the concerned genes, it was found that IL6, GM-CSF (CSF2), IL11, CCL3, S100A8 and S100A9 were significantly decreased in gut tumours of ApcMin/+ TLR4-/- mice compared with ApcMin/+ WT mice. In the functional study of core down-regulation factors, it was found that IL6, GM-CSF, IL11, CCL3 and S100A8/9 increased the viability of colon cancer cell lines and decreased the apoptosis rate of colon cancer cells with irradiation and chemical treatment.

Critical role of toll-like receptor 4 (TLR4) in dextran sulfate sodium (DSS)-Induced intestinal injury and repair.

Ulcerative colitis2 (UC) is an inflammatory bowel disease3 (IBD) that causes long-lasting inflammation and ulcers in the human digestive tract. The repair function of TLR4 in the intestinal epithelium is still unknown. Here, wild-type4 (WT) mice, TLR4-knockout mice5 (KO; TLR4-/-) and commensal-depleted mice were used as dextran sulfate sodium6 (DSS)-induced or radiation-induced colitis and injury models to explore the role of TLR4 signaling in intestinal injury. Exogenous lipopolysaccharide7 (LPS) promoted DSS-induced inflammatory cytokines and aggravated intestinal damage. TLR4 deficiency and commensal bacterial depletion inhibited the toxic effects of LPS, but these mice were more susceptible to DSS-induced and radiation-induced intestinal damage. Compared with WT mice, neither DSS nor radiation promoted production of more inflammatory cytokines in the guts of TLR4-KO and commensal-depleted mice. Introducing the cytokine repair factors, PGE2 and GM-CSF, increased the cytokine levels in the guts of DSS-induced colitis mice. We hypothesized that TLR4 and its ligands repaired the epithelium after DSS-induced and radiation-induced intestinal damage by upregulating PGE2 and GM-CSF. Transwell migration assays suggested that LPS, IL6, TNF, PGE2 and GM-CSF promoted intestinal cell migration, and cell viability analysis suggested that these factors protected against radiation-induced intestinal damage. Our data underscore the importance of the balancing role of TLR4 in intestinal injury and repair.

Toll-like receptor 4 (TLR4) deficiency aggravates dextran sulfate sodium (DSS)-induced intestinal injury by down-regulating IL6, CCL2 and CSF3.

BACKGROUND: Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) that causes long-lasting inflammation and ulcers in the human digestive tract. The repair role of TLR4 in the intestinal epithelium is still unknown. METHODS: By comparing to wild-type (WT) mice, Toll-like receptor 4 (TLR4)-knockout mice (TLR4-KO) were used as dextran sulfate sodium (DSS)-induced colitis models to explore the role of TLR4 signaling in intestinal injury. High-throughput RNA-Seq, RT-qPCR and ELISA were performed to screen and verify key differences in gut genes between WT and TLR4-KO mice. Functional study of core dysregulated factors was performed in intestinal cell lines. RESULTS: We found that DSS-induced intestinal injury was aggravated by LPS (TLR4 agonist) and TLR4-KO. When compared to WT mice, IL6, CCL2, CSF3, IL11, Ccnb1, Ccnd1 and TNF-α significantly decreased and Fas and FasL have increased in the gut of TLR4-KO mice. IL6, CCL2, CSF3, Fas and FasL have all increased in CT-26 cells treated with LPS. Combined with the above data and KEGG enrichment, it can be assumed that TLR4-KO might aggravate DSS-induced intestinal damage by attenuating cell cycle, cytokine-cytokine receptor interaction, and Toll-like receptor signaling pathway, and enhancing the apoptosis pathway. In the functional study of core dysregulated factors, it was found that LPS, IL6, IL11, CSF3, CCL2, S100A8, S100A9 and Mmp3 have improved viability of colon cancer cell lines and decreased apoptosis rate of mouse colon cancer cells when these were treated with DSS. However, Jo-2 (Fas agonistic monoclonal antibody) played the opposite role in colon cancer cells treated with DSS. CONCLUSIONS: TLR4 had a repairing effect on DSS-induced intestinal damage and it up-regulate IL6, CCL2 and CSF3. Fas and FasL enhanced DSS-induced colon injury in mice, but might have little to do with TLR4 signaling.

Exploring the potential relationship between indoor air quality and the concentration of airborne culturable fungi: a combined experimental and neural network modeling study.

Indoor airborne culturable fungi exposure has been closely linked to occupants' health. However, conventional measurement of indoor airborne fungal concentration is complicated and usually requires around one week for fungi incubation in laboratory. To provide an ultra-fast solution, here, for the first time, a knowledge-based machine learning model is developed with the inputs of indoor air quality data for estimating the concentration of indoor airborne culturable fungi. To construct a database for statistical analysis and model training, 249 data groups of air quality indicators (concentration of indoor airborne culturable fungi, indoor/outdoor PM2.5 and PM10 concentrations, indoor temperature, indoor relative humidity, and indoor CO2 concentration) were measured from 85 residential buildings of Baoding (China) during the period of 2016.11.15-2017.03.15. Our results show that artificial neural network (ANN) with one hidden layer has good prediction performances, compared to a support vector machine (SVM). With the tolerance of ± 30%, the prediction accuracy of the ANN model with ten hidden nodes can at highest reach 83.33% in the testing set. Most importantly, we here provide a quick method for estimating the concentration of indoor airborne fungi that can be applied to real-time evaluation.

CD25 siRNA induces Treg/Th1 cytokine expression in rat corneal transplantation models.

Corneal graft rejection is the major reason for transplant failure. CD25 plays an important role in the induction of corneal graft rejection by regulating CD4(+) T cell function. Furthermore, CD25-mediated signaling is closely associated with the expression of Treg cytokines (IL-10, TGF-ß) and Th1 cytokines (IFN-γ, IL-1ß, and TNF-α). In the current study, corneal transplantation was performed on Wistar rats as donors and Sprague-Dawley rats as recipients. The survival curves indicated that CD25 siRNA treatment significantly prolonged graft survival time (mean survival time [MST], 14.8 ± 0.7 days) as compared with controls (MST, 7.6 ± 0.7 days; n = 12, p < 0.01). HE staining showed that CD25 siRNA alleviated inflammatory cell infiltration. At days 3, 7, 14, and 21, the mRNA and protein expression of CD25 in the CD25 siRNA groups were less than those of the control group, although the most significant decrease of CD25 protein was at day 3. The expression of IL-10 and TGF-ß in the CD25 siRNA group increased, while IFN-γ, IL-1ß, and TNF-α expression decreased, as well as no significant changes in Foxp3 expression were observed at day 14 post-operation. In conclusion, CD25 siRNA gene therapy played a protective role in corneal graft rejection via up-regulation of Treg cytokine expression and down-regulation of Th1 cytokine expression.

[Effect of milk enriched with phytosterol ester on blood cholesterol of patients with hypercholesterolemia: a randomized controlled trial].

OBJECTIVE: To study the effect of milk enriched with phytosterol ester on blood cholesterol. METHODS: Participants with hypercholesterolemia were recruited from community health center and randomly assigned to 3 groups: milk enriched with phytosterol ester group( PS group, n = 59), normal milk group( n = 58) and non-dairy group( n = 62). The intervention lasted for 2 months. At baseline, all subjects in the 3groups received health education on prevention and control of hypercholesterolemia. For PS and normal milk groups, subjects consumed 500 g milk per day, the intake ofphytosterol in PS group was 1. 58 g / d. For non-dairy group, subjects did not consume any dairy products during the trial. Subjects were assessed on their physical activity level and blood cholesterols were measured during monthly follow-up. RESULTS: Finally 157 subjects completed the trial. By the end of the first month, the TC and LDL-C levels of PS group were significantly lower than that of normal milk group. After adjustment, there was no significant difference between baseline and 1-month TC levels in PS group. The levels of TG and HDL-C in PS group were significantly increased while the LDL-C level was significantly decreased after 1-month intervention. Compared with normal milk and nondairy groups, no differences were observed for these four indicators. After 2-month intervention, the TC and LDL-C levels of PS group were significantly lower than that of normal milk and non-dairy groups. The levels of TC and LDL-C in PS group were significantly reduced compared to baseline levels after adjustment. TG level was increased while HDL-C level was unchanged. Compared with normal milk and non-dairy groups, the levels of TC and LDL-C in PS group were significantly declined while no significant difference was observed for TG and HDL-C levels. CONCLUSION: s Milk enriched with phytosterol ester has a notable effect on lowering TC and LDL-C levels in subjects with hypercholesterolemia.

Effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection.

BACKGROUND: Corneal graft rejection is the major cause of corneal failures. Previous studies have shown that the CD25 monoclonal antibody can inhibit corneal graft rejection during the acute phase of rejection in rat models. In the current study, we evaluated the safety and efficacy of the topical Entranster™ vector in rat corneal applications and further investigated the effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection. METHODS: Fluorescence detection, clinical assessment, hematoxylin and eosin (HE), terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL) and CD11b assays were used to evaluate the safety and efficacy of CD25siRNA gene transfer in normal SD rat corneas. Orthotopic corneal transplants were performed in alkali burned SD rats. Corneal recipients were divided into four groups that received different treatments. Clinical assessment, western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry, HE and transmission electron microscopy (TEM) were performed on all grafts. RESULTS: Low toxicity, no immunogenicity and high transfection efficiency were observed in rat corneas treated with the Entranster™ vector. Reduced endothelial cell apoptosis, inflammatory cell infiltration and graft neovascularisation were observed in the CD25siRNA treatment groups. The graft survival curves showed that CD25siRNA treatment significantly prolonged graft survival time, with better graft transparency and less graft oedema. Lower CD25 and higher IL-10 expression were detected in the CD25siRNA treatment groups during the study period, and a higher FOXP3 level was found in the CD25siRNA group than in the CD25siRNA-twice group on days 14 and 21 post-operation. CONCLUSIONS: The Entranster™ vector is an effective vector for corneal gene therapy. CD25 siRNA gene transfer inhibits corneal graft rejection via upregulation of anti-inflammatory molecule expression.

Apoptosis of multiple myeloma cells induced by agonist monoclonal antibody against human CD28.

CD28 is expressed abnormally on human multiple myeloma (MM) cells but the significance had not been identified until now. In this paper, we are suggesting that abnormal expression of CD28 might be a marker of tumour progression. We therefore took the approach of generating a hybridoma cell line capable of secreting agonist monoclonal antibody directed against human CD28 (agonist anti-CD28 mAb) and then determined the expression of CD28 molecules on the MM cell lines U266 and XG1. The biological effects of agonist anti-CD28 mAb on cell growth and proliferation of U266 and XG1 cell lines were then analysed. Our results showed that the expression of CD28 on U266 and XG1 was significantly higher than that of PBTC or Jurkat cells. We found that by adding the agonist anti-CD28 mAb to cultures of U266 and XG1 cells their rate of growth and proliferation was obviously inhibited. Further morphological and molecular analyses found that U266 and XG1 incubated with agonist anti-CD28 mAb showed signs of nuclear condensation, chromatin marginal changes, cells membrane breaking, and cytoplasmic shrinkage. Vacuoles and apoptotic bodies were also observed using a transmission electron microscope and the development of typical DNA laddering patterns were found by the use of electrophoresis assays, suggesting that U266 and XG1 cells were undergoing apoptosis induced by agonist anti-CD28 mAb in vitro.