Label-free 3D molecular imaging of living tissues using Raman spectral projection tomography.

The ability to image tissues in three dimensions (3D) with label-free molecular contrast at the mesoscale would be a valuable capability in biology and biomedicine. Here, we introduce Raman spectral projection tomography (RSPT) for volumetric molecular imaging with optical sub-millimeter spatial resolution. We have developed a RSPT imaging instrument capable of providing 3D molecular contrast in transparent and semi-transparent samples. We also created a computational pipeline for multivariate reconstruction to extract label-free spatial molecular information from Raman projection data. Using these tools, we demonstrate imaging and visualization of phantoms of various complex shapes with label-free molecular contrast. Finally, we apply RSPT as a tool for imaging of molecular gradients and extracellular matrix heterogeneities in fixed and living tissue-engineered constructs and explanted native cartilage tissues. We show that there exists a favorable balance wherein employing Raman spectroscopy, with its advantages in live cell imaging and label-free molecular contrast, outweighs the reduction in imaging resolution and blurring caused by diffuse photon propagation. Thus, RSPT imaging opens new possibilities for label-free molecular monitoring of tissues.

[Multimodal imaging features of acute macular retinopathy in patients with COVID-19].

Objective: To investigate the multimodal imaging characteristics of acute macular retinopathy (AMR) and/or parafoveal acute middle maculopathy (PAMM) in patients with coronavirus disease 2019 (COVID-19). Methods: It was a cross-sectional study. Eight patients (15 eyes) diagnosed with AMN and/or PAMM, who presented for their initial visit at Kaifeng Eye Hospital between December 17 and December 31, 2022 and were also confirmed positive for COVID-19, were enrolled as the observation group. The patients were classified into four types based on swept-source optical coherence tomography (SS-OCT) findings. Fifteen healthy volunteers (15 eyes) without ocular or systemic diseases were recruited as the healthy control group, and one eye was randomly selected for analysis. All participants underwent detailed ophthalmic examinations, including best-corrected visual acuity (BCVA), slit-lamp biomicroscopy, fundus photography (FP), intraocular pressure measurement, fundus infrared imaging, OCT and OCT angiography (OCTA). The foveal avascular zone (FAZ) area of the macular center was measured. General information and multimodal imaging findings were collected and analyzed. The superficial capillary plexus vessel density (SCP-VD) and deep capillary plexus vessel density (DCP-VD) were measured in circular areas with diameters of 1.0 mm, >1.0 mm and ≤3.0 mm, and>3.0 mm and ≤6.0 mm centered on the foveal center, recorded as SCP-VD1.0, 3.0, 6.0 and DCP-VD1.0, 3.0, 6.0. Statistical analyses were performed using t-tests, Mann-Whitney U tests, and chi-square tests. Results: The observation group consisted of 6 males (11 eyes) and 2 females (4 eyes) with a mean age of (26.87±11.56) years. The healthy control group included 11 males (11 eyes) and 4 females (4 eyes) with a mean age of (28.75±12.30) years. There were no statistically significant differences in age and gender distribution between the two groups (all P>0.05). All patients in the observation group experienced high fever (≥39.0 ℃) and developed ocular symptoms during the febrile period or within 24 hours after fever resolution. Among all patients, there were 5 cases (7 eyes) of Type Ⅰ, 1 case (1 eye) of Type Ⅱ, 3 cases (4 eyes) of Type Ⅲ, and 2 cases (3 eyes) of Type Ⅳ. In Type Ⅲ and Ⅳ, 3 cases (4 eyes) exhibited weakly reflective cystic spaces in the outer plexiform or outer nuclear layers, and fundus photography revealed multiple gray or reddish-brown lesions in the macular region. One case (1 eye) showed retinal superficial hemorrhage. Cotton wool spots were observed in 2 cases (4 eyes). Fundus infrared imaging showed that Type Ⅰ manifested as weak reflectivity lesions in the parafoveal central zone, with the tip pointing towards the fovea. Type Ⅱ showed no apparent abnormalities in the macular region, while Type Ⅲ and Ⅳ displayed map-like weak reflective lesions spanning the foveal center. OCTA findings demonstrated that SCP-VD1.0 in the observation group was 6.93% (4.77%, 6.93%), significantly lower than the healthy control group's 10.66% (8.05%, 10.55%) (U=174.00, P=0.016). SCP-VD3.0 in the observation group was 37.14% (32.15%, 43.48%), also lower than the healthy control group's 43.06% (38.95%, 46.55%) (U=174.00, P=0.016). DCP-VD3.0 in the observation group was 48.20% (46.11%, 50.33%), lower than the healthy control group's 51.10% (50.04%, 53.02%) (U=188.00, P=0.009). DCP-VD6.0 in the observation group was 49.27% (47.26%, 51.67%), lower than the healthy control group's 52.43% (50.07%, 53.82%) (U=70.00, P=0.004). There were no significant differences in SCP-VD6.0 and DCP-VD1.0 between the two groups (both P>0.05). Conclusions: Acute macular retinopathy in patients with COVID-19 can involve all retinal layers and present as segmental hyper-reflectivity on SS-OCT. Fundus infrared imaging reveals weak reflectivity in the affected area, fundus photography shows multiple gray or reddish-brown lesions in the macular region, and OCTA demonstrates a decrease in SCP-VD and DCP-VD.

[Value of glycosylated hemoglobin A1c and apolipoprotein A-1 ratio on predicting outcome of patients with acute coronary syndrome].

Objective: To investigate the predictive value of glycosylated hemoglobin A1c/apolipoprotein A-1 (HbA1c/ApoA-1) ratio for major adverse cardiovascular events (MACEs) in patients with acute coronary syndrome (ACS). Methods: The present study is a retrospective cohort study. ACS patients who were hospitalized and underwent coronary angiography at Beijing Hospital from March 2017 to March 2019 were enrolled. Baseline information such as sex, age, previous history, Gensini score, HbA1c and ApoA-1 were analyzed. Patients were divided into two groups according to presence or absence of MACEs and the difference on HbA1c/ApoA-1 ratio was compared between the two groups. According to the tertiles of HbA1c/ApoA-1 levels, patients were divided into high (5.87-16.12), medium (4.50-5.83) and low (2.11-4.48) HbA1c/ApoA-1 groups. Cox proportional risk model was used to evaluate the differences in MACEs and all-cause mortality among the three groups. Kaplan-Meier survival analysis was used to compare the differences of MACEs between the various HbA1c/ApoA-1 groups. Results: A total of 366 ACS patients were included in this study. The mean age of the patients was (65.9±10.3) years. There were 59 MACEs and 10 all-cause deaths during the mean of (22.3±4.4) months follow-up. After adjusting for age, systolic blood pressure, history of diabetes and Gensini score, the incidence of MACEs was 2.45 times higher in the high HbA1c/ApoA-1 group than in the low HbA1c/ApoA-1 group (95%CI 1.16-5.18, P=0.019). There was no significant difference in all-cause mortality between the high and low HbA1c/ApoA-1 groups (P=1.000). Kaplan-Meier survival analysis showed that patients in the high HbA1c/ApoA-1 group had the highest risk of MACEs, while patients in the low HbA1c/ApoA-1 group had the lowest risk of MACEs (P<0.01). Spearman rank correlation analysis showed that HbA1/ApoA-1 ratio was positively correlated with Gensini score in ACS patients (r=0.274, P<0.01). Conclusion: High HbA1c/ApoA-1 ratio was an independent risk factor for MACEs in ACS patients. Patients with high HbA1c/ApoA-1 ratio had more severe coronary artery disease lesions. HbA1c/ApoA-1 ratio may be used as a potential risk stratification biomarker for ACS patients, it might be useful for the early identification of high-risk population and for predicting the incidence of MACEs among ACS patients.

Investigating the effect of graphene oxide in chitosan/alginate-based foams on the release and antifungal activity of clotrimazole in vitro.

Polyelectrolyte complexes (PECs) have been used as the matrix of solid foams for drug delivery. This study aimed at investigating the effect of graphene oxide (GO) and the composition of excipients in chitosan/alginate-based buccal foams on the clotrimazole release and antifungal activities. The investigation has been focused on the interactions of the drug with excipients in the foams, and the changes of ionization degree upon exposure to various media are discussed. The solid foams were prepared by mixing the excipients and clotrimazole via probe sonication, followed by a freeze-drying method. The pH values of the formulations were measured during the foam preparation process to estimate the ionization degree of clotrimazole and the other excipients. The foam matrix was the PECs between the cationic chitosan and anionic alginate. The mechanical strength of clotrimazole-loaded foams was lower than that of drug-free foams due to the positively charged clotrimazole interacting with the anionic alginate and interfering the PECs between chitosan and alginate. Addition of GO in the clotrimazole-loaded matrix made the foams mechanically stronger and contributed to a faster release of clotrimazole from the buccal foams by disrupting the electrostatic interactions between alginate and clotrimazole. However, addition of 1 wt% GO in the formulations didn't affect the antifungal activity of clotrimazole-loaded foams significantly. A lower amount GO in the formulation may be required for enhancing the antifungal effect, which should be further investigated in future.

[T lymphocyte subsets combined with inflammatory indicators for auxiliary diagnosis and prognosis evaluation of sepsis].

Objective: To explore the application value of T lymphocyte subsets combined with procalcitonin (PCT), C-reactive protein (CRP), neutrophil to lymphocyte ratio (NLR) and white blood cell count (WBC) in the auxiliary diagnosis and prognosis evaluation of sepsis. Methods: In a retrospective study, seventy-two patients with sepsis diagnosed and treated in Tianjin First Central Hospital from June 2018 to April 2021 were selected as the research objects, and included in the sepsis group were 46 males and 26 females, aged 68 (57.3, 80.3) years. In addition, 111 patients with local infection admitted to hospital during the same period were included in the local infection group, including 62 males and 49 females, aged 68 (51, 77) years. Sepsis patients were divided into survival group (43 cases) and death group (29 cases) according to the 28-day outcome. CD3+, CD4+, CD8+, CD4+/CD8+ ratio were detected by flow cytometry within 24 h after admission, PCT was detected by ELISA, CRP was detected by immunoturbidimetry, blood routine examination, blood lactic acid (Lac) and oxygen partial pressure (PO2) were detected by instrumental method. Multivariate Logistic regression analysis was used to evaluate the correlation between each indicator and sepsis, and receiver operating characteristic curve (ROC) was drawn to evaluate the diagnostic value of each indicator for sepsis. Multivariate Logistic regression analysis and Kaplan Meier survival analysis were used to evaluate the prognostic value of each index for patients with sepsis. Results: Peripheral blood CD3+, CD4+, CD8+, CD4+/CD8+ ratio and PLT in sepsis group were significantly lower than those in local infection group(Z=-8.184,P<0.001;Z=-7.210,P<0.001;Z=-5.936,P<0.001;Z=-2.700,P=0.007;Z=-6.381,P<0.001); PCT, CRP, NLR and Lac levels were significantly higher than those in local infection group(Z=-8.262,P<0.001;Z=-3.094,P=0.002;Z=-9.004,P<0.001;Z=-4.770,P<0.001). Multivariate Logistic regression model showed that PCT, NLR, CD3+, CD8+, CD4+/CD8+ were independent risk factors for sepsis. According to ROC curve analysis, AUC of sepsis patients diagnosed by each indicator were 0.862, 0.894, 0.858, 0.760 and 0.618, respectively. The cut-off values were 3.075 ng/ml, 10.715, 44.935×109/L, 27.463×109/L and 0.750, respectively. The NLR sensitivity was 80.6%, and the CD3+ specificity was 94.6%. The AUC of combined detection of PCT and NLR was 0.947, sensitivity was 87.5% and specificity was 91.9%. The combined detection AUC of PCT, NLR, CD3+, CD4+/CD8+ was 0.958, the sensitivity and specificity were 90.3% and 91.0% respectively(P<0.001). PCT and Lac in death group were significantly higher than those in survival group(Z=-2.302,P=0.021;Z=-3.095,P=0.002);Peripheral blood CD4+/CD8+ levels were significantly lower than those in survival group(Z=-3.691,P<0.001),Multivariate Logistic regression model showed that CD4+/CD8+ ratio was an independent risk factor for 28 d mortality in patients with sepsis (P<0.001). The ROC curve showed that the AUC was 0.758, and the Youden index reached the maximum when the cut-off value was 1.27, the sensitivity and specificity were 79.3% and 60.5%, respectively. Compared with patients with CD4+/CD8+ ≥1.27, 28-day mortality was significantly increased in patients with CD4+/CD8+<1.27 (P=0.032). Conclusion: The combined detection of PCT, NLR, CD3+ and CD4+/CD8+ can improve the auxiliary diagnostic efficiency of sepsis, and the ratio of CD4+/CD8+ in peripheral blood may have certain predictive value for the prognosis of sepsis.

Aberrant Expression of a Proliferation-Inducing Ligand Underlies Autoimmune Mechanisms in Immune Thrombocytopenia.

PURPOSE: To study the relationship between surface membrane-bound APRIL and ITP. METHODS: The peripheral blood of all subjects, 50 patients diagnosed with ITP and 25 healthy controls, was collected. Flow cytometry was used to detect the expression of membrane-bound APRIL on immune cells and platelets. ELISA was used to detect the content of soluble APRIL in plasma. RESULTS: Membrane-bound APRIL was only expressed on the surface of platelets in both ITP patients and controls. APRIL expression on the platelet surface was significantly lower in newly diagnosed (P < 0.001) and chronic (P < 0.001) ITP patients than in controls. Platelet surface APRIL level was significantly enhanced in patients with complete remission after treatment (P = 0.02) but not in those with no response after treatment. Platelet surface APRIL level in ITP patients was negatively correlated with serum APRIL level (r = -0.09765, P = 0.0424). CONCLUSIONS: Platelet surface APRIL may play a key immunoregulative role. Platelet surface APRIL is likely to be one source of the excessive serum APRIL in ITP patients. The effectiveness of treatment may be measured by determining the platelet surface APRIL levels in ITP patients.

[Role of imbalance of M1/M2 subsets of bone marrow macrophages in the pathogenesis of immune-mediated aplastic anemia in mice].

Objective: To investigate the role of macrophages (Mø) in the pathogenesis of modified immune-mediated aplastic anemia (AA) mice model. Methods: Before the establishment of the F1 AA mice model by total-body irradiation combined with allogeneic lymphocyte infusion, the mice of the CLO+AA group were treated with clodronate (CLO) liposomes to remove macrophages, and those of the PBS+AA group were treated with phosphate-buffered saline (PBS) liposomes and used as control. The severity of AA was observed by bone marrow (BM) pathological examination and peripheral blood cell count. Flow cytometry (FCM) was used to detect the CD4(+)/CD8(+) T lymphocyte subsets in the BM and Mø subsets in the BM and spleen of each group. The levels of IFN-γ, TNF-α, G-CSF, GM-CSF, EPO, and TPO in the peripheral blood were detected using enzyme-linked immunosorbent assay. Finally, the relationships between inflammatory factors and Mø subsets were analyzed. Results: The BM fatty conversion of mice in the CLO+AA group was significantly alleviated compared with the PBS+AA group. Hemoglobin counts were (91.50±31.63) and (110.65±24.15) g/L, respectively, and the platelet counts were (90.85±121.90) × 10(6)/L and (461.13±483.45) ×10(6)/L, respectively. The differences were all statistically significant (all P<0.05) . After removing macrophages, the proportions of CD4(+) and CD8(+) T lymphocytes in BM of mice in the CLO+AA group decreased, but the reduction of CD8(+) T cells was more significant. The proportions of CD4(+) T cells and CD8(+) T cells in BM of the PBS+AA group were (18.5±10.17) % and (36.23±6.40) %, respectively, and in the CLO+AA group were (7.58±8.00) % and (6.67±5.78) %, respectively. Similarly, the percentage of macrophages in the spleen and BM in the CLO+AA group was significantly reduced compared with the PBS+AA group, most of which were M1 macrophages (P<0.05) . The levels of IFN-γ in peripheral blood of the PBS+AA and CLO+AA groups were (602.37±104.62) ng/L and (303.01±87.22) ng/L, respectively, the levels of TNF-α were (34.46±1.42) ng/L and (23.25±4.21) ng/L, respectively, the levels of GM-CSF were (9.32 ± 2.00) ng/L and (64.85±12.25) ng/L, respectively, the levels of G-CSF were (5 891.78±2 632.39) ng/L and (17 784.16±488.36) ng/L, respectively, the levels of EPO were (9 667.31±4 501.95) ng/L and (2 078.02±897.56) ng/L, respectively, and the levels of TPO were (6.36±2.09) ng/L and (11.67±2.86) ng/L, respectively (all P<0.05) . Conclusions: This study confirmed that macrophages were involved in the pathogenesis of AA, and the degree of BM damage in AA mice was improved by removing macrophages in advance. The imbalance of M1/M2 macrophages and the changes of IFN-γ and TNF-α may be important mechanisms that eventually lead to AA.

Graphene oxide as a functional excipient in buccal films for delivery of clotrimazole: Effect of molecular interactions on drug release and antifungal activity in vitro.

Graphene oxide (GO) is an amphiphilic, high surface area material with great potential as a functional excipient in drug delivery. The present study aimed at incorporating GO in buccal polyelectrolyte films for delivery of antifungal drugs and investigating the effect of GO on the film properties and drug release profiles, as well as antifungal activities. Mucoadhesive excipients chitosan and alginate were selected to form polyelectrolyte films with the antifungal drug clotrimazole. The buccal formulations were prepared by mixing the excipients and clotrimazole via probe sonication, followed by film casting and drying. Inclusion of GO in the formulations increased clotrimazole release from the films in vitro (pH 6.8), possibly due to GO altering the electrostatic interactions between chitosan and alginate. An increase of in vitro activity against Candida albicans was observed when 0.04 wt% GO was added in the formulation containing clotrimazole. However, when the GO amount increased to 0.09 wt%, the films had similar antifungal ability to the films with 0.04 wt% GO, suggesting that the electrostatic and hydrophobic interactions between GO and clotrimazole also affects the antifungal effect of clotrimazole. In summary, GO has a great potential as a functional excipient for delivery of antifungal drugs.

MiR-182 affects renal cancer cell proliferation, apoptosis, and invasion by regulating PI3K/AKT/mTOR signaling pathway.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiR-182 affects renal cancer cell proliferation, apoptosis, and invasion by regulating PI3K/AKT/mTOR signaling pathway, by J.-H. Fu, S. Yang, C.-J. Nan, C.-C. Zhou, D.-Q. Lu, S. Li, H.-Q. Mu, published in Eur Rev Med Pharmacol Sci 2018; 22 (2): 351-357-DOI: 10.26355/eurrev_201801_14179-PMID: 29424922" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/14179.

Clinicopathological and prognostic value of hypoxia-inducible factor-1α in breast cancer: a meta-analysis including 5177 patients.

PURPOSE: Mounting studies have investigated the clinicopathological and prognostic value of hypoxia-inducible factor-1α (HIF-1α) in breast cancer (BC), yet conclusions remain controversial. Therefore, we conducted this meta-analysis to clarify this issue. METHODS: All relevant studies were searched using Cochrane Library, Web of Science, PubMed, and EMBASE online databases. Pooled odds ratios (ORs) and hazard ratios (HRs) with 95% confidence intervals (CIs) were applied to evaluate the clinicopathological and prognostic value of HIF-1α, respectively. Subgroup analysis and sensitivity analysis were performed to investigate heterogeneity and stability of the results. Begg's funnel plot and Egger's test were used to examine publication bias. RESULTS: A total of 31 eligible studies including 5177 subjects were enrolled. Of these, 25 studies assessed the prognostic role of HIF-1α and included 4546 individuals. Twenty-three studies involving 3277 individuals evaluated the clinicopathological significance of HIF-1α. High expression level of HIF-1α was correlated with poor overall survival (OS) (HR = 1.59, 95% CI = 1.40-1.80, P < 0.001), disease-free survival (DFS) (HR = 1.87, 95% CI = 1.53-2.28, P < 0.001), relapse-free survival (HR = 1.36, 95% CI = 1.07-1.73, P = 0.001), and cancer-specific survival (HR = 1.55, 95% CI = 1.10-2.19, P = 0.012). Pooled data from studies using multivariate survival analysis also showed that HIF-1α expression was associated with worse OS (HR = 1.59, 95% CI = 1.32-1.92, P < 0.001) and DFS (HR = 1.60, 95% CI = 1.39-1.84, P < 0.001). Additionally, high HIF-1α expression was associated with advanced tumor-node-metastasis stage, positive lymph-node status, negative ER status, ductal type, advanced histologic grade, high Ki67 expression, and strong VEGF expression. CONCLUSION: HIF-1α might serve as an independent prognostic biomarker and a promising therapeutic target for BC. Future large-scale prospective randomized trials are needed to confirm our findings.

MiR-130b/TNF-α/NF-κB/VEGFA loop inhibits prostate cancer angiogenesis.

BACKGROUND: Angiogenesis is a critical biological process essential for solid cancer growth and metastasis. It has been shown that microRNAs (miRNAs) play a vital role in a variety of biological processes in cancers. However, whether miR-130b is involved in prostate cancer angiogenesis remains ill-defined. METHODS: We performed the miRNA microarray to analyze miRNA expression in human prostate cancer specimens. In vitro gain-of-function assays and loss-of-function assays were conducted to explore the potential functions of miR-130b in human prostate cancer cells. Correlation analysis and dual-luciferase reporter assay were performed to validate whether tumor necrosis factor-α (TNF-α) was a direct target of miR-130b. The Matrigel plug and tumor vascular imaging assays were performed to confirm the anti-angiogenic activity of miR-130b in nude mice. RESULTS: We found that miR-130b was one of the miRNAs being most significantly downregulated. Subsequently, we found that miR-130b expression was markedly downregulated in human prostate cancer cell lines. Down-regulation of miR-130b in prostate cancer cells significantly promoted the proliferation, invasion and tubule formation of human umbilical vein endothelial cells (HUVECs), while ectopic expression of miR-130b blocked prostate cancer angiogenesis in vitro and in vivo. Mechanistic analyses indicated that tumor necrosis factor-α (TNF-α) was regulated by miR-130b directly. MiR-130b attenuated nuclear factor-κB (NF-κB) signaling and its downstream gene vascular endothelial growth factor-A (VEGFA) by directly inhibiting TNF-α expression. Additionally, subsequent investigations identified that the ectopic level of VEGFA markedly abrogated the anti-angiogenic effect induced by miR-130b. Interestingly, VEGFA could in turn decrease the expression of miR-130b, thus forming a negative feedback loop that drives the angiogenesis of prostate cancer. CONCLUSION: These findings show that miR-130b/TNF-α/NF-κB/VEGFA feedback loop is significantly correlated with angiogenesis in prostate cancer and miR-130b could be regarded as potential therapeutic target for prostate cancer anti-angiogenesis treatment.

[Evaluation of wear property of Giomer and universal composite in vivo].

OBJECTIVE: To observe the wear performance of Giomer and universal composite for posterior restorations by 3D laser scan method, in order to guide the material selection in clinic. METHODS: In this study, 48 patients (108 teeth) were selected according to the inclusion and exclusion criteria. All the patients in need of a minimum of 2 Class Ⅰ and/or Class Ⅱ restorations were invited to join the study. The teeth were restored with Giomer (Beautifil Ⅱ, BF) and universal composite (Filtek Z350, Z350) randomly. The restorations were evaluated at baseline and after 6-, 18-, 48-month using the modified United States Public Health Service (USPHS) criteria for clinical performance. The in vivo images and gypsum replicas were taken at each recall. A 3D-laser scanner and Geomagic Studio 12 were used to analyze the wear depth quantitatively. Statistical analysis was performed with SPSS 20.0. RESULTS: After 4 years, 89.6% patients were recalled. The survival rate of both materials was 95.8% (Kaplan-Meier survival analysis). Seven restorations of the two materials failed due to loss of restoration, bulk fracture, secondary caries and pulp necrosis. The wear patterns of restorations were divided into 2 classes. Pattern Ⅰ: occlusal contact areas showed the deepest and fastest wear depth; pattern Ⅱ: the wear depth was slow and uniform. Both materials showed a rapid wear in the first 6 months. Then the wear rate was decreased. The occlusal wear depth after 4 years were (58±22) µm and (54±16) µm for BF group and Z350 group respectively, which were in accordance with the American Dental Association (ADA) guidelines (wear depth for 3 years < 100 µm). No significant differences (P>0.05) were observed between the two groups. Regarding the restorations with wear pattern Ⅰ, the wear depth of BF group was higher than Z350 group at 6- and 48-month (P < 0.05), while there was no significant difference between restorations with wear pattern Ⅱ (P>0.05). CONCLUSION: Within the limitation of the study, after 4 years, the survival rate and wear resistance of Giomer met ADA guidelines for tooth-colored restorative materials for posterior teeth. When the two materials were applied in occlusal contact areas, wear resistance of Giomer was slightly lower than universal composite resin. No significant difference was found when they were applied in none of the occlusal contact areas.

Role of Hsa-miR-122-3p in steroid-induced necrosis of femoral head.

OBJECTIVE: To explore the clinical correlation between the hsa-miR-122-3p expression in bone marrow mesenchymal stem cells (BMSCs) and steroid-induced necrosis of femoral head (SONFH). PATIENTS AND METHODS: A total of 62 SONFH patients were selected as the experimental group, while another 72 patients with femoral neck fracture (FNF) were selected as the control group. The bone marrow was obtained from patients during operation and used to culture the BMSCs. The expression of hsa-miR-122-3p in BMSCs was detected via real-time quantitative polymerase chain reaction (qPCR) in both groups. The patients in experimental group were further divided into Ficat stage III group and stage IV group according to the Ficat stage, and the expression of hsa-miR-122-3p in BMSCs was also detected via qPCR in the two groups. RESULTS: The expression level of hsa-miR-122-3p in SONFH group was significantly lower than that in FNF group, and the difference was statistically significant (p<0.05). The expression level of hsa-miR-122-3p in Ficat stage IV group was significantly lower than that in stage III group (p<0.05). CONCLUSIONS: We demonstrated that the expression of hsa-miR-122-3p in BMSCs declined in SONFH group, indicating that hsa-miR-122-3p may be involved in the regulation of the pathological process of SONFH, and the expression level of hsa-miR-122-3p in BMSCs may be correlated with the progression of SONFH.

CD18 inhibits progression of kidney cancer by down-regulating Treg cell levels.

OBJECTIVE: Gene mutation is closely related to the occurrence of tumor. Renal cell carcinoma is a malignant tumor, seriously threatening patients' life quality. Regulatory T cells (Treg) play important roles in the development of several cancers. This study aimed to investigate whether CD18 affects renal carcinoma cell proliferation. MATERIALS AND METHODS: Thirty mice with renal cell carcinoma were constructed using gene-engineering mouse with CD18 deficiency, and another 30 normal C57 mice were used as control. Ki67 and micro-vessel density were detected by using immunohistochemistry (IHC) and immunofluorescence, respectively. The expression of CD3, CD4 and CD8 were detected in blood and spleen by quantitative PCR (q-PCR). Flow cytometry was used to detect the changes of Treg cells. RESULTS: The expression of Ki67 in C57 was significantly higher than that in CD18-/- mice (p<0.05). IHC results showed that CD31 was also significantly downregulated in CD18-/- group compared to control group (p<0.05). It was found that only high expression of CD4 in mesenteric lymph nodes of CD18-/- was considered as non-tumor-bearing. Flow cytometry results showed that Treg cells were significantly decreased in CD18-/- compared to C57 group (p<0.05). CONCLUSIONS: CD18-/- down-regulates Treg cells and inhibits the pathogenesis of renal cell carcinoma.

Synergistic antibacterial effects of Buddleja albiflora metabolites with antibiotics against Listeria monocytogenes.

Some plants of Buddleja genus possessed antimicrobial activity. In this study, the antibacterial activities of nine compounds from Buddleja albiflora were evaluated against Listeria monocytogenes, Staphyloccocus aureus, Escherichia coli and Pseudomonas aeruginosa, either alone or in combination with Erythromycin (Ery), Gentamicin (Gen) and Ampicillin (Amp). Checkerboard assay demonstrated that Kumatakenin elicited synergistic effects with all three antibiotics, and chrysin displayed synergistic effect with Ery against L. monocytogenes. Further experiments showed that the synergy effect was sufficient to eradicate biofilms formed by L. monocytogenes. Thus, our data highlighted that the combinations of secondary metabolites from B. albiflora and specific antibiotics were useful for the treatment of pathogens, which might help prevent the spread of antibiotic resistance through improving antibiotic effectiveness. SIGNIFICANCE AND IMPACT OF THE STUDY: This study measured the antibacterial activity of the metabolites from Buddleja albiflora and the synergistic interactions with antibiotics against Listeria monocytogenes on planktonic cells as well as on biofilms. The results of this study provide valuable evidence that Kumatakenin/antibiotic combination can be considered as a promising antimicrobial agent for the treatment of pathogens, which subsequently help prevent the spread of antibiotic resistance through improving antibiotic effectiveness.

[Association between fatty liver and type 2 diabetes in the baseline population of Jinchang Cohort].

Objective: To explore the association between fatty liver and type 2 diabetes mellitus (T2DM) in the baseline-population of Jinchang cohort study. Methods: Data from all the participants involved in the baseline-population of Jinchang cohort study was used, to compare the risks of T2DM in fatty liver and non fatty liver groups and to explore the interaction between family history or fatty liver of diabetes and the prevalence of T2DM. Results: Among all the 46 861 participants, 10 574 were diagnosed as having fatty liver (22.56%), with the standardized rate as 20.66%. Another 3 818 participants were diagnosed as having T2DM (8.15%) with standardized rate as 6.90%. The prevalence of T2DM increased in parallel with the increase of age (trend χ(2)=2 833.671, trend P<0.001). The prevalence of T2DM in the fatty liver group was significantly higher than that in the non-fatty liver group, both in men or women and in the overall population. Compared with the group of non-fatty liver, the risks of T2DM in fatty liver group were seen 1.78 times higher in males, 2.33 times in women and 2.10 times in the overall population, after adjustment for factors as age, levels of education, smoking, drinking, physical exercise, BMI, family history of diabetes and some metabolic indicators (pressure, TC, TG, uric acid, ALT, AST, gamma-glutamyl transferase). Date from the interaction model showed that fatty liver and family history of diabetes present a positive additive interaction on T2DM (RERI=1.18, 95%CI: 0.59-1.78; AP=0.24, 95%CI: 0.14-0.34; S=1.43, 95%CI: 1.21-1.69). Conclusions: Fatty liver could significantly increase the risk of T2DM and a positive additive interaction was also observed between fatty liver and family history of diabetes on T2DM. It was important to strengthen the prevention program on T2DM, in order to effectively control the development of fatty liver.

MiR-182 affects renal cancer cell proliferation, apoptosis, and invasion by regulating PI3K/AKT/mTOR signaling pathway.

OBJECTIVE: PI3K/AKT/mTOR signaling pathway plays a crucial role in tumorigenesis and development. It was shown that mTOR overexpression was associated with the pathogenesis of renal cancer. Down-regulation of MiR-182 was found in renal carcinoma tissue. This study thus aims to investigate the influence of miR-182 in regulating mTOR expression and renal carcinoma cell proliferation, invasion, and apoptosis. PATIENTS AND METHODS: The targeted regulatory relationship between miR-182 and mTOR was tested by dual luciferase assay. Renal carcinoma tissue and benign renal tissue were collected to detect miR-182 and mTOR expressions. MiR-182, mTOR, p-mTOR, and Survivin levels were compared between HK-2 and A498 cells. Renal carcinoma A498 cells were divided into four groups, including miR-NC, anti-miR-182 mimic, si-NC, and si-mTOR groups. Cell apoptosis and proliferation were evaluated by flow cytometry. Cell invasion was determined by transwell assay. RESULTS: Bioinformatics analysis revealed the complementary relationship between miR-182 and the 3'-UTR of mTOR mRNA. The level of miR-182 was significantly reduced, while mTOR expression was upregulated in renal carcinoma tissue compared with that in benign lesion, which was associated with TNM stage. MiR-182 expression was markedly declined, whereas mTOR, p-mTOR, and Survivin levels were apparently upregulated in A498 cells compared with that in HK-2 cells. The treatment of miR-182 mimic or si-mTOR transfection significantly downregulated mTOR, p-mTOR, and Survivin expressions, restrained cell proliferation and invasion, and enhanced cell apoptosis. CONCLUSIONS: The decreasing level of miR-182 plays a role in enhancing mTOR expression and promoting renal carcinoma pathogenesis. Overexpression of miR-182 inhibited mTOR expression and weakened cell proliferation and invasion, which provides leads to the future therapy of renal cancer.

Combined inhibition of ß-catenin and Bcr-Abl synergistically targets tyrosine kinase inhibitor-resistant blast crisis chronic myeloid leukemia blasts and progenitors in vitro and in vivo.

Tyrosine kinase inhibitor (TKI) resistance and progression to blast crisis (BC), both related to persistent ß-catenin activation, remain formidable challenges for chronic myeloid leukemia (CML). We observed overexpression of ß-catenin in BC-CML stem/progenitor cells, particularly in granulocyte-macrophage progenitors, and highest among a novel CD34+CD38+CD123hiTim-3hi subset as determined by CyTOF analysis. Co-culture with mesenchymal stromal cells (MSCs) induced the expression of ß-catenin and its target CD44 in CML cells. A novel Wnt/ß-catenin signaling modulator, C82, and nilotinib synergistically killed KBM5T315I and TKI-resistant primary BC-CML cells with or without BCR-ABL kinase mutations even under leukemia/MSC co-culture conditions. Silencing of ß-catenin by short interfering RNA restored sensitivity of primary BCR-ABLT315I/E255V BC-CML cells to nilotinib. Combining the C82 pro-drug, PRI-724, with nilotinib significantly prolonged the survival of NOD/SCID/IL2Rγ null mice injected with primary BCR-ABLT315I/E255V BC-CML cells. The combined treatment selectively targeted CML progenitors and inhibited CD44, c-Myc, survivin, p-CRKL and p-STAT5 expression. In addition, pretreating primary BC-CML cells with C82, or the combination, but not with nilotinib alone, significantly impaired their engraftment potential in NOD/SCID/IL2Rγ-null-3/GM/SF mice and significantly prolonged survival. Our data suggest potential benefit of concomitant ß-catenin and Bcr-Abl inhibition to prevent or overcome Bcr-Abl kinase-dependent or -independent TKI resistance in BC-CML.

[The preliminary study of stimulus input temporal changes on the visual cortex of rats at different ages].

Objective: To observe the impact of the input temporal changes on visual cortex of rats cells and the change of the synaptic efficacy, for the study of visual developmental plasticity mechanism. Methods: Experimental research. The brain slice of ten 14d healthy Wistar rats and ten 21d healthy Wistar rats were recorded with whole cell recording technique, using single stimulation and combined stimulate model, to observe the visual cortex of rats neurons changes in synaptic activity. Change the stimulus input sequence, and observe the visual cortex of rats changes of synaptic efficacy. Using the paired t test to compare the change of excitatory postsynaptic potential (EPSC) of visual cortex. Result: The difference between single stimulation and combined stimulate about EPSC was statistically significant, which was decreased (14.3±7.4) % (n=15) in single stimulation and (53.4±17.5) % (n=20) in combined stimulation for P14 rats which were long-term depression (ts2=3.9, ts1+s2=2.2; P<0.05) , and was increased (27.5±11.4) % (n=16) in single stimulation and (34.6±10.3) % (n=10) in combined stimulation for P20 rats which were long-term potentiation (ts2=2.3, ts1+s2=3.5; P<0.05) . Rats in different development period have a specific time window for input temporal changes. Combined stimulation patterns produced by the neuron cell reaction were not just an accumulation of simple response caused by single stimulation. For P14 rats, its time window was of about ±0.5ms. However, for P20 rats, scope of time window reduced to ±0.1ms. Conclusions: Different development period of rats, change the stimulation pattern can cause the change of the visual cortex synapses reaction, and stimulate the temporal change within a specific time window to producenonlinear results. (Chin J Ophthalmol, 2016, 52: 936-940).