ObjectiveTo detect the serum level of follistatin-like protein 1 (FSTL1) in patients with systemic lupus erythematosus (SLE) and its expression in renal biopsy tissues in lupus nephritis (LN) patients as well as its clinical significance were analyzed.MethodsThe serum concentration of FSTL1 in 54 SLE patients and 27 healthy controls was measured with enzyme-linked immunosorbent assay (ELISA).The distribution of FSTL1 in renal biopsy tissues was stained by immune-histochemical method.Mann-Whitney U test,t test,X2test and Pearson test were selected to compare the changes and data analysis.ResultsThe serum FSTLI level was significantly higher in SLE patients(26±21) μg/L than those of healthy controls ( 12± 14) μg/L (P＜0.01).The level of serum FSTL1 was significantly higher in SLE patients with hypertension than in patients without hypertension.The serum FSTL1 level had statistically significant changes between SLE patients with disease duration ≥ 5 years and ＜5 years.The level of serum FSTL1 correlated positively with SLEDAI score (r=0.319,P=0.022),age (r=0.700,P＜0.01),disease duration (r=0.513,P＜0.01),complement C4 level (r=0.443,P=0.004),and total serum cholesterol level (r=0.460,P=0.001 ).FSTL1 correlated inversely with platelet count (r=-0.422,P =0.001 ),anti-dsDNA antibody levels (r=-0.276,P=0.046).FSTL1 expression was evident in the cytoplasm of epithelial cells of kidney tubules.ConclusionThe level of serum FSTL1 is significantly increased in SLE patients.FSTL1 concentration correlats positively with disease activity.These data indicate that FSTL1 may play a role in the pathogenesis of SLE.

Objective To prepare human prostatic carcinoma-targeted ultrasound contrast agent and assess its targeted ability in vitro. Methods Human prostatic carcinoma-specific polyclonal antibody PSM(C-15) was attached to the surface of self-made liposome microbubbles by electrostatic attraction to prepare targeted microbubbles. Immunofluorescent staining assay was used to identify the combination of PSM(C-15) with liposome microbubbles and the targeted microbubbles were added to prostatic-carcinoma cells and then observed under the light and fluorescence microscope to evaluate the targeting ability of the targeted liposome microbubbles with prostatic carcinoma cells in vitro, while the common liposome microbubbles were controls. Results Targeted microbubbles were positive in immunofluorescent straining assay. In vitro study of the targeting ability showed the targeted microbubbles could actively adhere to LNCaP cell. While the control was negative. Conclusion The targeted liposome contrast agent with highly specific biological activity was prepared successfully. The contrast agent could bind to human prostatic carcinoma cells specially and effectively in vitro.

Objective To explore a promising system for tumor CD 44 receptor-targeted imaging and to investigate their physic-chemical properties and targeting effect on CD 44 abundant cancer cells in vitro.Methods The superparamagnetic iron oxide ( SPIO) nanoparticles were prepared by a coprecipitation in alkaline media starting from a mixed of the ferrous and ferric solution.And then the surface of the SPIO nanoparticles were modified with APTMS by a reaction with the hydroxyl groups.Finally, the hyaluronan-modified SPIO ( SPIO-HA) nanoparticles were prepared.Control and experimental groups were established after adding SPIO or SPIO-HA as agents respectively.Transmission electron microscopy ( TEM) and particle size analyzer were used to measure these nanoparticle sizes and the hydrodynamic diameters.Thermogravimetric analysis ( TGA) was carried out to evaluate the HA-content on the surface of SPIO-HA.The MRI T2 ralaxivities (1/T2 ) of the two groups at different Fe concentrations (0.09, 0.18, 0.27, 0.36, 0.45 mmol/L ) were measured on a 3.0T MR system.HepG2 cells and HL7702 cells were used for assessment of cells viability by methyl thiazolyl tetrazolium ( MTT ) assay.Prussian blue staining , immunoassay fluorescence image and flow cytometry were carried out to determine the targeted cellular uptake of SPIO-HA nanoparticles.MRI were performed to show the MR T 2 value changes after incubating with HepG2 cancer cells by using T 2 WI sequences at a clinical 3.0 T MR system.One-way analysis of variance was performed to determine significant changes in MR T 2 values of blank control , SPIO-HA and SPIO groups.Results The SPIO-HA and SPIO NPs were fairly homogeneous with an average core size of 18.2 and 22.4 nm, hydrodynamic diameter of 91.1 and 103.2 nm, Zeta potential of (-45.00 ±0.86) mV and (-18.50 ±0.73) mV, and magnetic relaxivity of 0.212 ×106 M-1 · s-1 and 0.191 ×106 M-1 · s-1.Based on the TGA data , HA accounted for 24%weight of each SPIO-HA.The internalization of the SPIO-HA was confirmed by prussian blue staining , while the cells showed no obvious blue stains with SPIO , incubation of SPIO-HA with tumor cells led to blue color inside the cells.After that, we examined cancer cell binding of FITC-SPIO-HA by immunoassay fluorescence image and flow cytometry.The green fluorescence resulting from FITC-SPIO-HA was observed inside the cells in both the cytoplasm and the plasmalemma.Tumor cells treated with SPIO-HA exhibited higher fluorescence signals with 7.97-fold enhancement observed for HepG 2 cells over control particles.In vitro MR, mean T2 values of blank control , SPIO and SPIO-HA groups were ( 115.20 ±0.36 ), ( 115.07 ±0.81 ) and ( 21.67 ±0.21 ) ms, respectively.There was significant difference among those three groups (F=31 703.339,P<0.01), MR T2 values of HepG2 cells treated with the SPIO-HA NPs were lower than blank and SPIO group.In comparison, SPIO did not generate any MRI signal changes compared with blank group.Conclusion The tumor CD44 receptor-targeted MR molecular probe SPIO-HA had a good physic-chemical property and well targeted HepG2 cells.

Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.

Objective To investigate the role of erlotinib in the expression of surfactant protein A (SP-A) in LPS-induced acute lung injury (ALI) of mice model.Methods C57BL/6 mice were randomly (random number)divided into control group (n=6),ER group (n=6),LPS group (n=6),and ER+LPS group (n=6).In the LPS group,2 mg/kg LPS was instilled into trachea of mice to induce lung injury.In control group,normal saline was instilled into trachea of mice instead.In the ER+LPS group and ER group,100 mg/kg of edotinib was instilled into stomach of mice,and one hour later.2 mg/kg LPS was instilled into trachea of mice in ER+PLS group to induce lung injury.Twenty-four hours later,bronchoalveolar lavage fluid (BALF) and lung tissue of mice in four groups were collected.HE staining were used for evaluating pathological changes of lung injury.Lung wet/dry weight ratio,protein concentrations and total cell numbers in the BALF were measured to determine the degree of pulmonary edema.Immunohistochemical staining and Western Blot were used for testing the protein expression of SP-A,Data of multiple groups were analyzed by one way variance (ANOVA) and inter-group comparisons were made by the least significant difference (LSD) tests.Results There was no significant difference in lung injury score (LIS) between control group (0.056±0.008) and ER (0.064±0.037) group,The LIS in LPS group (0.846-±0.047) was higher than that in control group,however the LIS in ER+LPS group (0.279±0.020) was significant lower than that in LPS group (P ＜ 0.05).Lung wet/dry weight,SP-A concentration and total cell numbers in the bronchoalveolar lavage fluid revealed that the degree of pulmonary edema in LPS group was higher than that in control group,and this pulmonary edema was reversed by erlotinib treatment.Immunohistochemical staining and Western blot showed that the expression of SP-A in LPS group was decreased compared with control group,but it was recovered after erlotinib treatment (P ＜ 0.05).Conclusions Erlotinib could protect the LPS-induced ALI,and it may be related to the regulation of SP-A.

We propose a novel palm-vein recognition model based on the end-to-end convolutional neural network. In this model, the convolutional layer and the pooling layer were alternately connected to extract the image features, and the categorical attribute was estimated simultaneously via the neural network classifier. The classification error was minimized via the mini-batch stochastic gradient descent algorithm with momentum to optimize the feature descriptor along with the direction of the gradient descent. Four strategies including data augmentation, batch normalization, dropout, and L2 parameter regularization were applied in the model to reduce the generalization error. The experimental results showed that for classifying 500 subjects form PolyU database and a self-established database, this model achieved identification rates of 99.90% and 98.05%, respectively, with an identification time for a single sample less than 9 ms. The proposed approach, as compared with the traditional method, could improve the accuracy of palm vein recognition in clincal applications and provides a new approach to palm vein recognition.
Algorithms ; Databases, Factual ; Hand ; blood supply ; diagnostic imaging ; Humans ; Neural Networks (Computer) ; Veins ; diagnostic imaging

Objective: To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P450 2C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology. Methods: Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results. Results: The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4% and 10.9%, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results. Conclusion A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.

Objective: Corydalis bungeana (CB) is a well-used medicinal herb in Mongolian folk medicine and has been traditionally applied as an antiobesity agent. However, the evidence-based pharmacological effects of CB and its specific metabolic alterations in the obese model are not entirely understood. This study aimed to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite alterations associated with weight loss and lipid metabolism in obese rats. Methods: A high-fat high-sugar (HFHS) diet was used to induce obese models in rats. CB extract was orally gavaged at 0.18, 0.9 and 1.8 g/kg doses for six weeks, and feed intake, body weight, fat pad weight, and blood indexes were measured. Blood serum metabolites were evaluated by gas chromatography/quadrupole time-of-flight tandem mass spectrometry (GC-TOF/MS). Results: The results showed that compared with the obese group, the administration of CB extract caused significant decreases in body weight (P < 0.05), feed intake, Lee's index, and perirenal, mesenteric, epididymal fat weight. CB extract also reduced blood triglyceride and total cholesterol levels (P < 0.05) of obese rats. Metabolomic findings showed that nine differential metabolites, including pyruvic acid, D-glucuronic acid, malic acid, dimethylglycine, oxoglutaric acid, pantothenic acid, sorbitol acid, fumaric acid and glucose 6-phosphate were identified under CB treatment and altered metabolic pathways such as TCA cycle, pantothenate and CoA biosynthesis, and glycolysis/gluconeogenesis. Conclusion: This study demonstrated weight loss and lipid lowering effects of CB on HFHS diet-induced obese rats and identified nine metabolites as potential biomarkers for evaluating the favorable therapeutic mechanism of CB via regulation of lipid and glucose metabolism.

Objective To investigate the expression level of serum miR-486-5p in patients with pancreatic cancer and the value of serum miR-486-5p combined with carbohydrate antigen 19-9 (CA19-9) in predicting the resectability of pancreatic cancer. Methods A total of 60 patients who were diagnosed with pancreatic cancer in Qingdao Municipal Hospital from September 2018 to December 2020 were enrolled, among whom 32 patients had resectable or borderline resectable pancreatic cancer (operable group) and 28 had unresectable pancreatic cancer (non-operable group), and a benign pancreatic disease group with 30 patients and a healthy control group with 44 individuals were also established. Quantitative real-time PCR was used to measure the serum level of miR-486-5p in each group, and the relative expression level of miR-486-5p was calculated to analyze its association with the clinical features of pancreatic cancer, including age, sex, tumor location, tumor size, TNM stage, lymphatic metastasis, and distant metastasis. The Mann-Whitney U test was used for comparison of non-normally distributed continuous variables between two groups, and the chi-square test was used for comparison of categorical variables. The receiver operating characteristic (ROC) curve was plotted, and a binary logistic regression analysis was used to calculate the combined predictive value and then investigate the value of serum miR-486-5p combined with CA19-9 in predicting the resectability of pancreatic cancer. Results The relative expression level of serum miR-486-5p in the operable group [2.16 (1.38~3.30)] and the non-operable group [4.65 (2.80~9.90)] was significantly higher than that in the benign pancreatic disease group [1.01 (0.52~1.53)] and the healthy control group [0.99 (0.24~1.01)] (all P < 0.001). There were significant differences in the number of patients with low or high expression of miR-486-5p between the patients with different TNM stages, presence or absence of lymphatic metastasis, and presence or absence of distant metastasis ( χ 2 =13.765, 5.157, and 6.638, all P < 0.05). Compared with CA19-9 alone, miR-486-5p+CA19-9 had a significantly better value in distinguishing the operable group from the benign pancreatic disease group (area under the ROC curve [AUC]=0.87, 95% confidence interval [ CI ]: 0.760-0.942; with a sensitivity of 81.3% and a specificity of 83.3%), distinguishing the operable group from the healthy control group (AUC=0.92, 95% CI : 0.836-0.970; with a sensitivity of 90.6% and a specificity of 86.4%), and distinguishing the operable group from the non-operable group (AUC=0.94, 95% CI : 0.884-0.998; with a sensitivity of 85.7% and a specificity of 93.7%) ( Z =2.841, 2.510, and 2.387, all P < 0.05), and the optimal cut-off values were 3.12, 3.21, and 6.63, respectively. Conclusion MiR-486-5p can be used as a serum biomarker for the diagnosis of pancreatic cancer, and miR-486-5p combined with CA19-9 has a better clinical value than CA19-9 alone in predicting the resectability of pancreatic cancer in the patients with benign pancreatic diseases and the healthy population.

Macrophages play an important role in material-related immune responses and bone formation, but the functionality of macrophage-derived extracellular vesicles (EVs) in material-mediated bone regeneration is still unclear. Here, we evaluated intracellular communication through small extracellular vesicles (sEVs) and its effects on endogenous bone regeneration mediated by biomimetic intrafibrillarly mineralized collagen (IMC). After implantation in the bone defect area, IMC generated more neobone and recruited more mesenchymal stem cells (MSCs) than did extrafibrillarly mineralized collagen (EMC). More CD63
Biomimetics ; Bone Regeneration ; Cell Differentiation ; Collagen ; Extracellular Vesicles ; Macrophages ; Osteogenesis