BACKGROUND: Cancer stem-like cells (CSCs) are a small subset of cells in tumors that exhibit self-renewal and differentiation properties. CSCs play a vital role in tumor formation, progression, relapse, and therapeutic resistance. B7-H3, an immunoregulatory protein, has many protumor functions. However, little is known about the mechanism underlying the role of B7-H3 in regulating gastric cancer (GC) stemness. Our study aimed to explore the impacts of B7-H3 on GC stemness and its underlying mechanism. METHODS: GC stemness influenced by B7-H3 was detected both in vitro and in vivo . The expression of stemness-related markers was examined by reverse transcription quantitative polymerase chain reaction, Western blotting, and flow cytometry. Sphere formation assay was used to detect the sphere-forming ability. The underlying regulatory mechanism of B7-H3 on the stemness of GC was investigated by mass spectrometry and subsequent validation experiments. The signaling pathway (Protein kinase B [Akt]/Nuclear factor erythroid 2-related factor 2 [Nrf2] pathway) of B7-H3 on the regulation of glutathione (GSH) metabolism was examined by Western blotting assay. Multi-color immunohistochemistry (mIHC) was used to detect the expression of B7-H3, cluster of differentiation 44 (CD44), and Nrf2 on human GC tissues. Student's t -test was used to compare the difference between two groups. Pearson correlation analysis was used to analyze the relationship between two molecules. The Kaplan-Meier method was used for survival analysis. RESULTS: B7-H3 knockdown suppressed the stemness of GC cells both in vitro and in vivo . Mass spectrometric analysis showed the downregulation of GSH metabolism in short hairpin B7-H3 GC cells, which was further confirmed by the experimental results. Meanwhile, stemness characteristics in B7-H3 overexpressing cells were suppressed after the inhibition of GSH metabolism. Furthermore, Western blotting suggested that B7-H3-induced activation of GSH metabolism occurred through the AKT/Nrf2 pathway, and inhibition of AKT signaling pathway could suppress not only GSH metabolism but also GC stemness. mIHC showed that B7-H3 was highly expressed in GC tissues and was positively correlated with the expression of CD44 and Nrf2. Importantly, GC patients with high expression of B7-H3, CD44, and Nrf2 had worse prognosis ( P = 0.02). CONCLUSIONS B7-H3 has a regulatory effect on GC stemness and the regulatory effect is achieved through the AKT/Nrf2/GSH pathway. Inhibiting B7-H3 expression may be a new therapeutic strategy against GC.
Humans ; Cell Line, Tumor ; Neoplasm Recurrence, Local ; NF-E2-Related Factor 2/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Signal Transduction ; Stomach Neoplasms

Background: The incidence of ulcerative colitis (UC) has gradually increased in China in recent years. The pathogenesis of UC is related to the dysfunction of immune system. B7-H5 is an important immune checkpoint molecule and is significant for the regulation of immune function. Ainis: To investigate the expression and clinical significance of B7-H5 in UC. Methods: A total of 65 UC tissue specimens were collected from Jan. 2010 to Dec. 2020 at the First Affiliated Hospital of Soochow University, and 5 healthy subjects were served as controls. Immunohistoehemistry and immunofluorescence were used to detect the expression of B7-H5, and its relationship with elinieopathologieal characteristics of UC patients was analyzed. Results: Expression of B7-H5 was significantly increased in UC patients than in controls (P < 0. 001). B7-H5 expression in UC patients was positively correlated with ESR and CRP (P < 0. 01), but not related to gender, age, extent of lesion, Mayo score and UCEIS score (P > 0. 05). Conclusions; The expression of B7-H5 in UC patients is significantly increased and is correlated with ESR and CRP, and can be used as a new marker for reflecting the severity of inflammation in UC patients.

Yuebawei refers to 8 kinds of genuine medicinal materials in south China area，namely Citri Grandis Exocarpium， Citrus reticulata， Amomum villosum， Pogostemon cablin， Morinda officinalis， Aquilaria sinensis， Citrus medica and Polygonum multiflorum. The results of this review show that， the pharmacological effects of Yuebawei are very extensive. It has significant effects in relieving cough and phlegm， anti-bacterial and anti-inflammatory， enhancing immunity， anti-oxidation， antidepressant， immune regulation and regulating gastrointestinal function. Its development and application prospects are broad， but most of the existing pharmacological studies stay on pharmacodynamic studies. In the aspect of the safety research of Yuebawei， there are few studies on the safety of other medicinal materials， except for the liver toxicity of P. multiflorum； at the same time， there are few comparative studies on the therapeutic advantages of Yuebawei and other local medicinal materials in clinical application. It is necessary to strengthen the research on the material basis of pharmacological action mechanism， and safety and efficacy observation of clinical medication for Yuebawei， so as to provide scientific basis for the development of new drugs and clinical promotion and application of Yuebawei.

Rutin,a flavonoid found in fruits and vegetables,is a potential anticancer compound with strong anti-cancer activity.Therefore,electrochemical sensor was developed for the detection of rutin.In this study,CoWO4 nanosheets were synthesized via a hydrothermal method,and porous carbon(PC)was prepared via high-temperature pyrolysis.Successful preparation of the materials was confirmed,and character-ization was performed by transmission electron microscopy,scanning electron microscopy,and X-ray photoelectron spectroscopy.A mixture of PC and CoWO4 nanosheets was used as an electrode modifier to fabricate the electrochemical sensor for the electrochemical determination of rutin.The 3D CoWO4 nanosheets exhibited high electrocatalytic activity and good stability.PC has a high surface-to-volume ratio and superior conductivity.Moreover,the hydrophobicity of PC allows large amounts of rutin to be adsorbed,thereby increasing the concentration of rutin at the electrode surface.Owing to the syn-ergistic effect of the 3D CoWO4 nanosheets and PC,the developed electrochemical sensor was employed to quantitively determine rutin with high stability and sensitivity.The sensor showed a good linear range(5-5000 ng/mL)with a detection limit of O.45 ng/mL.The developed sensor was successfully applied to the determination of rutin in crushed tablets and human serum samples.

We developed a high-efficiency microfluidic chip for extracting exosomes from human plasma. We collected peripheral blood from normal human, designed and fabricated a microfluidic chip based on nanoporous membrane and agarose gel electrophoresis to isolate exosomes. The extracted exosomes were characterized by transmission electron microscopy, nanosight and Western blotting, the morphology, concentration and particle size of exosomes were identified and analyzed. Meanwhile, we used ultracentrifugation and microfluidic chip to isolate exosomes separately. The particle size and concentration of the exosomes extracted by two methods were compared and analyzed, and their respective extraction efficiency was discussed. Finally, the expression level of miRNA-21 in exosomes was analyzed by RT-PCR. The microfluidic chip isolated (in 1 hour) high-purity exosomes with size ranging from 30-200 nm directly from human plasma, allowing downstream exosomal miRNA analysis. By comparing with ultracentrifugation, the isolation yield of microfluidic chip was 3.80 times higher than ultracentrifugation when the volume of plasma sample less than 100 μL. The optimized parameters for exosome isolation by gel electrophoresis microfluidic chip were: voltage: 100 V; concentration of agarose gel: 1.0%; flow rate of injection pump: 0.1 mL/h. The gel electrophoresis microfluidic chips could rapidly and efficiently isolate the exosomes, showing great potential in the research of exosomes and cancer biomarkers.
Exosomes ; Humans ; MicroRNAs/genetics* ; Microfluidics ; Plasma ; Ultracentrifugation

Objective:To evaluate the feasibility of high resolution MRI for the measurement of anterior cartilaginous acetabulum-head-index (A-CAHI) and the value of A-CAHI for predicting hip clinical function after treatment in developmental dysplasia of the hip (DDH).Methods:The imaging data of 92 hips from 61 children with treated DDH were retrospectively reviewed in Shandong Medical Imaging Research Institute from January 2019 to January 2020. All children underwent conservative treatments or surgical interventions 3 years ago. Hip function after treatment was evaluated clinically based on the modified MacKay criteria. The hips were divided into satisfactory clinical function group (McKay rating excellent or good, n=46) and unsatisfactory group (McKay rating fair or poor, n=46). All patients were imaged with conventional MRI, high resolution fat suppressed proton density weighted image (FS-PDWI) of the unilateral hip joint in oblique sagittal view, and anteroposterior hip radiographs. A-CAHI and lateral cartilaginous acetabulum-head-index (L-CAHI) were measured respectively on high-resolution oblique sagittal PDWI and conventional coronal T 1WI. Acetabulum head index (AHI) was also measured on anteroposterior hip radiograph. Mann-Whitney U test or independent-samples t test was used to compare the difference of A-CAHI, L-CAHI and AHI between satisfactory and unsatisfactory clinical function groups. The diagnostic value using A-CAHI, L-CAHI, AHI, or A-CAHI combined with L-CAHI for unsatisfactory clinical function were investigated by the ROC curve. The area under the curve (AUC) and the Z statistic were used to compare diagnostic performance. Results:The values of A-CAHI, L-CAHI and AHI were significantly higher in satisfactory clinical function group compared with the unsatisfactory group ( Z=-7.746, -7.735, t=-7.199, all P<0.001).A-CAHI combined with L-CAHI had the significant highest diagnostic accuracy compared with A-CAHI, L-CAHI and AHI (AUC were 0.994, 0.969, 0.968, 0.861, respectively), with significant differences ( Z=1.975, 2.006, 3.553, P=0.048, 0.051,<0.001). The sensitivity and specificity of A-CAHI combined with L-CAHI for the diagnosis of prognosis were 95.7% and 97.8%, respectively. Conclusions:A-CAHI measured by high resolution MRI was found to have the highest diagnostic accuracy for prediction of hip clinical function in the treated DDH, and combined with L-CAHI can improve the diagnostic accuracy significantly.

Lung cancer is a malignant tumor with high incidence and mortality in the world, which seriously threatens people's safety and health. Early diagnosis of lung cancer is the key part in the process of prevention and treatment of lung cancer. It can improve the survival of patients with lung cancer. Exosomes are closely related to the invasion and metastasis process of tumor, it plays an important role in the development of lung cancer. Biomarkers based on exosomes have become a powerful diagnostic tool of lung cancer. Exosomes are lipid bilayer vesicles with uniform size and diameter of 30 nm-200 nm secreted by cells. Exosomes contain different types of nucleic acids and proteins. These nucleic acids and proteins are derived from their parent cells (including parent cancer cells), which have a wide range of physiological functions, including immune regulation, intercellular communication and other physiological activities. Biomacromolecules in exosomes, such as single-stranded RNA, long noncoding RNA, microRNA, protein and lipids, which can provide valuable genetic information for early clinical diagnosis of lung cancer. This review described the origin, structural characteristics, extraction methods, biological characteristics of exosomes and the relationships of exosomes in the early diagnosis of lung cancer.
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Objective To evaluate the diagnostic performance of the automated quantitative analysis by coronary computed tomography angiography (CCTA) for lesion specific hemodynamic significance assessed by fractional flow reserve(FFR). Methods One hundred and fifteen patients with one hundred and fifty?one vessels,who successively underwent invasive coronary angiography with evaluation of FFR(values≤0.8 were defined as lesion specific hemodynamically significant), were analyzed by coronary CT angiography. FFR≤0.80 was found in 54(35.76％) of the 151 vessels, which was divided into two groups (group of hemodynamically significant and group of hemodynamically non-significant). CCTA images were quantitatively analyzed with automated software to obtain the following index:minimal lumen diameter(MLD), maximum diameter stenosis(MDS％), minimal lumen area(MLA), maximum area stenosis(MAS％), lesion length (LL), total plaque volume(TPV), total plaque burden(TPB), calcified plaque volume(CPV), calcified plaque burden (CPB), non-calcified plaque volume(NCPV), non-calcified plaque burden(NCPB), lipid plaque volume(LPV), lipid plaque burden(LPB), fibrous plaque volume(FPV), fibrous plaque burden(FPB), napkin-ring sign(NRS), remodeling index(RI) and eccentric index(EI). Logistic regression and area under the receiver operating characteristics were used for statistical analysis. Results MDS％(65.04％± 8.20％), MAS％(73.91％± 7.58％), TPB(57.96％± 11.17％), CPB[4.32％(0.11％, 5.34％)], LPB[14.89％(9.30％, 19.23％)], CPV[30.68 (0.29, 33.36)mm3], LPV[(81.72(33.92, 94.68)mm3]in the group with hemodynamic significance were larger than those in group with normal hemodynamic status[58.27％± 9.50％, 64.83％± 8.31％, 53.88％± 11.77％, 2.05％(0.00％, 3.42％), 11.83％(6.34％, 16.8％), 12.53(0.00, 13.24)mm3, 60.71(24.1, 75.11)mm3, respectively], which was statistically significant(t=4.41,P<0.01;Z=6.63,P<0.01;t=2.08,P<0.05;Z=-2.47,P<0.01;Z=-2.30,P<0.05;Z=-2.48, P<0.01;Z=-2.55, P<0.01, respectively). MLD[1.24(1.04, 1.46)mm]and MLA[3.61(2.40, 4.80) mm2]in the group with hemodynamic significance were smaller than those in group with normal hemodynamic status[1.53(1.32,1.72)mm, 5.28(4.00,6.40)mm2],which was statistically significant[Z=-4.82,-5.40, respectively;P<0.01].In logistic regression analysis, only MAS％(OR:1.08,95％CI:1.01-1.15,P=0.02), CPB (OR:1.16,95％CI:1.02-1.33,P=0.02) and LPB(OR:1.10,95％CI:1.01-1.19,P=0.02), MLA(OR:0.69, 95％CI:0.49-0.98,P=0.04)were significant predictors of hemodynamic significance. For predicting lesion specific hemodynamic significance, compared with MLA(0.76), MDS％(0.71), CPB(0.62) and LPB(0.61), except for MLA(Z=0.77, P=0.44), the AUC of MAS％(0.79) was significantly increased(Z=2.54, P=0.01;Z=2.91, P<0.01;Z=2.94, P<0.01, respectively). However, combination of other index to MAS％[MAS％+MLA％(0.81), MAS％+MDS％(0.80), MAS％+TPB(0.80), MAS％+CPB(0.80), MAS％+LPB(0.81)] did not show significantly difference over MAS％(Z=1.10, 0.71, 0.40, 0.54, 1.07, respectively;P>0.05). Conclusion Compared with diameter stenosis, area stenosis substantially improves the prediction of lesion specific hemodynamic significance.

Objective To establish a rapid and sensitive liquid chromatograph-mass spectrometry (LC-MS/MS) method for the determination of thiabendazole in honeysuckle flowers. Methods The acetic ether was selected as extraction solvent. The mass spectrometer analysis was conducted in the positive ionization electrospray mode using SIM. The transitions m/z 202→175 was used to quantify thiabendazole. Results The satisfactory linearity was obtained in the range of 0.1×10-5-2×10-5mg for thiabendazole (r=0.999 5), and the limit of detection (LOD) of 10.0 ng/ml and the mean recovery of 93.70% were obtained by this LC-MS/MS method. Conclusions The method of LC-MS/MS is sensitive, simple and accurate, and it proved to be suitable for the determination of thiabendazole in Honeysuckle flowers.