Objective:To establish a method for determination of total flavonoids,eriodictyol-7-methylether and isosakuranetin in Artemisia halodendron seeds.Method:The content of total flavonoids was determined by ultraviolet spectrophotometry at 506 nm,and compared with eriodictyol-7-methylether as a reference substance;the content of eriodictyol-7-methylether and isosakuranetin was determined by RP-HPLC,the analytical column was YMC-Pack ODS-A column(5?m,4.6 mm?250 mm),the mobile phase consisted of acetonitrile-0.2% phosphoric acid aqueous(35:65) at a ow rate of 1.0 ml?min-1,and the detection wavelength was 288 nm.Results:The linear range of total avonoids was 0.1228--0.7368mg(r=0.9991) and the average recovery(n=6) was 100.5%(RSD=1.41%);the linear ranges of eriodictyol-7-methylether and isosakuranetin were 3.016--30.16?g/ml(r=0.9999)and 3.304--33.04?g/ml(r=0.9999),respectively;the average recoveries(n=6) were 98.9%(RSD=1.20%)and 96.8%(RSD=1.38%),respectively.Conclusion:The methods of UV and HPLC are simple,accurate with good reproducibility and suitable for determination of total avonoids,eriodictyol-7-methylether and isosakuranetin.

Objective:To investigate the value of counting microvessel density (MVD) and lymphatic vessel density (LVD) in predicting distant metastasis of pancreatic cancer within 1 year after surgery.Methods:The clinicopathological data of 47 patients with pancreatic cancer who underwent surgery in Laibin People's Hospital from January 2020 to December 2021 were retrospectively analyzed. The patients were divided into non-metastasis group( n=24) and metastasis group( n=23) according to whether distant metastasis occurred during 1-year follow-up. Immunohistochemistry was used to detect the CD 34 expression in microvascular epithelial cells and D2-40 level in lymphatic epithelial cells from pancreatic cancer tissues. MVD and LVD in cancer tissues and adjacent normal tissues were counted. The relationship between MVD and LVD in cancer tissues and clinicopathological characteristics such as gender, age, tumor diameter, tumor differentiation, lymph node metastasis, vascular invasion, nerve invasion and tumor stage were analyzed. The receiver operating characteristic curve (ROC) was drawn and the area under the curve (AUC) was calculated to evaluate the value of MVD and LVD in predicting distant metastasis of pancreatic cancer within 1 year after surgery. The effects of MVD and LVD on the distant metastasis rate of pancreatic cancer within one year after operation were evaluated. Univariate and multivariate logistic regression were used to analyze the independent influencing factors for distant metastasis of pancreatic cancer within 1 year after surgery. Results:MVD and LVD in metastatic cancer tissues were higher than those in adjacent normal tissues [(72.52±9.73) vs (51.73±7.95)/400 times field of view, (23.78±6.87) vs (14.00±5.66)/400 times field of view]. MVD and LVD in the non-metastasis group were also higher than those in the adjacent normal tissues [(63.20±6.52) vs (54.79±5.80)/400 times field of view, (16.25±5.15) vs (13.62±5.03)/400 times field of view], and all the differences were statistically significant ( P<0.05). MVD in cancer tissue was significantly increased in patients with tumor diameter ≥2 cm, lymph node metastasis, vascular invasion and high TNM stage ( P<0.05), and LVD was significantly increased in patients with tumor diameter ≥2 cm, lymph node metastasis, moderate and low differentiation, vascular invasion, nerve invasion and high TNM stage ( P<0.05). The AUC values of MVD and LVD in predicting distant metastasis of pancreatic cancer within 1 year after surgery were 0.799 (95% CI 0.659-0.939) and 0.803(95% CI 0.676-0.929), and the cut-off values were 70.5 and 20.5/400 times field of view, respectively. The sensitivity was 73.9% and 69.6%, and the specificity was 87.5% and 83.7%. The cumulative distant metastasis rate within 1 year after operation in high MVD and high LVD groups was significantly higher than that in low MVD and low LVD groups ( P<0.05). Multivariate logitic regression analysis showed that tumor diameter ≥2 cm ( OR=1.757, 95% CI 1.536-3.846, P<0.05), lymph node metastasis ( OR=2.364, 95% CI 1.036-4.175, P<0.05), high MVD ( OR=4.345, 95% CI 1.245-3.736, P<0.05) and high LVD ( OR=3.637, 95% CI 1.426-4.035, P<0.05) were independent risk factors for distant metastasis of pancreatic cancer within 1 year after surgery. Conclusions:Increased MVD and LVD in pancreatic cancer tissues are independent influencing factors for distant metastasis within 1 year after surgery, which can be used to predict whether patients have distant metastasis within 1 year after surgery.

Chitinase AO-801 is a hydrolase secreted by Arthrobotrys oligospora during nematode feeding, while its role remained elusive. This study analyzed the molecular characteristics of recombinant chitinase of Arthrobotrys oligospora (reAO-801). AO-801 belongs to the typical glycoside hydrolase 18 family with conserved chitinase sequence and tertiary structure of (α/β)8 triose-phosphate isomerase (TIM) barrel. The molecular weight of reAO-801 was 42 kDa. reAO-801 effectively degraded colloidal and powdered chitin, egg lysate, and stage I larval lysate of Caenorhabditis elegans. The activity of reAO-801 reached its peak at 40˚C and pH values between 4-7. Enzyme activity was inhibited by Zn2+, Ca2+, and Fe3+, whereas Mg2+ and K+ potentiated its activity. In addition, urea, sodium dodecyl sulfate, and 2-mercaptoethanol significantly inhibited enzyme activity. reAO-801 showed complete nematicidal activity against C. elegans stage I larvae. reAO-801 broke down the C. elegans egg shells, causing them to die or die prematurely by hatching the eggs. It also invoked degradation of Haemonchus contortus eggs, resulting in apparent changes in the morphological structure. This study demonstrated the cytotoxic effect of reAO-801, which laid the foundation for further dissecting the mechanism of nematode infestation by A. oligospora.

Cystic echinococcosis (CE) in sheep is a hazardous zoonotic parasitic disease that is caused by Echinococcus granulosus (Eg). At present, serological test is an important diagnostic method for Eg infection in domestic animals. Here, a fusion protein Eg mefAg-1 harboring 8 dominant B-cell epitopes of Eg such as antigen B, tetraspanin 1, tetraspanin 6, reticulon and Eg95 was produced in E. coli and evaluated for CE in sheep by indirect ELISA. Eg mefAg-1 showed in ELISA a high sensitivity (93.41%) and specificity (99.31%), with a coincidence rate of 97.02%. Overall, it is suggested that the Eg mefAg-1 could be a potential antigen candidate for CE serodiagnosis in sheep.
Animals, Domestic ; Echinococcosis ; Echinococcus granulosus ; Enzyme-Linked Immunosorbent Assay ; Epitopes, B-Lymphocyte ; Methods ; Parasitic Diseases ; Sensitivity and Specificity ; Serologic Tests ; Sheep

Objective:To investigate the effect of irradiation on the expression of miR-150-5p and miR-23a-3p in human peripheral blood serum by collecting peripheral blood of tumor patients before and after radiotherapy, so as to provide scientific basis for finding radiation biomarkers.Methods:A total of 63 tumor patients treated with radiotherapy from October 2021 to March 2022 were enrolled in this study. The relative expression levels of miR-150-5p and miR-23a-3p in peripheral blood serum in these patients were detected using the real-time fluorescence quantitative PCR (qPCR) before and after radiotherapy. The differential changes in the expression levels of the two miRNAs in the peripheral blood serum of the patients before and after radiotherapy were compared, and their relationships with factors such as cancer types were analyzed.Results:The relative expression levels of miR-150-5p and miR-23a-3p in peripheral blood serum of the patients after radiotherapy were significantly lower than those before radiotherapy ( t = 4.97, Z = -2.77, P < 0.05). Among different cancer types, the relative expression level of miR-150-5p in the patients with breast cancer, esophageal cancer, or other digestive tract cancer decreased after radiotherapy ( t = 3.47, 2.47, 2.87, P < 0.05), and the relative expression level of miR-23a-3p in the patients with digestive tract cancer decreased after radiotherapy ( Z = -1.99, P < 0.05). The changes in the expression level of miR-150-5p before and after radiotherapy were not affected by gender, age, chemotherapy, and cancer type ( P > 0.05). By contrast, the changes in the expression level of miR-23a-3p before and after radiotherapy were significantly affected by gender, age, and chemotherapy ( t=2.04, -3.34, -2.29, P<0.05). Conclusions:The expression of miR-150-5p in the serum of tumor patients may be affected by radiotherapy, which has the potential to be used as a biological indicator of radiation.

Integrase plays a critical role in the recombination of viral DNA into the host genome. Therefore, over the past decade, it has been a hot target of drug design in the fight against type 1 human immunodeficiency virus (HIV-1). Bovine immunodeficiency virus (BIV) integrase has the same function as HIV-1 integrase. We have determined crystal structures of the BIV integrase catalytic core domain (CCD) in two different crystal forms at a resolution of 2.45 Å and 2.2 Å, respectively. In crystal form I, BIV integrase CCD forms a back-to-back dimer, in which the two active sites are on opposite sides. This has also been seen in many of the CCD structures of HIV-1 integrase that were determined previously. However, in crystal form II, BIV integrase CCD forms a novel face-to-face dimer in which the two active sites are close to each other. Strikingly, the distance separating the two active sites is approximately 20 Å, a distance that perfectly matches a 5-base pair interval. Based on these data, we propose a model for the interaction of integrase with its target DNA, which is also supported by many published biochemical data. Our results provide important clues for designing new inhibitors against HIV-1.
Animals ; Catalytic Domain ; genetics ; Cattle ; DNA ; genetics ; DNA, Viral ; HIV-1 ; genetics ; metabolism ; Humans ; Immunodeficiency Virus, Bovine ; enzymology ; genetics ; Integrases ; chemistry ; genetics ; metabolism