1.Separation of Triacylglycerols in Edible Oil by Reversed Phase Ultra Performance Liquid Chromatography-Mass Spectroscopy
Rong HE ; Xiaolin SHAN ; Fangyuan DONG ; Xu XU
Chinese Journal of Analytical Chemistry 2015;(9):1377-1382
A reversed phase ultra performance liquid chromatography-mass spectrometric method was developed for the separation and analysis of triglycerides in edible oils. The samples were separated by using three ultra performance C18 columns in series with a total length of 40 cm (10 cm + 15 cm + 15 cm) at high pressure with acetonitrile-isopropanol (50:50, V/V) as mobile phase at a flow rate of 0. 2 mL/min and at col-umn temperature of 25℃, and detected by APCI ionization-mass spectrometry. The edible oil sample was dis-solved in isopropanol and injected in LC-MS directly. The triglycerides in edible oils were distinguished to their better fine components which included corn oil, peanut oil, sunflower seed oil, rice oil, olive oil, sesa-me oil and soybean oil. The chromatograms of different edible oils showed that the same kind of edible oil was composed of similar triglyceride composition and content, while the different kind of edible oils differed. The experimental result showed that the method could be use for identifying 5% lard adulterated in soybean oil. The method suggests a significant research way for identifying adulteration in edible oil.
2.Clinical research on prophylactic pelviureteric instillation chemotherapy after endoscopic management of upper tract urothelial carcinoma
Fangyuan ZHANG ; Dong WEI ; Pengjie WU ; Xin CHEN ; Jianye WANG
Chinese Journal of Geriatrics 2015;34(8):878-880
Objective To investigate the effect of endoscopic management of upper tract urothelial carcinoma (UTUC),and evaluate the indication and clinical value of postoperative prophylactic pelviureteric instillation chemotherapy.Methods Patients who met the inclusion criteria included elderly patients,patients who could not tolerate radical surgery,patients with renal insufficiency or needed dialysis after removal of the kidney,patients with bilateral UTUC,patients with tumor≥3 cm in diameter but could be completely resectted under ureteroscopy who required to preserve renal function.Surgical procedure was successful.The ureteral stents were placed,and prophylactic ureteral infusion chemotherapy via the retrograde transvesical ureteric catheterisation was conducted 1-2 times one week at 2 days after surgery.40 milligram of pirarubicin hydrochloride or epirubicin hydrochloride was dissolved in 40 milliliter sterile water for the prophylactic infusion chemotherapy.These instillations were completed within 40 minutes.Results Totally,9 patients (3 males and 6 females) aged 60-86 years,mean age of (69.7±6.4) years,were enrolled in this study.The carcinoma of the renal pelvis was found in 2 cases and ureteral tumors in 7 cases.In pathological results,ureteral polyps was found in 1 case,papillary ureteral neoplasm with low malignant potential in 2 cases,urinary tract epithelial carcinoma in 4 cases (1 case with poorlydifferentiation,1 case with well-differentiation,2 cases with urinary tract epithelial cancer),and 2 cases were lack of pathological reports because of too few pathologic specimens.4 patients received adjuvant chemotherapy by instillation successfully,and lumbago,fever or other complications were not found.Patients were followed up for a mean period of 15 moths (4-31 months).Only 1 patient had recurrence of bilateral ureteral tumor 15 months after surgery,and no tumor recurrence was found in other patients.Conclusions Endoscopic management is a safe and effective in treating UTUC,which can preserve renal function by avoiding nephrorectomy in some patients.The instillation chemotherapy after endoscopic management is safe,which has a satisfactory effect,but further validation in a large clinical sample is needed.Patients with tumor resection under ureteroscopy who cannot obtain the pathology results need to be closely followed up.
3.Value of 18F-FDG PET/CT in diagnosis of prosthetic vascular graft infection after endovascular aortic repair
Fangyuan REN ; Changjing ZUO ; Jianhua WANG ; Gaofeng SUN ; Chao CHENG ; Aisheng DONG ; Jian YANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2019;39(3):138-141
Objective To study the features and clinical value of 18F-fluorodeoxyglucose (FDG) PET/CT in prosthetic vascular graft infections (PVGIs) after endovascular aortic repair (EVAR).Methods Data of 27 patients (22 males and 5 females,age range:21-77 years,average age:42 years) who underwent PET/CT imaging after EVAR from October 2011 to January 2017 were studied retrospectively.A total of 11 cases were finally diagnosed as PVGIs (PVGIs group),and the remaining 16 cases were defined as the negative group.PET/CT imaging features were compared between two groups.The detection rates of PET/CT and aortic CT angiography (CTA) for PVGIs were calculated and compared.Two-sample t test and Fisher exact test were used for data analysis.Results Significant uptake of FDG around the stents was detected by PET/CT in all patients in PVGIs group.The maximum standardized uptake value (SUVmax) of 11 patients in early imaging was 14.9±3.3 (10.8-21.8) and that of 9 patients in delayed imaging was 13.8±3.4 (10.6-19.1).Six patients of the negative group underwent the delayed imaging.No uptake or mild uptake of FDG around the stents was observed in negative group,with the SUVmax of 1.7±0.8(1.0--2.9) in early imaging and 1.6±0.7(1.1-2.5) in delayed imaging.SUVmax in negative group was significantly lower than that in PVGIs group (t values:12.6 and 11.8,both P<0.001).Five patients in PVGIs group were diagnosed as graft infections by the aortic CTA,while the remaining 6 cases showed no definitive infection signs on aortic CTA.The detection rate of aortic CTA was 5/11,which was significantly lower than that of PET/CT (P<0.05).In negative group,6 patients had abnormal FDG uptakes in other areas and were finally confirmed as infectious lesions (n=3) or malignant tumors (n=3).Conclusion Compared with aortic CTA,18F-FDG PET/CT is more sensitive and accurate in detection and diagnosis of PVGIs after EVAR.
4.Effect and mechanism of Acyl-CoA: lysocardiolipin acyltransferase 1 knockdown on hepatocyte steatosis and oxidative stress
Xiaona HU ; Fangyuan DONG ; Xin JIANG ; Xueying JI ; Jie CHEN ; Xiaofeng YU ; Zhijun BAO
Chinese Journal of Digestion 2020;40(8):546-554
Objective:To investigate the effect and mechanism of Acyl-CoA: lysocardiolipin acyltransferase 1 (ALCAT1) on hepatocyte steatosis and oxidative stress in fatty liver cell model.Methods:A fatty liver cell model was established and induced by free fatty acids (FFA). The expression of ALCAT1 in fatty liver cell model was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The empty siRNA plasmid and ALCAT1 siRNA plasmid were constructed. For the fatty liver cell model group, human normal hepatocytes (L-02 cells) were transfected with empty siRNA plasmid for 24 hours, and then cultured with FFA for 24 hours. For the ALCAT1 interfering group, L-02 cells were transfected with ALCAT1 siRNA plasmid for 24 hours, and then cultured with FFA for 24 hours. And L-02 cells cultured in common medium were used as as blank control group. Lipid droplet deposition and mitochondrial morphology were observed under transmission electron microscopy. The expression levels of autophagy-associated proteins (microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ and Beclin1) and key proteins of autophagy signal pathway (mammalian target of rapamycin (mTOR) and serine/threonine kinase (AKT)) were measured by Western blotting. The expression levels of oxidative stress products (malondialdehyde, 4-hydroxynonenal (4-HNE) and reactive oxygen species (ROS)) and inflammatory factors (interleukin-6(IL-6) and tumor necrosis factor (TNF)-α) were detected by enzyme-linked immunosorbent assay (ELISA) kits. Independent sample t test was used for statistical analysis. Results:The mRNA and protein expression levels of ALCAT1 of the fatty liver cell model group were both higher than that of negative control group (9.26±0.83 vs. 1.02±0.12, 0.35±0.02 vs. 0.17±0.01), and the differences were statistically significant ( t=9.82 and 6.83, both P<0.05). The results of electron microscopy indicated that the deposition of lipid droplets of the fatty liver cell model group and ALCAT1 interfering group were both higher than that of blank control group (17.67±3.52 and 7.67±0.33 vs. 4.33±0.33), the quantity of lipid droplets deposition of ALCAT1 interfering group was lower than that of fatty liver cell model group (7.67±0.33 vs. 17.67±3.52), and the differences were statistically significant ( t=3.76, 7.07 and 2.82, all P<0.05). The degree of mitochondria swelling of fatty liver cell model group was higher than that of blank control group and the degree of mitochondria swelling of ALCAT1 interfering group was lower than that of fatty liver cell model group. The results of Western blotting showed that the expression level of LC3-Ⅱof the fatty liver cell model group was higher than that of the blank control group (0.43±0.01 vs. 0.28±0.02), and the difference was statistically significant ( t=7.32, P<0.05). However there was no significant difference in the expression level of Beclin1 between fatty live cell model group and blank control group (0.93±0.05 vs. 0.98±0.05, P>0.05). The expression levels of LC3-Ⅱ and Beclin1 of the ALCAT1 interfering group were both higher than those of the fatty liver cell model group and blank control group (0.95±0.04 vs. 0.42±0.01 and 0.28±0.02, 2.07±0.06 vs. 0.93±0.05 and 0.98±0.05), and the differences were statistically significant ( t=13.30, 15.63, 14.05 and 13.02, all P<0.05). The expression levels of mTOR of the fatty liver cell model group and ALCAT1 interfering group were both lower than that of the blank control group (1.44±0.02 and 0.74±0.01 vs. 1.93±0.10), the expression level of mTOR of the ALCAT1 interfering group was lower than that of the fatty liver cell model group (0.74±0.01 vs. 1.44±0.02), and the differences were statistically significant ( t=4.83, 12.04 and 32.14, all P<0.05). The expression levels of phosphorylated AKT of the fatty liver cell model group and ALCAT1 interfering group were both lower than that of the blank control group (0.14±0.01 and 0.07±0.01 vs. 0.28±0.01), while the expression level of phosphorylated AKT of the ALCAT1 interfering group was lower than that of the fatty liver cell model group (0.07±0.01 vs. 0.14±0.01), and the differences were statistically significant ( t=8.59, 14.10 and 5.96, all P<0.05). The results of ELISA indicated that the expression levels of ROS, malondialdehyde, 4-HNE, IL-6 and TNF-α of the fatty liver cell model group and the ALCAT1 interfering group were all higher than those of the blank control group ((11.44±0.30) and (5.84±0.36) g/L vs. (1.72±0.38) g/L; (19.94±2.47) and (11.95±1.55) μmol/L vs. (1.47±0.18) μmol/L; (5.00±0.43) and (2.99±0.37) ng/L vs. (1.46±0.23) ng/L; (203.40±5.16) and (92.07±11.98) ng/L vs. (23.32±3.33) ng/L; (123.70±8.38) and (67.42±4.88) ng/L vs. (47.18±4.57) ng/L), and the differences were all statistically significant ( t=19.86, 7.86, 7.45, 6.74, 7.22, 3.49, 29.34, 5.53, 8.02 and 3.03, all P<0.05). While the expression levels of ROS, 4-HNE, IL-6 and TNF-α of the ALCAT1 interfering group were all lower than those of the fatty liver cell model group ((5.84±0.36) g/L vs. (11.44±0.30) g/L, (2.99±0.37) ng/L vs. (5.00±0.43) ng/L, (92.07±11.98) ng/L vs. (203.40±5.16) ng/L and (67.42±4.88) ng/L vs. (123.70±8.38) ng/L), and all the differences were statistically significant ( t=11.99, 3.51, 8.54 and 5.81, all P<0.05). There was no statistically significant difference in the expression of malondialdehyde between ALCAT1 interfering group and fatty liver cell model group ((11.95±1.55) μmol/L vs. (19.94±2.47) μmol/L, P>0.05). Conclusions:The expression of ALCAT1 is up-regulated in fatty liver cell model. Knockdown of ALCAT1 can inhibit the expression of mTOR pathway proteins, activate autophagy, alleviate hepatocyte steatosis, oxidative stress and inflammatory response.
5.Clinical characteristics and treatment prognosis of ocular toxocariasis
Yongwei ZHOU ; Qiuming LI ; Jiahui WU ; Xiaoyan LU ; Ge YANG ; Zhirou HU ; Fangyuan ZHEN ; Shuqian DONG
Chinese Journal of Ocular Fundus Diseases 2023;39(6):483-488
Objective:To observe the clinical characteristics and treatment prognosis of patients with ocular toxocariasis (OT).Methods:A retrospective clinical trial. From March 2018 to September 2021, 40 eyes of 40 OT patients diagnosed by ophthalmic examination in the First Affiliated Hospital of Zhengzhou University were included in the study. All patients underwent best corrected visual acuity (BCVA) and scanning laser ophthalmoscope (SLO) examination. Color Doppler ultrasound flow imaging (CDFI), fluorescein fundus angiography (FFA) and optical coherence tomography (OCT) were performed in 25, 26 and 26 eyes, respectively. Among the 40 patients, there were 23 males (57.5%, 23/40) and 17 females (42.5%, 17/40). All patients were monocular. Thirty patients (75.0%, 30/40) were younger than 18 years old, with the mean age of (9.60±0.60) years. Ten patients (25.0%, 10/40) were great than or equal to 18 years old, with the mean age of (34.60±4.52) years. Thirty-three patients (82.5%, 33/40) lived in rural areas for a long time. There were 27 patients (67.5%, 27/40) with a history of contact with dogs and cats. In 40 eyes, peripheral granuloma (peripheral type), posterior pole granuloma (posterior pole type), vitreous opacity similar to endophthalmitis (turbid type) and hybrid type were 18(45.0%, 18/40), 11(27.5%, 11/40), 6(15.0%, 6/40) ang 5(12.5%,5/40), respectively. All patients were treated with drugs and/or surgery after definite diagnosis. There were 28 eyes of peripheral type, posterior pole type and hybrid type, 17 eyes were treated with surgery and 11 eyes with drug treatment, respectively. Five eyes with turbid type were only treated with drugs. In 40 patients, 33 patients participated in follow-up. The follow-up time after treatment was (18.78±9.44) months. The improvement of BCVA was observed. The number of eyes with different BCVA before and after treatment was compared by χ2 test or Fisher's test. Results:At the first visit, the BCVA ranged from light perception to 0.6, including 20 eyes with BCVA <0.1, 13 eyes with BCVA 0.1-0.3, and 7 eyes with BCVA >0.3. The posterior vitreous anterior limiting membrane was thickened in 24 eyes (60.0%, 24/40). There were 27 eyes (67.5%, 27/40) with lamellar vitreous opacity and 22 eyes (55.0%, 22/40) with peripheral/posterior pole granulomas. Among 25 eyes examined by CDFI, 14 eyes (56.0%, 14/25) showed characteristic stratified or diffuse opacity in vitreous body. Of the 26 eyes examined by FFA, 15 eyes (57.7%, 15/26) had "fern-like" leakage of retinal capillaries, and the lesion had a patchy non-perfused area. In 26 eyes examined by OCT, epiretinal membrane, cystoid macular edema and vitreoretinal traction were 8 (30.8%, 8/26), 5 (19.2%, 5/26) and 2 (7.7%, 2/26) eyes, respectively. At the last follow-up, compared with before treatment, the BCVA of 5 eyes with turbid type increased, and the difference was statistically significant ( P<0.05). In 28 eyes with peripheral type, posterior pole type and hybrid type, 17 eyes with surgical treatment improved BCVA, and the difference was statistically significant ( χ2=6.258, P<0.05). In 11 eyes only treated with drugs, BCVA remained unchanged, and the difference was not statistically significant ( χ2=0.594, P>0.05). Conclusions:OT patients are mostly children; retinal granulomas, gray-white hyperplastic membrane behind lens or vitreous stratified opacity are specific characteristics. OT is mainly treated by glucocorticoid drugs and vitrectomy.
6.Anti-cancer Effect of Traditional Chinese Medicine in Regulating Cellular Senescence: A Review
Shuang ZHAO ; Dong WANG ; Haoyang SHI ; Fangyuan WANG ; Jingyan MENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):265-271
In recent years, the incidence and mortality rates of cancer have been increasing, posing a serious threat to human health. Western medicine mainly uses treatments such as surgical resection, chemotherapy, immunotherapy and targeted therapy, but they are prone to complications, drug resistance and adverse reactions. A growing number of studies have shown that traditional Chinese medicine has obvious advantages in the treatment of cancer, reducing the recurrence rate of cancer and improving the quality of survival of patients. Cellular senescence refers to a state of irreversible cell cycle growth arrest when cells cease to proliferate after a limited number of divisions, resulting in a decline in cell proliferation and differentiation capacities and physiological functions, accompanied by morphological changes such as flattening and multinuclear morphology. At the molecular level, it shows increased expression of DNA damage-related genes, reduced expression of cell cycle-related factors and significant secretory activity. The malignant development of cancer is closely related to cellular senescence. With the increasing number of cancer cell proliferation, cancer-related genes undergo continuous mutations, freeing them from cellular senescence and thus achieving unlimited proliferation. Through recent studies, it has been found that induction of tumor cell senescence, possibly through modulation of cellular DNA damage, cell cycle arrest and senescence-associated secretory phenotype (SASP), which converts the suppressive immune tumor microenvironment to an activated immune tumor microenvironment and thus reverses the escape of tumor cell senescence, is a promising strategy for cancer therapy. However, the mechanism of cellular senescence in cancer progression is not fully understood, especially the anti-cancer role played by traditional Chinese medicine in regulating cellular senescence. This article summarized and concluded the specific molecular mechanisms of cellular senescence, the role of cellular senescence in cancer progression, and the mechanism of anti-cancer effects of traditional Chinese medicine based on cellular senescence from the perspective of regulating cellular senescence, with a view to providing ideas and methods for the anti-cancer effects of traditional Chinese medicine and the development of new drugs.
7.Exploration of Spectrum-effect Relationship of Zhuriheng Dropping Pills Against Macrophage Foaming Based on UPLC-Q-Exactive Orbitrap MS
Qiong ZHAI ; Fangyuan LIANG ; Yuewu WANG ; Ren BU ; Xin DONG ; Jingkun LU ; Peifeng XUE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):176-186
ObjectiveThrough the correlation analysis between intestinal absorption profile and inhibition of macrophage foaming, the pharmacodynamic components of Zhuriheng dripping pills(ZRH) were explored to provide a basis for establishing its quality standard. MethodIntestinal absorption fluids with 0, 5, 10, 15, 20 times clinical equivalent doses were prepared by a rat everted gut sac(EGS), and the oxidized low density lipoprotein(ox-LDL)-induced RAW264.7 macrophage foaming model was used to investigate the effect of intestinal absorption fluid with different doses on the accumulation of lipids in RAW264.7 cells by oil red O staining and cholesterol content determination, and to screen for the optimal dose. Ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used to analyze and identify intestinal absorption fractions of ZRH intestinal absorption fluids, and partial least squares-discriminant analysis(PLS-DA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were performed on different doses of ZRH intestinal absorption fluids using SIMCA 13.0 with peak area as the independent variable and the pharmacodynamic indicators as the dependent variables to screen the compounds with variable importance in the projection(VIP) value>1.0 as contributing components, and Pearson correlation analysis was used to determine the spectral effect relationship, determined the compounds and positive correlation with pharmacodynamic were as active ingredients. Molecular docking was used to verify the binding energy of peroxisome proliferator-activated receptor α(PPARα), PPARγ, PPARβ, human retinoid X receptor α(RXRA) and nuclear transcription factor-κB(NF-κB) with the active ingredients in ZRH intestinal absorption fluids. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was performed to detect the mRNA levels of PPARγ, scavenger receptor A1(SRA1) and adenosine triphosphate-binding cassette transporter A1(ABCA1) in RAW264.7 cells, Westen blot was used to detect the expression level of PPARγ protein in RAW264.7 cells, and enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of interleukin(IL)-1β and NF-κB in RAW264.7 cells. ResultAccording to the results of oil red O staining and cholesterol content determination, the ZRH intestinal absorption fluids could significantly reduce macrophage foaming, and intestinal absorption fluids with 15, 20 times clinical equivalent doses had the best effect, the 15-fold ZRH intestinal absorption fluid was finally determined as the study subject. Spectral effect relationship showed that 52 corresponding peaks in the ZRH-containing intestinal fluid were positively correlated with the efficacy, including organic acids, phenylpropanoids, iridoids, flavonoids, bile acids, coumarins and chromones. Target validation results showed that 86.9%-96.2% of the total components processed good binding activities with the key targets of PPARα, PPARγ, PPARβ, RXRA and NF-κB, and the docking energy values were all less than -6.0 kcal·mol-1(1 cal≈4.19 J). The results of validation showed that, compared with the normal group, the model group showed a significant increase in the levels of SRA1 and PPARγ mRNA expression, a significant decrease in ABCA1 mRNA expression, a significant increase in the level of PPARγ protein expression, and a significant increase in the levels of IL-1β and NF-κB(P<0.01), compared with the model group, the 15-fold intestinal absorption fluid group showed a significant decrease in the levels of SRA1 and PPARγ mRNA expression(P<0.05, P<0.01), ABCA1 mRNA expression level was significantly up-regulated, the levels of IL-1β and NF-κB were significantly reduced(P<0.01), and PPARγ protein expression level was significantly reduced(P<0.05). ConclusionThis study identifies 52 components and their metabolites in ZRH intestinal absorption fluid that are positively correlated with the inhibition of macrophage foaming, which may be related to the regulation of the PPARs pathway in cells and the reduction of the levels of inflammatory factors, and can provide a reference for the quality control and clinical application of ZRH.
8.Influencing factors of the TEG maximum amplitude reduction in emergency trauma patients
Changfu JI ; Dong LAI ; Yan TIAN ; Fei LAI ; Shuting LIN ; Min LONG ; Fangyuan LI ; Fangling LIAN
Chinese Journal of Blood Transfusion 2024;37(6):638-642
【Objective】 To investigate the causes of abnormal decrease in maximum amplitude(MA) of thromboelastography(TEG) and its effect on prognosis by monitoring the changes of coagulation-related indexes in emergency trauma patients. 【Methods】 A total of 319 cases of trauma patients admitted to our hospital from September 2020 to September 2023 were retrospectively analyzed, and the coagulation-related indexes of 0 h and 24 h after admission were observed. According to the MA results, they were divided into normal MA group(>50 mm) and reduced MA group(≤50 mm) to compare the hemoglobin(Hb), platelets count(Plt), activated partial thromboplastin time(APTT), prothrombin time(PT), fibrinogen(Fib), thrombin time(TT), D-dimer(D-D), coagulation reaction time(R), clot formation kinetics(Angle), 30 min clot dissolution rate(Ly30), MA, thrombine-antithrombin complex(TAT) and plasminase-α2 plasminase inhibitor complex(PIC). The correlation between MA and fibrinolysis indexes in 319 trauma patients was analyzed. According to whether tranexamic acid(TXA) was used, the reduced MA group was divided into a TXA group and a non-drug group. The differences in the change of the above coagulation-related indexes, mortality rate and changes in blood product dosage were compared between the two groups. 【Results】 Compared with the normal MA group, Hb, Plt, Fib, diastolic blood pressure and GCS scores decreased, while heart rate, ISS score and mortality increased significantly in the reduced MA group(P<0.05). The R, PT and TT were prolonged significantly(P<0.05), and PIC and D-D increased significantly(P<0.05) in the reduced MA group. Correlation analysis found that MA had no correlation with Ly30, TAT and APTT, but was correlated with Angle(r=0.803), Plt(r=0.544), Fib(r=0.581), PIC(r=-0.443) and D-D(r=-0.343). Compared with the non-drug group, the change of Angle, MA and FIB in the TXA group increased significantly(P<0.05), while the change of PIC decreased(P<0.05). Cryoprecipitate and platelet transfusion in the TXA group reduced significantly(P<0.05), and red blood cell transfusion had a decreasing trend, but the difference was not significant(P>0.05). The mortality rate in the TXA group was reduced significantly(P<0.05). 【Conclusion】 Hyperfibrinolysis may be an important factor in the abnormal decrease of MA in emergency trauma patients. Treatment with TXA can improve its effect on MA, and reduce the transfusion of blood products and the patient mortality.