1.Protective effects of cordycepin on renal proximal tubular cells injury induced by lipopolysaccharide
Yongli YANG ; Hui XIANG ; Zhiying XIAO
Chinese Critical Care Medicine 2021;33(2):203-206
Objective:To investigate the protective effect and potential mechanism of cordycepin on renal proximal tubular cells injury induced by lipopolysaccharide (LPS).Methods:Renal proximal tubular cells NRK-52E were incubated on a cell culture plated at a density of 1×10 5/mL for experiment, then divided into control group (Ctrl group), LPS group (cells were stimulated with 1 mg/L LPS), 10 μmol/L or 20 μmol/L cordycep in intervention groups (LPS+C 10 group and LPS+C 20 group). Cell viability was measured using cell counting kit-8 (CCK-8) reagent. The level of intracellular reactive oxygen species (ROS) was detected by 2',7'-dichlorofluorescin diacetate (DCFH-DA) staining. The protein expressions of inflammatory factors intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), interleukin-1β (IL-1β), and nuclear factor-κB (NF-κB) were detected by Western blotting. Results:Compared with the Ctrl group, LPS significantly inhibited NRK-52E cell viability, increased intracellular ROS, and up-regulated the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB. Compared with LPS group, after treated with 10 μmol/L or 20 μmol/L cordycepin, NRK-52E cell viability was significantly increased (Ctrl group as 1: 0.717±0.017, 0.916±0.036 vs. 0.554±0.046) and intracellular ROS level was significantly decreased (Ctrl group as 1: 1.527±0.165, 1.098±0.168 vs. 2.543±0.127), meanwhile the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB were significantly down-regulated [Ctrl group as 1, ICAM-1/GAPDH: 2.364±0.097, 1.561±0.074 vs. 3.101±0.121; VCAM-1/GAPDH: 2.866±0.135, 1.920±0.098 vs. 4.170±0.119; IL-1β/GAPDH: 2.358±0.107, 1.563±0.179 vs. 3.301±0.210; phosphorylation NF-κB p65 (NF-κB p-p65)/GAPDH: 2.559±0.166, 1.596±0.148 vs. 3.183±0.098], the differences were statistically significant (all P < 0.05). Compared with the LPS+C 10 group, the cell activity of LPS+C 20 group was more significant (0.916±0.036 vs. 0.717±0.017, P < 0.01), and the expressions of ICAM-1, VCAM-1, IL-1β, NF-κB were down-regulated more significantly (ICAM-1/GAPDH: 1.561±0.074 vs. 2.364±0.097, VCAM-1/GAPDH: 1.920±0.098 vs. 2.866±0.135, IL-1β/GAPDH: 1.563±0.179 vs. 2.358±0.107, NF-κB p-p65/GAPDH: 1.596±0.148 vs. 2.559±0.166, all P < 0.05).Conclusion:Cordycepin could significantly increase the survival rate of NRK-52E cells, reduce intracellular ROS level, and inhibit inflammation, and the anti-inflammation effect can be related with NF-κB pathway.
3.Preparation of monoclonal antibody against human tissue kallikrein and development of an ELISA kit
Xiao RAN ; Qin ZHANG ; Zhixiong XIANG ; Hui YANG
Chinese Journal of Microbiology and Immunology 2011;31(2):178-182
Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.
4.Evaluation of parameters for predicting the response to clomiphene citrate in women with polycystic ovary syndrome
Hui MAO ; Wenyan XI ; Yongkang YANG ; Xiang XUE ; Shengyu FU
Journal of Chinese Physician 2015;17(12):1823-1826
Objective To investigate whether clinical,endocrine,and sonographic characteristics of infertile women with polycystic ovary syndrome (PCOS) could predict the ovarian response to clomiphene citrate (CC) medication.Methods A prospective study was conducted,85 women with PCOS were given CC 50 mg per day for 5 days.In the case of an absent response,doses were increased to 100 daily in subsequent cycles.Clinical,endocrine,and sonographic characteristics were compared between responders and nonresponders.Results The cycle history (oligomenorrhea or amenorrhea) and antral follicle count (AFC) were significantly different in responders from those in nonresponders (P < 0.05).There were no differences in age,serum testosterone,follicle stimulating hormone (FSH),luteotropic hormone (LH),body mass index (BMI),insulin resistant index (HOMA-IR),and glucose and insulin levels between the two groups (P > 0.05).In addition,multivariate logistic regression analysis indicated that cycle history and AFC were the predictors of ovarian response to CC treatment.Conclusions The cycle history and AFC can be used as the simple and effective parameters to predict ovarian response to CC treatment in PCOS patients.
5.Changes of Plasma Brain Natriuretic Peptide and N-Terminal Pro-B-Type Natriuretic Peptide of Sepsis Combined with Myocardial Injury in Newborns
ying-chun, HUI ; xiang-yu, DONG ; yang, SHEN ; qian, NI
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To explore the changes of plasma brain natriuretic peptide(BNP) and N-terminal pro-B-type natriuretic peptide (NT-proBNP)of sepsis combined with myocardial injury in newborns.Methods According to neonatal sepsis treatment program,45 cases of sepsis newborns in NICU of the Second Hospital in Lanzhou University from Jul.2007 to Jun.2008 were collected.According to the myocardial injury diagnostic criteria,45 cases neonatal sepsis were divided into myocardial injury group(n=22) and non-myocardial injury group(n=23).Myocardial injury group was also divided into congenital heart disease group and non-congenital heart disease group accor-ding to echocardiography.At the same time,30 healthy newborns were collected as healthy control group.Every newborns were tested the level of plasma BNP,NT-proBNP,creatine kinase-MB(CK-MB) and cardiac-troponin I (cTnI).Results There were significant difference between myocardial injury group,non-myocardial injury group and healthy control group in the levels of plasma BNP,NT-proBNP,CK-MB and cTnI,those in congenital heart disease group were higher than those in non-myocardial injury group and the healthy control group(Pa0.05).Conclusions BNP and NT-proBNP can be early used to diagnose myocardial injury and heart failure of neonatal sepsis associated with CK-MB and cTnI.In NICU,infants with sepsis should normally test BNP and NT-proBNP in order to early diagnose myocardial injury of neonatal sepsis.
6.Clinicopathological characteristics of inflammatory myofibroblastic tumor: analysis of 173 cases
Qin XIANG ; Yingan JIANG ; Hui SUN ; Guifang YANG ; Jun LIN
Chinese Journal of General Practitioners 2012;(10):769-771
The clinicopathological data of 173 cases of inflammatory myofibroblastic tumor (IMT)were retrospectively analyzed.Among them 125 were males and 48 females with a mean age of 47.9 y (5-78 y).The lesions of 117 cases were located in lungs,41 cases in eyes,8 in ileocecum,2 in liver,2 in spleen,1 in abdominal wall,1 in maxillary sinus and 1 in face.The average diameter of lesions were 3.5 cm,ranging from 1.0 cm to 7.0 cm.The clinical manifestations were not specific,depending on the locations of tumor.The imaging examinations were helpful for diagnosis.The prcsence of slender-spindled myofibroblasts and proliferation of fibroblast cells,with infiltration of chronic inflammatory cells were the basic histopathological features of the disease.Combined with the histological characteristics and immunohistochemical staining IMT can be differentiated from other spindle cell tumors.Appropriate surgical resection is the main treatment for IMT.
7.Application of the cell-free plasma DNA in the diagnosis and treatment of malignancies
Xiang ZHU ; Hui WU ; Aihua YUAN ; Kunxing YANG ; Hongyong CAO
Journal of International Oncology 2014;41(3):180-183
The cell-free plasma DNA (cfpDNA) has been suggested as a useful tumor marker for its quantitative and qualitative tumor-specific alterations that reflect the biological characteristics and the progression and outcomes of tumors.Therefore,it has been used as liquid biopsy to detect cfpDNA in peripheral blood for the diagnosis,monitoring of clinical effects,and prognosis of malignancies
8.Analysis of Hemorheological Examination of 3483 Healthy Adults in Xi'an
Yong-ying LIU ; Ji-qing YANG ; Hui WANG ; Zhen XIANG
Chinese Journal of Rehabilitation Theory and Practice 2006;12(8):721-722
ObjectiveTo analyze the relationship between result of hemorheology and non-pathological factors such as sex, age and life habit.Methods3483 healthy adults who had health examination were divided into different groups according to sex and age, and results of hemorheological test of them were analyzed and compared with reference values.ResultsAll hemorheological indexes of men were higher than that of women. The whole blood viscosity of female had an increasing trend along with the age increasing. However, the result of hemorheology of male showed that the index of the age of 30~49 was higher than the age of more than 50, and had a decreasing trend along with the age increasing after the age of 50. The index of high shear viscosity, low shear viscosity and hematocrit of both male and female were all higher than the reference values offered by apparatus.ConclusionEffect of non-pathological factors such as age, sex and life habit on index of hemorheology should be considered.
9.Advances in antiviral research of adaptor-associated protein kinase 1 (AAK1) inhibitors
Xiang QI ; Song-wei JIANG ; Ying-hui YUAN ; Li XU ; Zi HUI ; Xiang-yang YE ; Tian XIE
Acta Pharmaceutica Sinica 2022;57(7):1991-2002
As one of the major sources of infection, viruses could infect all organisms including bacteria, plants, animals, and humans. Infectious diseases caused by viruses pose a great threat and damage to human health and economic activities all over the world. Adaptor-associated protein kinase 1 (AAK1) is a member of the Ark1/Prk1 family of serine/threonine kinases and a specific key kinase regulating the phosphorylation of AP-2 protein μ2 subunit T156. In the past, AAK1 has been regarded as a feasible biological target for the treatment of nerve pain. Recently, scientists have found that inhibiting AAK1 can regulate endocytosis and inhibit virus invasion into cells. Therefore, AAK1 could be the potential target of anti-virus therapy. This paper reviews the research progress of small molecule AAK1 inhibitors in the field of antiviral, analyzes the future research directions and challenges, and provides new ideas for the development of antiviral drugs targeting AAK1.
10.Research progress of ATR kinase-targeted inhibitors in the cancer therapy
Ying-hui YUAN ; Ji-long DUAN ; Zi HUI ; Tian XIE ; Xiang-yang YE
Acta Pharmaceutica Sinica 2022;57(3):593-604
Cancer, also known as malignant tumor, is the second largest disease after heart disease, which is characterized by genomic instability and mutagenicity. Ataxia telangiectasia and RAD3-related kinase (ATR) are members of phosphatidylinositol 3-kinase (PIKK) family, belonging to serine/threonine kinase, one of the key kinases in DNA damage response (DDR) and DNA repair pathway. This paper reviews the latest progress in the ATR inhibitor field including mechanism of action (MOA), therapeutic applications, and the combination therapy from the perspective of medicinal chemistry. It also discusses the possible challenges and future directions of developing ATR inhibitor antitumor drugs, which could provide the scientists in this field the convenience for access the information and application guidance for clinical studies.