1.Inhibitory effects of jiajianzhujing decoction containing serum on proliferation of rat choroidal vascular endothelial cells
Yanjiang, LIU ; Zefeng, KANG ; Kai, XING ; Jian, LIU
Chinese Journal of Experimental Ophthalmology 2016;34(12):1065-1071
Background Choroidal neovascularization (CNV) of macular zone is one of blinding eye diseases.Conventional treatment methods include photodynamic therapy and intravitreal injection of anti-vascular endothelial growth factor (VEGF) drugs.However,there are some adverse effects following these treatments.Researches showed that fiajianzhujing decoction,a Chinese herbal medicine,can inhibit CNV.Objective This study was to investigate the impact of drug serum offiajianzhujing decocation (drug serum) on the proliferation of rat choroidal vascular endothelial cells (RCVECs).Methods Jiajianzhujing decoction (0.525 g/ml) was used by intragastric administration with the dosage 20 ml/kg for 3 days,and the abdominal aortic blood was collected 2 hours after last dosage to prepare the drug serum.Cultured RCVECs were divided into 4 groups.CoCl2 with the concentration of 100 μmol/L (100 μl) was added in the medium to establish the anoxic models in the CoCl2 group,and 20% drug serum was added in the anoxic culture medium in the drug serum+CoCl2 group.Regular culture medium was used in the blank control group,and 20% pure serum was added in the anoxic culture medium in the pure serum+CoCl2 group.The cells were incubated for consecutive 24 hours,and the growth of the cells was detected by monotetrazolium (MTT) assay as absorbance (A).The expressions of gene and protein of hypoxia-inducible factor-1 α (HIF-1 α) and VEGF in the cells were detected by reverse transcription PCR and Western blot,respectively.This study was approved by the Ethics Committee of Eye Hospital of China Academy of Chinese Medical Sciences.Results Cultured cells grew well with fusiform shape.Hypoxic cell models were created by adding CoCl2.The mean A values were 0.659± 0.051,0.757±0.553,0.683±0.037 and O.731 ±O.038 in the blank control group,CoCl2 group,drug serum+CoCl2 group and pure serum+CoCl2 group,respectively,and the A value in the CoCl2 group was significantly elevated in comparison with the blank control group and drug serum+CoCl2 group (both at P<0.O1).The significant differences were found in the relative expressions of HIF-1 α and VEGF among the blank control group,CoCl2 group,drug serum+ CoC12 group and pure serum+CoCl2 group (HIF-1 α:F =3.100,P<0.05;VEGF mRNA:F =3.420,P<0.05;HIF-1 α protein:F=470.600,P =0.000;VEGF protein:F =146.700,P =0.000),and the relative expressions of HIF-1α mRNA,VEGF mRNA,HIF-1α protein and VEGF protein in the CoCl2 group were significantly higher than those in the blank control group and drug serum+CoCl2 group (all at P<0.05).Conclusions Drug serum ofjiajianzhujing decoction can inhibit the growth and proliferation of hypoxic RCVECs by down-regulating the secretion of HIF-1α and VEGF.
2.Association between T(-1340)G polymorphism of ALOX5AP gene and coronary artery disease in the Han population of North China
Zhidong LI ; Xiaolin ZHANG ; Yaling HAN ; Chenghui YAN ; Jian KANG ; Zefeng WU
Chinese Journal of Tissue Engineering Research 2009;13(50):9974-9978
OBJECTIVE:To investigate the possible association between the gene ALOX5AP encoding 5-lipoxygenase activating protein (FLAP)and coronary artery disease(CAD)in the Han population of North China.METHODS:A total of 680 cases underwent selective coronary angiography(SCA)from Shenyang General Hospital of Chinese PLA was recruited from January 2006 to September 2007.According to the results of SCA.680 cases were divided into CAD group with angiography positive(n=336)and control group with angiography negative or the stenosis of coronary arteries<50%(n=344)without evidence of cardiac ischemia.Single nucleotide polymorphisms of ALOX5AP gene was screened in 48 unrelated Han individuals of North China by polymerase chain reaction fPCR)-Re-sequencing method and 7 polymorphisms were found.The genotype and allele distribution of T(-1340)G polymorphism between two groups was determined by polymerase chain reaction and restriction fragment Iength polymorphism(PCR-RFLP)analysis in CAD and controI subjects.RESULTS:The genotype frequencies of TT,TG and GG in the ALOX5AP T(-1 340)G polymorphism were 26.79%,51 179%and 21.43%in CAD patients,33.72%,47.38%and 18.90%in the controls,respectively(x~2=3.90,P>0.06).The genotype distribution between two groups was in accordance with hardy-weinberg equilibrium.There are no significant differences in the distribution of three genotypes between the two groups.The frequencies of ALOX5AP G allele in cases and controls were 47.32%,42.59%,respectively(x~2=3.08,P>0.05).Subsequent stratified analysis by gender also showed no statistical significance in the genotype frequencies and allele frequencies between the two groups.CONCLUSION:The result suggests that T(-1340)G polymorphism of the ALOX5AP gene might not be associated with CAD in the Han population of North China.
3.Effect of Jiajian-Zhujing Decoction on the AKT/mTOR signaling pathway in ARPE-19 cells after AKT transfection
Wenli CHU ; Zefeng KANG ; Shuiling CHEN ; Xuelian HAO ; Jian LIU ; Weiyi LI ; Fangfang TAO
International Journal of Traditional Chinese Medicine 2020;42(4):347-352
Objective:To study the mechanism of the effect of Jiajian-Zhujing Decoction on the expression of VEGF on ARPE-19 cells after AKT transfection. Methods:To prepare the serum and blank serum of Jiajian-Zhujing Decoction and divide ARPE-19 cells into the normal group, model group, blank serum group, medicated serum group, Conbercept group and combined group. Except normal group, this research established AKT transfected cell model. Then cultured the normal group and model group with conventional method, and the blank serum group was cultured with 10% blank serum, the medicated serum group was cultured with 10% medicated serum, the Conbercept group was cultured with 20 μg/ml Conbercept, the combined group was cultured with 10% medicated serum and 20 μg/m Conbercept. The proliferation of ARPE-19 cells in each group was detect by the CCK-8 method. The levels of AKT, mTOR and VEGF mRNA were detected by real-time quantitative PCR. Western blot was used to detect the expression of AKT, mTOR and VEGF. Results:After being cultured for 24, 48 and 72 hours, compared with the model group, the cell proliferation rate in blank serum group, medicated serum group, Conbercept group and combined group significantly decreased ( P<0.05). Compared with the model group, the expression of AKT mRNA (24 h: 3.10 ± 0.48, 1.97 ± 0.14, 1.26 ± 0.24 vs. 4.77 ± 0.68; 48 h: 3.52 ± 0.82, 2.62 ± 0.77, 1.10 ± 0.19 vs. 6.12 ± 1.21), mTOR mRNA (24 h: 3.02 ± 0.26, 2.45 ± 0.75, 1.13 ± 0.15 vs. 4.48 ± 0.80; 48 h: 1.29 ± 0.30, 1.30 ± 0.57, 0.65 ± 0.19 vs. 2.54 ± 0.62), VEGF mRNA (24 h: 3.33 ± 0.62, 2.18 ± 0.20, 1.55 ± 0.28 vs. 5.53 ± 1.02; 48 h: 2.35 ± 0.54, 1.23 ± 0.28, 0.93 ± 0.25 vs. 3.59 ± 0.40), AKT protion (24 h: 0.45 ± 0.09, 0.25 ± 0.05, 0.14 ± 0.04 vs. 0.62 ± 0.04; 48 h: 0.36 ± 0.06, 0.23 ± 0.04, 0.14 ± 0.03 vs. 0.54 ± 0.08), mTOR protion (24 h: 0.35 ± 0.05, 0.24 ± 0.02, 0.18 ± 0.02 vs. 0.52 ± 0.09; 48 h: 0.23 ± 0.04, 0.29 ± 0.04, 0.14 ± 0.03 vs. 0.40 ± 0.10), VEGF protion (24 h: 0.14 ± 0.03, 0.33 ± 0.04, 0.24 ± 0.03 vs. 0.54 ± 0.10; 48 h: 0.24 ± 0.03, 0.17 ± 0.02, 0.11 ± 0.02 vs. 0.42 ± 0.10) significantly decreased ( P<0.05), and the combined group was significantly lower than that of the Conbercept group ( P<0.05). Conclusions:AKT transfection can promote the proliferation of ARPE-19 cells, and Jiajian-Zhujing Decoction can significantly inhibit this proliferation. Jiajian-Zhujing Decoction may inhibit the activity of AKT/mTOR signaling pathway to reduce the expression of VEGF.
4.Efficacy of chemical thoracic sympathetic nerve modulation combined with pulsed radiofrequency in treating upper limb postherpetic neuralgia
Keyue XIE ; Qianying LIU ; Kang AN ; Bing HUANG ; Ming YAO ; Yanbao SUN ; Jianbing MA ; Zefeng ZHU
Chinese Journal of Anesthesiology 2018;38(8):929-932
Objective To evaluate the efficacy of chemical thoracic sympathetic nerve modulation combined with pulsed radiofrequency in treating upper limb postherpetic neuralgia ( PHN). Methods Forty-two patients of both sexes with upper limb PHN, aged 48-75 yr, were divided into 2 groups ( n=21 each) using a random number table method: chemical thoracic sympathetic nerve modulation combined with pulsed radiofrequency group ( TSNM+PR group) and pulsed radiofrequency group ( PR group) . TSNM+PR group was treated using chemical thoracic sympathetic nerve modulation combined with pulsed radiofrequen-cy, and PR group received pulsed radiofrequency alone. The occurrence of treatment-related adverse reac-tions was recorded. Numeric rating scale scores were recorded preoperatively and at 1 day and 1 and 3 months after operation, and the efficacy was graded. The effective treatment and pain recurrence were re-corded 3 months after operation. Quantitative sensory nerve tests were performed to record the current per-ception threshold before operation and on 1 day, 1 month and 3 months after operation. Results Compared with PR group, numeric rating scale score was significantly decreased, the therapeutic effect was en-hanced, the rate of effective treatment was increased, the recurrence rate of pain was decreased at 1 and 3 months after surgery, the current perception threshold at 250 and 5 Hz on the ipsilateral side was increased at 1 and 3 months after surgery in TSNM+PR group ( P<0. 05) . No treatment-related adverse reactions were found in two groups. Conclusion Chemical thoracic sympathetic nerve modulation combined with pulsed radiofrequency provides reliable therapeutic effect and higher safety for upper limb PHN.
5.Effect of Parkinson′s disease factor on sedative efficacy of dexmedetomidine
Zefeng FAN ; Qiuyue FU ; Xiang LIU ; Chengwei YANG ; Mingming HAN ; Wanhong LI ; Juan LI ; Fang KANG
Chinese Journal of Anesthesiology 2022;42(11):1285-1288
Objective:To evaluate the effect of Parkinson′s disease factor on the sedative efficacy of dexmedetomidine.Methods:The patients of either sex, aged 45-64 yr, of American Society of Anesthesiologists Physical Status classification Ⅱor Ⅲ, with body mass index of 18.5-30.0 kg/m 2, undergoing non-intracranial space-occupying lesions in neurosurgery, were selected.Patients were divided into control group (group C) and Parkinson′s disease group (group P) according to whether they had Parkinson′s disease or not.The ED 50 of dexmedetomidine was determined by using the Dixon′s up-and-down method.The initial dose of dexmedetomidine was 0.5 μg/kg in both groups, and each time the concentration increased/decreased by 0.05 μg/kg in the next patient, which was repeated until 7th independent crossover pair (loss of consciousness) appeared, and then the test was ended.The ED 50 and 95% confidence interval of dexmedetomidine inducing loss of consciousness were calculated using the probit test in a Logistic regression model.Hypertension, hypotension, bradycardia and nausea and vomiting were recorded. Results:Compared with group C, the ED 50 of dexmedetomidine inducing loss of consciousness was significantly increased in group P ( P<0.05), and no significant change was found in the incidence of adverse reactions in group P ( P>0.05). Conclusions:Parkinson′s disease factor can decrease the sedative efficacy of dexmedetomidine.
6.Risk Factors for Duodenal Stump Leakage after Laparoscopic Gastrectomy for Gastric Cancer
Lihu GU ; Kang ZHANG ; Zefeng SHEN ; Xianfa WANG ; Hepan ZHU ; Junhai PAN ; Xin ZHONG ; Parikshit Asutosh KHADAROO ; Ping CHEN
Journal of Gastric Cancer 2020;20(1):81-94
PURPOSE:
Duodenal stump leakage (DSL) is a potentially fatal complication that can occur after gastrectomy, but its underlying risk factors are unclear. This study aimed to investigate the risk factors and management of DSL after laparoscopic radical gastrectomy for gastric cancer (GC).
MATERIALS AND METHODS:
Relevant data were collected from several prospective databases to retrospectively analyze the data of GC patients who underwent Billroth II (B-II) or Roux-en-Y (R-Y) reconstruction after laparoscopic gastrectomy from 2 institutions (Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, and HwaMei Hospital, University of Chinese Academy of Sciences). The DSL risk factors were analyzed using univariate and multivariate analysis regression.
RESULTS:
A total of 810 patients were eligible for our analysis (426 with R-Y, 384 with B-II with Braun). Eleven patients had DSL (1.36%). Body mass index (BMI), elevated preoperative C-reactive protein (CRP) level, and unreinforced duodenal stump were the independent risk factors for DSL. DSL was diagnosed in 2–12 days, with a median of 8 days. Seven patients received conservative treatment, 3 patients received puncture treatment, and only 1 patient required reoperation. All patients recovered successfully after treatment.
CONCLUSIONS
The risk factors of DSL were BMI ≥24 kg/m², elevated preoperative CRP level, and unreinforced duodenal stump. Nonsurgical treatments for DSL are preferred.
7.Advances in research on relationship between indoor artificial light at night and human health
Fengrui ZHU ; Zefeng KANG ; Furong DENG
Journal of Environmental and Occupational Medicine 2024;41(2):214-218
With the acceleration of global urbanization, the intensity and coverage of artificial light at night (ALAN) are increasing, and its service duration is obviously prolonged. ALAN exposure is not only related to the occurrence and development of cardiovascular, metabolic, sleep, myopia, and mental diseases, but also may induce cancer. Previous studies have focused on the health effects of outdoor ALAN, but people spend more than 80% of their lives indoors, hence it is of great significance to understand the relationship between indoor ALAN and population health to create a healthy indoor environment and protect the health of the population.
8.Development and application of regulatory T cells in corneal transplant rejection
Xin DU ; Ruijuan GUAN ; Zefeng KANG ; Ling LI ; Xin YAN ; Xiaoying ZHANG
International Eye Science 2024;24(8):1246-1249
Corneal transplantation is an effective treatment for corneal blindness, and it is the only hope for patients with corneal blindness. Cornea has no blood vessels and no lymphatic vessels, which is called immune privilege organ, so the success rate of corneal transplantation is significantly higher than that of other organ transplantation, but the rejection reaction after corneal transplantation is still the main reason for the failure of corneal transplantation. The directional movement of immune cells to lymphoid tissue and inflammatory sites is the mainly immune response after organ transplantation. And the regulatory T cells(Treg)play a key role in immune regulation, which can induce immune tolerance by regulating and inhibiting the activation of effector T cells and reduce the rejection reaction after corneal transplantation. In addition, this review also discussed the effectiveness of applying cordyceps sinensis extract FTY720 to enhance the function of Treg. Based on this, we briefly reviewed the sources, mechanism of action and treatment of Treg after corneal transplantation, so as to provide some reference for the subsequent clinical application transformation and basic research.
9.Regulatory role of PI3K/AKT pathway in diabetic retinopathy
Baohua LI ; Zefeng KANG ; Xinyue HOU ; Jianquan WANG ; Man SONG ; Danyu LI ; Mengyu LIU ; Xin YAN
International Eye Science 2024;24(9):1426-1431
Diabetic retinopathy(DR)is one of the major complications of diabetes mellitus, characterized by neurodegeneration and microangiopathy. Currently, the treatment of DR is mainly focused on the management of late complications and has not achieved the desired clinical outcome. Evidence suggests that the PI3K/AKT pathway, as one of the important intracellular signaling pathways during the cell cycle, is involved in the whole process of DR pathogenesis. This article focuses on the structural composition, activation and blocking pathways, conduction pathways, regulatory mechanisms and biological functions of the PI3K/AKT signaling pathway to review its role in DR and to explore the potential of targeting the PI3K/AKT pathway for the treatment of DR.
10.Pressure-induced differentiation of rabbit retinal stem cells into retinal ganglion cells
Min DAI ; Zefeng KANG ; Zhulin HU ; Yan LI
Chinese Journal of Experimental Ophthalmology 2022;40(12):1134-1140
Objective:To investigate the effect of pressure on the differentiation of rabbit retinal stem cells (RSCs) co-cultured with retinal ganglion cells (RGCs).Methods:SPF grade New Zealand rabbits on the day 22 of gestation were selected, and embryos were removed to obtain retinal ciliary margin pigment epithelial tissue and culture primary RSCs.Six SPF grade newborn New Zealand rabbits were selected, and retinal neuroepithelial layer tissues were isolated to culture primary RGCs.Rabbit RSCs cultured in vitro were identified by immunofluorescence staining of nestin antibody, bromodeoxyuridine (BrdU) cell proliferation assay kit, RSCs spontaneously differentiated cells immunofluorescence detection and flow cytometry.RGCs were identified through immunofluorescence staining of Brn3b antibody and Thy1.1 antibody.A co-culture system of RGCs and RSCs cultured in the upper and lower layers of a transwelll plate respectively was constructed.The mRNA and protein expression levels of nestin and Thy1.1 in RSCs and differentiated cells under pressures of 0, 20, 40, 60, 80 mmHg (1 mmHg=0.133 kPa) were detected by real-time fluorescence quantitative PCR and Western blot.The feeding and use of laboratory animals were in accordance with the Regulations on the Administration of Laboratory Animals promulgated by the State Science and Technology Commission.The study protocol was approved by the Ethics Committee of Yunnan University Affiliated Hospital (No.KPRC-IACUC17008). Results:RSCs cultured in vitro were nestin-positive.The percentage of BrdU-positive isolated RSCs was (92.26±3.28)%.Some cells differentiated from RSCs were Brn3b-positive, accounting for (13.00±3.06)%, and some were GS-positive, accounting for (31.60±3.67)%.RGCs cultured in vitro were Brn3b- and Thy1.1-positive.There were statistically significant differences in the relative mRNA and protein expressions of nestin and Thy1.1 between RSCs and differentiated cells under different pressures (mRNA: F=127.600, 137.400; both at P<0.01; protein: F=82.480, 158.700; both at P<0.001). The relative mRNA and protein expressions of nestin were significantly reduced in RSCs, and relative mRNA and protein expressions of Thy1.1 were significantly increased in differentiated cells at 20, 40, 60 and 80 mmHg in comparison with 0 mmHg (all at P<0.05). When the pressure was 40 mmHg, the relative mRNA and protein expressions of nestin were lowest in RSCs, and the relative mRNA and protein expressions of Thy1.1 in differentiated cells were highest. Conclusions:Within a certain range, pressure can promote the differentiation of RSCs co-cultured with RGCs into ganglion-like cells, and excessive pressure can inhibit the differentiation of RSCs.