1.Influence of RNAi inhibiting JMJD3 gene expression on growth,proliferation and invasion of hepatocellular carcinoma cells
International Journal of Laboratory Medicine 2017;38(10):1333-1335
Objective To study the influence of small interfering RNA(siRNA) inhibiting Jumonji-containing JMJD3 gene on growth,proliferation and invasion of hepatocellular carcinoma cells.Methods JMJD3 was silenced by RNA interference(siR-JMJD3),non-specificity sequence pSilercer 2.1 was transfected into liver cancer cells QGY 7703 and HepG2 cells as the negative control.MTT assay,colony formation and Transwell invasion assays were used to detect the cell growth,proliferation and invasion ability of hepatocellular carcinoma cells.Results The Western Blot detection results showed that the siR-JMJD3 plasmid transfected experiment group successfully inhibited the JMJD3 protein expression level in liver cancer cells.The MTT results showed that after transfecting siR-JMJD3,the growth activity of QGY-7703 and HepG2 were decreased by 25% or 17% compared with the control group,.The colony formation results showed that in the colony formation number of two cell lines were decreased by 31% and 25% respectively.The Transwell invasion test results indicated that transmembrane cells number was decreased by 44% and 47% respectively.Conclusion Using siRNA technique can effectively inhibit JMJD3 gene expression,meanwhile effectively inhibits the in vitro growth,proliferation and invasion of liver cancer cells QGY-7703 and HepG2 and provides a new thinking for the biological therapy of liver cancer.
2.Quality Standard for Biyankang Granule
China Pharmacist 2016;19(9):1788-1790,1791
Objective: To establish a method for the quality standard of Biyankang granule. Methods: Scutellaria baicalensis Georgi and Puerariae Iobatae radix were identified by TLC. The content of baicalin was determined by HPLC with a Dubhe C18 column (250 mm × 4. 6 mm, 5 mm). The mobile phase was methanol and 0. 4% phosphoric acid with gradient elution. The flow rate was 1. 0 ml· min-1 and the detection wavelength was 280 nm. The column temperature was 30℃ and the injection volume was 10 ml. Re-sults:The spots in TLC were clear without any interference. A good linear relationship was shown within the range of 15. 02-300. 43 m g·ml-1 for baicalin (r=1.000 0). The average recovery of baicalin was 102.62% (RSD =0.72%, n =6). Conclusion: The method is specific with high accuracy and good reproducibility, which can be used for the quality control of Biyankang granule.
3.Influences of notoginsenoside R1 on the injury of alveolar epithelial cells infected by Klebsiella pneumoniae by regulating JAK2STAT3 signaling pathway
LIU Huixue ; ZHANG Peipei ; AN Jiangke
China Tropical Medicine 2023;23(7):754-
Abstract: Objective To investigate the influences of notoginsenoside R1 (NGR1) on cell injury and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway of alveolar epithelial cells infected by Klebsiella pneumoniae (Kp). Methods A549 cells were grouped into five groups: control group (C group), infection group (Infect group), infection + low NGR1 group (Infect + L-NGR1 group), infection + high NGR1 group (Infect + H-NGR1 group), and infection+high NGR1+JAK2/STAT3 pathway inhibitor group (Infect+H-NGR1+SD-1029 group). Cell proliferation was measured using CCK8; ELISA kits were applied to detect the contents of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interferon γ (IFN-γ) in the culture medium; flow cytometry was applied to detect apoptosis; RT-qPCR was applied to detect the expressions of JAK2/STAT3; Western blot was applied to detect JAK2/STAT3 pathway, autophagy protein microtubule-associated protein 1 light chain 3 (LC3), autophagy-relatedgene5 (Atg5), autophagy-related gene (Atg) 6 (Beclin-1), apoptosis protein B-cell lymphoma 2 (Bcl-2), Bcl-2-accociated protein (Bax), cysteinyl aspartate specific proteinase (cleaved-caspase-3) proteins expression. Results Compared with the C group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect group were obviously decreased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Compared with Infect group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect+L-NGR1 group and Infect+H-NGR1 group were obviously increased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-Caspase-3 were obviously decreased (P<0.05). Compared with Infect+H-NGR1 group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the protein phosphorylation levels of JAK2, STAT3 in the Infect+H-NGR1+SD-1029 group were obviously decreased (P<0.05), and the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Conclusions NGR1 can activate the JAK2/STAT3 signaling pathway, promote autophagy of alveolar epithelial cells, and inhibit Kp-induced inflammatory injury and apoptosis of alveolar epithelial cells.
4.Development of surgical first aid kits of for combat readiness
Suqin MIAO ; Qiong ZHANG ; Peipei SHEN
Journal of Regional Anatomy and Operative Surgery 2016;(2):87-89,90
Objective To develop new surgical first aid kits for combat readiness ,and increase the ability of field first aid .Methods On the basis of disposable surgical kits ,the surgical towels were improved .The water absorbing layer was made of SPA materials which inte-grated optimization for surgical use .Results The new surgical first aid kits for combat readiness have the surgical towels with strong water absorption,complete sterile materials,and short time for ready of supplies.Conclusion The new surgical first aid kits for combat readiness have the advantages of small volume ,practicability,available for surgery ,at the same time,which could keep the surgical incision dry and the temperature of patients stability and be helpful to decrease the risk of pressure ulcer and infection ,simplify work procedure .
5.The clinical value of multi-slice spiral CT in diagnosis of non-angle caused nutcracker syndrome
Peipei HAO ; Xuehua YANG ; Yonggao ZHANG
Journal of Practical Radiology 2014;(4):631-634
Objective To evaluate the diagnostic value of multi-slice spiral CT for non-angle caused nutcracker syndrome(NCS). Methods A total of 177 cases of clinical suspected nutcracker syndrome patients recevied multi-slice spiral CT examination,,inclu-ding 33 patients with negative results,144 cases patients with positive results.The analysis was performed by measuring the angle between superior mesenteric artery(SMA)and the abdominal aorta(AA),and the ratio of the diameter of the narrowest and expan-sion in the left renal vein.Results Within 33 cases of nutcracker-negative patients,the angle of SMA and AA was (46.06 ± 24.46)°;the ratio was (2.11±0.76).144 cases was diagnosed as nutcracker-positive patients,the angle of SMA and AA was (23. 10±9.63)°;the ratio was (3.83±1.24).There was positive correlation between angle and ratio in NCS (P<0.0001).Among9 ca-ses of nutcracker syndrome caused by non-angle factors three were oppressed by the uncinate process pancreas,two cases were caused by the malformations of the left renal vein,one case was posterior nutcracker syndrome,one case was oppressed by portal veinOne case of pressured renal vein due to tortuosity renal vein coiled renal artery.One case of the left renal vein compression lumen narrowing,causing significant downward pressure on the left renal vein.Conclusion Spiral CT angiography reconstruction can clear-ly show the cause and the location of the left renal vein compression,which may provide helpful information in diagnosing nutcracker syndrome caused by non-angle factors.
6.Inhibition of laser-induced choroidal neovascularization by anti-osteopontin antibody
Yan, SU ; Peipei, ZHANG ; Fang, LIU
Chinese Journal of Experimental Ophthalmology 2014;32(9):813-818
Background It has been proved that as an important adhesion protein of extracellular matrix,osteopontion (OPN) can affect tumor neovascularization.Some new researches showed that anti-OPN antibody plays a role in regulating the neovascular vessel formation.Choroidal neovascularization (CNV) has the same structure with tumor neovascularization,but whether anti-OPN antibody restricts new vessel formation is unclear.Objective This study was to investigate the inhibitory effect of anti-OPN antibody on CNV.Methods Laser-induced CNV models were created in 40 eyes of 40 male SPF C57BL/6J mice by Argon laser photocoagulation of retinas,with the wavelength 514 nm.Thirty-six successful models were randomly divided into anti-OPN antibody group,mouse-IgG group and PBS group by the randomized number table.On the second day after photocoagulation,anti-OPN antibody of 400 μg was intraperitoneally injected in the anti-OPN antibody group,and the equivalent amount of mouse IgG and PBS were used in the same way in the mouse IgG group and PBS group.The CNV was evaluated by fundus fluorescein angiography (FFA) on the seventh days after photocoagulation.The mice were immediately sacrificed and the eyeballs were enucleated on the fourteenth day after photocoagulation,and 4 eyeballs in each group were used to observe the areas of CNV on the retinal pigmental epithelium-choroid-sclera fiat mounts,and the other 8 eyeballs of each groups were used to analyze the expression levels of OPN mRNA and vascular endothelial growth factor(VEGF) mRNA using quantitative fluorescence-PCR (QF-PCR).Results FFA showed fluorescein leakage areas around laser spots 7 days after photocoagulation,indicating that CNV appeared.The CNV areas were ([16.98±0.70] × 103) μm2,([27.13 ± 0.81] × 103) μm2 and ([35.39±2.14] ×103) μm2 respectively in the anti-OPN antibody group,mouse IgG group and PBS group,with a significant difference among the 3 groups (F =533.76,P =0.00),and the CNV area was significantly smaller in the anti-OPN antibody group compared with those of the mouse IgG group and PBS group (q =-3.95,-4.40,both at P<0.05).No significant difference was found in the OPN mRNA expression between the antiOPN antibody group and mouse IgG group (t =-5.26,P =0.66).However,the expression of VEGF mRNA in choroidal tissue was significantly declined in the anti-OPN antibody group than that in the mouse IgG group (t =-6.74,P<0.01).Conclusions Anti-OPN antibody suppresses the formation of CNV in laser-induced mouse model by down-regulating VEGF.
7.Clinical Observation of Tiaozhi Decoction for Treatment of Coronary Heart Disease Induced Stable Angina Complicated with Hyperlipemia
Ting LIU ; Shirong ZHANG ; Peipei LI
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(1):35-39
Objective To investigate the therapeutic effect of Tiaozhi Decoction for the treatment of coronary heart disease induced stable angina complicated with hyperlipemia, and to observe its effect on serum inflammatory factors. Results One hundred qualified patients were evenly randomized into treatment group and control group. Both groups were given conventional western medical treatment with reference to Guide for Diagnosis and Treatment of Chronic Stable Angina, and additionally, the control group was given oral use of Simvastatin and the treatment group was given Tiaozhi Decoction orally. The treatment of the two groups covered 8 weeks. Before and after treatment, blood lipid levels of total cholesterol ( TC) , triglyceride ( TG) , low-density lipoprotein cholesterol ( LDL-C) , high- density lipoprotein cholesterol ( HDL-C) , apolipoprotein A (ApoA) and apolipoprotein B (ApoB) were observed. The frequency of angina pectoris attack and the dosage of Nitroglycerin Tablets per week in both groups were recorded during the treatment. Therapeutic effect on lowering blood lipid and on improving electrocardiogram was evaluated after treatment. Serum levels of hypersensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), homocysteic acid (Hcy), adiponectin (APN), and oxidized low-density lipoprotein (ox-LDL) were detected before and after treatment. Results(1) The results of Ridit analysis showed that the treatment group had better therapeutic effect on lowering blood lipid and on improving electrocardiogram than the control group ( P<0.05) . ( 2) After treatment, TG, HDL-C, ApoB and ApoA levels were much improved in the treatment group compared with those in the control group (P<0.01) . ( 3) The frequency of angina pectoris attack and the dosage of Nitroglycerin Tablets per week were reduced in the treatment group compared with those in the control group (P<0.01) . (4) After treatment, the treatment group had lower hs-CRP, IL-6, Hcy and ox-LDL levels, and higher APN level than the control group (P<0.01) . Conclusion Tiaozhi Decoction is effective for the treatment of coronary heart disease induced stable angina complicated with hyperlipemia through improving lipid metabolism and reducing angina pectoris attack, and the mechanism may be related with the regulation of inflammatory factors.
8.Determination of catechin,epicatechin,gaIIic acid and procyanidin B2 in procy-anidins by RP-HPLC
Lu ZHANG ; Peipei PAN ; Saizhen CHEN
Journal of China Pharmaceutical University 2016;(1):54-57
To develop a RP-HPLC method for the determination of catechin (C),epicatechin (EC),gallic acid (GA)and procyanidin B2 (PCB2 )in procyanidins and compare the contents of C,EC,GA and PCB2 in procyani-dins purchased from different manufacturers.A RP-HPLC method was developed and the determination was car-ried out on a Hypersil ODS2 column (4.0 mm ×200 mm,5 μm).The mobile phase consisted of methanol and 2% acetic acid with gradient elution and the detection wave-length was at 280 nm.There was a good linear rela-tionship between concentration and the peak area in the range of 0.1-50 μg/mL (r =0.998 6)for catechin,0.1-50 μg/mL (r =0.994 5)for epicatechin,0.05-50 μg/mL (r =0.999 9)for gallic acid and 0.1-50 μg/mL (r =0.992 2)for procyanidin B2 ,respectively.The average recoveries of catechin,epicatechin,gallic acid and PCB2 were 98.36%,98.21%,89.60% and 98.47%,respectively and the RSDs were 1.39%,0.84%,2.12% and 2.46%,respectively.The method is simple,accurate,reproducible and can be used for assay of C,EC,GA,PCB2 in procyanidins.There was a great difference in the content of four substance in procyanidins purchased from dif-ferent manufacturers.
9.Quantitative evaluation of Ki-67 labeling index in glioma with transfer constant derived from dynamic contrast-enhanced MRI
Fei DONG ; Peipei ZHANG ; Biao JIANG ; Qian LI ; Qiang ZENG ; Weiwei WANG ; Peipei PANG ; Minming ZHANG
Chinese Journal of Radiology 2017;51(8):568-571
Objective To study the value of transfer constant(Ktrans)derived from dynamic contrast-enhanced MRI (DCE-MRI) for quantitative evaluation of Ki-67 labeling index (Ki-67 LI) in glioma. Methods Twenty patients with glioma who underwent DCE-MRI and operation were retrospectively reviewed. The Ktrans value and Ki-67 LI were acquired and correlated using the Spearman correlation test. Also, the Ktrans values were compared between high(larger than 10%)and low(no more than 10%)Ki-67 LI group with Mann-Whitney U test, receiver operating characteristic curves was performed to evaluate the diagnostic value. Results The Ktrans value(0.0165 to 0.8048, median 0.1252)was significantly associated with Ki-67 LI(5%to 50%, median 20%) (r=0.721,P<0.001), and the Ktrans value was significantly higher in high Ki-67 group(0.0810 to 0.8048, median 0.1810)than that in low Ki-67 LI group(0.0165 to 0.1456, median 0.0697)(Z=-3.209, P=0.001). The most predictive Ktrans value differentiated high Ki-67 LI and low Ki-67 LI with an area under the curve(AUC) of 0.945 at a sensitivity of 92.3% and specificity of 85.7%. Conclusion Ktrans value could be used for quantitative evaluation of Ki-67 LI in glioma.
10.Imaging features of malignant external otitis
Peipei ZHANG ; Jianhua YUAN ; Zhongxiang DING ; Fanghong CHEN ; Fuxing ZHANG
Journal of Practical Radiology 2014;(10):1634-1637
Objective To explore the value of computed tomography (CT)and magnetic resonance imaging (MRI)in the diagno-sis and delineation of the extent of malignant external otitis (MEO).Methods Clinical manifestations and imaging features of 10 pa-tients with definite diagnosis of MEO were collected and analyzed.Plain and contrast-enhanced CT of the ear were performed on all 10 patients,of which 6 patients also received MRI scans.Results Abnormal soft tissue opacity within the external auditory canal and involvement of surrounding structures were found in all 10 patients.Bone erosion of the external auditory canal was showed in 7 pa-tients.Other CT findings included bone erosion of skull base and intracranial involvement.Abnormal soft tissue of the external audi-tory canal,effusion in the mastoid cavity and medullary abnormalities were showed very well on MR images in 6 patients while the cortical bone erosion was not well showed.Conclusion Imaging features of the lesions in the external auditory canal,bone erosion of skull base and intracranial involvement play crucial roles in the diagnosis and delineation of the extent of the lesion in patients with MEO.