1.The study of using siRNA technology to silence expression of LT gene of Enterotoxigenic Escherichia coli
Rui FU ; Hua LIU ; Yong DUAN ; Yuming WANG ; Bin SHAN
International Journal of Laboratory Medicine 2015;(8):1009-1011,1014
The distinctive LT siRNAs were designed according to the LT sequence .During the process of cultivation ,siRNA targeting the LT gene ,non‐specific control siRNA ,negative control siRNA and culture medium were added into siRNA group (siRNA‐LT1 group , siRNA‐LT2 group) ,siRNA‐coa3 group ,siRNA‐NC group and blank control group ,respectively ,and three times in each group (1 nmol each time) .After siRNA added at the first time ,bacteria was collected in 45 min (A) ,90 min (B) and 135 min (C) time points .The expression of mRNA in three time points (A ,B and C) were detected by real‐time fluorescence quantitative PCR .The protein level of LT in siRNA‐LT1 group ,siRNA‐LT2 group and blank control group were detected by Western blot in three time points .Results The results of real‐time fluorescence quantitative PCR showed that inhibition of siRNA‐LT1 on the expression of LT mRNA at the three time points(A ,B and C)were 70 .9% ,70 .1% ,72 .5% respectively ,and inhibition of siRNA‐LT2 on the ex‐pression of LT mRNA at the three time points(A ,B and C)were 70 .1% ,69 .2% and 70 .5% respectively .In the three time points (A ,B and C)the inhibition rate of the expression of LT mRNA in siRNA‐LT1 group and siRNA‐LT2 group were statistically lower than that in the siRNA‐NC group ,siRNA‐coa3 group and blank control group (P<0 .05) .The results of Western blot showed that in siRNA‐LT1 group the inhibitory rate of expression of LT protein in the three time points were 43 .1% ,18 .4% and 5 .0% ,re‐spectively ;in the siRNA‐LT2 group were 38 .2% ,15 .4% and 30 .1% ,respectively .Conclusion The specific siRNA could inhibit the expression of LT gene in vitro .
2.Clinical Study on Compound Sophora Flavescentis Radix (Kushen) Injection Combined with Gamma Knife for Treatment of Locally Advanced Pancreatic Carcinoma
Lei WANG ; Lingzhao LI ; Xiaofang WANG ; Guoyong SHAN ; Rui LIU ; Song ZHANG ; Xiaodong WU
Chinese Journal of Information on Traditional Chinese Medicine 2015;(6):21-24
Objective To evaluate the clinical efficacy of Compound Sophorae Flavescentis Radix (Kushen) Injection combined with gamma knife for treatment of locally advanced pancreatic carcinoma. Methods Totally 65 patients of locally advanced pancreatic cancer were randomly divided into two groups:32 cases of simple gamma knife group (group A), 33 cases of Compound Kushen Injection combined with gamma knife treatment (group B). Patients received enhancement computed tomography scanning and CA199 review at 1, 3, 6, 9, 12 months after treatment, and once every six months, a total of 36 months of follow-up. Short-term and long-term efficacy was observed after treatment, and survival curve analysis was carried out. Results Clinical benefit response was 53.13% in group A and 84.85% in group B (P=0.006);median survival time of two groups was 11.00 months in group A and 23.00 months in group B;the average survival time in group A was 16.37 months, and 22.28 months in group B (P=0.017). In the two groups, treatment of the 65 patients was completed smoothly as planned, and no one had serious complications, such as pancreatic leakage, skin burns, viscera perforation and radioactive enteritis. Conclusion Compound Kushen Injection combined with gamma knife can treat locally advanced pancreatic carcinoma safely and efficiently, which can improve patients’ life quality, especially in cancer pain control.
3.Effect of hydrogen sulfide on oxidative stress and endoplasmic reticulum stress in diabetic cardiomyopathy.
Rui YANG ; Qiang JIA ; Xiao-fen LIU ; Qin GAO ; Lei WANG ; Shan-feng MA
Chinese Journal of Applied Physiology 2016;32(1):8-12
OBJECTIVETo investigate the effects of hydrogen sulfide (H₂S) on oxidative stress and endoplasmic reticulum stress (ERS) in a rat model of diabetic cardiomyopathy (DCM).
METHODSThirty male SD rats were randomly divided into control group, diabetes group and treatment group( n = 10). Intraperitoneal injection of streptozotocin was utilized to establish a rat model of DCM. The rats with DCM in treatment group were intraperitoneally injected with NaHS solution. After treated for 12 weeks, the hearts isolated from rats were perfused on a langendorff apparatus. The ventricular hemodynamic parameters were measured. The ultrastructures of myocardium were observed using electron microscopy. The content of malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in myocardial tissue were determined by spectrophotometry. The expressions of C/EBP homologous protein( CHOP), glucose-regulated protein 78 (GRP78) and Caspase 12 at mRNA level in myocardium were detected using RT-PCR.
RESULTSCompared with control group, the cardiac function and myocardial ultrastructure were damaged obviously in diabetic rats. In myocardial tissue, the content of MDA was increased, while the activities of SOD and GSH-Px were decreased. CHOP, GRP78 and Caspase 12 mRNA expressions were increased significantly. Compared with diabetes group, cardiac function and myocardial ultrastructure damage were improved in treatment group. The content of MDA was decreased, while the activities of SOD and GSH-Px were increased significantly. The mRNA levels of CHOP, GRP78 and Caspase 12 were increased.
CONCLUSIONH2S can protect myocardium in diabetic rats, maybe it is related to reduce oxidative stress damage and inhibition of the ERS-induced apoptosis pathway.
Animals ; Apoptosis ; Caspase 12 ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Cardiomyopathies ; drug therapy ; Endoplasmic Reticulum Stress ; Glutathione Peroxidase ; metabolism ; Heat-Shock Proteins ; metabolism ; Hydrogen Sulfide ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Myocardium ; ultrastructure ; Oxidative Stress ; Rats ; Streptozocin ; Superoxide Dismutase ; metabolism ; Transcription Factor CHOP ; metabolism
4.Data collection of signals in the multi-channel sEMG system of masticatory muscles and development and preliminary clinical application of an analytic system.
Hongliang DU ; Xin LI ; Shan LI ; Rui ZHANG ; Rong SONG ; Lan LI ; Wei WANG ; Hong KANG
Journal of Biomedical Engineering 2014;31(1):23-28
The aim of this study was to design a simple, economic, with high Common Mode Rejection Ratio (CMRR), preamplifier and multi-channel masticatory muscle surface electromyography (sEMG) signal acquisition system assisting to diagnose temporomandibular disorders (TMD). We used the USB interface technology in the EMG data with the aid of the windows to operate system and graphical interface. Eight patients with TMD and eight controls were analyzed separately using this system. In this system, we analyzed sEMG by an optional combination of time domain, frequency domain, time-frequency, several spectral analysis, wavelets and other special algorithms under multi-parameter. Multi-channel sEMG System of Masticatory Muscles is a simple, economic system. It has high sensitivity and specificity. The sEMG signals were changed in patients with TMD. The system would pave the way for diagnosis TMD and help us to assess the treatment effect. A novel and objective method is provided for diagnosis and treatment of oral-maxillofacial disease and functional reconstruction.
Algorithms
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Computer Graphics
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Data Collection
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Electromyography
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Humans
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Masticatory Muscles
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physiology
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physiopathology
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Sensitivity and Specificity
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Signal Processing, Computer-Assisted
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Temporomandibular Joint Disorders
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diagnosis
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User-Computer Interface
5.A study on the mechanism of iodine-induced thyroid epithelial cell injury in the induction of autoimmune thyroiditis
Xiaochun TENG ; Na MAN ; Zhongyan SHAN ; Chenling FAN ; Hong WANG ; Rui GUO ; Weiping TENG
Chinese Journal of Internal Medicine 2008;47(3):193-196
Objective To investigate the effect of iodine excess on thyroid follicle epithelial ultrstructure and the relationship between thyroid iniury and autoimmune thyroiditis. Methods NOD.H-2h4 mice and Kunming mice were randomly divided into four groups receiving plain water,5 fold,10 fold,and 100 fold excessive iodine water.4,8 and 24 weeks after receiving iodine water,the mice were killed.After fixation with osmic acid and dual staining with uranyl chloride and citrate lead,thyroid gland ultrstructure was examined with electron microscopy.Resuits Iodine treated NOD.H-2h4 mice exhibited marked accumulation of peroxisome and secondary lysosomes,apoptosis and necrosis of thyroid epithelial cell.damage of thyroid follicles and lymphocvtic infiltration.The observed changes induced by iodine were in a dose dependent way.Conclusion The oxidative iniury on the thyroid epithelial cells induced by iodine excess might be the prerequisite for the creation of autoimmune thyroiditis.
7.Effects of ursolic acid from loquat leaves on proliferation inhibition andexpression of PPAR-γ, TGF-β1 in rat hepatic stellate cells
Yangwu ZHANG ; Weisheng LUO ; Shan CHEN ; Rui HUANG ; Quanxiao TAN ; Shiyan WANG ; Xia ZHANG ; Chuanfeng XUAN
Chinese Pharmacological Bulletin 2017;33(4):517-521
Aim To investigate the effects of ursolic acid from loquat leaves on proliferation inhibition and expression of PPAR-γ 、TGF-β1 in rat hepatic stellate cells, so as to explore the mechanism of anti-hepatic fibrosis of UA.Methods HSC-T6 cells were randomly divided into blank group, rosiglitazone control group, the low, medium and high concentration of UA group to detect the cell proliferation inhibition by CCK-8 after 24,48,72 h.The content of type collagen Ⅰ in cell culture supernatant of each group was examined by ELISA.The expression of PPAR-γ mRNA, TGF-β1 mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR.Effects on HSC-T6 PPAR-γ and TGF-β1 protein from each group were detected by immunocytochemical method.Results CCK-8 results showed that the inhibitory rate of UA on cell proliferation increased with the prolongation of drug action time(P<0.01).ELISA results showed that with the increase of the concentration of UA, the content of type Ⅰ collagen content decreased(P<0.01).Real-time PCR results showed that with the increase of the concentration of UA, and the expression of PPAR-γ mRNA increased, the expression of TGF-β1 mRNA decreased(P<0.01).The results of immunocytochemistry showed that the expression of PPAR-γ protein was increased, and the expression of TGF-β1 protein was decreased with the increase of the concentration of UA(P<0.01).All effects mentioned above were dose-dependent.Moreover, the effects in the high concentration groups were stronger than those in control group.Conclusion UA can inhibit the proliferation of HSC-T6 cells, Which may be associated with the up-regulation of PPAR-γ expression and the down-regulation of TGF-β1 expression.
8.Implication of MAPK in sodium salicylate-induced heat shock protein 27 expression in human lens eplthelial Cells in vitro
Rui-ying, GAO ; Zhi, WANG ; Yi-zhuang LI ; Hui, CHEN ; Shan-hua, LU ; Bo, TIAN
Chinese Journal of Experimental Ophthalmology 2011;29(5):412-417
Background Heat shock proteins (HSPs) are highly conserved proteins that are induced in cells when confronted with a wide variety of proteotoxic stresses.HSP27 has a high degree of similarity with α-crystallin protein.The abnormality of HSP27 structure and expression are closely related to the formation of cataracts.Our previous study showed sodium salieylate has the protective effect on H2O2-induced lens damage.Objective This study was to investigate the roles of MAPK signal pathway in sodium salicylate-induced the expression of HSP27 in human lens epithelial cells (LECs) in vitro.Methods Human LECs were incubated in the fresh media containing sodium salicylate at different concentrations (0-55 mmol/L) for different times (1-5 hours) and allowed to be recovered in fresh medium without sodium salicylate for 1-24 hours with or without pretreatment with P38MAPK inhibitor (SB203580), ERK1/2 inhibitor (PD98059) and JNK/SAPK inhibitor ( SP600125). The expressions of P38MAPK, EBK1/2, JNK/SAPK, phosphorylated P38MAPK, phosphorylated ERK1/2, phosphorylated JNK/SAPK and HSP27 were detected by Western blot. HSP27 mRNA was detected by RT-PCR. The expression of HSP27 was also detected by immunohistochemistry. Results There was only weak expression of HSP27 in normal human LECs.After stimulation of 35-55 mmol/L sodium salicylate was removed and human LECs were cultured again for 6 hours,the expression of HSP27 in LECs were significantly increased ( F= 509. 953,P<0. 01). HSP27 was absent expressed in human LECs in 55 mmol/L sodium salicylate stimulation for 1-5 hours groups, but LECs were re-cultured for 3,6 hours after removed the stimulation, the expression of HSP27 was elevated (F = 452. 534, P<0. 01). Activation of P38 M APK occurred after sodium salicylate stimulation 30 minutes and 1 hour ( F = 865.68, P<0. 01). However, ERK 1/2 was expressed after sodium salicylate was eliminated for 1-6 hours ( F = 388.84, P<0. 01). JNK/SAPK was inactived by sodium salicylate. The expression of HSP27 could be down-regulated with the pretreatment of SB203580 and PD98059. Conclusion Sodium salicylatc can induce the expression of HSP27 in human (LECs) . The effects are mediated,at least in part ,through the activation of P38MAPK and ERK1/2 signaling pathway .
9.The effect of scalp acupuncture combined with rehabilitation training on the regulation of neuro-plasticity-associated protein MAP-2 in gerbils modelling cerebral ischemia and reperfusion
Xiaode GUO ; Ying ZHOU ; Rui WANG ; Xiang ZHOU ; Yinfeng GUO ; Jun TANG ; Shan GAO
Chinese Journal of Physical Medicine and Rehabilitation 2015;37(1):7-10
Objective To observe the effect of scalp acupuncture and rehabilitation training on the regulation of neuro-plasticity-associated protein MAP-2 in gerbils modelling cerebral ischemia and reperfusion.Methods Cerebral ischemia and reperfusion were modelled into thirty-six male gerbils.They were then randomly divided into a model group (A),a scalp acupuncture and rehabilitation training group (B) and a rehabilitation training group (C),each of 12 cases.The scalp acupuncture and rehabilitation training were administered daily to the gerbils in group B beginning 24 h after modeling,for 14 d.Any changes in Bederson scores were observed after one,7 and 14 days.On the 14th day the expression of MAP-2 around the infarct focus was detected using western blotting in all 3 groups.Results On the 7th day the average Bederson score in group B (1.81 ± 0.52) was not significantly different from that in group A (2.13 ± 0.49) or group C (2.00 ± 0.31) (P > 0.05).At the end of the treatment,however,there were significant differences between the groups,with group B (0.47 ± 0.31) scoring significantly better than groups C (1.04 ± 0.63) and A (1.46 ± 0.72) (P < 0.05).Group C was also significantly better than group A (P <0.05).The expression of MAP-2 as measured as integrated optical intensity (IOD) in group B (0.91 ±0.18) was significantly higher than that in group A (0.43 ± 0.21) and group C (0.67 ± 0.24) (P < 0.05) ; so was group C compared to group A (P <0.05).Conclusion The scalp acupuncture combined with rehabilitation training can improve the recovery of motor function and enhance the expression of neuro-plasticity-associated protein MAP-2 in gerbils with after cerebral ischemia and reperfusion.
10.Posterior reversible encephalopathy syndrome:diagnosis with CT and MRI
Guang-Bin WANG ; Rui-Qin SHAN ; Bin ZHAO ; Lee C Chiu ; Hao SHI ;
Chinese Journal of Radiology 2001;0(09):-
Objective To evaluate the role of CT and MR/in the diagnosis of posterior reversible encephalopathy syndrome(PRES).Methods Eight women with PRES(6 pregnant women,1 case after chemotherapy,and 1 patient with hypertension)were enrolled in our study.All of them had MR imaging (T_1WI,T_2WI,FLAIR,DWI),and five cases underwent post-contrast T_1WI and three dimensional contrast enhanced MR angiography(3D CEMRA).Two cases also had CT scan.Results MRV in all 8 patients showed no evidence of stenosis,dilation,or thrombosis in cranial veins and sinuses.MRI demonstrated multiple lesions located in bilateral parieto-occipital lobes(8 cases),bilateral basal ganglia(2 cases),and bilateral frontal lobes(4 cases).The lesions were prominent within white matter,some of them involved gray matter(3 cases).Lesions appeared as hyperintense signals on FLAIR and T_2-weighted images, isointense or mildly hypointense signals on T_1-weighted images,normal or decreased intensity on DWI,and isointensity or hyperintensity on apparent diffusion coefficient(ADC)maps.Post-contrast T_1WI showed mild reversible enhancement and 3D CEMFdisplayed numerous reversible“grape-like”enhancements in terminal arterial branches along the middle cerebral artery(MCA),anterior cerebral artery(ACA)and posterior cerebral artery(PCA).Follow-up scan showed decreased abnormal signals.Conclusion Lesions of PRES are usually located in parieto-occipital lobes,especially in white matter,but they can also be seen in frontal lobes and basal ganglia bilaterally.Post-contrast T_1WI and 3D enhanced MRA can provide useful information in the manifestation of reversible enhancement.MRI has advantages to display lesion in PRES,