1.Effects of interleukin-1 ,interleukin-6 and tumor necrosis factor-α on migration of human lens epithelial cells
Chinese Journal of Experimental Ophthalmology 2012;(10):873-876
Background Lens epithelial cells (LECs)play an important role in maintaining the lens transparency.Inflammatory cytokines have been clarified to participate in the formation of traumatic and after cataracts.However,the influence of inflammatory cytokines on the migration of LECs is still studying.Objective This study was to investigate the effects of interleukin-1 alpha (IL-1α),interleukin-1 beta (IL-lβ),IL-6,tumor necrosis factor alpha(TNF-α) and their mixture on the migration of human LECs in vitro.Methods HLE-B3 cells were cultured in DMEM containing 0.5% fetal bovine serum.10 μg/L of IL-1α,IL-1β,IL-6,TNF-α or the mixture was added in the medium for 24 hours,respectively.Wound-scratch assay and transwell monolayer permeability assay were used to evaluate the effects of different cytokines on LECs migration.The migrating cell numbers at the scratch zone and the transmembrane cell numbers at 12:00,3:00,6:00,9:00 and the center areas on the transwell chamber were calculated under the inverted microscope.In the control group,LECs were cultured in DMEM with 0.5% fetal bovine serum only.Results Wound-scratch assay revealed that 24 hours after cytokine incubation,the LECs numbers in the scratch zone were significantly less in the control group compared with the IL-1β group and TNF-αgroup (P =0.000,0.000),and those in the mixture group were higher than the IL-1β group and TNF-α group (P=O.000,0.000).However,no significant change was found in migrating LECs number between IL-1o group or IL-6 group and the control group (P =0.600,0.098).Transwell assay displayed that after 24-hour culture with cytokines,the largest density of transmembrane LECs was obtained in the mixture group,with a significant difference in comparison with the IL-1β group or TNF-α group (P=0.000,0.000).However,the transmembrane LECs were much more in the IL-1 β group or TNF-α group compared with the control group (P =0.000,0.000).Only a few transmembrane LECs were seen in the IL-1α group and IL-6 group in comparison with the control group(P =0.056,0.327).Conclusions IL-lα and IL-6 can not stimulate the migration of human LECs,but TNF-α and IL-1β can enhance their migration markedly.Mixture of these cytokines may play a much powerful influence on LECs migration.
2.Relationship between plasma TRAIL before 20 weeks′gestation and pregnancy induced hypertension
Cheng ZHOU ; Hongling YANG ; Fangling ZENG ; Min JIANG ; Juan WU
Chinese Journal of Laboratory Medicine 2015;(8):522-527
Objective To assess the relationship between maternal plasma tumor necrosis factor-related apoptosis-inducing ligand ( TRAIL ) before 20 weeks′gestation and hypertensive disorder complicating pregnancy (HDCP);and to evaluate the predictive value of plasma TRAIL for HDCP.Methods A 2-phase screening/validation study was designed.In the screening phase , a nested, case-controlled study was performed , the plasma samples collected before 20 weeks′gestation from 20 women who later developed HDCP and 20 age-and gestation week-matched controls were tested in prospective screening test for protein expression profiling during pregnancy and HDCP.Plasma samples were analyzed by a human protein microarray technology designed to detect 507 proteins simultaneously.Differently expressed proteins′functional annotation and clustering were performed by using of Database for Annotation , Visualization and Integrated Discovery ( DAVID) and Gene Ontology ( GO) database.The TRAIL level of plasma samples obtained before 20 weeks′gestation from 53 women who later developed HDCP and 106 similarly matched controls were further validated by ELISA and 62 clinical risk factors were investigated.Logistic regression and ROC analysis were used to evaluate the relationship between TRAIL and HDCP and its predictive value for HDCP.Results In protein microarray analysis , 23 proteins expressed differently before 20 weeks′gestation between the two groups.Further validation results showed that TRAIL levels in HDCP patients were lower significantly (45.7 ±13.1) pg/ml than those in healthy pregnant controls (51.2 ±14.7)pg/ml, P=0.021.Multiple factor logistic regression analysis of 159 pregnancies showed that three features were finally entering the logistic model, they were:anemia (OR=4.87, 95% CI 1.05-24.26), pre-pregnancy BMI (OR=1.72, 95% CI 1.35 -2.19) and TRAIL (OR=0.96, 95% CI 0.92 -0.99).The predictive accuracy of logistic model was 81.8%.The model significantly increases the predictive value (AUC=0.81, 95%CI 0.73-0.87) compared to TRAIL as independent predictor (AUC=0.59, 95%CI 0.51-0.67).Conclusions Totally 23 proteins were expressed differentially before 20 weeks′gestation in plasma of women who later developed HDCP , confirming that HDCP is a heterogeneous disease with different biological changes.The data suggests that plasma TRAIL levels relate with the development of HDCP and its combination with pre-pregnancy BMI and anemia have a high predictive value for HDCP before 20 weeks′gestation.
3.The value of tumor type M2 pyruvate kinase in the diagnosis of lung cancer
Dong WANG ; Juan ZHOU ; Min LIU ; Hong-Xu XU ;
Chinese Journal of Laboratory Medicine 2001;0(05):-
Objective To investigate the diagnostic values of tumor M2-PK in lung cancer.Methods The concentration of Tumor M2-PK in plasma was detected by ELISA in 106 health controls,77 benign lung disease patients and 92 lung cancer patients.Results TuM2-PK concentration in plasma and the positive rate were singificantly higher in lung cancer(22.1 U/m1,71%)than that in benign lung disease and in health controls(10.5 U/ml,4% and 8 U/ml,3%)(P
4.HPLC simultaneous determination of contents of 5 saponin constituents in Ophiopogonis Radix.
Fa-ming WU ; Xiao-yang CAI ; Pan WANG ; Xiao-hong BAO ; Min LI ; Juan ZHOU
China Journal of Chinese Materia Medica 2015;40(20):4022-4025
This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Ophiopogon
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chemistry
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Plant Roots
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chemistry
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Saponins
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chemistry
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isolation & purification
5.Comparative study of radiation dose and image quality between spectral CT scanning and conventional scanning on neck
Tan GUO ; Cheng ZHOU ; Juan CHEN ; Guogeng WU ; Zhenghan YANG ; Zhengyin SHEN ; Min CHEN
Chinese Journal of Radiology 2015;49(4):279-282
Objective To investigate the radiation dose and image quality of spectral and conventional CT scan in neck.Methods Sixty patients with enhanced neck CT scan were analyzed retrospectively.The 30 patients with spectral CT scan were included in spectral CT group,and the 30 patients with conventional CT scan were included in conventional CT group.The tube voltage,tube current and rotation speed of the spectral CT group were fast switching with 80 and 140 kVp,630 mA and 0.5 s,respectively.The scanning parameters of conventional CT group were 120 kVp,auto tube current (100 to 600 mA),and 0.6 s,respectively.The objective evaluation [noise and (contrast to noise ratio) CNR] and subjective scores in the upper,middle and lower neck were evaluated.The radiation dose was also evaluated in the two groups.The independent-samples t test was used in comparison of the radiation dose.The independent-samples t test and the rank sum test were used to compare the objective and subjective image quality.Results The CT dose index of spectral and conventional scan in the neck were 17.77 mGy and (17.26±2.18) mGy,respectively without significant difference (t=-1.26,P=0.218).The noises of 65 keV images in upper,middle and lower neck were (4.5 ± 0.8),(4.5 ± 0.9),(5.2 ± 1.0) HU,and the noises of conventional CT images in above-mentioned regions were (4.5± 1.1),(4.1± 1.0),(5.0± 1.7) HU.There was no significant differences (t=0.102,-1.362,-0.621;P>0.05).The subjective scores of 65 keV images in upper,middle and lower neck were (4.1 ± 0.3),(4.7 ± 0.5),(3.8 ± 0.7),while the subjective scores of conventional CT images in associated region were (4.1±0.4),(4.6 ±0.5),(3.5 ±0.6),wihtout significant differences (Z=-0.286,-0.531,-1.568;P>0.05).The noises of 55 keV images in upper,middle and lower neck were (5.4±0.9),(5.6± 1.1),(6.6± 1.6) HU,which were significantly higher than noise of conventional images (t=-3.614,-5.560,-3.784;P<0.05).The subjective scores of 55 keV images in upper and middle neck were (3.7±0.5),(4.2±0.4),which were significantly lower than those of conventional images (Z=-2.541,-3.136;P<0.05).The subjective score of 55 keV images in lower neck was (3.3±0.8),which was no significant difference in comparison with conventional image (Z=-1.318,P>0.05).There was no significant difference between the CNR of conventional images and CNR of spectral images with 65 keV and 55 keV in upper,middle and lower neck (P>0.05).Conclusion The radiation dose and the image quality of spectral CT scan are same to the conventional CT scan in the neck,the 65 keV monochromatic images can be used routinely.
6.Kneading and dispersing manipulation in treatment of early-stage acute mastitis: a randomized controlled trial.
Min ZHOU ; Xin LI ; Yiqin CHENG ; Rong SHEN ; Ying ZHAO ; Huaizhi ZHAO ; Juan WANG ; Dongmei SHI
Journal of Integrative Medicine 2009;7(12):1130-3
Background: Acute mastitis is an acute infectious disease of breast. Antibiotic treatment is often unable to obtain a good effect, and we should actively look for a safe and effective non-drug therapy. Objective: To validate the clinical efficacy of kneading and dispersing manipulation in treatment of early-stage acute mastitis. Design, setting, participants and interventions: According to the multicenter randomized controlled trial design, 198 cases of acute mastitis from Yueyang Hospital of Integrated Traditional Chinese Medicine, Longhua Hospital, and Shanghai Yangpu Maternity and Child Health Hospital were randomly divided into treatment group and control group. There were 99 cases in each group. Patients in the treatment group were only treated with manipulation, and cefradine was orally administered to patients in the control group. Main outcome measures: The local breast lump size, clinical symptoms and the adverse reactions in the two groups were observed before and after the treatment. Results: The total response rates in the treatment and control group were 95.92% (94/98) and 80% (76/95) respectively. There was a significant difference in the total response rate between the two groups (P<0.05). There were significant differences in the score of breast lump size, and the score of signs and symptoms between the two groups (P<0.05). Conclusion: Kneading and dispersing manipulation has certain effects on early-stage acute mastitis, and the therapy is safe and repeatable.
7.The value of carotid plaque magnetic resonance imaging and sequence optimization in preoperative assessment in elderly patients with carotid atherosclerosis
Yan SONG ; Min CHEN ; Cheng ZHOU ; Juan HUANG ; Nan LUO ; Yuhui DENG ; Yuan FU
Chinese Journal of Geriatrics 2011;30(6):455-459
Objective To study the value of carotid plaque magnetic resonance imaging (MRI) in pre-operation assessment in the elderly patients with carotid atherosclerosis and explore the possibility of minimizing the contrast weightings to gain sweeptime. Methods Totally 70 elderly patients with cerebral ischemia (average age of 68.8 years) underwent carotid MRI and digital subtraction angiography (DSA) due to the appearance of carotid plaque detected by ultrasound. Carotid plaque MRI was acquired with 3.0T MR scanner and 8 channel surface coil. The standard carotid plague MRI program included pre-and post-contrast T1 weighted imaging (T1WI), T2 weighted imaging, proton density weighted imaging and 3D time of flight MR angiography (3D TOF MRA). All these program were divided into two combinations: the 5-sequence MRI (all the sequences) and 2-sequence MRI (T1WI and TOF MRA). Digital subtraction angiography (DSA) in coronal and lateral views of carotid artery was performed with GE Advantx LCN+. The software SPSS 13.0 was used to statistically analyze the difference between MRI and DSA, and that of two sequence combinations was used in the detection of luminal stenosis and fibrous cap (FC) rupture. Results Totally 135 arteries were analyzed while 3 arteries in one patient were excluded due to the poor quality image and stent placement. The degree of luminal stenosis were (38.3±31.0)% and (38.5±30.9)%, respectively, detected by the two MRI sequence-combination with no significant difference (t=2.447, P>0.05) and was (35.1±31.8)% by DSA. There was a good concordance between MRI and DSA in luminal stenosis detection (Kappa value: 0.773). No statistical difference was found between two MR sequence combinations in detecting FC rupture (both in 36 vessels). DSA detected FC rupture of 16 vessels, showing remarkably difference contrast to MRI(χ2=12.0, P<0.01). Conclusions MRI can accurately detect the luminal stenosis and FC rupture. The short time scanning resulting from sequence optimization could make MRI much more suitable than DSA to do the pre-operation assessment for senile carotid atherosclerotic patients.
8.Comparison of MRI and DSA in assessment of carotid artery stenosis and atherosclerotic plaque
Yan SONG ; Min CHEN ; Cheng ZHOU ; Juan HUANG ; Jiachun LIU ; Yuan FU
Chinese Journal of Radiology 2011;45(11):1004-1007
Objective To compare the efficacy of MRI and DSA in assessment of carotid artery stenosis and atherosclerotic plaque.Methods Forty-six patients with carotid plaque detected by ultrasound were enrolled in this study,and 89 carotid arteries were evaluated by MRI and DSA.MRI examination was acquired with 3.0 T MR scanner and 8 channel phase-array surface coil.The MRI sequences consisted of pre-and post-contrast T1WI,T2WI,PDWI,TOF.Anterior-posterior and lateral views of carotid artery were performed on DSA.The degree of carotid artery stenosis was evaluated by the NASCET standard.Fibrous cap rupture,intraplaque hemorrhage,and calcification were also evaluated on MRI and DSA.Statistical comparison was performed with the Kappa value and paired Chi-square test.Results The degree of carotid artery stenosis was 50% (16%-78%) on MRI and 47% (7%-73%) on DSA.Two imaging modalities were in good consistency in evaluation of the degree of stenosis ( Kappa =0.882,P < 0.01 ).There was statistical difference in detecting fibrous cap rupture by MRI and DSA (34 vessels vs 10 vessels,respectively,x2 =20.346,P < 0.01 ).Furthermore,thirty-seven vessels with intraplaque hemorrhage and 71 vessels with calcification in the plaque were found on MRI but none on DSA.Conclusion MRI is a reliable tool in assessment of the degree of the carotid stenosis and it is superior to DSA in detecting fibrous cap rupture,intraplaque hemorrhage,and calcification.
9.Eukaryotic Expression of ?ig-h_3 Gene and Its Effects on Secretion of MMPs in the Human Hepatoma 7721 Cells
Juan TANG ; Jian-Li JIANG ; Hong-Wei ZHOU ; Hua XIONG ; Xiang-Min YANG ; Zhi-Nan CHEN ;
China Biotechnology 2006;0(09):-
?ig-h_3 was first identified as a transforming growth factor-beta1-inducible gene in human lung adenocarcinoma cell line. It encodes for a secreted extracellular matrix (ECM) protein, which is thought to act on cell attachment and ECM composition. Previous study showed that ?ig-h_3 were highly expressed in human hepatoma cell lines and lowly expressed in human normal hepatic cells. The present study aimed to transfect ?ig-h_3 into 7721 cells to investigate its effect on secretion of MMPs in the transfected human hepatoma cells. Full-length ?ig-h_3 gene,cloned by reverse transcription polymerase chain reaction (RT-PCR) was inserted into the eukaryotic expression vector pEGFP-C_2. The recombinant plasmid was transfected into 7721 cells with Lipofectamine2000 and Gelatin-Zymography were adopted to detect the production of MMPs in the transfected cells. Results showed that ?ig-h_3/pEGFP-C_2 recombinant expression plasmid was successfully constructed and achieved high transfection efficiency. MMPs expression of the transfected cells was promoted significantly. These results suggest that overexpression of ?ig-h_3 promoted the production of MMPs, indicating that ?ig-h_3 may play roles in the invasive and metastatic processes of hepatoma.
10.Effect of radix salviae militiorrhizae on acute lung injury caused by two hits in rats
Li-Juan WU ; Xiao-Min HUANG ; Yu-Zhou HE ; Yun-Kai WANG ;
Chinese Journal of Emergency Medicine 2006;0(09):-
Objective To investigate the effects of radix salviae militiorrhizae (RSM) on acute lung injury induced by"two hits"and to study its probable mechanism.Method Thirty Wister rats were randomly divided into three groups:namely normal control group,model group and RSM treatment group.The model was created by"two-hits"in which 0.2 ml/kg oleic acid was injected into tail vein first,and then 2 mg/kg lipopolysaccharide was administered four hours later.After model rats sacrificed,the pathological changes of lung were observed,and lung wet/dry weight ratio,protein content,and the ratio of neutrophiles in brochoalveolar lavage fluid (BALF) were calculated.In addition,the expression of Fas,FasL protein and apoptosis were evaluated by immunohistochemical studies and TUNEL technique.Results The acute lung injury rat model was successfully induced by"two hits".The gross and micrographic injury of lung was milder in RSM treatment rats than in model rats.The W/D ratio,protein contents and the ratio of neutrophiles in BALF were also markedly reduced in comparison with model rats,while the expression of Fas and Fasl,and the apoptosis index in model rats were significantly increased compared with other two groups.Furthermore,it showed a positive correlation between the expression of Fas,FasL,and the number of cell with apoptosis.Conclusions RSM shows a protective effect on ALT rats caused by"two hits"likely reaulted from inhibiting the expressions of Fas and Fasl,which are associated with the cell apoptosis of lung tissue.