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.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.
6.Drug-induced pulmonary injury: CT findings in hemopathic patients
Juan ZHOU ; Min JIANG ; Gongjie LI ; Xudong XING ; Fugeng SHENG ; Penggang QIAO ; Hongtao ZHANG
Chinese Journal of Radiology 2012;46(6):505-507
Objective To investigate the spiral CT findings in hemopathic patients with druginduced pulmonary injury.Methods CT images obtained in 11patients with drug-induced pulmonary injury were retrospectively analyzed.Six patients had antineoplastic agent-induced pulmonary injury and 5 patients had non-neoplastic agent-induced pulmonary injury (immunosuppressor in 2 patients,antifungal in 2 patients,antineoplastic immunomodulators in 1 patient).CT findings were reviewed by a chest radiologist.Results All 11patients had parenchymal abnormalities on MSCT scans,including ground-glass opacities( n =8 ),consolidation( n =5 ),interlobular septal thickening( n =3 ) and focal fibrosis ( n =2 ).The abnormalities were bilateral and asymmetric in all patients.They were mainly in the peripheral lung regions in 6 patients,in the central lung regions in four,and irregularly located in one.The abnormalities involved mainly the lower lung zones in six patients,the upper lung zones in two,and all lung zones homogeneously in three.One patient had fluid in bilateral pleural cavities.Three patients were given the same agent once more after the imaging turned to normal,and they presented with same clinical symptoms and similar but more serious imaging findings.Conclusions Drug-induced pulmonary injury usually manifests as areas of ground-glass opacity and consolidation,which most commonly involves the peripheral lungs and lower lung zones.Drug-induced pulmonary injury shows reproducible but more serious lesions when the patient is given the same agent once more.
7.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.
8.Construction of Saccharomyces cerevisiae whole-cell biocatalyst system for conversion miltiradiene.
Yuan CAI ; Juan GUO ; Yongjin ZHOU ; Zhiwei ZHU ; Wenyan WU ; Luqi HUANG ; Min CHEN ; Zongbao ZHAO
Acta Pharmaceutica Sinica 2013;48(10):1618-23
Tanshinones are the bioactive components of the Chinese medicinal herb Salvia miltiorrhiza, while its biosynthetic pathway remains to be characterized. Rapid identification and characterization of the genes correlated to tanshinones biosynthesis is very important. As one of the intermediates of tanshinones biosynthesis, the ferruginol content is relative low in both root and engineered bacteria. It is urgent to construct an efficient system for conversion of miltiradiene to ferruginol to obtain large amount of ferruginol as the substrates for further identifying other downstream genes involved in tanshinones biosynthesis. In this study, we constructed the whole-cell yeast biocatalysts co-expressing miltiradiene oxidase CYP76AH1 and cytochrome P450 reductases (SmCPR1) from Salvia miltiorrhiza, and then characterized it with RT-PCR. After permeabilization, the yeast whole-cell could catalyze turnover of miltiradiene to ferruginol efficiently through single-step biotransformation with a conversion efficiency up to 69.9%. The yeast whole-cell biocatalyst described here not only provide an efficient platform for producing ferruginol in recombinant yeast but also an alternative strategy for identifying other CYP genes involved in tanshinones biosynthesis.
9.A 2-year follow-up study on 166 leprosy patients treated with uniform multidrug therapy
Jianping SHEN ; Wenzhong LI ; Min ZHOU ; Hongjiang MOU ; Xia BAO ; Rongde YANG ; Juan WANG
Chinese Journal of Dermatology 2010;43(2):75-78
Objective To assess the efficacy of 6-month uniform multidrug therapy in various types of leprosy. Methods A field trial was conducted among 166 patients with different types of leprosy. All patients were treated with uniform multidrug therapy for 6 months, then followed up for 2 years. Clinical and bacterio-logical improvements were evaluated. Results Among the 166 patients, 31 dropped out due to various reasons,and 135 completed the 6-month treatment and 2-year follow-up. Among the 135 patients, 45 (33.3%) were skin smear negative, and the other smear-positive 90 had an average bacterial index (BI) of 2.91±1.45 (range: 0.1-6.0) before treatment. At the end of the 2-year follow-up, the 45 skin smear-negative patients showed 93.3% improvement in skin lesions and 80.0% improvement in nerve impairments, and the smear-posi-tive 90 patients showed 95.6% improvements in skin lesions and 77.8% improvement in nerve impairments.Skin smear turned negative in 49 (54.4%) out of the smear-positive 90 patients with the average BI declining to 0.66±0.99. The annual decrease in BI reached 0.9 during the first 2.5 years after the beginning of treat-ment. Twenty-five patients developed leprosy reaction during the follow-up, including 13 cases of type Ⅰ leprosy reaction and 12 cases of type Ⅱ leprosy reaction. Relapse was noted in 1 patient with muhibacillary leprosy 13 months after the termination of treatment. Conclusions The short-term efficacy of uniform multidrug therapy is similar to that of 2-year treatment with routine multidrug therapy. However, further studies are required to survey the incidence of leprosy reaction and relapse in patients treated with uniform multidrug therapy.
10.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.