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.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
3.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.
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.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.
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.Clinical observation on the treatment of locally advanced cervical cancer by recombinant human adenovirus-p53 combined with neoadjuvant chemotherapy
Jie XIAO ; Jian ZHOU ; Fumin LIU ; Min FU ; Juan LIU ; Peng GENG
Cancer Research and Clinic 2013;(1):32-35
Objective To evaluate the efficacy and safety of recombinant adenovirus-p53 (rhAd-p53) combined with neoadjuvant chemotherapy in treatment of locally advanced cervical cancer.Methods Forty patients with stage Ⅰ R2-Ⅲ A locally advanced cervical cancer were randomly divided into 2 groups,gene therapy + neoadjuvant chemotherapy group (rhAd-p53+PVB group,n =20.They received one course of chemotherapy consisting of PVB.rhAd-p53 solution 1 ×1012 VP was injected intratumorally every three days for three circles since the 3rd day of PVB chemotherapy) and chemotherapy group (PVB group,n=20,the above course of chemotherapy was conducted).The volums of tumors was observed.Patients were monitored for adverse event.The expression of VEGF,p53 pertein and MVD in tumor tissue was detected by immunohistochemistry.Results The evaluation was performed three weeks after the completion of chemotherapy.The PVB group response rate (CR+PR) was 75 %,while the effective rate was 95 % of the PVB combined with gene group.After using of the PVB chemotherapy,the tumor was shrunk by (11.42±2.78) cm2.However,the volums of tumor were significantly shrunk by (15.25±4.00) cm2 using the PVB combined with gene therapy,and P < 0.05.The positive expression rate of VEGF,p53 protein and MVD were reduced respectively in PVB group and rhAd-p53 + PVB group with statistic significance.There were no additional adverse events by recombinant adenovirus-p53 combined with neoadjuvant chemotherapy.Conclusion A potentially gene therapeutic agent for cervical cancer treatment,intratumoral injection of rhAd-p53 is effective.
8.Efficacy comparison of uniform versus routine multidrug therapy for the treatment of multi-bacillary leprosy patients based on bacterial index and leprosy reactions
Meiwen YU ; Jianping SHEN ; Liangbin YAN ; Min ZHOU ; Hongjiang MOU ; Xia BAO ; Rongde YANG ; Juan WANG
Chinese Journal of Dermatology 2012;45(8):553-556
Objective To compare the efficacy of uniform multi-drug therapy (UMDT) versus routine multi-drug therapy (RMDT) for the treatment of multi-bacillary (MB) leprosy patients based on bacterial index changes and frequencies of leprosy reaction.Methods This study recruited newly diagnosed leprosy patients after taking informed consent in three districts of Guizhou province as well as in one district of Yunnan province from November 2003 to June 2005.The patients received 6-month UMDT or 2-year RMDT.Clinical follow up and bacterial reexamination were carried out once a year.Changes of bacterial index (BI) and frequencies of leprosy reaction were compared between the patients receiving RMDT and UMDT.Results A total of 166 patients received UMDT and 170 received RMDT in this study.Among the UMDT-treated patients,114 were skin smear positive,and 83 had been followed up for 42 months; of the RMDT-treated patients,149 underwent all the bacterial examinations during a 48-month follow up.The mean bacterial index decreased from 2.84 before treatment to 0.33 at the end of the 42-month follow up in the 83 patients,and from 2.55 to 0.26 at the end of the 48-month follow up in the 149 patients,with no significant difference in the changes of bacterial index between the two groups (t =0.77,P > 0.05).Bacterial index became negative in 73.5% (61/83) of the UMDT-treated patients and in 77.2% (115/149) of the RMDT-treated patients (x2 =0.40,P> 0.05)at the end of follow up.During the follow up peroid,the incidence of type Ⅰ leprosy reaction was 14.6% (13/89) in the UMDT group,significantly higher than that in the RMDT group (3.4% (5/149),x2 =10.08,P< 0.01 ).Conclusions There is no significant difference in mean bacterial index changes and bacterial clearance rate during the follow up peroid between UMDT- and RMDT-treated patients.The incidence of type Ⅰ leprosy reaction is higher in the UMDT group than in the RMDT group,and further investigation is needed to clarify the mechanisms underlying the phenomenon.
9.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.
10.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.