1.The relationship between complement 3 and IgAN in clinic and pathology
Ping WU ; Jun ZOU ; Shunjie CHEN ; Bo ZHOU ; Gengru JIANG
Journal of Chinese Physician 2015;17(6):883-887
Objectives By Oxford pathological classification and analysis of circulating complement complement 3 (C3),renal C3 deposition,and clinical laboratory tests,we discussed the correlation between complement C3 and immunoglobin A nephropathy (IgAN) in pathogenesis.Methods A retrospective study of 558 IgAN cases at Xinhua Hospital from January of 2000 through December of 2013 was performed.All 558 IgAN diagnoses were made and confirmed by renal needle biopsy.Results Compared to patients who had circulating C3 < 0.9,patients with circulating C3 level > 0.9 showed statistically significant decreases in serum creatinine [(100.92 ± 105.31) μmol/L vs (157.58 ± 208.39) μmol/L,t =-2.283,P =0.025],blood urea nitrogen [(5.69 ± 2.88) mmol/L vs (7.69 ± 5.90) mmoL/L,t =-2.81,P =0.006];besides,other parameters like IgA,body weight,body mass index (BMI),cholesterol,triglyceride,serum IgA/C3 ratio,albumin,and estimated glomeruli filtrate rate (eGFR) also presented statistically significant differences between two patient groups;no statistically significant differences were observed between two groups in glomerular mesangial cell proliferation,capillary endothelial proliferation,segmental glomerular sclerosis or adhesions,renal tubule atrophy,tubulointerstitial fibrosis,and formation of glomerular crescent;meanwhile,no statistically significant differences were observed between two groups in mesangial depositions of IgA,IgG,IgM,and complement C3;meanwhile the blood level of C3 between C3 deposition negative group,deposition 2 + and 3 + subgroup showed statistically significant differences (2.493 and 2.782;0.013 and 0.006),nevertheless,prognostic indices in Oxford classification,such as mesangial cell proliferation,capillary endothelial proliferation,segmental glomerular sclerosis or adhesions,renal tubule atrophy and tubulointerstitial fibrosis,were also statistically different between two patient groups (Pearson Square test result was 50.782,35.141,21.105,30.182,respectively;P <0.01).Conclusions Renal deposition of complement C3 or decrease in circulating C3 level may be associated with a poor prognosis of IgA nephropathy,and alteration in C3 dynamics may be implicated in the pathogenesis of IgAN through its involvement in humoral immunity.
2. Extraction kinetics of volatile components from Foeniculum vulgare by steam distillation
Chinese Traditional and Herbal Drugs 2018;49(12):2855-2865
Objective To reveal the dissolution rule of volatile oil in the extraction processfrom Foeniculum vulgare by GC-MS analysis. Methods The volatile oil of F. vulgare was extracted by steam distillation; The volatile oil/perfume system was collected at fixed time interval (30 min); The volatile oil part and perfume part were separated. Using N-docosane and methyl myrisate as double internal standard, GC-MS was selected for analysis and quantification. The main components were extracted by thermography, and the distribution regularity of components was definited. The principal components with the impact on the composition distribution were investigated according to the physical and chemical properties of the molecular weight, melting point, boiling point, and density of the compounds. Results The GC-MS analysis results concluded 123 volatile components. The main components were estragole, anethole, and (R)-(+)-limonene. There were 60 and 27 endemic components in aromatic aqueous solution and volatile oil, respectively, in which 27were common composition. The content of anethole in water was positively related to the endemic components in aromatic water, and even had a positive correlation with anethole artemisia content and (R)-(+)-limonene content. The specific components in the oil were positively related to the content of the main components (R)-(+)-limonene in the volatile oil. The principal component analysis showed that the physical and chemical properties of the compounds were important factors affecting the distribution of components; PC1 (molecular weight and positive correlation of melting point), PC2 (refractive index positive correlation), and PC3 (water-soluble negative correlation) were the principal components that lead to differences in component distribution. Conclusion In the process of extracting volatile oil from F. vulgare, steam distillation is affected by the physical and chemical properties of volatile components. Some components are specifically distributed in aromatic perfume and volatile oil system. The endemic components of aromatic water increase the content of the main components in the water system, which may lead to theproduction of "emulsification" during the extraction process of volatile oil and reduce the yield and quality of volatile oil.
3.Effectiveness of premedication with pronase for visualization of the mucosa during endoscopy: a double-blinded, multi-center, prospective, randomized study
Dong WANG ; Jun FANG ; Youming LI ; Liping HE ; Bo JIANG ; Xiaoping ZOU ; Zhaoshen LI
Chinese Journal of Digestive Endoscopy 2013;30(11):604-607
Objective To assess the efficacy and safety of premedicaton with pronase before upper gastrointestinal endoscopy (UGI).Methods A total of 440 outpatients from 5 centers were randomly assigned to receive endoscopy with one of three premedications as follows:dimethylpolysiloxane (DMPS) and pronase (group A,n =170) ; DMPS and sodium bicarbonate (group B,n =170) ; DMPS,pronase and sodium bicarbonate (group C,n =100).Six endoscopists,who were unaware of the premedication types,calculated the visibility scores (range,1 to 4) of the antrum,gastric body,and fundus.The sum of the scores from the three locations was defined as the total visibility score.Results With regards to routine white light endoscopy,the total visibility score of group C was significantly higher than that of group A (P =0.0001),and the score of group A was significantly higher than that of group B (P =0.0019).Concerning chromoendoscopy,the total visibility score of group C was significantly higher than that of group A (P =0.0054),and the score of group A was also significantly higher than that of group B (P < 0.0001).Conclusion Combined application of pronase,dimethylpolysiloxane,and sodium bicarbonate before UGI endoscopy significantly improves endoscopic visualization.
4.A Novel RT-LAMP Assay for Rapid and Simple Detection of Classical Swine Fever Virus
Lei CHEN ; Xuezheng FAN ; Qin WANG ; Lu XU ; Qizu ZHAO ; Yuanchen ZHOU ; Jun LIU ; Bo TANG ; Xingqi ZOU
Virologica Sinica 2010;25(1):59-64
A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV.PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.
5. The investigation of key factors influencing the extraction process of chuanxiong oil by steam distillation
Chinese Pharmaceutical Journal 2019;54(18):1468-1474
OBJECTIVE: To investigate the key process parameters that affect the extraction of essential oil from Ligusticum chuanxiong by steam distillation. METHODS: Volatile oil and emulsified aromatic water are the products of the extraction process of volatile oil. The nature of volatile oil will directly affect the formation of emulsified aromatic water. The influence of extraction temperature, the ratio of liquid to medicine, medicinal material granularity process parameters on the stability of the emulsified fragrance water and the yield of volatile oil were analyzed, furtherly the corresponding extraction process control model of volatile oil was established to reveal the key technology and parameter during the process of volatile oil extraction and separation. RESULTS: The relative viscosity, relative density, surface tension and contact angle of Ligusticum chuanxiong oil decreased gradually with the increase of temperature, while the interfacial tension increased with the increase of temperature. The liner model of aromatic water TSI and process parameters was TSI = 0. 877 + 0. 230 × extraction temperature - 0. 024 × comminution particle size + 0. 010 × liquid ratio + 0. 292 × condensation temperature + 0. 776 × collection temperature. The liner model of the physicochemical properties of TSI and volatile oil was TSI = 0. 877 - 0. 170 × density + 0. 098 × viscosity - 0. 301 × surface tension + 0. 695 × interfacial tension - 0. 060 × contact angle. The liner model of volatile oil yield and process parameters was that yield = 5. 065 + 0. 258 × extraction temperature + 0. 127 × particle size + 0. 016 × liquid ratio + 0. 264 × condensation temperature + 0. 264 × collection temperature + 0. 502 × TSI. The liner model with the physicochemical properties of volatile oil was the yield = 5. 065 - 0. 196 × density - 0. 167 × viscosity - 0. 201 × surface tension + 0. 153 × interfacial tension - 0. 065 × contact angle. CONCLUSION: TSI and yield of aromatic water of Ligusti-cum chuanxiong are directly proportional to the extraction temperature and comminution particle size of medicinal materials. The yield of volatile oil shows a significant positive correlation with the TSI value, which can reflect the yield of volatile oil and is a specific index that can evaluate the process parameters objectively.
6.Efficacy of autologous bone marrow-derived cells transfer for patients with chronic ischemic heart disease: a meta-analysis.
Chun-yu ZHANG ; Ai-jun SUN ; Jun-bo GE ; Shu-ning ZHANG ; Ke-qiang WANG ; Yun-zeng ZOU
Chinese Journal of Cardiology 2010;38(7):656-661
OBJECTIVEWe aimed to perform a meta-analysis of clinical trials on the efficacy of autologous bone marrow-derived cells (BMCs) transfer for patients with chronic ischemic heart disease.
METHODSWe searched MEDLINE, EMBASE, and Cochrane database through September 2009. Eligible studies were randomized controlled trials of autologous BMCs infusion in patients with chronic ischemic heart disease. We gathered information about left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV) and death, and did a random-effect meta-analysis to obtain summary effect estimates for outcomes. The pooled analyses were performed and forest plots were generated with RevMan 5.0 software. Heterogeneity was assessed by meta-regression with STATA 10.0 software. Additionally, subgroup analysis was performed to compare the effect of intracoronary BMCs transfer with intramyocardial cell injection on LVEF.
RESULTSEleven trials with 490 participants were identified. There were 268 patients in BMCs group, and 222 in control group. In control group, the patients received saline injection or autologous plasma injection or no injection. BMCs transfer was performed via intracoronary transfer or intramyocardial injection. Compared with controls, BMCs transfer significantly improved LVEF by 4.63% (95%CI 2.42 to 6.84; P < 0.01). BMCs transfer was also associated with significant reductions in LVEDV (standardized mean difference -0.55, 95%CI -0.94 to -0.17, P = 0.005) and LVESV (standardized mean difference -0.45, 95%CI -0.73 to -0.17, P = 0.002). In addition, BMCs treatment was associated with a significant effect on death (OR 0.42, 95%CI 0.18 to 1.01, P = 0.05). Subgroup analysis indicated that intramyocardial cell injection was preferred due to its more significant improvement of LVEF than intracoronary cell therapy. Meta-regression suggested the existence of a negative association between baseline LVEF and LVEF change.
CONCLUSIONBMCs infusion is associated with a significant improvement in LVEF, and an attenuation of left ventricular remodeling.
Bone Marrow Transplantation ; Humans ; Myocardial Ischemia ; surgery ; Randomized Controlled Trials as Topic ; Transplantation, Autologous
7.In vitro and in vivo MR imaging of SHU 555A-labelled swine bone marrow mesenchymal stem cells
Zhe-Yong HUANG ; Jun-Bo GE ; Shan YANG ; Shao-Heng ZHANG ; Rong-Chong HUANG ; Meng-Su ZENG ; Hang JIN ; Ai-Jun SUN ; Ju-Ying QIAN ; Yunzeng ZOU ;
Journal of Interventional Radiology 1994;0(02):-
Objective To detect the feasibility of magnetically labeled swine bone marrow mesenehymal stem cells(MSCs)with SHU 555A combined with poly-L-arginine(PLL),under MR imaging in vitro and in vivo.Methods Swine mesenehymal stem cells were isolated and culture-expanded 3 passages in vitro,then magnetically labeled by incubation with SHU 555A(25?g Fe/ml,Resovist,Schering)for 24 hours with 750 ng/mL poly-L-lysine(PLL;average MW_275 kDa)added 1 hour before incubation.Cellular iron incorporation and detention at 0 d,4 d,8 d,12 d,16 d,20 d after labeling was qualitatively assessed using Prussian blue and quantified at atomic absorption spectrometry.Cell viability was assessed by trypan-blue exclusion test.Cell suspensions underwent MR imaging with T_1-and T_2-weighted spin-echo and fast field-echo sequences on a clinical 1.5 T MR system.At last,1?10~6 SHU 555A labeled and unlabeled MSCs were transextracardially implanted into the infracted and normal myocardium approximately 2 week following the ligation of left anterior descending coronary artery in 1 swine respectively,and finally performed 1.5-T MRI within 1 week after infarction.Results①Intracytoplasmic particles stained with Prussian blue stain were detected for all cells with mean cellular iron content of(13.13?2.30)pg per cell.With division of stem cells, the stained particles decreased gradually with iron content(0.68?0.20)pg per cell.at 16 days after labeling, approximately to the prelabeled baseline values.(0.21?0.06)pg per cell(P>0.05).The viability of the labeled cells at various time points were not significantly different with that of nonlabeled cells(P>0.05).②MR images showed signal intensity changed most obviouly in T2*WI in vitro.The percentage change of signal intensity increased with increasing cell numbers,and decreased with the time.As few as 5?10~4-1?10~5 cells could be detected by using this approach.③Two injected sites containing MR-MSCs were detected in vivo,presentingas low signal intensity areas with the T_2*WI scanning sequence.Conclusion Swine bone marrow MSCs can be labeled with SHU555A-PLL and depicted with a standard 1.5-T MR imager in vitro and in vivo.(J lntervent Radiol,2007,16:115-121)
9.Apoptosis of Fas(+) Yac-1 cells induced with Fas ligand-transfected COS-7 cells.
Ling-Bo LIU ; Ping ZOU ; Yan CHEN ; Shan-Jun SONG
Journal of Experimental Hematology 2002;10(4):327-331
The possibility of immunotherapy for lymphoma by single FasL-Fas way was investigated. After pBillneo-mFasL was transformed into competent E. coli DH5alpha and amplified, the plasmid DNA was prepared and purified from the DH5alpha. To determine the primary structure and inserting direction of mFasL cDNA gene in pBillneo-mFasL, the plasmid DNA was cleaved by restriction enzyme, and the mFasL cDNA of pBillneo-mFasL was amplified by polymerase chain reaction (PCR), the DNA sequence of the PCR product was analysed by automatic DNA sequencing. After pBillneo-mFasL was transfected into COS-7 cells by liposome, the COS-7 cells were selected with G418 selective medium, and the expressing levels of mFasL cDNA on the COS-7 cell membrane was assayed by Western Blot. After the COS-7 cells higher expressing mFasL protein and mouse lymphoma cell line Yac-1 expressing Fas were cocultured for 5 hours, the suspending Yac-1 cells were collected and labeled by annexin V/PI kit. The apoptosis rate of the Yac-1 cells was tested by flow cytometry. The EcoRI cleaving products of pBillneo-mF asL included 920 bp and 7227 bp fragments. Its Hind III cleaving products included 1293 bp and 6807 bp fragments. These results showed: (1) the length of DNA sequence containing mFasL cDNA within pBillneo-mFasL is the same as theoretical length; (2) the inserting of mFasL cDNA in pBillneo-mFasL was in positive orientation. The expected 890 bp DNA fragments of mFasL cDNA (from ATG to +36 bp following TAA) emerged in PCR product with pBillneo-mFasL as a template. The sequencing result of the PCR product equaled the known mFasL cDNA sequence in the gene bank. The COS-7 cells transfected by pBillneo-mFasL and selected with G418 culture medium expressed more mFasL membrane protein assayed by Western Blot. After the COS-7 cells were cocultured with Fas(+) Yac-1 cells in different E:T ratios (1:1, 5:1 and 10:1) for 5 hours, the apoptosis rates of Yac-1 cells were (22 +/- 4.8)%, (32.18 +/- 7.8)%, and (51.8 +/- 5.4)%, respectively. These were obviously different from the control group (P < 0.01), in which the COS-7 cell was transfected by pBillneo (not carrying mFasL gene). It was concluded that lymphoma cells highly expressing Fas can be effectively killed through single Fas-FasL way in vitro.
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Apoptosis
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COS Cells
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Fas Ligand Protein
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Immunotherapy
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Lymphoma
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pathology
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therapy
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Membrane Glycoproteins
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genetics
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physiology
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Mice
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Transfection
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Tumor Cells, Cultured
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fas Receptor
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analysis
10.Protection mechanisms of ATP-sensitive K channels on hippocampal CA1 neurons during chronic severe hypoxia.
Lian-Yan HUANG ; Wen-Jun LI ; Bo-Xing LI ; Fei ZOU
Chinese Journal of Applied Physiology 2007;23(3):257-261
AIMTo study the protection mechanisms of K(ATP) channels on hippocampal CA1 neurons during chronic severe hypoxia.
METHODSp53 expression, DNA fraction, and cell apoptosis were examined in cultured hippocampal neurons in control group, hypoxia group, hypoxia group treated with K(ATP) channels antagonist and hypoxia group treated with K(ATP) channels agonist.
RESULTSIn the group of a 12 h long exposure to oxygen concentration of 0%, diazoxide (100 micromol/L), the K(ATP) channels agonist, reduced p53 expression and the hypoxia-induced apoptosis. In contrast, tolbutamide (100 micromol/L), the K(ATP) channels antagonist, significantly rose p53 expression and the hypoxia-induced apoptosis, which could be reversed by p53 inhibitor TSA.
CONCLUSIONK(ATP) channels protect hippocampal neurons against chronic severe hypoxia by suppressing p53 expression.
Adenosine Triphosphate ; metabolism ; Animals ; Apoptosis ; Cell Hypoxia ; Cell Survival ; Diazoxide ; pharmacology ; Genes, p53 ; Hippocampus ; cytology ; metabolism ; KATP Channels ; antagonists & inhibitors ; metabolism ; Neurons ; cytology ; metabolism ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Tolbutamide ; pharmacology