1.The maturation induction of human monocyte-derived dendritic cells
Dongyin LI ; Chuan GU ; Jun MIN ; Zhonghua CHU ; Qingjia OU
Chinese Journal of Immunology 1985;0(06):-
Objective:To investigate the most effective strategy for mature induction of dendritic cells.Methods:Human monocyte-derived dendritic cells were induced in the presence of cytokines GM-CSF and IL-4. On day 6, the immature DCs were pulsed with each of CD40L, LPS, TNF-? or a cocktail of cytokines(TNF-?, IL-6, IL-1?, PGE2). DCs were harvested after 24 h induction. The surface markers for maturation CD80,CD83,CD86 and HLA-DR were detected by FCM. FITC-Dextra endocytic activity was measured by FCM. IL-12 production was detected by ELISA. The capacity of DCs for T cell activation was detected by MTT assay.Results:CD40L,LPS,TNF-? and the cocktail of cytokines all could induce DCs’ maturation. The most effective scheme for induction of maturation was the cocktail of cytokines, and the expression rate of CD83 was up to 66.91%(P
2.The influence of Survivin in drug resistance of breast cancer cell.
Ping XIE ; Qiu-rong RUAN ; Jun-chuan LI
Chinese Journal of Pathology 2005;34(6):367-368
Antimetabolites, Antineoplastic
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pharmacology
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Apoptosis
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Breast Neoplasms
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genetics
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metabolism
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pathology
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Cell Line, Tumor
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Drug Resistance, Neoplasm
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Female
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Fluorouracil
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pharmacology
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Gene Expression Regulation, Neoplastic
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Humans
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Inhibitor of Apoptosis Proteins
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Microtubule-Associated Proteins
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biosynthesis
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genetics
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physiology
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Neoplasm Proteins
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biosynthesis
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genetics
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physiology
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Oligodeoxyribonucleotides, Antisense
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genetics
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RNA, Messenger
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biosynthesis
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genetics
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Transfection
3.Clinical study of autologous tumor tissue lysate loading dendritic cells for the treatment of hepatocellular carcinoma
Dongyin LI ; Chuan GU ; Jun MIN ; Zhonghua CHU ; Qingjia OU
Clinical Medicine of China 2008;24(7):693-696
Objective To investigate the feasibility and safety of autologous tumor tissue lysate loading den-dritic cells(DC) for the treatment of hepatocellular carcinoma (HCC). Methods The monocytes-derived DC were induced and antigen loaded with tumor tissue lysate to produce DC vaccine. Vaccination and clinical observation were conducted in 12 HCC patients for 41 times. Results The average output of DC was 1.69×107(1.69×107±9.44×106>) from 90 ml peripheral blood. 63.41% (26/41)patients appeared to develop delayed-type hypersensitivity after intradermal injection. After an average of 9 months follow up, 1 patient out of 4 recurrence and metastasis pa- tients survived for 17 months. The other three patients progressed. Out of 8 patients undergoing immunotherapy post- operatively,6 patients had no signs of recurrence and the others were found to have liver rceurrence and progression. Conclusion DC based immunotherapy is safe and feasible,with no side effects,which can be applied in the immu- notherapy strategy of HCC patients.
5.Effects of sample digestion conditions on urinary iodine measurement results
Wei, MA ; Xiu-wei, LI ; Shi-chuan, LIU ; Shu-hua, LI ; Lie-jun, LIU
Chinese Journal of Endemiology 2013;32(6):694-696
Objective To study the effects of sample digestion conditions on measurement results of urinary iodine determined by As(Ⅲ)-Ce4+ catalytic spectrophotometry with ammonium persulfate digestion,and to promote the application of newly revised (the 2012 edition) national standard method for determination of urinary iodine.Methods According to the newly revised national standard method,various digestion conditions,such as ammonium persulfate concentration (0.8-1.3 mol/L,group interval 0.1),digestion instruments (heating block and drying oven) and standing time after digestion(0.5,1.0,2.0,4.0 and 22.0 h),were studied.The samples included 3 standard materials,which were GWB09108k,GWB09109f and GWB09110m containing iodine of (68.2 ± 9.0),(138.0 ± 10.0) and (221.0 ± 10.0) μg/L,and 5 urine samples with iodine concentration of 100-300 μg/L.Results Measurement results among the three groups of 0.9,1.0 and 1.1 mol/L ammonium persulfate digestion fluid showed no significant difference(P > 0.05).The digestive effect showed no significant difference between heating block and drying oven (P > 0.05) except one standard material in low concentration (GBW09108k).After digestion,samples were placed 0.5-22.0 h,the measurement results between groups showed no significant difference (P > 0.05).Conclusions Appropriate concentrations of ammonium persulfate are from 0.9 mol/L to 1.1 mol/L.Heating block is recommended for the digestion,however,when absent,drying oven can be used alternatively.The standing times from 0.5 h to 22 h after digestion have not affected the measurement results.
6.Free vascularized leg perforator flap for the wounds of the extremities
Yongqing XU ; Yueliang ZHU ; Xinyu FAN ; Ji SHI ; Liangbin MEI ; Jun LI ; Hui TANG ; Chuan LI
Chinese Journal of Microsurgery 2012;(6):450-452,后插3
Objective To evaluate the locating effects of a portable ultrasound for leg perforator flaps,and the clinical effects of leg perforator flaps for the wounds of the extremities.Methods Since December 2009 to March 2011,thirty-one cases of soft tissue defects of the extremities were treated with free vascularized leg perforator flaps in our center.A portable ultrasound were used for the locating of the perforator arteries of the leg before the operations.These flaps include 24 cases of the fibular artery perforator flaps (4 with simultaneous fibula transfer),three cases of the posterior tibia artery perforator flap,two cases of the medial gastrocnemius artery perforator flap,and 2 cases of the lateral gastrocnemius artery perforator flap.The flap size ranged from 15.0 cm × 8.0 cm to 1.0 cm × 1.5 cm.The caliber diameter of the perforator artery ranged from 0.2 mm to 1.8 mm.The wounds included 21 cases of the hand,five cases of the forearm,and 5 cases of the leg.Results The coincidence rate intraoperatively of the portable ultrasound was 96.8%.All the flaps survived and the wounds healed uneventfully.The donor site of the flaps were either closed directly or closed with partial split-thickness skin graft.Conclusion The portable ultrasound is an effective,reliable and accurate instrument for locating the leg perforator flaps.The leg perforator flaps have the advantages of convenient harvest,satisfactory thickness,less donor site morbidity,and high survival rate.
7.Keratocytoma of submandibular gland:report of a case.
Ji-man LI ; Ying WANG ; Jin-chuan FAN ; Mei TANG ; Jun HOU ; Lun LI
Chinese Journal of Pathology 2013;42(12):838-839
Carcinoma, Squamous Cell
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pathology
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Child
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Cysts
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pathology
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Diagnosis, Differential
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Epithelium
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pathology
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Female
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Follow-Up Studies
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Humans
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Keratin-19
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metabolism
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Keratin-5
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metabolism
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Keratin-6
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metabolism
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Keratin-7
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metabolism
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Keratins
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metabolism
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Neoplasm Recurrence, Local
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surgery
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Reoperation
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Submandibular Gland
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surgery
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Submandibular Gland Neoplasms
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metabolism
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pathology
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surgery
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Transcription Factors
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metabolism
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Tumor Suppressor Protein p53
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metabolism
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Tumor Suppressor Proteins
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metabolism
8.A report of atypical hypomyelinating leukodystrophy with atrophy of the basal ganglia and cerebellum caused by a de novo mutation in tubulin beta 4A (TUBB4A) gene and literature review
Ying DU ; Chuan LI ; Jun GUO ; Peng GUO ; Zhuyi LI ; Wei ZHANG
Chinese Journal of Internal Medicine 2017;56(6):433-437
Objective To explore the clinical symptoms and neuroimaging features of a patient with atypical hypomyelinating leukodystrophy with atrophy of the basal ganglia and cerebellum (H-ABC) caused by a novel TUBB4A mutation.Methods We analyzed the clinical data, imaging features and the result of genetic testing of a case diagnosed as atypical H-ABC.Results The initial symptoms were progressive spasticity, mild cerebellar ataxia and mild cognitive impairment.MRI showed regional blurring of slight high signal on T2-weight and FLAIR image in white matter of the bilateral midbrain ventral, internal capsule, posteior horn of lateral ventricle and centrum semiovale, with normal bilateral cerebellar and caudoputamen nucleus.Compared with normal subjects of the same age and gender, hypometabolism was found by 18F-FDG-PET in brainstem, cerebellar and caudoputamen nucleus in the patient.Genetic testing revealed a de novo pathogenic exome missense heterozygous mutations c.70G>A in TUBB4A, which was not reported in the human gene mutation database (HGMDpro) and was assessed to be a pathogenic mutation by pathogenic mutation prediction software.Conclusions The diversity of TUBB4A gene mutations may cause different functional and/or structural impairment in subcortical white matter, cerebellar and caudoputamen nucleus, leading to atypical symptoms and neuroimaging features.Genetic testing for pathogenic mutation in TUBB4A gene is a key for the diagnosis of H-ABC.
9.Characteristics of proliferation and myotube cell formation of rat skeletal muscle satellite cells cultured in vitro
ke, XU ; zu-jun, FANG ; ying-chuan, LI ; jie, ZHENG ; qiang, DING
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
Objective To establish a method of isolation and purification of rat skeletal muscle satellite cells,and observe the characteristics of proliferation and myotube cell formation of skeletal muscle satellite cells cultured in vitro. MethodsPurified skeletal muscle satellite cells were obtained by improved two-step enzymatic digestion method and pre-plating technique.Immunohistochemical staining was employed to identify the skeletal muscle satellite cells cultured in vitro.The growth of skeletal muscle satellite cells cultured in vitro was examined by MTT assay.The differentiation of skeletal muscle satellite cells was observed by inverted microscopy. Results The skeletal muscle satellite cells with higher purity were obtained and confirmed by the high expression of desmin.When cultured in vitro,the latent phase of skeletal muscle satellite cells was the first to the second day,and the platform phase was the fifth to the sixth day.Myotube cells gradually formed when cell confluence was more than 60% to 70% or differential medium with lower fetal bovine serum was used.Conclusion The combination of improved two-step enzymatic digestion method and pre-plating technique serve as an easy and practical way to obtain skeletal muscle satellite cells with higher purity.Skeletal muscle satellite cells can form myotube cells with contraction characteristics without any special induction.
10.Effects of Adenovirus-mediated Angiotensin-Converting Enzyme-shRNA on Spontaneously Hypertensive Rats
Jun-Hua HE ; Chuan-Shi XIAO ; Mao-Lian LI ; Yun-Fei BIAN ;
Chinese Journal of Hypertension 2006;0(08):-
Background Hypertension is a multigenetic inheritable disease.Gene therapy with long-term effects and less side effects by regulating gene expression has been shown to be a potential and exciting prospect. Objective To investigate the effects of RNA interference(RNAi)targeting angiotensin-converting enzyme(ACE)on the blood pressure and ACE expression in kidney of spontaneously hypertensive rats(SHR).Methods SHR were randomly to receive placebo(n=12)or control adenovirus Ad5-EGFP)or a single injection of recombinant adenovi- ral vectors,Ad5-EGFP-ACE-shRNA(n=12,iv).Normotensive Wistar-Kyoto rats(WKY)were served as normal control group.SBP was measured before and after the intervention.Aorta,lung,myocardium and kidney were studied using fluorescence microscope to identify the sites of Ad5-EGFP-ACE-shRNA.Expressions of ACE mRNA and protein in kidney were evaluated by RT-PCR and Western blot.Results SBP of the treat group was effectively reduced by 19.0?3.2 mmHg at the 3rd day,and 22.1?3.3 mmHg at the 13th day of the experiment.The anti- hypertensive effect significant remained at least for 14 days.On the contrary,increase in BP was shown in placebo and the adenovirus control group.Compared with placebo or adenovirus control rats,ACE mRNA expression level in kidney of the treated rats was lower by 61.1% and 62.3% respectively,with ACE protein expression level lower- ing by 56.2% and 53.30% as well(ail P0.05). Conclusion RNA interference targeting ACE gene inhibits the expressions of ACE mRNA and protein.A single dose injection resulted in a prolonged decrease in BP.The evidence of strong antihypertensive effect by genetic therapy justifies efforts for further investigation.