1.The Teaching Experience of Nuclear Medicine for the International Medical Students
Chinese Journal of Medical Education Research 2003;0(02):-
Medical education for the international medical students is an important part of home medical colleges. This article focuses on the training of teacher’s ability,the understand- ing of the international medical students,the preparation for lessons and the agile use of many teaching ways during the teaching process of Nuclear Medicine for the international medical stu-dents. Up to now,we still have no textbooks of Nuclear Medicine for the international medical students. This is the problem that exists in the Nuclear Medicine teaching.
3.Construction of and screening for phage antibody library of human lung adenocarcinoma
Hua PANG ; Yi LUO ; Shaolin LI
Journal of Third Military Medical University 2003;0(22):-
Objective To construct human single-chain antibody gene library associated with lung adenocarcinoma, and screen lung adenocarcinoma cell A549-specific antibody. Methods The lymphatic tissue near lung adenocarcinoma was used to construct human single-chain antibody gene library. The specific antibody to lung adenocarcinoma cell A549 was screened from the antibody library. Positive clone bacteria were transformed to E. coli HB2151, and their dissolvability was observed. The soluble scFv was purified by affinity chromatography and its relative molecular mass was determined by SDS-PAGE and Western blotting. The specificity of scFv to human lung adenocarcinoma cells was identified with ELISA. Results The phage antibody library of 4.6?107 was constructed successfully. Single-chain antibody of human lung adenocarcinoma was enriched. The ratio of yield was increased gradually, 181 times higher after 5 rounds of panning than after the first round of panning. SDS-PAGE and Western blotting results showed soluble scFv' molecular mass was about 30 000. ELISA showed dissolved antibody had high specificity to bind A549 cells, not MDA-MB-435. Conclusion The single-chain antibody gene library of human lung adenocarcinoma was constructed successfully, and specific antibodies to lung adenocarcinoma were screened, providing the basis for single-chain antibody radionuclide imaging and treatment.
4.Screening and identification of human anti-HIF-1? scFv of lung adenocarcinoma
Jie WANG ; Jianguo LI ; Hua PANG ; Zhiping PENG ; Shaolin LI
Journal of Third Military Medical University 2003;0(23):-
Objective To screen human anti-hypoxia-inducible factor (HIF)-1? scFv of lung adenocarcinoma from large phage antibody library and identify the positive clones. Methods Panning of large phage antibody library against lung adenocarcinoma cell line A549 and HIF-1? was conducted respectively to select specific antibodies. E. coli HB2151 was infected to induce the expression of soluble scFv. The binding activity and specificity were tested by ELISA and immunocytochemical assay. The expression and relative molecular weight of the soluble scFv was detected by SDS-PAGE and Western blot analysis. Results After panning,the target scFv was enriched,and ELISA results showed that positive reactions to HIF-1? were detected in 5 of 10 random clones with a positive ratio of 50%. Immunocytochemical analysis showed the specific affinity of the antibodies to A549 cells. The soluble human anti-HIF-1? scFv fragments of lung adenocarcinoma were expressed in E. coli HB2151 and then confirmed by SDS-PAGE. The result of Western blotting showed that the relative molecular weight of the soluble scFv was about 30?103. The binding activity and specificity were confirmed by ELISA. Conclusion Human anti-HIF-1? scFv of lung adenocarcinoma is successfully obtained with large phage antibody library technique.
5.Dysfunction of epidermal barrier in psoriasis
Hua GU ; Na LI ; Ying TU ; Qin PANG ; Li HE
Chinese Journal of Dermatology 2012;45(2):134-135
ObjectiveTo seek experimental evidence of epidermal barrier dysfunction in psoriasis,and to provide a basis for adjuvant therapy of psoriasis.MethodsPhysiometric methods were used to determine the value of sebum content,transepidermal water loss(TEWL) and water content of stratum corneum in 60 patients with psoriasis and 48 normal human controls.The ultrastructure of lamellar bodies was observed with transmission electron microscopy,and the expression of acid ceramidase in normal skin and psoriatic lesions was detected by using immunohistochemical techniques.ResultsCompared with the normal skin,TEWL value was increased(P < 0.01),but water content of stratum corneum decreased(P < 0.01 ) in psoriatic lesions,and sebum content was similar between normal skin and psoriatic lesions.As electron microscopy showed,lamellar bodies in keratinocytes were reduced in number with a disorganized arrangement and irregular size in psoriatic lesions.The expression of acid ceramidase also decreased in psoriatic epidermis.Conclusions The function of epidermal barrier in psoriasis is impaired,and to restore epidermal barrier function and enhance hydration may serve as an important adjuvant therapy of psoriasis.
6.Effects of Prinsepia utilis Royle oil on the synthesis of ceramide and expression of ceramidase
Ying TU ; Hua GU ; Na LI ; Qin PANG ; Li HE
Chinese Journal of Dermatology 2012;45(10):718-722
Objective To evaluate the effects of Prinsepia utilis Royle oil (PURO) on the synthesis of ceramide and expression of acid ceramidase N-acylsphingosine amidohydrolase 1 (ASH1),and to explore the mechanisms underlying its moisturizing and skin barrier-repairing effects.Methods Keratinocytes from human foreskin tissue were classified into 2 groups to be cultured in keratinocyte-serum free medium (K-SFM) with or without the presence of PURO.Enzyme linked immunosorbent assay (ELISA) was performed to measure the level of ceramide in the culture supernatant of keratinocytes at 0,3,8,24 and 48 hours.The back of nude mice was divided into 4 areas,i.e.,test area,matrix area,blank control area and negative control area.Acetone and ether were used to destroy the epidermal barrier in the test,matrix,and blank control areas,then,the former 2 areas were topically treated with emulsions containing 1% PURO and matrix,respectively,and the blank control area remained untreated.The epidermal barrier remained intact and untreated in the negative control area.Noninvasive methods were used to determine transepidermal water loss (TEWL),epidermal moisture content and skin lipid content in these areas on day 0,1,3,and 7.Skin tissue was obtained from these areas on day 0 and 7 followed by an immunohistochemical study for the quantification of ASH1 expression.Results The level of supernatant ceramide increased with time in the PURO-treated keratinocytes,which was significantly higher at 24 hours and 48 hours than at 0 hour (1.3817 ± 0.100 and 1.3737 ± 0.047 vs.0.7630 ± 0.143,both P < 0.05).The supernatant ceramide was also elevated in the PURO-treated keratinocytes compared with untreated keratinocytes at 24 and 48 hours (both P < 0.05).Noninvasive skin tests showed a gradual decrease in the TEWL,but an increase in the epidermal moisture content and skin lipid content with time in the 3 epidermal barrier-destroyed areas.As far as the test area was concerned,TEWL value was significantly lower on day 3 and 7 than on day 0 (10.85 ± 0.64 and 8.01 ± 0.58 vs.12.65 ± 0.71,both P < 0.05),while a significant increment was observed in the skin lipid content on day 3 and 7 compared with day 0 (29.14 ± 0.40 and 31.30 ± 0.88 vs.27.02 ± 0.65,both P < 0.05),as well as in the epidermal moisture content on day 1,3 and 7 compared with day 0 (13.98 ± 0.28,15.00 ± 0.38 and 15.86 ± 0.18 vs.11.74 ± 0.62,all P< 0.05).On day 7,there was a statistical decline in TEWL value,but an elevation in epidermal moisture content,skin lipid content and ASH1 expression in the test area compared with the matrix area and blank control area (all P < 0.05).Also,the expression of ASH1 was upregulated on day 7 compared with day 0 in the 3 barrier-destroyed areas (all P < 0.05).Conclusion PURO may exert skin-moisturizing and barrier-repairing effects by enhancing the synthesis of ceramide and expression of acid ceramidase ASH1.
7.Dietary Patterns and Cardiovascular Risk Factors in Essential Hypertensives
Li-Song LIU ; Qi HUA ; Bei-Lei PANG ;
Chinese Journal of Hypertension 2007;0(03):-
Background Although great advances have been made in prevention and treatment of CVD through drug therapies and other procedures,diet and lifestyle modification remain the foundamental aspects in clinical inter vention for prevention and treatment of hypertension.Objective The purpose of this research is to investigate the impact of variour dietary patterns on traditional cardiovascular risk factors in essential hypertensives in Beijing area. Food frequency questionnaire (FFQ) survey was carried out in a cohort of 424 patients of hypertension.Methods All participants completed an FFQ.Baseline clinical data (height,weight,waist circumference and biochemical da- ta) were collected by physical examination and biochemical assay.According to their dietary patterns,three dietary pattern were delineated:vegetarian diet(n=95),meat dominant diet(n=133) and balanced diet(n=196).Clinical and biochemical data were compared between them,and analyzed by multivariate logistic regression models.Results 1)The morbidity of obesity in meat dominant people was significant higher than vegetarians (46.6% vs 21.7%,P
8.Construction of human phage display antibody ScFv library and identification of antibody ScFv against lung adenocarcinoma
Yi LUO ; Hua PANG ; Shaolin LI ; Hui CAO ; Shujie LI ; Shubin WANG ; Chunbo FAN
Basic & Clinical Medicine 2009;29(11):1155-1160
Objective To construct a human phage single chain-antibody library, and to sieve out the antibody ScFv against lung cancer from the library. Methods Total RNA was abstracted from lymph node tissue of the lung cancer, and was used to amplify V_H and V_L gene by RT-PCR. V_H and V_L were joined by a DNA linker by SOE-PCR to form the single chain variable fragment ( ScFv) gene. ScFv gene was coloned into the phage vector pCANT-AB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression. Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning, the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis. Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.
9.Expressions of RGC-32 and E-cadherin in pancreatic cancer and their clinicopathological significance
Liang ZHU ; Huizhen ZHAO ; Huifang PANG ; Hua QIN ; Peiyuan LI ; Demin LI ; Qiu ZHAO
Chinese Journal of Pancreatology 2012;12(3):173-176
Objective To investigate the expressions of RGC-32 and E-cadherin in pancreatic cancer and analyze their clinicopathological significance and the correlation with each other.Methods SP immunohistochemistry was used to detect the expressions of RGC-32 and E-cadherin in 42 cases of pancreatic cancer tissues,12 cases of chronic pancreatitis tissues and 8 cases of normal pancreatic tissues.Results The positive staining for RGC-32 was predominantly observed in the cytoplasm of pancreatic acinar cells.The positive staining for E-cadherin was mainly observed in the cytomembrane of normal pancreatic and chronic pancreatitis acinar cells,but aberrant expression ( cytoplasm expression and ( or ) weaker expression) could be found in pancreatic cancer cells.The positive expression rate of RGC-32 and aberrant expression rate of E-cadherin were 78.6% (33/42) and 54.8% (23/42),respectively,in pancreatic cancer tissues,which were significantly higher than those in normal pancreatic tissues [37.5% (3/8) and 0] and chronic pancreatitis [41.7% (5/12)and 8.3% (1/12) with statisticai significance,P <0.05].The expression of RG C-32 in pancreatic cancer was associated with lymph node metastasis and TNM staging (P =0.016,0.025,respectively),but not with age,gender and differentiation degree ( P =0.831,1.000,0.629,respectively).The aberrant expression of E-cadherin was associated with differentiation degree,lymph node metastasis and TNM staging ( P =0.024,0.004,0.004,respectively),but not with age and gender ( P =0.970,1.000,respectively).A significantly positive correlation was found between positive expression rate of RGC-32 and aberrant expression rate of E-cadherin (r =0.458,P <0.01 ).Conclusions Both positive expression rate of RGC-32 and aberrant expression rate of E-cadherin are up-regulated significantly in pancreatic cancer tissues and RGC-32 may be involved in the invasion and metastasis of pancreatic cancer by regulating epithelial mesenchymal transition.
10.Effects of cobalt chloride mimetic hypoxia on the proliferation, apoptosis and migration of human pancreatic cancer cell line PANC1
Liang ZHU ; Shengnan XU ; Huifang PANG ; Huizhen ZHAO ; Hua QIN ; Peiyuan LI ; Demin LI ; Qiu ZHAO
Chinese Journal of Pancreatology 2012;12(2):111-114
ObjectiveTo investigate the influence of cobalt chloride ( CoCl2 )-mimetic hypoxia on theproliferation,apoptosis and migration of human pancreatic cancer cell fine PANC1.MethodsPANC1 cells were treated with 0(control),100,200,400,800 μmol/L CoCl2 respectively for 24 h.Real-time RT-PCR and Western blot were used to determine hypoxia induced factor ( HIF)-1o mRNA and protein expression respectively,and cell counting kit-8(CCK-8) assays,flow cytometry and cell scratch test were used to examine the proliferation,apoptosis and migration of PANC1 cells,respectively.ResultsIn the control group and 100,200,400 and 800 μmol/L CoCl-2 groups,the expressions of HIF-1t mRNA were 1,1.08 ±0.12,1.12 ± 0.09,1.04±0.11,0.66 ±0.07,and the expressions of VEGF mRNA were 1,2.69±0.35,4.81 ±0.54,2.19 ± 0.21,0.79 ± 0.08,while the expressions of HIF-1 α protein were 0.23 ± 0.03,0.36 ± 0.04,1.15 ± 0.11,1.08 ± 0.09,0.44 ± 0.04; and the expressions of VEGF protein were 0.14 ± 0.02,0.12 ± 0.01,0.95 ±0.09,0.87 ±0.09,0.55 ±0.06; and cell viability rates were 100%,(98.43 ±2.88)%,(76.15 ± 0.70)%,(53.87 ±0.77)%,(35.23 ±0.67)% ; while cell apoptotic rates were (5.2 ±1.12)%,(5.74 ± 1.07)%,(6.82 ± 1.85)%,(12.09 ±3.53)%,(31.88 ±6.95)% ; the cell migration distance of PANC1 cells were (43.24 ±3.67)%,(59.46 ±5.39)%,(80.56 ±8.05)%,(63.89 ±5.96)%,(9.09 ± 1.59 ) %.Compared with those of control group,the expressions of VEGF mRNA,VEGF and HIF-1 α protein,cell migration distance showed a two-way variation ( ascending first and descending later) (P <0.05 ),and the expression of HIF-1α mRNA and cell proliferation rate was decreased in a dose-dependent manner,while the cell apoptosis was increased in a dose-dependent manner.Conclusions CoCl2 significantly inhibits the proliferation and promotes apoptosis of PANC1 cells at certain level.CoCl2 has a two-way effect on the migration of PANC1 cells,and it may be related to the direct injury of high concentration of CoCl2 on cells.