1.Expression and activity detection of novel humanized anti-CD20 monoclonal antibodies
Ru JIA ; Yanwen JIN ; Ping LI ; Xuan LIU ; Ting GAO ; Zijing LIU ; Cheng CAO
Military Medical Sciences 2014;(11):875-878
Objective To construct humanized monoclonal antibodies against CD 20 and check their affinity to CD 20 antigen and their anti-tumor activity.Methods Based on the computer model , human IgG1 candidates closest to rituximab in crystal structure were selected in the Protein Data Bank ( PDB) .With the selected human IgG 1 candidates as the frame , we modified and transplanted the complementarity determining region ( CDR) of rituximab .First,the target gene fragments were obtained by overlapping PCR.Then, the sequences of the light chains(L) and the heavy chains(H) were inserted in-to the pcDNA3.3 and pOptiVEC vectors.Next, the constructed clones were transfected into 293F cells through transient transfection.After a large-scale cell culture, the mAb was purified by affinity chromatography rProtein A column.The puri-ty and expression level of the humanized antibodies was tested by sodium dodecyl sulfate ( SDS)-polyacrylamide gelelectro-phoresis(PAGE).The affinity of the humanized antibodies to CD20 was assessed with Fortebio assay.Finally, the anti-tumor activity of the constructed antibodies was detected by checking the tumor growth inhibition of the nude mice transplan-ted with tumor .Results Three humanized monoclonal antibodies against CD 20 were expressed and purified successfully . In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25 ×103 and 55 ×103 , respectively.The band size of the antibodies matched the expected value.Fortebio assay revealed that the humanized antibodies could bind to CD20 with high affinity (rituximab:6.48 ×10 -9mol/L, L4H7:1.91 ×10 -9mol/L, L5H5:7.35 ×10 -10mol/L,and L5H7:1.91 ×10 -9mol/L).The tumor growth inhibition experiment showed that the anti-tumor activity of L5H7 mAb was better than that of rituximab .Conclusion Three humanized monoclonal antibodies against CD 20 have been successfully construc-ted and expressed.L5H7 mAb possesses high affinity for CD20 and a good ability to kill tumor cells.
2.ECG characteristics of left and right ventricular origin in outflow tract premature ventricular contraction with V3 migration of precordial lead
Chenglong MIAO ; Jue WANG ; Lu XU ; Ru XING ; Yan JIA ; Liu HUANG ; Yanwei WANG ; Suyun LIU
Clinical Medicine of China 2021;37(3):243-249
Objective:To investigate the electrocardiographic characteristics of left and right ventricles origin of premature ventricular contractions(PVCs) during V3 transition of precordial leads, right ventricular outflow tract (RVOT) anterior septum and right coronary sinus (RCC), and RVOT middle-posterior septum and left coronary sinus (LCC).Methods:From January 2017 to September 2019, 91 patients with ventricular extrasystole of outflow tract who had V3 transition in precordial lead and had successful radiofrequency ablation in RVOT anterior septum, middle posterior septum, LCC and RCC were selected for retrospective case control study.The electrocardiography measurements of PVCs were compared between the anteroseptal RVOT group and RCC group, as well as the middle-posterior septal RVOT group and the LCC group, respectively.The measurements included the R-wave amplitude in lead Ⅰ, Ⅱ, Ⅲ and aVF, R amplitude ratio in leads Ⅲ to Ⅱ, Q-wave amplitude in lead aVL and aVR, Q amplitude ratio in leads aVL to aVR, R-wave and S-wave amplitude from leads V1 to V3, the V2S/V3R index, the transition zone index, and the V2 transition ratio.Results:Thirty-six cases originated from the anteroseptal RVOT, and 11 from the LCC.Lead I R-wave amplitude in anterior septal RVOT was higher than LCC group((0.22±0.25) mV vs.(-0.17±0.33) mV; P=0.003). R-wave amplitude in lead Ⅱ was lower than that in the LCC group((1.59±0.35) mV vs.(1.76±0.27) mV; P=0.035). R-wave amplitude in lead aVF was lower compared with the LCC group((1.53±0.35) mV vs.(1.78±0.39) mV; P=0.050). The V2S/V3R index showed a significant difference between these two groups(1.99±0.66 vs.0.76±0.38; P<0.001). The V2 transition ratio also appeared a significant difference between the two groups(0.69±0.43 vs.1.05±0.35; P=0.005). PVCs arose from the middle-posterior septal RVOT in 32 cases, and from the RCC in 12 cases.Compared with RCC group, lead Ⅰ R-wave amplitude showed lower ((0.25±0.31) mV vs.(0.57±0.12) mV; P<0.001); R amplitude ratio in leads Ⅲ to Ⅱ higher (0.89±0.14 vs.0.72±0.18; P=0.002); Q amplitude in lead aVL((0.72±0.24) mV vs.(0.51±0.16) mV; P=0.002)higher, and Q amplitude ratio in leads aVL to aVR higher in the middle-posterior septal RVOT(0.76±0.23 vs.0.50±0.21; P=0.002). Conclusion:Among the cases with lead V3 transition, PVCs originated from the anteroseptal RVOT show significantly different R wave in lead Ⅰ, Ⅱ, aVF, V2S/V3R index, and the V2 transition ratio compared with those from the LCC.The PVCs from the middle-posterior septal RVOT and the RCC have different R wave in lead Ⅰ, R amplitude ratio in leads Ⅱ and Ⅲ, Q amplitude ratio in leads aVL and aVR.Combined with its different characteristics, it can help to identify the origin of left and right ventricles.
3.TWO PSEUDOMONAS ACT ON HYDROCARBON AND THEIR SYNERGISTIC EFFECT
Jinyu SHAN ; Ying JIA ; Jianhuang LIU ; Liyu-Ping YING ; Zhi-Yu CHEN ; Ru-Lin LIU ;
Microbiology 1992;0(04):-
Two Pseudomonas strains, K80 - B and K80 - A, can use hydrocarbon as carbon source by two different mechanisms. Different effects can be observed when they are applied to 69-8 oil of Dagan Oil Field. The strain K80-B producing both hydrocarbon pseudosolubilizing factor and hydrocarbon emulsifying factor can decrease the viscosity and solidifying point of the 69-8 oil 51.6% and 2℃ respectively. Whereas the other can only decrease 43.5% and 3℃ respectively. When the two strains are mixed, synergistic effect on crude oil is obvious.
4.Acute tonsillitis treated by blood-letting at Shaoshang (LU 1).
Xi-Ru LIU ; Yun-Jia ZHANG ; Wei-Dong SHEN
Chinese Acupuncture & Moxibustion 2011;31(12):1126-1126
Acute Disease
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therapy
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Adolescent
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Adult
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Bloodletting
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Child
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Female
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Humans
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Male
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Middle Aged
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Tonsillitis
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therapy
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Young Adult
5.The Changes of Glutathione and Ergosterol in Saccharomyces cerevisiae Under High Pressure
Chang-Sheng QIAO ; Bo-Ning LIU ; Xu XU ; Shi-Ru JIA ;
China Biotechnology 2006;0(01):-
The growth changes of glutathione (GSH) and ergosterol in Saccharomyces cerevisiae (CICC1447 and CICC1339) were detected under 0.5Mpa pressure with compressed high-pure air (O-2∶N-2=21∶79). The results showed that logarithmic phases of the two strains were delayed; their biomass and special growth rate were lower than those of control sample (0.1MPa) and the double time were prolonged under 0.5MPa. High-pressure could increase the content of GSH obviously, compared to ambient atmosphere control samples. When the holding time was 3h, the content of GSH and ergosterol in CICC1447 increased 42.6% and 20.1%, respectively. However, the content of GSH in CICC1339 increased 58.7% when the holding time was 6h, while ergosterol content reduced. The results indicated that different yeast strains have different stress-response mechanism to copy with high-pressure shock.
6.Empirical Study on the Effect of Osmotic Opening of Blood Brain Barrier on the Facilitation of Nerve Growth Factor Passing
tian-ming, JIA ; ru, BAO ; yu-feng, LIU ; kai-xian, DU ; bin, GAN
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To explore the change of nerve growth factors(NGF) through the blood brain barrier(BBB) after distal intravenous injection of mannitol into the experimental rats and the effect of exogenous NGF on the expression of growth associated protein-43 in hypoxic-ischemic brain.Methods One hundred cases of 7 days rats were divided into 2 units.One unit was divided into 3 groups:treatment group,control group and sham operated group,20 rats in each group.The other unit was divided into 4 groups:mannitol and NGF treated group,NGF treated group,control group,and sham operated group,there were 10 rats in each group.The model rats with perinatal hypoxic-ischemic brain damage(HIBD) rats were prepared by ligation of left common carotid artery with a temporary systemic hypoxia(inhaling 80 mL/L O2 and 920 mL/L N2).The sections of brains were processed by immunochemistry with antibodies against GAP-43,and the study and memory ability of rats were tested by maze test.The effect of osmotic opening of BBB on the facilitation of NGF′s passage was tested by ELISA.Results The expression of GAP-43 increased after NGF treatment,and the differences were remarkable(P
7.Influence of beta-amyloid protein and cholesterol on the pathological changes of Alzheimer's disease and expression of nicotinic acetylcholine receptors in rats.
Ru-yu LIU ; Ran GU ; Xiao-lan QI ; Jia CHEN ; Jia-liu LIU ; Zhi-zhong GUAN
Chinese Journal of Pathology 2007;36(3):184-189
OBJECTIVETo study the influence of beta-amyloid protein (Abeta) and cholesterol on the pathological changes of Alzheimer's disease (AD) and on the expression of nicotinic acetylcholine receptor (nAChR) subunits in the brains of rats.
METHODThe rats were treated by intracerebroventricular injection of Abeta1-42 and fed with a diet containing 5% cholesterol to establish animal model of AD. The pathological changes, learning and memory, and expression of nAChRs of rats were analyzed by Bieoschowsky staining, immunohistochemistry, water-labyrinth, Western blot, and RT-PCR.
RESULTSAbeta intracerebroventricular injection induced Abeta deposition in rat brains and high-cholesterol diet resulted in hypercholesterolemia in the animals. Injection of Abeta caused a reduction of learning and memory of rats and modifications of the expression of nAChRs. Cholesterol enhanced these effects of Abeta on neuropathology and expression of nAChRs.
CONCLUSIONSAbeta can induce marked neuropathological changes, influence the learning and study ability, and modify the expression of nAChRs. Cholesterol can enhance the neurotoxicity of Abeta.
Alzheimer Disease ; chemically induced ; metabolism ; pathology ; physiopathology ; Amyloid beta-Peptides ; metabolism ; Animals ; Cerebral Cortex ; metabolism ; pathology ; Cholesterol ; blood ; Drug Synergism ; Female ; Hypercholesterolemia ; blood ; Learning ; drug effects ; Male ; Peptide Fragments ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Nicotinic ; biosynthesis ; genetics
8.Breast ptosis correction with a knitted polypropylene mesh.
Jia-ming SUN ; Qun QIAO ; Zhi-fei LIU ; Ru ZHAO
Chinese Journal of Plastic Surgery 2003;19(1):27-29
OBJECTIVETo evaluate a knitted polypropylene mesh used for mammapexy in correcting the breast ptosis.
METHODSTwenty-one patients with the mild or moderate breast ptosis were undergoing the mammapexy with a knitted polypropylene mesh. The operation was performed through the periareolar incision and a monofilament knitted polypropylene mesh was subcutaneously implanted on the surface of the gland and fixed upward.
RESULTSAll of the patients were satisfactory of the appearance with the follow-ups from 2 to 16 months. No secondary ptosis, hypertrophic scars and foreign body reactions were found.
CONCLUSIONThe above-mentioned technique could be a good, safe and reliable method for correcting breast ptosis.
Humans ; Mammaplasty ; instrumentation ; methods ; Polypropylenes ; Surgical Mesh
9.Establishment of chronic heart failure rat model of Xin-qi deficiency complicated blood stasis and edema syndrome and judgment of diagnosis information integration.
Jia-Mao LIN ; Wei-Xing GUO ; Ying WANG ; Ying LIU ; Ya-Ru LI ; Heng LI ; Jie ZHOU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(12):1457-1462
OBJECTIVETo establish and evaluate chronic heart failure (CHF) rat model of Xin-qi deficiency complicated blood stasis and edema syndrome (XQD-BSES).
METHODSTotally 40 SD rats were randomly divided into the normal control group (Control), the propylthiouracil (PTU) group, the adriamycin (ADR), and the ADR + PTU group. Normal saline was used as equivalent solvent of each group. Rats in the Control group were intragastrically and intraperitoneally injected with normal saline. Rats in the PTU group were intragastrically injected with PTU suspension and intraperitoneally injected with normal saline. Rats in the ADR group were intragastrically injected with ADR solution and intraperitoneally injected with normal saline. And rats in the ADR + PTU group were intragastrically injected with PTU suspension and intraperitoneally injected with ADR solution. The dose of PTU was 0.2% of daily forage weight, once daily. The dose of ADR was 3.5 mg/kg, once per week. The modeling lasted for 6 weeks. Left ventricular ejection fraction (LVEF), left ventricular fraction shortening (LVFS), brain natriuretic peptide (BNP), heart rate (HR), respiratory rate (RR), urine output, ear temperature, exhaustive swimming test (EST), Tri-iodothyronine (T3), tetra-iodothyronine(T4), thyroid stimulating hormone (TSH) as well as heart, lung, liver weight indices and their pathological sections were integrated and compared.
RESULTSCompared with the Control group, LVEF, LVFS, BNP, HR, RR, heart, lung, liver weight indices, urine output, ear temperature, EST, and T3, T4, and TSH changed significantly in the ADR group, the PTU group, and the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure occurred in pathological sections of heart, lung, and liver. Compared with the ADR group, LVEF, LVFS, BNP, and lung, liver weight indices, urine output, ear temperature, T3, T4, and TSH changed significantly in the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure were more serious in pathological sections of heart, lung, and liver. Compared with the PTU group, LVEF, LVFS, BNP, HR, RR, urine output, EST, T4, heart and lung weight indices changed significantly in the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure were quite serious in pathological sections of heart, lung, and liver.
CONCLUSIONADR + PTU was an appropriate method to establish CHF rat model of XQD-BSES.
Animals ; Edema ; Heart Failure ; diagnosis ; Heart Ventricles ; Humans ; Judgment ; Models, Animal ; Qi ; Rats ; Ventricular Function, Left
10.Pharmacodynamical study on Danqi capsule.
Fu-huo WU ; Xue-mei LIU ; Ru JIA
China Journal of Chinese Materia Medica 2005;30(23):1869-1873
OBJECTIVETo assess the pharmacodynamical actions of Danqi capsule which was reported to promote blood circulation by removing blood stasis, regulation Danqi and relieving pain so as to be used to treat thoracic obstruction, headache, menstrual pain in clinic.
METHODTo compare the pharmacologic effects of Danqi capsule Danqi tablet in the rats with acute myocardial ischemia and the mice with the increased oxygen-consumption induced by isoproterenol injected subcutaneously. The pain models were prepared by injection of acetic acid and uterospasm model in the female mouse was induced by diethylstilbestrol and pitocin. A hyperlipidemia model was also made in the rats.
RESULTDanqi capsule could significantly improve ECG in myocardial ischemia of rats induced by isoproterenol and prolong the mice survival time under hypoxic situation. In the experiment to observe the pain response with body twist as a index induced by acetic acid and uterospasm induced by diethylstilbestrol and pitocin, Danqi capsule could significantly shorten the latency and the time course of body twist. The contents of triglyceride(TG) and total cholesterol(TC) were decreased, while the level of high-density liporotein-cholesterol(HDL-c) was increased after treatment of Danqi capsule in in the rats with hyperlipidemia. Hemorheological data showed that the blood viscosity, blood reductive viscosity, erythrocyte rigidity index and electrophoresis time were significantly decreased by Danqi capsule in the animal model mentioned above as compared with control group (P < 0.05 or P < 0.01). In addition, the dose of Danqi capsule is less than that of Danqi tablet for producing equivalent effect.
CONCLUSIONDanqi capsule plays a good role in improving myocardial ischemia, increasing the tolerant ability against oxygen-deficiency, alleviating pain and descending the levels of blood fat and blood viscosity.
Animals ; Blood Viscosity ; drug effects ; Capsules ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Electrocardiography ; drug effects ; Erythrocyte Aggregation ; drug effects ; Female ; Hyperlipidemias ; blood ; Male ; Mice ; Mice, Inbred ICR ; Myocardial Ischemia ; physiopathology ; Pain ; physiopathology ; Panax notoginseng ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Triglycerides ; blood