Objective: Overexpression of the HER2/neu oncogene is a frequent molecular event in multiple human cancers. Being a cancer antigen, p185 erbB2 is an ideal target for immunotherapy. In order to decrease the immunogenicity of mouse anti-p185 erbB2 monoclonal antibody in human cancer therapy, we constructed the eukaryotic expression vector of anti-p185 erbB2 chimeric monoclonal antibody and verified expression of the chimeric antibody in CHO-dhfr - cell. Methods: The variable regions of light chain and heavy chain were amplified with RT-PCR and inserted into the chimeric antibody vector pWSD2. After CHO-dhfr - cells were transfected with recombination plasmid by lipofectAMINE, the chimeric antibody expressing level was identified with RT-PCR, indirect-ELISA, and Western blot. The specificity of the anti-p185 erbB2 chimeric antibody was testified with ELISA assay and immunoprecipitation. Moreover, the effects of chimeric antibody on the proliferation of breast cancer cell line SKBR3, which is overexpressing p185 erbB2 , were measured with MTT assay in vitro. Results: The anti-p185 erbB2 chimeric antibody eukaryotic expression vector was constructed successfully and the expression of the chimeric antibody in CHO-dhfr - was verified by RT-PCR, indirect-ELISA, and Western blot. ELISA assay showed that chimeric antibody reacted with cells overexpressing p185 erbB2 specifically, but did not react with that non-overexpressing p185 erbB2 . Immunoprecipitation test confirmed that the chimeric antibody could bind to p185 erbB2 specifically. The MTT assay demonstrated that the chimeric antibody could inhibit the growth of SKBR3 cells overexpressing p185 erbB2 . Conclusion: The anti-p185 erbB2 mouse/human chimeric antibody that was expressed in CHO-dhfr - cells can bind to p185 erbB2 specifically and inhibit proliferation of SKBR3 cells overexpressing p185 erbB2 . It has a potential application in biotherapy of cancer.

Suitable pretreatment of biological samples can truly reflect the role of law of the measured components played in the body and will provide experimental evidence for the studies on metabolic process, material basis of efficacy, mechanism of action, pharmacology, toxicology and the others. Biological samples include blood, urine, hair, tears, etc. There are also many samples processing methods, such as the direct protein precipitation, liquid-liquid extraction and solid phase extraction and so on. These methods could be used alone or combined.
Animals ; Body Fluids ; chemistry ; Chemical Precipitation ; Chemistry Techniques, Analytical ; methods ; Humans ; Liquid-Liquid Extraction ; Proteins ; isolation & purification ; Solid Phase Extraction

Objective To investigate the effect of acupuncture at Sanyinjiao with functional magnetic reso- nance imaging(fMRI),analyse the regulating effect of the acupoint Sanyinjiao on various brain areas,explore the mechanisms of Sanyinjiao(SP.6) acupuncture in treating diseases.Methods Eight right-handed healthy volun- teers were scanned at 1.5 Tesla magnetic resonance imaging scanner while they were aeupunctured at the right Sany- injiao acupoint.Image data were co-registered to correct head motion,spatial normalization and deconverlution analy- sis were conducted.Functional activation maps were generated.Results There observed certain activation in both sides of medial frontal gyrus,right middle frontal gyrus,left inferior frontal gyrus,both sides of paracentral lobule, precentral gyrus and postcentral gyrus,left inferior parietal lobule,both sides of preeuneus,right middle temporal gy- rus,right cingulated gyrus,posterior cingulated gyrus,both sides of superior temporal gyrus,transverse temporal gy- rus,insula and thalamus.Conclusion Acupuncturing the Sanyinjiao can lead to the functional changes in parietal lobe,frontal lobe,temporal lobe and insula,precuneus,cingulated gyrus,thalamus,which are closely related to the therapeutical effects of Sanyinjiao.The phenomenon that needling Sanyinjiao can activate certain brain areas proved the correlation between acupoint and corresponding brain cortices.

ZHU Lian is the founder of international communication of acupuncture-moxibustion after establishment of New China. This article discussed ZHU Lian's great deal of pioneering work and historic contributions from the aspects of acupuncture-moxibustion treatments for foreign people, international training and communication, and the international influence of Xin Zhenjiu Xue. Our research showed that ZHU Lian's early work didn't only enlarge acupuncture's worldwide influence and spreading pace, especially in Soviet Union, India and Korea, but also accumulated precious experiences for each subject of Chinese medicine attending into international medical care and academic communication in the future.
Acupuncture ; education ; history ; Acupuncture Therapy ; history ; instrumentation ; Asia ; China ; Europe ; History, 20th Century ; Humans ; Internationality ; Moxibustion ; history ; methods

AIM: To analyze the genotype of the allele distribution of a polymorphic G-954C within the 5 upstream promoter region of the nitric oxide synthetase 2A gene (NOS2A) in samples of diabetic retinopathy in patients with cystoid macular edema in the mainland of China.METHODS: Eighty-nine patients with diabetic retinopathy and cystoid macular edema and 90 healthy controls were enrolled in this study. Nest polymerase chain reaction (PCR)was performed, and restriction endonudease digestion and gene fragments sequence were examined to detect the genotype of NOS24 G-954C.RESULTS: The genotypes of the sample population of 89 cases and 90 healthy controls were all detected as GG.CONCLUSION: The distribution of G-954C of NOS2A polymorphism are at a lower frequency in China, with little relevancy to the frequency of diabetic retinopathy combined with cystoid macular edema.

Deficiency Diseases ; prevention & control ; Goiter ; chemically induced ; Humans ; Iodine ; administration & dosage ; adverse effects ; deficiency

OBJECTIVETo explore the association between polymorphism in the ACE I/D gene and blood pressure-lowering response to hydrochlorothiazide (HCTZ) in 829 patients.

METHODSHCTZ 12.5 mg was taken once a day for six weeks. The blood pressure reduction and ratio reaching target blood pressure were compared in different ACE genotype groups.

RESULTSThe reduction in SBP of patients carrying DD was greater than that in other groups carrying II or ID (12.2 mmHg versus 5.4 mmHg, 12.2 mmHg versus 4.4 mmHg, respectively, P<0.05). The reduction in MAP of patients carrying DD was also greater than that in other groups carrying II or ID (6.9 mmHg versus 3.9 mmHg, 6.9 mmHg versus 3.6 mmHg, respectively, P<0.05). The ratio reaching target blood pressure in DD groups was significantly higher than that in II or ID groups (P<0.05). The pre-treatment SBP, DD genotype, aldosterone levels entered the multi-linear regression model significantly and might affect the reduction of SBP. The pre-treatment DBP, aldosterone levels, DD genotype entered the multi-linear regression model significantly and might affect the reduction of DBP. The pre-treatment MAP, DD genotype, aldosterone levels entered the multi-linear regression model significantly and might affect the reduction of MAP.

CONCLUSIONACE genotyping is associated with blood pressure-lowering response to HCTZ. Specific genotypes might be associated with the response to specific antihypertensive treatment.

Aged ; Alleles ; Antihypertensive Agents ; therapeutic use ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Hydrochlorothiazide ; therapeutic use ; Hypertension ; drug therapy ; genetics ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; genetics

OBJECTIVETo study the expression of lipopolysaccharide (LPS) receptor CD14 and Toll-like receptor 4 (TLR4) on Kupffer cells and its role in ischemia-reperfusion injury (IRI) on rat liver graft.

METHODSThe Kupffer cells following IRI were isolated and divided into control, ischemia-reperfusion (IR), and anti-CD14 antibody group. The mRNA and protein expression of CD14 and TLR4, nuclear factor kappa B (NF-kappaB) activity and TNF-alpha level in supernatant were measured.

RESULTSThe mRNA and protein expression of CD14 and TLR4 in IR group were significantly higher than those in control group (P < 0.01). The NF-kappaB activity and TNF-alpha level in IR group were significantly higher than those in control group (P < 0.01), and they greatly decreased after anti CD14 antibody treatment (compared with IR group, P < 0.05), but were still significantly higher than those in control group (P < 0.01).

CONCLUSIONSLPS following IRI could up-regulate CD14 and TLR4 gene and protein expression on Kupffer cells, and subsequently activate NF-kappaB to produce cytokines, but other signal transduction pathways might also participate in the IRI.

Animals ; In Vitro Techniques ; Kupffer Cells ; pathology ; physiology ; Lipopolysaccharide Receptors ; genetics ; metabolism ; Liver Transplantation ; pathology ; physiology ; Male ; NF-kappa B ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology ; physiopathology ; Signal Transduction ; Toll-Like Receptor 4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo study the chemical constituents of radix and rhizome of Ligularia sagitta.

METHODIsolation and purification were carried out on the column chromatography of silica gel and sephadex LH -20. The structures were identified by spectral analysis.

RESULTSeven terpenoid compounds were isolated from the radix and rhizome of L. sagitta. Five of them are sesquiterpenoids, the structures were determined as 7alpha-hydroxy-9(10) -ene-1, 8-dioxo-6, 7-dihydrofuranoeremophilane (1), 1p, 10P3-epoxy-6beta, 8 beta-dihydroxy-eremophil7 (11) -en-12, 8alpha-olide (2) , 1-oxo-9-desoxycacalol (3), benzofuranoeremophil-l-ene (4) and bakkenolide (5); other two compounds belong to triterpenoid. 3p, 16P3-dihydroxy-12-oleanen-28-al (6) and lupeol (7).

CONCLUSIONCompoundsl-3 and 6 were: isolated from L. sagitta for the first time, and compounds 3 and 6 were obtained from Ligularia for the first time.

4-Butyrolactone ; analogs & derivatives ; chemistry ; isolation & purification ; Asteraceae ; chemistry ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Sesquiterpenes ; chemistry ; isolation & purification ; Terpenes ; chemistry ; isolation & purification

Objective Clinical data of 19 Chinese patients with 21 hydroxylase deficiency (21OHD) were analyzed to improve the diagnosis and treatment level. Methods Clinical features and laboratory data were collected from 19 patients with 21OHD before and after treatment. Results In male patients, the average age of early appearance of secondary sexual character was (9.3?2.8)yrs, and excess androgen resulted in phallic enlargement. Primary amenorrhea was the most common complaint in female(87.5%), and the signs included a varying degree of labioscrotal fusion and clitoral enlargement. The average level of 17-hydroxy progesterone(17OHP) was (63.42?35.07) ?g/L, and adrenocorticotrophic hormone(ACTH), dehydroepiandrosterone(sodium) sulfate(DHEAS) and testosterone(T) were obviously elevated. CT scan showed bilateral adrenal hyperplasia. The level of 17OHP was significantly decreased after treatment[(63.42?35.07) ?g/L vs (3.15?2.71) ?g/L](P