Ever from FDA approved the Rituxan in 1997,an Anti-CD-20 chimeric antibody,there have been more than 20 antibodies approved and they have got into more than 56 countries.About half approvied antibodies are curing kinds of cancers and seven of them have got a excellent sale targets to be "Blockbuster Drug".As the cell culture technology improving,the large scale cell expression titer comes from 1g/L to 5g/L.At the same time,the culture scale is coming form 12,000L to 15,000L,even 20000L and it can run several bioreactors at the same time.All these are make the improvement of the antibody purification.The new edition of media,system,chromatography technology and filtration technology can product the protein to several tens kilogram antibody per cycle and tons per year,although the productivity is only several kilograms per cycle many years ago.Also the new combined technology makes a two-step chromatography production protocol come into been.This will highly improve the antibody productivity and save the investment cost.

Cassiae Semen is a common traditional Chinese medicine, and contents of anthraquinones of Cassiae Semen different significantly from area to area. According to Chinese Pharmacopoeia (2010 edition), only contents of aurantio obtusin and chrysophanol were used to evaluate the quality of Cassiae Semen, another data could be added later. Ten batches of Cassiae Semen from different areas were determined, and total anthraquinones, total free anthraquinones and total combined anthraquinones contents were assessed by ultraviolet visible spectrophotometer, contents of aurantio obtusin, rhein, aloe emodin, emodin, chrysophanol and physcion were determined by HPLC. After that, principal components analysis was used to evaluate these data determined previous by dimension reduction analysis. At last, the result suggests that three main components were found out, it shows that content of aloe emodin could be used to evaluate the quality of Cassiae Semen as well as contents of aurantio obtusin and chrysophanol. And Cassiae Semen from Hebei province posseses higher quality than Cassiae Semen from other different areas. All these results can provide a good reference for quality evaluating of Cassiae Semen medicinal materials at a certain extent.
Anthraquinones ; analysis ; Cassia ; chemistry ; Chromatography, High Pressure Liquid ; Principal Component Analysis

AIM:To study the effects of water decoction of Platycladus orientalis on reducing high blood glucose and lipids in diabetic and hyperlipidemia nice,respectively.METHODS:The mouse model of hyperlipidemia was established.The increasing rate of weight, hepatic index, lipid content of liver and atherosclerosis index (AI) were meas-ured.Blood levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), adiponec-tin (ADP) and interleukin-6 (IL-6) were detected.According to these indexes, the function of water decoction of Platy-cladus orientalis for reducing high blood lipids caused by hyperlipidemia was evaluated .At the same time, the model of dia-betes mellitus was established .The levels of fasting plasma glucose , fasting plasma insulin and blood tumor necrosis factor-α( TNF-α) were measured .The glucose tolerance test was performed .The insulin resistance index was calculated .The function of water decoction of Platycladus orientalis to reduce high blood glucose caused by diabetes was observed by these above indexes .RESULTS:Water decoction of Platycladus orientalis lowered the increasing rate of weight , hepatic index and lipid content of liver , reduced the blood content of TC , TG and IL-6, and elevated the blood HDL-C and ADP content caused by high-fat diet apparently (P<0.05).The effect of water decoction of Platycladus orientalis at high dose was more effective than that at low dose .Water decoction of Platycladus orientalis obviously alleviated the abnormity of glucose tole-rance test, reduced blood TNF-αand insulin levels, and decreased insulin resistance index caused by diabetes apparently (P<0.05).The effect of the drug at high dose was stronger than that at low dose .CONCLUSION:The water decoction of Platycladus orientalis dose-dependently reduces blood glucose caused by diabetes and blood lipids caused by hyperlipi -demia in the mice.The mechanism to reduce blood lipids may be related with elevating blood ADP content and reducing blood IL-6 content, and the mechanism to reduce blood glucose may be related with lowering blood TNF-αcontent.

OBJECTIVETo observe the effects of Danlou Tablet (DT) on inflammatory reaction, and expressions of lipoprotein-associated phospholipase A2 (LP-PLA2), secretory phospholipase A2 (sPLA2), and to analyze potential mechanisms.

METHODSForty male Wistar rats were randomly and equally divided into five groups, i.e., the normal control group, the model group, the Western medicine (WM) group, the low dose DT group, the high dose DT group, 8 in each group. Rats in the normal control group were fed with basic forage for 12 successive weeks, while AS rat model was established in rats of the other four groups by feeding high fat and sugar forage plus intraperitoneal injection of vitamin D₃. Normal saline, atorvastatin calcium suspension (at the daily dose of 1.8 mg/kg), low dose DT suspension (at the daily dose of 450 mg/kg), and high dose DT suspension (at the daily dose of 900 mg/kg) were administered to rats in the model group, the WM group, the low dose DT group, the high dose DT group respectively by gastragavage for 8 successive weeks. The general condition of all rats was observed. Rats were sacrificed after gastric administration and their serum collected. Serum levels of lipids (TC, TG, HDL-C, LDL-C) and inflammatory factors [IL-6, TNF-α, monocyte chemoattractant protein 1 (MCP-1), oxidized low-density lipoprotein (ox-LDL), lipoprotein-associated phospholipase A2 (LP-PLA2), secretory phospholipase A2 (sPLA2)] were detected. Pathological changes of thoracic aorta were observed by HE staining. Protein and gene expressions of LP-PLA2 and sPLA2 in thoracic aorta were measured by Western blot and real-time fluorescent quantitative PCR respectively.

RESULTSCompared with the normal control group, rats in the model group were in low spirits and responded poorly. Typical atherosclerotic plaque could be seen in thoracic aorta of rats in the model group. Serum levels of TC, TG, LDL-C, IL-6, TNF-α, MCP-1, ox-LDL, LP-PLA2, and sPLA2 significantly increased (P < 0.05); protein and gene expressions of LP-PLA2 and sPLA2 in rat thoracic aorta increased (P < 0.05) in the model group. After 8 weeks of intervention, rats in 3 medication groups appeared active, and HE staining showed subsidence of plaque in rat thoracic aorta. Compared with the model group, serum levels of TC, TG, LDL-C, IL-6, TNF-α, MCP-1, ox-LDL, and LP-PLA2 decreased in 3 medication groups (P < 0.01, P < 0.05); serum sPLA2 level decreased, protein and mRNA expressions of LP-PLA2 and sPLA2 in rat thoracic aorta decreased in the WM group (P < 0.01, P < 0.05); protein and mRNA expressions of LP-PLA2 in rat thoracic aorta significantly decreased in the low dose DT group (P < 0.01, P < 0.05), and those of LP-PLA2 and sPLA2 decreased in the high dose DT group (P < 0.01, P < 0.05).

CONCLUSIONDT could fight against inflammatory reaction and AS possibly through inhibiting LP-PLA2 expression and reducing ox-LDL production.

1-Alkyl-2-acetylglycerophosphocholine Esterase ; blood ; Animals ; Aorta, Thoracic ; pathology ; Atherosclerosis ; drug therapy ; Chemokine CCL2 ; blood ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; drug therapy ; Interleukin-6 ; blood ; Lipids ; blood ; Lipoproteins, LDL ; blood ; Male ; Phospholipases A2 ; blood ; Plaque, Atherosclerotic ; Random Allocation ; Rats ; Rats, Wistar ; Tablets ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo explore the characteristics of integrative medicinal diagnosis and treatment on chronic heart failure (CHF) by retrospectively analyzing the clinical data of 134 patients.

METHODSData of CHF patients hospitalized in Department of Cardiology, Xiyuan Hospital from Jan. 2003 to Dec. 2004 were collected and their characteristics of TCM syndromes, condition of treatment (both Western and Chinese medicine), and fatality rate were investigated and analyzed using the data system of Chinese medicine clinical research for coronary heart disease.

RESULTSIn the 134 CHF patients, the commonly encountered basic diseases were mainly the coronary heart disease, diabetes mellitus and hypertension; the main TCM syndrome types were qi deficiency and blood-stasis type, phlegm-stasis inter mingling type, both qi-yin deficiency type, inner water-dampness stagnancy type and yang deficiency type; showing a certain regularity between syndrome type distribution and cardiac functional NYHA grading. The total fatality rate was 6.87%, which was positively correlated with the age of patients, existence of old myocardial infarction and impairing of heart function. The Chinese medicinal treatment applied was mainly the methods of supplementing qi, activating blood circulation, dissolving phlegm and nourishing yin, and the preparations used for intravenous perfusion were mainly injections of Shengmai, Salvia mitrorrhiza, Sanchi total saponin, and acanthopanax.

CONCLUSIONThe syndromes in CHF patients of Xiyuan Hospital mainly covered qi deficiency, blood stasis, phlegm blocking, and yang deficiency, who were mostly treated with integrative medicinal treatment. However, its superiority needs the support from evidence-based medicine.

Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Heart Failure ; diagnosis ; drug therapy ; Humans ; Integrative Medicine ; methods ; Medicine, Chinese Traditional ; Qi ; Retrospective Studies

OBJECTIVETo prepare a novel floating micropellets containing hydroxy propyl-beta-oyclodextrin(puerarin-HP-beta-CD) for gastroretentive dosage form and evaluate its pharmaceutical characteristics in vitro.

METHODThe puerarin HP-beta-cyclodextrin inclusion complex was prepared by freeze-drying method. Puerarin and its HP-beta-CD inclusion complex were incorporated into alginate beads, respectively. The effect of methyl cellulose (MC), Mg-Stearate and chitosan on buoyancy and cumulative release rate of puerarin were investigated in simulated gastric fluid.

RESULTThe spectrums of FTIR and profiles of X-ray powder diffraction of samples proved the formation of inclusion complex between puerarin and HP-beta-CD. Magnesium stearate had a significant effect on the buoyancy of micropellets, and satisfied results were gained by the content with 2%. The solubility of puerarin was increased 65.6-fold by the formation of inclusion complex, the dissolution rate and cumulative release percentage also improved significantly although with the burst release phenomena. The micropellets showed sustained release properties by using puerarin-HP-beta-CD inclusion complex mixed with puerarin (1:1) and treated thoroughly under homogenizer.

CONCLUSIONThe solubility and release rate of puerarin are increased by the formation of inclusion complex with HP-beta-CD and its gastroretentive dosage forms displayed satisfied floating and sustained release characteristics.

2-Hydroxypropyl-beta-cyclodextrin ; Dosage Forms ; Freeze Drying ; Isoflavones ; chemistry ; Solubility ; Spectroscopy, Fourier Transform Infrared ; X-Ray Diffraction ; beta-Cyclodextrins ; chemistry

OBJECTIVETo evaluate the usefulness of contrast-enhanced ultrasonography (CEUS) in the diagnosis of biliary diseases.

METHODSCEUS was performed in 57 patients with biliary diseases. The contrast enhancement characteristics and the morphologic features were observed. The ultrasonographic results were compared with those obtained through conventional 2-D ultrasound (2D-US), color Doppler flow ultrasound (CDFI), and clinical, surgical, and laboratory findings. In some cases, the ultrasonographic results were also compared with those obtained from contrast-enhanced computed tomography (CECT).

RESULTSThe diagnostic accuracy of 2D-US combined with CEUS was significantly higher than that of 2D-US combined with CDFI 87.7% vs 71.9%; chi(2) = 4.41, P < 0.05). CEUS clearly showed the presence/absence of blood supply in biliary lesions and offered real-time imaging of the microcirculation perfusion in the lesions. It also offered useful information to differentiate biliary tumors from stones, bile mud, and/or blood clots. It distinctly displayed the size and contour of the lesions as well as the infiltrated range, depth, and the involved area. However, CEUS is most useful in reflecting blood perfusion patterns; it had limited value in differentiating the malignancies of polypoid lesions. The diagnostic accuracy (87.0% vs 91.3%;chi(2) = 0. 45, P > 0.05) and the size and range of the lesions displayed (0.4-6.2 cm vs 0.4-6.0 cm, P = 0.721) were not significantly different between CEUS and CECT.

CONCLUSIONCEUS is a useful tool in the routine ultrasonography of biliary diseases.

Adult ; Aged ; Aged, 80 and over ; Biliary Tract Diseases ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Ultrasonography ; Young Adult

Objective To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneously determining five compounds in Curcumae Aromaticae Radix.Methods An HPLC method was developed as QAMS to determine curcuma diol,ocathydro-1,4-dihydroxy-1,4-dimethyl-7-(propan-2-ylidene)azulen-5(1H)-one,original curcumol and curcumin in Curcumae Aromaticae Radix,using curdione as intermal reference substance,and the relative correction factor (RCF) of the four components was determined by HPLC with good reproducibility.Their contents in 10 batches of samples,collected from different areas,were determined by both external standard method and QAMS.Result No significant differences were found in the quantitative results of four compounds in 10 batches of Curcumae Aromaticae Radix determined by external standard method and QAMS.Conclusion It is feasible and suitable to evaluate the quality of Curcumae Aromaticae Radix by QAMS.

OBJECTIVEThe phosphatidylinositol-3-kinase (PI3K)-AKT signaling pathway is considered to play an important role in tumorigenesis. Frequent somatic mutations in the PI3K subunit p110a (PIK3CA) occur in a variety of cancer types. The purpose of this study was to determine the relationship between PIK3CA mutation in breast cancer and pathological features and outcome of patients.

METHODSThe PIK3CA mutations in exons 7, 9, 20 were screened in 250 primary breast cancers using PCR and fluorescent (F)-SSCP, and the results were analyzed according to their cliniopathological data.

RESULTSThe frequency of PIK3CA mutations among the 250 cases was 35.2% (88/250), point mutations in exon 7 were found in 8 (3.2%) cases,40 (16.0%) cases in exon 9 and 47 (18.8%) cases in exon 20. No significant correlation between PIK3CA mutation and age, histological type, differentiation, and lymph node metastasis was observed. Mutations were associated with larger tumor size (P = 0.004) and positive estrogen receptor status (P = 0.008). Patients with PIK3CA mutations showed a significantly worse survival (P = 0.004), particularly in those with positive estrogen receptor expression or non-amplified HER-2 (both P = 0.002).

CONCLUSIONSPIK3CA mutations may play an important role in the carcinogenesis and development of breast cancer. The association with large tumor size, ER+ and poor survival indicates that PIK3CA mutation could be an independent factor for tumor malignant phenotype and prognosis.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Carcinoma, Lobular ; genetics ; metabolism ; pathology ; Carcinoma, Medullary ; genetics ; metabolism ; pathology ; Class I Phosphatidylinositol 3-Kinases ; Exons ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Middle Aged ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Point Mutation ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Survival Rate ; Tumor Burden ; Young Adult

Background and Objective:Adenovirus vector has been widely used in tumor gene therapy.ING4 is a member of growth inhibiting factors and a potent anti-tumor gene which could induce apoptosis of many tumor cells.This study was to investigate the inhibitory effects of adenovirusmediated ING4(Ad-ING4)gene on the proliferation of human prostate cancer PC-3 cells in vitro and in vivo,and to explore its mechanisms.Methods:Ad-ING4 was obtained by virus-amplification technique.After transfection of purified Ad-ING4 into PC-3 cells,the expression of ING4 was detected by reverse transcription-polymerase chain reaction(RT-PCR);the influence of Ad-ING4 transfection on cell proliferation was evaluated using MTT assay.Cell apoptosis was assessed using Hoechst33258 staining and flow cytometry. RT-PCR was performed to detect the mRNA levels of the transcription of apoptosis-related genes such as bcl-2,bax,p53,and caspase-3.Athymic nude mice bearing PC-3 tumors were intratumorally injected with Ad-ING4 (100 μL,1×10~9 pfu/mL). Tumor growth was recorded.All nude mice were killed at the end of the experiment to observe the growth of xenografts. The expressions of Bcl-2, Bax, Caspase-3, and CD34 proteins in tumor tissues were detected by immunohistochemistry. Results: Human ING4 gene was successfully transcribed in PC-3 cells and induced apoptosis by up-regulating p53,bax,caspase-3 expression and down-regulating bcl-2 expression. Inhibition of cell proliferation was significant in PC-3 cells. Tumor growth was significantly inhibited in the Ad-ING4 group as compared with that in the Ad-GFP group and the PBS group (P＜0.05). The weight inhibitory rate was 37.0% in the Ad-ING4 group. The expressions of Bax and Caspase-3 were up-regulated,and the expressions of Bcl-2 and CD34 were downregulated in the Ad-GFP group. Conclusions: Adenovirus-mediated ING4 gene exhibits anti-tumor ability in human prostate cancer PC-3 cells in vitro and in vivo,and induces apoptosis. This may be related to the upregulations of p53, bax, Caspase-3 and down-regulation of bcl-2.