Quality control of biotech drugs has attracted increasing attentions in recent years. Deamidation reaction is one of the major concerns in quality control of biotech drugs, due to the generation of isoaspartic acid(isoAsp) . This paper describes the deamidation of asparagine(Asn) residues and its effects on the biological drugs. The detection methods currently used in China and overeas for this reaction, including pretreatment protocols and instrumental analysis were described. The identification and determination of isoaspartyl sites were also described in detail, along with the positive impact on the development of biotech drugs in China by the studies on deamidation reactions.

Objective To probe into effect of Dracocephalum heterophyllum Benth flavonoids(DHBF) on noradrenaline (NE) -induced myocardial cell hypertrophy, and to study the mechanism. Methods SD neonatal rat myocardial cells were cultured in vitro. The experiment was divided into normal control group, model control group (NE 2 μmol·L-1) , prazosin group (prazosin 50 μmol·L-1) ,low-,medium-and high-dose DHBF group (DHBF 10,25,50 μmol·L-1) .Cardiomyocyte hypertrophy were induced by NE(2 μmol·L-1) . DHBF and prazosin were intervened respectively. CCK-8 method was used to observe the activity of myocardial cells. RT-PCR technique was used to detect the expression of mRNA of cardiac hypertrophy c-jun and brain natriuretic peptide (BNP) ,and Western blotting was used to detect the protein expression of CaN and NFAT-3 in myocardial cells. Confocal laser scanning was used to detect the surface area of myocardial cell. Results Compared with the normal control group, survival rate of cardiomyocytes was significantly decreased, expression of mRNA of c-jun and BNP significantly upregulated, protein expression of CaN and NFAT-3 decreased, and surface area increased in model control group (P<0.05) . Compared with the cell of model control group, low-,medium-and high-dose DHBF significantly reversed NE-induced decrease of the survival rate, increase of surface area, increase of c-jun and BNP mRNA, and increase of CaN and NFAT-3 protein expression (P<0.05) . Conclusion DHBF can improve the survival rate of cardiac hypertrophy patients, down-regulate c-jun and BNP mRNA expression, decrease CaN and NFAT-3 protein expression, and decrease NE-induced surface area of cardiomyocytes.

Objective To evaluate the safety of morphine hydrochloride injection. Methods Ear verin injection was used to evaluate the vascular irritation using the comparison of left side with right side in rabbits. Quadriceps femoris injection was used to evaluate the muscle irritation using the comparison of left side with right side in rabbits. Guinea pigs were intravenously injected with morphine hydrochloride injection at a dose of 2.8 mg·kg-1 once daily 3 times, stimulation was performed on 14 d after the last sensitization and the booster dose was 2 times the sensitization dose. The allergic reactions were observed. The different concentrations of morphine hydrochloride injection were placed in 2％ rabbit erythrocyte suspension, and then the hemolyzation and agglutination were observed. Results There were no significant vascular or muscular irritation and injury effects of morphine hydrochloride injection in rabbits. There were no evidenceof hemolyzation and agglutination in rabbit erythrocytes in vitro. No allergic reactions on guinea pigs in vivo were observed. Conclusion After treatment of morphine hydrochloride injection, neither obvious vascular /muscle stimulation or sensitization, nor hemolyzation or agglutination appeared in rabbits. The research results provide basic reference for the clinical rational and safe application of morphine hydrochloride injection.

Objective To observe the effect of Kadsura coccinea on the expression of Bcl-2, Bax and proliferating cell nuclear antigen (PCNA) in liver tissue of experimental hepatic fibrosis rats, to further discuss the mechanism of anti-liver fibrosis. Methods Sixty SD rats were randomly divided into normal control group, model control group, colchicine group, low-dose Kadsura coccinea group (2.5 g·kg-1) and high-dose Kadsura coccinea group (5 g·kg-1) (n = 12) . All groups except the normal control group were treated with CCl4, rich fat and poor protein to establishexperimental hepatic fibrosis animal model. The second day after modeling, drug treatmentwas started, till the end of the sixth week. Pathological section of the rat' s liver was examined in order to observe its tissue under a optical microscope. Liver tissues were taken to examine the degree of liver fibrosis by HE and Masson staining, and the expression of Bcl-2, Bax and PCNA protein were detected by immunohistochemistry. Results Kadsura coccinea relieved the degree of necrosis of liver cells, liver fat' s degeneration and collagen fiber hyperplasia significantly. Compared with the model control group, expression of Bax in low-,high-dose Kadsura coccinea group and colchicine group were significantly decreased, and the expression of PCNA in hepatic fibrosis rats were enhanced(P<0.05 or P<0.01) . Conclusion Kadsura coccinea has a certain inhibitory effects on experimental hepatic fibrosis in rats, and its mechanism may be related to inhibiting the expression of Bax protein and promoting the expression of PCNA protein in liver tissues.

Objective To investigate the effect of baicalin on CT26.WT cells of colon cancer in mice, and to discuss the cell death form. Methods CT26.WT cells were divided into four groups including of control group , routine cultured in fresh medium, the baicalin group, added with concentration of 100 μmol·L-1 baicalin, the z-VAD-fmk group, was added with final concentration of 20 μmol·L-1 z-VAD-fmk, and the combination group, added final concentration of 20 μmol·L-1 z-VADfmk,1 h before adding 100 μmol·L-1 baicalin. Then the inhibitory effect of baicalin on cell proliferation and cell viability were detected by CCK-8 method. The changes of nucleus were detected by DAPI staining, the ultrastructure of cells was observed by TEM, and the effect of baicalin on the expression of RIP3 gene and protein in cells was detected by QPCR method and Western blotting. Results Compared with control group, the differences of baicalin group and combination group had statistically significance (P<0.05) . cell death rate for control group was (10.54±0.19) ％ ,for baicalin group was (34.93±0.16) ％ ,for z- VAD group was (11.23±0.59) ％, and combination group was (23.27±1.20) ％ (P<0.01) . Compared with the normal control group, baicalin group showed nuclear concentration and fragmentation. there was obvious nuclear fragmentation in the combination group against baicalin group. The results of electron microscopy showed that the cells of baicalin were necrotic, cell swelling, mitochondria swelling and contents leaking. Baicalin group significantly up - regulated RIP3 mRNA expression (P < 0. 01) and enhanced RIP3 protein expression (P < 0. 05) . Conclusion Baicalin induces the necrosis of ct26. WT cells, and can significantly increase the gene and protein expression of RIP3.

Objective To investigate the effects of disulfiram combined with copper (DSF /Cu) on proliferation and migration of brain metastases from lung cancer. Methods The cell were divided into blank control group, valproate group (1.0 mmol·L-1 of valproate) , DSF /Cu (1,2,3,4,5 mg /0.17 mg) . Brain metastases from lung adenocarcinoma were incubated 72 hours.The changes of proliferative ability of cancer cells were detected by CCK-8 method, the effect of DSF /Cu on the migration ability of cancer cells was detected by Transwell chamber, and the degree of apoptosis of cancer cells was detected by Annexus V-PI double staining method. Results The ability of proliferation was significantly inhibited, the ability of migration was significantly reduced,and the rate of cell apoptosis was significantly increased in each dose group of brain metastases from lung adenocarcinoma compared with the blank control group. With the increase of DSF /Cu dose, its ability to inhibit the proliferation and migration of metastases cancer cells and induce apoptosis of metastases cancer cells has increased gradually. Conclusion DSF /Cu can inhibit the proliferation and migration of brain metastases from lung adenocarcinoma and induce apoptosis. DSF is expected to be a new method of the combined treatment of brain metastases cancer.

Polymorphism of chemical drugs has become a hot topic in pharmaceutical research at home and abroad. In this review, the phenomena, causes and significance of polymorphism were introduced briefly. The international drug development process and characteristics of the polymorphic drug development in our country were analyzed. Finally, the present situation and development in China was summarized from four aspects including the basic theory, technical methods, intellectual property rights and supervision. This paper can provide a reference for correct understanding the research level of polymorphic drugs in China and clarifying the direction of polymorphic drug research.

Objective To study the gastrointestinal absorption process of three letrozole polymorphs in rats, and evaluate the different pharmacokinetics parameters of different polymorphs. Methods A total of 18 SD rats were given the different letrozole polymorphs. Then the high-performance liquid chromatographic method was used for the determination of plasma concentration of letrozole in these SD rats.Finally the pharmacokinetic parameters among the different polymorphs were calculated. Results Cmax of letrozole crystal form I, crystal form II and crystal form III were (9.247± 4.612) ,(23.387± 9.049) and (15.682±1.589) mg·L-1, respectively, and AUC0→t were(198.115±47.014) ,(476.641±125.467) and (271.817±41.068) mg·L-1·h,respectively. Conclusion The different crystal forms of letrozole result in different plasma concentration in SD rats. Crystal form II may be its preponderant polymorphs which deserves further research and development.

The pharmaceutical co-crystal has attracted a lot of attention in recent years as a new direction in the research of polymorphism drugs. The research on pharmaceutical co-crystal has scientific significance for improving the solubility, bioavailability and physical or chemical stability of drugs. In this paper, from the perspective of drugs for the treatment of cardiovascular diseases(including five major types: heart failure, hypertension, coronary heart disease and arrhythmia, stroke) , the latest research results of pharmaceutical co-crystal reported in recent years are reviewed, hope to provide reference for the follow-up research and promote the development of pharmaceutical co-crystal in China.

Objective According to the clinical medicinal crystal form———form γ of levonorgestrel, to establish the quantitative analysis method for levonorgestrel form γ by powder X-ray diffraction (PXRD) . Methods Firstly, single crystal X-ray diffractometry and powder X-ray diffractometry were used to confirm that the prepared levonorgestrel form γ was 100％ polymorphic purity, which provided a standard sample for quantitative analysis by single peak method; then, the standard samples of different quality levonorgestrel form γ for powder X-ray diffraction were weighed, the peak intensity values of characteristic diffraction peaks d = 6.4 , d = 6. 1  and d = 5. 6  of form γ as quantitative parameters selected, a linear relationship between the peak intensity value and the quality of form γ was established; finally, the content of levonorgestrel form γ was quantitatively analyzed. Results The peak intensity values of characteristic diffraction peaks d = 6.4 , d = 6.1  and d = 5.6  of levonorgestrel form γ and the quality showed a good linear relationship.In the range of form γ masses of 5 mg to 50 mg, the regression linear equations wereY = 459.59X+5 536.5, R2 = 0.993 0, Y = 430.03X+6 867.6, R2 = 0.990 5,Y = 615.95X+ 6 209.5, R2 = 0.990 8,respectively. Conclusion The method is simple, rapid, accurate and reliable, it can be used as quality control method for levonorgestrel polymorphs.