Objective: To establish a determination method of chrysophanol in Kuiyanqing Xiangkouye . Methods:HPLC with DiamondTM C18 column(4.6?200mm,5?m)was used . Methanol: 0.1 %phosphoric acid(85:15)was used as mobile phase . The flow rate was 1.0mL/min . The detection wavelength was at 254nm . Results:The linearity of chrysophanol was in the range of 0.06462～1.03392 ?g . The recovery was 97.67 %and RSD was 1.37 %. Conclusion:This method is convenient and with a good resolution and can be used for the quality control of Kuiyanqing Xiangkouye .

AIM: To compare N-LOK with ?-CD on inclusion ability for the volatile oil from Duhuo Jisheng Granula (Radix Angelicae, Pubescentis, Herba Asari, Radix Saposhni-koviae, etc). and its process. METHODS: Colloidal mill and mixer were adopted, and analysis was on use of DSC and GC/MS. RESULTS: N-LOK inclusion complex included oil ratio was twice as large as the ?-CD, GC/MS analysis showed that the main components of volatile oil before and after being covered had no significant difference. CONCLUSION: N-LOK inclusion complex included oil ratio was far higher than the ?-CD. N-LOK inclusion complex could spray to dry directly, but ?-CD inclusion complex needed refrigeration and low temperature were necessary. The method of covered with N-LOK is suitable for mass production in the traditional Chinese patent medicine in the size of manufacture.

BACKGROUND: Because of the non-homology of protein and gene between human and animals, to promote osteogenesis or spinal fusion of animals by construction of tissue-engineered bone with the human gene has influenced the experimental validation.OBJECTIVE: To construct the seed cell line for bone tissue engineering with stable expression of human bone morphogenetic protein 2 (hBMP2).METHODS: The full-length hBMP2 gene was cloned from human muscle tissues by nested RT-PCR and transfected to human bone marrow mesenchymal stem cells (hBMSCs) with lipidosome. The transfected hBMSCs were cultured with G418 in vitro to screen and purify the cells. A series of analyses such as RT-PCR, dot-ELISA, immunohistochemstry and alkaline phosphatase activity analysis were performed to evaluate the situation of hBMP2 expression and secretion at 48 hours and 3 weeks after the transduction. hBMSCs transduced with empty plasmid and the normal hBMSCs served as positive control and blank control groups, respectively, which were used for observation of cell growth, proliferation and biological characteristics of transfected cells. RESULTS AND CONCLUSION: The transfected hBMSCs appeared in small groups or clusters, and had a good proliferation after subculture in vitro. Some G418-resistance cell clones and calcium nodules were found when cultured with G418 in vitro. No significant difference was noted in the cell proliferation between the hBMP2 transfection group and two control groups. The ALP activity in the hBMP2 transfection group remained significantly higher than that in the two control groups (P < 0.01). At 48 hours and 3 weeks after transduction, hBMSCs could express actively hBMP2 by RT-PCR monitoring, and had a positive reaction of dot-ELISA and immunohistochemical analysis. The expression of hBMP2 gene in the experiment group at 48 hours was significantly higher than that at 3 weeks after transduction while there was no expression of hBMP2 gene in the two control groups. The above results show that the hBMSCs transfected by hBMP2 gene not only have potentials of normal proliferation and osteogenic differentiation, but also can stably express hBMP2.

AIM To establish an HPLC-DAD method for the simultaneous content determination of four constituents in Compound Kendir Leaves Tablets Ⅰ (Apocyni veneti Folium,Chrysanthemi indici Flos,Stephaniae tetrandrae Radix,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Shimadzu VP-ODS column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of methanol-0.5％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 260 nm and 325 nm.RESULTS Chlorogenic acid,hydrochlorothiazide,buddleodide and promethazine hydrochloride showed good linear relationships within the ranges of 24.91-498.2 ng (r =0.999 9),286.33-5 726.7 ng (r =0.999 9),10.04-200.9 ng (r =0.999 9) and 154.80-3 096.1 ng (r =0.999 9),whose average recoveries were 98.3％ (RSD =1.3％),99.1％ (RSD =0.6％),98.5％ (RSD =1.0％) and 99.3％ (RSD =1.2％),respectively.CONCLUSION This simple,accurate and reliable method can be used for the quality control of Compound Kendir Leaves Tablets Ⅰ.

Aim To investigate the effect of plumbagin on invasion and apoptosis of human hepatocellular car-cinoma ( HepG2 ) . Methods HepG2 cells were cul-tured in vitro with different concentrations of plumba-gin, then cell proliferation was observed by MTT as-say; cell invasion was observed by transwell invasion assay; cell apoptosis was detected by flow cytometry, and the protein expression of Bax, Bcl-2 was detected by immunocytochemistry. Results MTT results showed that plumbagin could significantly inhibit cell proliferation compared with the control group, and in a dose-dependent manner ( P <0. 05 ) . Transwell inva-sion assay showed that cell invasion was significantly decreased with increasing concentrations of plumbagin ( P < 0. 01 ) . Flow cytometry showed that apoptosis rate was significantly higher in 4 , 8 μmol · L-1 group of plumbagin compared with that of the control group ( P<0. 01 ) . Immunocytochemistry showed that, with the increasing concentration of plumbagin, Bax protein expression increased, Bcl-2 protein expression was de-creased, both in a dose-dependent manner ( P <0. 01 ) . Conclusion Plumbagin can inhibit HepG2 cell proliferation and accelerate apoptosis of HepG2 cells, but also has the ability to inhibit HepG2 cell in-vasion.
( P < 0. 01 ) . Flow cytometry showed that apoptosis rate was significantly higher in 4 , 8 μmol · L-1 group of plumbagin compared with that of the control group ( P<0. 01 ) . Immunocytochemistry showed that, with the increasing concentration of plumbagin, Bax protein expression increased, Bcl-2 protein expression was de-creased, both in a dose-dependent manner ( P <0. 01 ) . Conclusion Plumbagin can inhibit HepG2 cell proliferation and accelerate apoptosis of HepG2 cells, but also has the ability to inhibit HepG2 cell in-vasion.

AIM: To study the determination of dl-tetrahydropalmatine (THP) in Chi nese traditional patent medicines by TLC-Scanning from the viewpoint of the pro cess control of manufacture. METHODS: To select the 5 kinds of representative drugs and its prep arations contained, THP was extracted with solvent method. The mobile phase contains diethylamine. THP w as separated with high-efficiency silica gel plate or handcrafted silica gel p late. Iodine was used for chronogenic agent and fluorescence defectiion for dete rmination. TLC-Scanning results were compared with HPLC. RESULTS: The method is practicable, the result is correlative with the HPLC. CONCLUSION: TLC-Scanning is fit for the quality control of the Chi nese traditional patent medicine in the process of manufacture.

N 6-methyladenosine (m 6A) is the most abundant RNA base modification in mammals, especially in eukaryotic messenger RNA (mRNA). N 6-methyladenosine modification can regulate RNA splicing, translocation, stability and ultimately affect protein synthesis. m 6A modification is catalyzed by RNA writers, reduced by erasers and also be recognized by readers. Abnormal changes ofm 6A levels are closely related to tumor occurrence and development, including proliferation, growth, invasion and metastasis. In the process of tumor radiotherapy, m 6A modification affects the efficacy of radiotherapy by affecting DNA damage, tumor stem cell generation and tumor cell radiation sensitivity. This article reviews the role of m 6A-modified epigenetic regulation in malignant tumors and the research progress of its mechanism in tumor radiotherapy, in order to provide new ideas for the development of clinical tumor molecular targeted therapies and radiosensitizers.

OBJECTIVE:To prepare total flavonoids of Hippophae rhamnoides(TFH)-PVP K30 solid dispersion,and to char-acterize and study its in vitro dissolution. METHODS:Solvent method was used to prepare TFH-PVP K30 solid dispersion with dif-ferent drug-loading ratio of 1:1,1:2,1:3,1:4,1:5;single factor test was designed to screen drug-loading ratio using dissolution parameter Td as index;orthogonal test was designed to optimize ultrasonic time,temperature of water bath and drying time for prep-aration technology using in vitro dissolution rate as index,and then validated. SEM,DSC and FT-IR were used to characterize sol-id dispersion. RESULTS:Td of TFH-PVP K30 solid dispersion was the lowest when drug-loading ratio was 1:3. Optimal technolo-gy was ultrasonic time 10 min,temperature of water bath 60 ℃ and drying time 12 h. 90 min accumulative dissolution rate of pre-pared TFH-PVP K30 solid dispersion was 90.22% in average(RSD=1.74%,n=3). The results of SEM,DSC and FT-IR showed that the drug as amorphous form dispersed in the PVP K30,the formation of hydrogen bond of the both. CONCLUSIONS:TFH-PVP K30 solid dispersion is prepared successfully,and in vitro dissolution rate of it is improved significantly.

Objective: To investigate the effect of zinc supplementation on expression of MT-1 and MT-2 mRNA in placenta and small intestine of pregnant rats. Method: Eighteen healthy pregnant SD rats were randomly divided into two groups,normal control (ZC) and zinc supplement group (ZS). Rats in both groups were fed the same normal diet. Rats in ZC group drank deionized water and those in ZS group drank the water supplemented with 1.26 mmol/L Zn. At gestational 9.5d and 17.5d, serum zinc was determined by atomic absorption spectrophotometer and the expression of MT-1 and MT-2 mRNA of placenta and small intestine was examined by RT-PCR. Results: Serum zinc levels at gestational 9.5d and 17.5d were higher in ZS group. Relative expression of both MT-1 and MT-2 mRNA in placenta, MT-1 mRNA at gestational 17.5d and MT-2 mRNA at each time point were higher in ZS group. At gestational 9.5d, there was no difference between two groups in expression of MT-1 mRNA. Conclusion: The expression of MT-1 and MT-2 mRNA in placenta and small intestine was up-regulated by dietary zinc supplementation during pregnancy.