Arsenic trioxide albumin microspheres (As2O3-BSA-NS) were prepared by using methods of chemical cross-linking. The desirability function (DF), calculated according to the size (<1 microm) distribution, drug loading and drug trapping efficiency, was introduced as a total index for the microspheres formulation. Four factors, inculding W/O ratio, decentralization speed, BSA concentration and stirring stabilization time, were selected and arranged in an orthogonal experimental table. The release characteristic was studied by the drug release experiment in vitro. The four factors affected DF differently. Decentralization speed behaved as the maximum (P<0.01), followed by BSA concentration (P<0.05) and the W/O ratio dose (P<0.05). Stirring stabilization time did not influence DF (P>0.05). The release experiment in vitro showed that As2O3 in As2O3-BSA-NS was released more slower than pure As2O3. It was concluded that regular As2O3-BSA-NS may be prepared by the methods of chemical cross-linking, which was optimized by orthogonal experimental analysis of different factors, and the microspheres can release As2O3 slowly.
Arsenicals/*chemistry ; Cross-Linking Reagents/pharmacology ; Delayed-Action Preparations ; Drug Carriers/*chemistry ; Drug Delivery Systems ; Microspheres ; Oxides/*chemistry ; Serum Albumin, Bovine/*chemistry ; Technology, Pharmaceutical

OBJECTIVE: To establish a quality standard for Jieyinling lotion. METHODS: Cortex Phellodendri and Epimedium Herb were identified by TLC, and the content of Icariin was determined by HPLC. RESULTS: The TLC spots were fairly clear and a good reproducibility was obtained. The linear range of Icariin was 2.0 ～10.0?g(r=0.999 6),with average recovery at 100.1%,RSD =1.92%. CONCLUSION: The established method is reliable and accurate, and suitable for the quality control of Jieyinling lotion.

Objective To study the antitumor activity of steroid saponins,such as Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin,extracted from Paris polyphylla var.yunnanensis against lung adenocarcinoma cells.Methods Taking mouse lung adenocarcinoma LA795 cell line and its T739-bearing mice as models,the antitumor activity of steroid saponins of P.polyphylla var.yunnanensis against lung adenocarcinoma cells was carried out in vitro and in vivo.The inhibitory activity of Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin on the LA795 cell line was studied by means of MTT assay and their antitumor effects in vivo were carried on inbred strains of mice(T739) affected by LA795 metastatic lung cancer.Results The MTT assay of in vitro cytotoxicity of Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin showed strong cytotoxicity against the growth of LA795 cells.IC50 Values were 24.33 ?g/mL,1.85,and 149.75 ?mol/L,respectively.The tumor weight in all treated groups was significantly lower than that in the control group and the inhibition of tumor growth were 41.82%,29.44%,and 33.94% in their high-dose groups.The metastasis of tumor cells was inhibited by different degrees in treated groups.Conclusion Steroid saponins of P.polyphylla var.yunnanensis have tumoricidal activity on lung adenocarcinoma cell line,both in vitro and in vivo.

Objective:To construct a RNAi lentiviral vector targeting rat CD80 gene and detect its effect of gene silencing in NRK and IEC6 cells.Methods:The effective sequence of siRNA targeting rat CD80 gene was confirmed in our previous work.Oligo-DNA fragment containing short hairpin frame was synthesized and reannealed,and then cloned into pGCSIL-GFP lentiviral expression vector.PCR and sequencing analysis were made for verifying the positive clones.The virus packaging plasmids were transfected into 293T cells to harvest shRNA lentivirus.After infection in NRK and IEC6 cells,Real-time PCR was performed to determine the expressing level of CD80.Results:PCR and sequencing revealed that shRNA plasmids was correctly constructed.Virus with a titer of 4×10~8 TU/ml was successfully packaged.CD80 expression in NRK and IEC6 cells could be knockdown by virus infection as characterized by 66.9% and 63.5% decrease of CD80 mRNA in NRK and IEC6 cells respectively,compared with negative control lentivirus.Conclusion:The recombinant lentiviral shRNA expressing vector targeting rat CD80 gene has been successfully constructed and packaged.CD80 mRNA could be down-regulated availably in NRK and IEC6 cells.

OBJECTIVETo compare steroidal saponins-main active ingredients of Paris polyphylla from different areas in Guizhou province, in order to provided basis for further proving Guizhou province to be the planting base of P. polyphylla.

METHODThe contents of nine steroidal saponins, namely paris-VII, (25R)-5-en-spirost-3beta,17alpha-diol-3-O-alpha-L-rhamnopyranosyl-(1-->2) [alpha-L-rhamnopyranosyl(1-->4)]-beta-D-glycopyanoside (PGRR), paris-H, paris-VI, paris-II , paris-III, gracillin, paris-I and paris-V of P. polyphylla from eight areas of Guizhou province were determined by HPLC-UV.

RESULTFive steroidal saponins-paris-VII, PGRR, paris-H, paris-VI and gracillin were detected in all the drugs, which provided basis for distinguishing P. polyphylla. P. polyphylla from Dushan showed the highest content of steroidal saponins (9.62%), followed by Tongren (6.39%), and the lowest was Zunyi (0.99%).

CONCLUSIONSaponins from different areas of Guizhou province show significant difference in variety and content, therefore Dushan is suitable for planting P. polyphylla.

China ; Chromatography, High Pressure Liquid ; methods ; Liliaceae ; chemistry ; Phytosterols ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Rhizome ; chemistry ; Saponins ; analysis ; isolation & purification

OBJECTIVETo investigate the influence of interferon-alpha on the variation of cytotoxic T lymphocytes (Tc) and suppressor T lymphocytes (Ts) in the peripheral blood of 32 patients with chronic hepatitis B, and to analyse the relationship between the efficacy of interferon-alpha and the variation of Tc and Ts cells.

METHODSThe peripheral blood Tc and Ts cells were detected by the double-staining immunocytochemistry technique.

RESULTSAt the end of the treatment with interferon-alpha, there were 9 complete responders (CR), 12 partial responders (PR) and 11 non-responders (NR). Tc cells significantly increased and Ts cells markedly decreased in CR or PR group compared with the healthy control group. There was no significant difference in the level of Tc and Ts cells between CR and PR groups, and no significant difference in the level of Tc cells in NR, CR and PR groups, The Ts cells was significantly higher in NR group than in CR or PR group.

CONCLUSIONSThe treatment of interferon-alpha can result in the change of Tc and Ts cells in patients with chronic hepatitis B. The variation of Ts cells may play an important role in the efficacy of interferon-alpha against hepatitis B virus.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Lymphocyte Count ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; cytology ; drug effects ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; Time Factors ; Treatment Outcome

OBJECTIVETo study the chemical constituents in herbs of Paris verticillata.

METHODThe compounds were isolated by column chromatography with silica gel and purified by Sephadex LH-20 and RP-HPLC. The structures were identified by means of NMR analysis.

RESULTNine compounds were isolated from the EtOAc extract and the n-BuOH extract of P. verticillata. Their structures were identified as beta-sitosterol (1), stigmasterol (2), daucosterol (3), beta-ecdysterone (4), 4-hydroxymethyl-gamma-butyrolactone (5), diosgenin-3-O-alpha-L-arabinofuranosyl (1-->4)-[alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glycopyranoside (6), pennogenin-3-O-alpha-L-arabinofuranosyl(1-->4)-beta-D-glycopyranoside (7), pennogenin-3-O-alpha-L-arabinofuranosyl (1-->4)-[alpha-L-rhamnopy-ranosyl (1-->2)]-beta-D-glycopyranoside (8), and pennogenin-3-O-alpha-L-rhamno-pyranosyl (1-->4)-alpha-L-rhamnopyranosyl (1--4)-[alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glycopyranoside (9).

CONCLUSIONCompounds 1-9 are isolated from P. verticillata for the first time, and compounds 3, 5 are isolated from the genus Paris for the first time. The compounds 6-9 showed certain inhibition activeness of LA-795 cells, especially, the effects of compounds 6, 8 and 9 were more significant.

Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Magnetic Resonance Spectroscopy ; Magnoliopsida ; chemistry ; Mice

Summary: Arsenic trioxide albumin microspheres (As2O3-BSA-NS) were prepared by using methods of chemical cross-linking. The desirability function (DF), calculated according to the size (<1 μm) distribution, drug loading and drug trapping efficiency, was introduced as a total index for the microspheres formulation. Four factors, inculding W/O ratio, decentralization speed, BSA concentration and stirring stabilization time, were selected and arranged in an orthogonal experimental table. The release characteristic was studied by the drug release experiment in vitro. The four factors affected DF differently. Decentralization speed behaved as the maximum (P<0.01), followed by BSA concentration (P<0.05) and the W/O ratio dose (P<0.05). Stirring stabilization time did not influence DF (P>0.05). The release experiment in vitro showed that As2O3 in As2O3-BSA-NS was released more slower than pure As2O3. It was concluded that regular As2O3-BSA-NS may be prepared by the methods of chemical cross-linking, which was optimized by orthogonal experimental analysis of different factors, and the microspheres can release As2O3 slowly.

OBJECTIVE:To establish the method for simultaneous determination of 9 components in Huichun yuzi granules. METHODS:LC-MS/MS method was adopted. The determination was performed on Shim-pack XR-ODS C18column with column temperature of 30 ℃. The sample size was 5 μL,and ion source as electrospray ion source;MRM mode was adopted. The acetonitrile-water was used as mobile phase for ferulic acid,rutin,paeonol,icariin and schisandrin(gradient elution);positive ion mode monitoring was conducted. The methanol-0.1% fomic acid water was used as mobile phase for naringin,verbascoside, amygdalin and protocatechuic acid(gradient elution);negative ion mode monitoring was conducted. RESULTS:The linear ranges of ferulic acid,rutin,paeonol,icariin,schisandrin,naringin,verbascoside,amygdalin and protocatechuic acid were 0.499 5-500, 25-1 000,0.245 9-250,5.185 1-1 000,0.981 9-1 000,0.248 1-125,2.510 4-250,10-2 500,4.999 7-1 000 ng/mL(r≥0.997 8), respectively. The limits of detection were 0.075,0.30,0.072,0.015,0.015,0.075,0.15,0.30,0.15 ng/mL,respectively;the limits of quantitation were 0.25,1.00,0.24,0.51,0.49,0.25,0.50,0.99,0.49 ng/mL,respectively. The recoveries rate were 91.39%-103.56%(RSD=1.03%-3.67%,n=6).RSDs of stability test ranged 1.4%-3.4%(n=6)within 48 h at room temperature. CONCLUSIONS:The method is rapid,sensitive and reproducible,and can be used for content determination of 9 components in Huichun yuzi granules.It can be used for quality control of Huichun yuzi granules.