[Analysis of projects funded by NSFC in field of pharmacology of traditional Chinese medicine between 2010 to 2013 fiscal years].

Projects which supported by National Natural Science Foundation of China (NSFC) in discipline of pharmacology of Chinese medicine between 2010 to 2013 financial years were reviewed. Based on these research items, new features and problems were summarized in this field.

Growth curve of murine xenotropic leukemia virus-related virus grown in Chinese hamster ovary cells.

BACKGROUND: Murine xenotropic leukemia virus-related virus (XMulV) is used as a model virus in the evaluation of viral inactivation in Chinese hamster ovary (CHO) cell-derived pharmaceutical proteins. Mus dunni cells and mink lung cells are used to produce XMulV particles. In consideration of the characteristics of XMulV, we tried to propagate the viruses on CHO cells, a nonmurine cell line. METHODS: The viruses were harvested from CHO cells from Day 2 to Day 7 postinfection, and reverse transcription-quantitative polymerase chain reaction was performed to quantify the viruses on different days. A cell-based infectivity assay was used to evaluate the XMulV titers. RESULTS: The content of the XMulV virions began to increase on Day 5 and grew exponentially from Day 6 to Day 7 postinfection. The growth curve was a typical single-step growth curve. Titers of the viral stock harvested on Day 7 were assayed on PG-4 cells, and the titers were 8.78 ± 0.25 log10 PFU/mL. CONCLUSION: Based on these data, we conclude that CHO cells could be a host cell line for XMulV particles. XMulV produced on Day 7 in CHO cells could be used at a laboratory scale for the evaluation of XMulV clearance in pharmaceutical proteins derived from CHO cells.

[Applications and approved projectsof general program, young scientist fund and fund for less developedregion of national natural science funds in discipline of Chinese materia medica, NSFC in 2012].

The applications accepted and approved by general program, young scientist fund and fund for less developed region of national natural science funds in the discipline of Chinese materia medica, NSFC in 2012 have been introduced. The research contents of the funded projects in the popular research areas have been summarized and the problems in the applications have been analyzed to give a reference to the scientists in the field of Chinese materia medica.

[National Natural Science Foundation of China promotes the development of Chinese medicine in China--an analysis of Chinese medicine project funded by National Natural Science Foundation of China in the last decade].

In recent years, projects funded by the Division V III of Health Sciences of the National Natural Science Foundation of China (NSFC) increased steadily, which enhanced the process of modernization of Chinese medicine (CM). We analyzed CM projects funded by NSFC during 2003 -2012, which aimed to provide reference to experts in the CM field.

[Effect of icariin on learning and memory abilities and activity of cholinergic system of senescence-accelerated mice SAMP10].

OBJECTIVE: To investigate the effect of icariin (ICA) on learning and memory abilities and cholinergic system in senescence-accelerated mice SAMP10. METHOD: The 8-month-old senescence-accelerated mice were randomly divided into the model SAMP10 group and the positive Donepezil group (1 mg x kg(-1)) and ICA groups (50, 100, 200 mg x kg(-1)), with 12 mice in each group. Another 12 8-month-old mice SAMR1 were selected as the normal control group. After 30 days of oral administration, Morris water maze, SMG-2 water maze and experimental platform were used to test the effects of ICA on learning and memory abilities of SAMP10 groups. By colorimetric determination of AChE activity in the cortex, enzyme-linked immunosorbent assay detection of ACh, ChAT, MCBC of the cerebral cortex, the effect of ICA on the cholinergic system of SAMP10 was observed. RESULT: ICA could improve the abilities of space exploration and positioning navigation of SAMP10, shorten the latency in SMG-2 water maze, enhance their jumping ability in response to the passive test, and increase levels of ACh, ChAT, MCBC in the cerebral cortex of SAMP10. But its active effect on AChE in SAMP10 cortex was not obvious. CONCLUSION: Different doses of icariin can improve learning and memory abilities of SAMP10 to varying degrees, which may be related to its effect on the cholinergic system.

JR6, a new compound isolated from Justicia procumbens, induces apoptosis in human bladder cancer EJ cells through caspase-dependent pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Numerous efforts have been conducted in searching for effective agents against cancer, in particular from herbal medicines. Justicia procumbens is a traditional herbal remedy which was produced in the south-western and southern provinces of China and Taiwan province used to treat fever, pain, and cancer. Here, we identified a new compound 6'-hydroxy justicidin A (JR6) from Justicia procumbens, which showed obvious anti-cancer effects. MATERIALS AND METHODS: The cytotoxicity activity was assayed using MTT and SRB. Intracellular ROS visualization and quantification were acquired by using a laser scanning confocal microscopy. Apoptosis was measured using a propidium iodide (PI) apoptosis detection kit by flow cytometry. Activation of caspases (caspase-3, caspase-8, and caspase-9) was evaluated respectively using GloMax luminescence detector and Caspase-Glo 3,8,9 assay kits. Loss of mitochondrial membrane potential was observed by microscopy using JC-1 dye. Quantitative real-time PCR analysis was employed to detect the expression of protein associated with cell death. RESULTS: JR6 remarkably inhibited growth in human bladder cancer EJ cells by decreasing cell proliferation, reduced the SOD activity, increased the content of reactive oxygen species (ROS), and induced apoptosis. Activation of caspase-8, caspase-9, and the subsequent activation of caspase-3 indicated that JR6 may be inducing intrinsic and extrinsic apoptosis pathways. Caspase-3, caspase-8, and caspase-9 inhibition rendered this extract ineffective, thus JR6-induced apoptosis is caspase-dependent. JR6 also disrupted the mitochondrial membrane potential (Δψm) and unregulated the Bax and p53 expressions in EJ cells. CONCLUSION: These observations suggest that JR6 induce apoptosis through caspase-dependent pathway in human bladder cancer EJ cells, emphasizing the importance of this traditional medicine and thus presents a potential novel alternative to bladder cancer therapy.

[Applications and spproved projects of general program, young scientist fund and fund for less developed region of national natural science funds in discipline of Chinese materia medica, NSFC in 2011].

The applications accepted and approved by general program, young scientist fund and fund for less developed region of national natural science funds in the discipline of Chinese materia medica, NSFC in 2011 have been introduced. The character and problems in these applications have been analyzed to give a reference to the scientists in the field of Chinese material medica.

Total synthesis of 6'-hydroxyjusticidin A.

The first total synthesis of 6'-hydroxyjusticidin A, isolated from Justicia procumbens L. with good inhibitory activity against cancer cells, has been accomplished. The structure was confirmed by ¹H NMR, ¹³C NMR, and HR-ESI-MS. The key steps involved a Diels-Alder cycloaddition reaction and a reduction in NaBH4.

Chrysin improves cognitive deficits and brain damage induced by chronic cerebral hypoperfusion in rats.

Chronic cerebral hypoperfusion, induced by permanent occlusion of bilateral common carotid arteries (2VO), is related to neurological disorders and contributes to cognitive decline. Chrysin (5,7-dihydroxyflavone) is an important member of the flavonoid family. The aim of this study is to investigate the effects of chrysin on cognitive deficits and brain damage in this rat 2VO model. At 52days after ligation, the escape latency in Morris water maze was significantly increased in rats subjected to 2VO, the neuronal damage was also increased accompanied by a large proliferation in glial fibrillary acidic protein (GFAP) immunoreactivity with marked white matter lesions, and neuronal cell apoptosis, all of which were significantly alleviated by long treatment of chrysin (30mg/kg). Biochemical examinations revealed that chrysin decreased lipid peroxide, reduced the increased activities of superoxide dismutase, and attenuated the decreased activities of glutathione peroxidase in 2VO rats. The results suggest that chrysin may have therapeutic potential for the treatment of neurodegeneration and dementia caused by decreased cerebral blood flow, which is most likely related, at least in part, to its anti-inflammatory and antioxidant properties.

[Effects of Yinqiao detoxifcation oral liquid on NK cell viability and serum concentration of IFN-gamma in SCID mouse infected by influenza virus FM1].

OBJECTIVE: To explore the dynamic effects of Yinqiao detoxifcation oral liquid (YQD) on NK cell lysis in spleen and serum concentration of IFN-gamma in SCID mouse infected by influenza A virus FM1. METHOD: The mice were divided into six different groups randomly. Except normal control, other mice were intranasally instilled with 15 TCID of virus. Three dosage groups of YQD (5.0, 10.0, 20.0 g x kg(-1)) were respectively fed with YQD. Positive control group was administrated orally with 0.07 g x kg(-1) of ribavirin. Normal control and model group were fed with physiological saline. After 1, 3, 5 and 7 days' infection, spleens and serum were collected. Then the NK cell lysis was detected by LDH release kit and the concentration of IFN-gamma was examined by ELISA assay. RESULT: Contents of IFN-gamma reached to peak value on the 3th day and until the 5th day. Later, the level of IFN-gamma returned to normal level. The variation tendency of activity of NK cell in spleen was according with that of IFN-gamma. But it reached the maximum value until the 5th day. The activity of NK cell lysis in three groups of YQD was well above that i n model group. In addition, therapeutic action of both 10.0 g x kg(-1) and 20.0 g x kg(-1) of YQD treatment groups was better than that of 5.0 g x kg(-1). CONCLUSION: The data showed that serum concentration of IFN-gamma and NK cell lysis were improved by YQD at different time, which was demonstrated YQD could perform well in immune system.

[Experiment study about effect of Yinqiao detoxifcation oral liquid on natural killer cells and TNF-alpha, TGF-beta1 of BALB/C nude mouse infected by influenza virus].

OBJECTIVE: To explore the effect of Yinqiao detoxifcation oral liquid on activity of natural killer cells (NK) and serum content of TNF-alpha, TGF-beta1 of BALB/C nude mouse infected by influenza virus. METHOD: To establish infected mice model by FM1 followed by intragastric administration of Yinqiao detoxifcation oral liquid for treatment. LDH method was used to observe NK cells. ELISA method was used to determine the levels of TNF-alpha, TGF-beta1, in serum on 1st, 3rd, 5th, 7th days after infection. RESULT: Comparing to the normal group, the NK activity of the model group was significantly increased on 1 dpi (day post infection), and significantly decreased on 3, 5, 7 dpi. The NK activity of three dosage groups (5, 10, 20 g x kg(-1)) of Yinqiao detoxifcation oral liquid were respectively higher than that of the model on 3, 5, 7 dpi, especially with high dose (P < 0.01). The serum level of TNF-alpha and TGF-beta1 of model group is higher than that of normal group on 1, 3, 5, 7 d. Compared with model group, the serum level of Yinqiao detoxifcation oral liquid groups (5, 10, 20 g x kg(-1)) were decreased in different degree on every time point, especially the serum level of the higher dose of Yinqiao detoxifcation oral liquid decreased on 3 dpi (P < 0.05), Yinqiao detoxifcation oral liquid inhibit the serum level of TGF-beta1 in a dose-dependent manner. CONCLUSION: Yinqiao detoxifcation oral liquid could enhance the activity of NK cell and decrease the serum level of TNF-alpha and TGF-beta1 of the mice infected by influenza virus.

Neuroprotective effects of icariin on memory impairment and neurochemical deficits in senescence-accelerated mouse prone 8 (SAMP8) mice.

The senescence-accelerated mouse prone 8 (SAMP8) is a novel aging model characterized by early onset and rapid advancement of senescence. In the present study, 6-month-old male SAMP8 mice were orally administered icariin (75, 150mg/kg) for 15weeks. Mice were submitted to passageway water maze test and step-down passive avoidance test for evaluating cognitive impairments. The HPLC-EC technique was used to determine the monoamine contents in the brain. The effects of icariin on oxidative stress and the acetylcholinesterase (AChE) activity of SAMP8 mice were also investigated. We found that icariin treatment significantly prevented learning and memory impairments of SAMP8 mice in passageway water maze test and step-down test. Icariin could partly reverse alterations of monoamines and metabolites levels in the cortex and hippocampus of SAMP8 mice. Furthermore, icariin-treated SAMP8 mice had significantly decreased malondialdehyde (MDA), nitric oxide (NO) contents, lowered nitric oxide synthase (NOS) activity and higher glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities in the brain homogenates and serum. Meanwhile, the acetylcholinesterase activity was markedly inhibited after icariin administration. However, the positive control piracetam did not show significant beneficial effects. In conclusion, the present findings demonstrated that the improvement of icariin on cognitive impairments in SAMP8 mice may be due to increasing monoamines levels, inhibiting oxidative damage and decreasing acetylcholinesterase activity.

Two new triterpenoids from Photinia serrulata.

Two new triterpenoids, 2alpha,3beta,11alpha,13beta-tetrahydroxy-12-ketooleanan-28-oic acid(1) and 3beta-hydroxy-12-keto-9(11)-ursen-28,13beta-olide (2) were isolated from the leaves ofPhotinia serrulata. Their structures were identified by spectral methods. Compounds 1 and2 were assessed for cytotoxic activity against three human tumor cell lines (A-549, HCT-8,and BEL-7402), and they showed no cytotoxic effects at concentrations up to 5microg/mL.