To investigate the neuroprotective of ligustilide (LIG) against glutamate-induced apoptosis of PC12 cells, cell viability were examined by MTT assay. Flow cytometry was applied to assay cell apoptosis rate. Intracellular calcium concentration was measured by using fluorescent dye Fluo-3/AM. Cytochrome C (Cyt C), Caspase-3, Bax and Bcl-2 protein expression were assayed by western blot. The results showed that glutamate is cytotoxic with an inhibitory concentration 50 (ID50) of 15 mmol · L(-1). Pretreatment with LIG (1, 5, 15 μmol · L(-1)) significantly improved cell viability. The apoptosis rate in glutamate-induced PC12 cells was 13.39%, and decreased in the presence of LIG (1, 5, 15 μmol · L(-1)) by 9.06%, 6.48%, 3.82%, separately. Extracellular accumulation of Ca2+ induced by glutamate were significantly reduced by LIG. The results of western blot manifested that pretreatment LIG could decrease the release of Cyt C from mitochondria, down-regulate Caspase-3 protein expression and up-regulate Bcl-2/Bax ratio, thereby protects PC12 cells from apoptosis. In summary, LIG had protective effect on glutamate-induced apoptosis in PC12 cells through attenuating the increase in intracellular Ca2+ concentration, and inhibiting the release of Cyt C from mitochondria to cytoplasm.
4-Butyrolactone ; analogs & derivatives ; pharmacology ; Aniline Compounds ; Animals ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; Cytochromes c ; metabolism ; Glutamic Acid ; adverse effects ; Mitochondria ; metabolism ; PC12 Cells ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Xanthenes ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the immunological activity of Streptomyces polysaccharide (SMP) on normal and immunosuppressed mice.

METHODSThe effect of SMP on the proliferating activity of normal mouse splenocytes was tested in the mixed lymphocyte culture, and the changes of peripheral blood T lymphocytes were evaluated with acid a-naphthyl acetate esterase (ANAE) method. The ratio of Lyt2+ and L3T4+ T cell subsets was measured by flow cytometry.

RESULTSSMP stimulated obvious proliferation of mixed lymphocytes, showed protective effects on T lymphocyte and increased the ratio of Lyt2+ and L3T4+ cell subsets to nearly normal level in immunosuppressed mice.

CONCLUSIONSSMP can regulate the immune function in mice.

Animals ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Immunocompromised Host ; immunology ; Lymphocyte Culture Test, Mixed ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Polysaccharides ; immunology ; Spleen ; cytology ; Streptomyces ; chemistry ; immunology ; T-Lymphocytes ; cytology ; immunology

Evaluation of the permeability mainly focuses on intestinal absorption in biopharmaceutics classification system (BCS). It is more complicated that the absorption and metabolism under multicomponent environment in biopharmaceutics classification system of Chinese materia medica (CMMBCS) compared with single component environment, which needs suitable mathematical models to be described. Therefore, with full consideration of existing single component mathematical algorithm combining with the characteristics of intestinal absorption and metabolism, we explored and designed a new mathematical algorithm of intestinal absorption and metabolism of multicomponent drug. Then we put forward a new coefficient, P (influence), the relative change rate of the single component's intestinal absorption and metabolism under multicomponent environment compared with single component environment, which described the influences of intestinal absorption and metabolism of the component under multicomponent environment. Moreover, P (influence) highlights the distinctive characteristics of multicomponent drug's intestinal absorption and metabolism, and lays the foundation for the construction of CMMBCS.
Algorithms ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Humans ; Intestinal Absorption ; Intestines ; chemistry ; metabolism ; Models, Theoretical ; Solubility

Sanjie Zhentong capsules were scanned by using a near infrared spectra probe with different drug mass fraction and the spectral information of capsule shells and contents in it were obtained. Then partial least squares (PLS) models were developed for the prediction of mass fraction of Fritillariae Thunbergii Bulbus and Resine draconis in Sanjie Zhentong capsules. The correlation coefficient (r9c)) and root mean standard error( RMSEC) of 0.949 5, 0.958 2 and 4.742 4, 4.135 7. The models obtained correlation coefficient (r(v)) of 0.919 2, 0.936 7 and root mean square error (RMSECV) of 6.158 9, 5.037 3 respectively in the training set. The paired T test analysis of statistics showed that there were no significant difference between predictive values and measure values. The established models reflected a strong prediction performance and can meet the needs of the production.
Capsules ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Least-Squares Analysis ; Spectroscopy, Near-Infrared ; methods

OBJECTIVETo prove the therapeutic effects of Sihuang powder (composed by four traditional Chinese herbs: root of baikal skullcap, bark of amur corktree, root of sorrel rhubarb, fruit of cape jasmine, which were mixed with wild Chrysanthemum flower solution)in treating acute synovitis in experimental rabbit knee osteoarthritic models induced by papain injection and to explore its mechanism.

METHODSThirty-two New-Zealand white rabbits were divided into 6 groups: blank group, model group, Sihuang powder with high dosage group (2 g/kg), Sihuang powder with low dosage group (1 g/kg), Yingtaiqing group and wild Chrysanthemum flower group. The latter four groups were treated respectively with low and high dose Sihuang powder synovium and cartilage were tested concentrations of nitrogen monoxide (NO) and IL-1 level and then were prepared for pathologic and histologic observation 10 days later. Cartilage pathologic changes were record and synovium pathologic changes were valued by means of Mankin's value system.

RESULTSThe NO concentration of synovium in Sihuang powder with high dosage group was lower than that of model group, and there was significantly differences between the two groups (P < 0.01). The IL-1 level of synovium was failed after treated with Sihuang powder with high dosage (P < 0.05). Sihuang powder with low dosage and Yingtaiqing also could restrain IL-1's release (P < 0.05). In Mankin's value system, Sihuang powder with high dosage almost eliminated inflammatory cells infiltrating in synovium, which was seldom found in other groups. The value of Sihuang powder with high dosage group was the lowest in treatment groups (P < 0.005). Sihuang powder with low dosage group and wild Chrysanthemum flower group also decreased the degree of inflammatory in synovium (P < 0.05).

CONCLUSIONSihuang powder can reduce the concentration of NO and IL-1 and improve inflammatory cell infiltrate in lining cells of synovium. Moreover, it can alleviate swelling and pain of joint, improve joint movement and postpone degeneration of the cartilage.

Acute Disease ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Interleukin-1 ; analysis ; Nitric Oxide ; analysis ; Osteoarthritis ; drug therapy ; pathology ; Papain ; toxicity ; Powders ; Rabbits ; Synovitis ; chemically induced ; drug therapy ; pathology

OBJECTIVETo investigate the protective effect of phosphodiesterase type 5 inhibitors (tadalafil) on the testis following testicular ischemia-reperfusion injury in rats.

METHODSEighty-four healthy adult male SD rats were randomly and equally divided into groups A (sham operation), B (testicular torsion + low-dose tadalafil), C (testicular torsion + high-dose tadalafil), and D (testicular torsion + placebo). Models were established in the latter three groups by 7200 torsion of the right testis for 2 hours. The animals in groups A and B were treated by gavage with tadalafil at the dose of 0. 5 mg per kg per day, those in group C at 2 mg per kg per day, and those in group D with saline at the same dose. After 3, 7, and 14 days of treatment, the torsioned testes were harvested for evaluation of the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the testis tissue. The pathological changes in the testis were observed under the light microscope.

RESULTSAt 3, 7, and 14 days, the SOD activity was (254.46 +/- 7.43), (278.49 +/- 8.33), and (317.99 +/- 3.31) nU/mg prot in group B, and (277.12 +/- 8.80), (309.40 +/- 2.14), and (320.39 +/- 4.72) nU/mg prot in group C, all obviously higher than in D ([223.21 +/- 4.65], [231.45 +/- 4.16] and [248.28 +/- 5.74] nU/mg prot), while the MDA content was lower in the former two groups than in the latter. At 3 and 7 days, the SOD activity was significantly higher and the MDA level significantly lower in group C than in B (both P < 0.01) , while at 14 days, neither showed any remarkable differences between the two groups (P > 0.05). No obvious histopathological change was observed in the testis tissue of group A. At 3 and 7 days, pathological examination of the testis tissue revealed significant differences in the number of seminiferous epithelial layers, testicular histological score, and seminiferous tubule diameter in group B (P < 0.01), but the three indexes at 14 days in group B and at 7 days in group C exhibited no remarkable differences from those at 14 days in group A.

CONCLUSIONTadalafil can alleviate testicular ischemia-reperfusion injury following testis torsion/detorsion in a time- and dose-dependent manner.

Animals ; Biomarkers ; metabolism ; Carbolines ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Male ; Malondialdehyde ; metabolism ; Phosphodiesterase 5 Inhibitors ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Seminiferous Tubules ; pathology ; Spermatic Cord Torsion ; complications ; Superoxide Dismutase ; metabolism ; Tadalafil ; Testis ; blood supply ; metabolism ; pathology ; Time Factors

China's remote medical insurance policy included 5 different types of network subjects(policy community,intergovernmental network,producer network,professional network and issue network).The separate interests among competing stakeholders were the main obstacles to the implementation of remote medical insurance policy.The problems mainly reflected as sector interest disputed in policy community;the fragmentation of intergovernmental network;misallocation of the power and responsibilities between policy community and intergovernmental network;interest alliances and conflicts between intergovernmental network and producer network;the weak voice and insufficient action of issue network.Therefore,countermeasures were put forward to implement the remote medical insurance policy:firstly,adjust the distribution of responsibility between policy community and intergovernmental network,public interest oriented;secondly,eliminate local protectionism among the intergovernmental network and unify the relevant medical insurance policies;thirdly,improve the public policy participation system and strengthen the discourse power of issue network;lastly,build an open policy network and promote policy actors to collaborate with each other.

Objective To investigate the protective effect of modified Qianjin Weijing Decoction(MQJWJD)on TNF-α and NF-κB in rats with lung injury induced by particulate matter; To discuss relevant mechanism of action. Methods A tracheal drip 15 mg/kg fine particles of saline solution was used to establish modeling, every other day, three times. Thirty-two Wistar rats were randomly divided into normal group, model group and MQJWJD high-dose and low-dose groups, with eight rats in each group. Medication groups were given relevant medicine for gavage. The level of TNF-α in bronchoalveolar lavage fluid (BALF) was measured by ELISA. The expression of NF-κB protein in lung tissues was measured by immunohistochemistry. The histopathology of the lung injury was observed by light microscope. Results Compared with normal group, the level of TNF-α and the expression of NF-κB protein in the model group were higher than those in the normal group (P<0.01). Compared with model group, the level of TNF-α and the expression of NF-κB protein in MQJWJD low-dose and high-dose groups were lower than those in the model group (P<0.05, P<0.01). Pathological observation showed that, compared with normal group, model group showed intratracheal, alveolar and interstitial bacteria within a large number of fine particles calm, alveolar and pulmonary interstitial visible large amounts of phagocytic fine particles of macrophages and accompanied by more neutrophils and lymphocyte infiltration; Lung tissue pathological changes were significantly lighter in MQJWJD high-dose and low-dose groups than the model group. MQJWJD high-dose group showed mild inflammation, alveolar and pulmonary interstitial visible phagocytic fine particles of macrophages, a small amount of neutrophils and lymphocyte infiltration. Conclusion MQJWJD can reduce the pulmonary injury in rats induced by particulate matter and has protective effects on the rat model through decreasing the levels of TNF-α and the expressions of NF-κB protein in injured lung tissues.

OBJECTIVETo study the variation of TFAR-19 protein expression and Annexin-V apoptosis protein proportion in the mice thymus cell apoptosis procedure induced by water platform environment or electric emotional stress and the regulatory effect of Modified Xiaoyao Pill (MXP).

METHODSThe mouse emotional stress model was established by water platform equipment or electrical stimulation. The serum glucocorticoid was detected by radioimmunoassay, the TFAR-19 protein was detected by flow cytometry analysis, and Annexin-V apoptosis protein proportion was calculated by immunofluorescence technique.

RESULTSIn the groups of mouse stressed by water platform environment, the level of serum glucocorticoid, the TFAR-19 protein expression and the Annexin-V apoptosis protein proportion increased in the thymus cell along with the stress time prolonging (P<0.05 or P <0.01). The serum glucocorticoid level in mice treated with MXP was lower than that in the untreated group (P <0.05). In the groups of emotional stressed mouse established by electrical stimulation, the above-mentioned variations also revealed. All these variations could be alleviated with MXP (P<0.05).

CONCLUSIONThe water platform environment stress is a chronic continuous stress and electrical stimulation is an acute smooth stress, both of them could damage thymus function through neuro-endocrineo-immune network, but different in duration for causing severe injury. Chinese medicine MXP can alleviate the damage of thymus induced by either of them to certain degree.

Animals ; Annexin A5 ; biosynthesis ; Apoptosis Regulatory Proteins ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Female ; Glucocorticoids ; blood ; Male ; Mice ; Neoplasm Proteins ; biosynthesis ; Stress, Psychological ; blood ; physiopathology ; Thymus Gland ; cytology ; drug effects ; metabolism

Objective:To investigate the central mechanism of moxibustion in treating chronic inflammatory visceral pain(CIVP)and its analgesic effect from the perspective of the p38 mitogen-activated protein kinase(MAPK)/Ets-like transcription factor 1(ELK1)signaling pathway in the spinal cord. Methods:Clean-grade male Sprague-Dawley rats were randomly divided into a normal group,a model group,a herb-partitioned moxibustion(HPM)group,a sham-HPM group,a p38 MAPK inhibitor group,and a dimethyl sulfoxide(DMSO)group.CIVP rat models were prepared using an enema mixture of 2,4,6-trinitrobenzene sulfonic acid solution and 50%ethanol.The HPM group was treated with HPM;the sham-HPM group was treated the same as the HPM group,but the moxa cones were not ignited;rats in the p38 MAPK inhibitor group received L5-L6 intrathecal injection of p38 MAPK inhibitor(SB203580);rats in the DMSO group received L5-L6 intrathecal injection of 2%DMSO.Abdominal withdrawal reflex(AWR),mechanical withdrawal threshold(MWT),and thermal withdrawal latency(TWL)were used to observe pain-related behaviors in each group.Hematoxylin-eosin staining was used to observe the morphological changes in rat colon tissue.Western blotting and real-time quantitative reverse-transcription polymerase chain reaction were used to detect the phosphorylated protein and mRNA expression of apoptosis signal-regulating kinase 1(ASK1),MAPK kinase(MKK)3/6,p38 MAPK,ELK1,and mitogen and stress-activated protein kinase 1(MSK1)in the spinal cord. Results:Compared with the normal group,CIVP rats had severe colonic inflammatory injuries,and the pathological injury scores increased significantly,along with increased AWR scores under different colorectal distension(CRD)stimulation pressures and decreased MWT and TWL;the mRNA and phosphorylated protein expression of p38 MAPK,ELK1,MSK1,ASK1,MKK3,and MKK6 all increased in the spinal cord(P<0.01).After HPM treatment,the colon injuries were repaired,and the pathological injury scores decreased;under different CRD stimulation pressures,the AWR scores decreased,and the MWT and TWL increased;the mRNA and phosphorylated protein expression of p38 MAPK,ELK1,ASK1,and MKK3 in the spinal cord also decreased,with statistically significant differences compared with the model group and the sham-HPM group(P<0.01).There were no significant differences in the above indicators between the HPM group and the p38 MAPK inhibitor group(P>0.05),and the same was true regarding the comparisons between the model group and the DMSO group. Conclusion:HPM exerted analgesic effects via downregulating the mRNA and phosphorylated protein expression of ASK1,MKK3,p38 MAPK,and ELK1 in the spinal cord of CIVP rats.The inhibition of spinal p38 MAPK/ELK1 signaling pathway activation may be one of the mechanisms by which HPM relieves pain in CIVP.