Objective: To optimize the purification technology of total flavonoids from Astragali Companati Semen (ACS) by macroporous adsorption resin. Methods: The process parameters of water decoction extraction of ACS and purification of total flavonoids extracted from ACS by D-101 macroporous adsorption resin were optimized by orthogonal experiment design method. Results: The optimal extraction process were as follows: water decoction extract for three times, each time 0.5 h, water addition of 10, 6, and 6 times amount of the medicine respectively. The optimized technology conditions of D-101 macroporous adsorption resin were as follows: The flavonoids concentration of sample liquid of ACS was about 1.0 mg/mL, the diameter ratio was 1∶5, the sample flow rate was 1 BV/h, the rate of sample weight was 0.6 g/mL (herbs/resin); The velocity of water elution was 1 BV/h and its elution volume was 4 BV; The elution concentration, elution velocity, and elution volume were 70%, 1 BV/h, and 5 BV, respectively. The content paste rate of made ACS of total flavonoids was 2.65%, the mass fraction of total flavonoids was 56.24%, and the process transfer rate was 68.37%. Conclusion: The method is simple and feasible for purification of total flavonoids extracted from ACS.

Objective To discuss the less invasive stabilization system(LISS)in treatment of fractures around the knee joint.Methods Between February 2002 and December 2005,103 cases of periarticular fractures of knee joint were treated with LISS in our department.There were 58 cases of comminuted distal femoral fracture and 45 cases of comminuted proximal tibia fracture.Follow-ups were conducted once per week during the first four weeks and once per month in the following months to observe whether such complications as loosening of internal fixators and nail breakage would occur.Results All the cases were followed up for a mean time of 16.5 months(range, 6 to 27 months).Loosening and breakage of internal fixation was found in only one case,while others obtained bony union.In fractures of types A and C,the knee range of motion varied from 95?to 145?(average,130?)and 85?to 140?(average,110?)in flexion respectively.HSS score:excellent in 84 cases(81.6%),good in 15 cases (14.6%),fair in three cases(2.9%),and poor in one case(0.9%).Conclusions The LISS can be the best choice for the treatment of periarticular fractures around the knee joint,but we should take care to implant the plate and screws in a proper position and direction and avoid early weight-bearing.In application of the 13-hole plate, large incision should be made to approach the distal tibia for fear of damaging the superficial peroneal nerve.

0.05 indicating no statistical difference.However,the noise measurements for the L and C groups were 30.05 and 27.80,respectively,with P

Objective To evaluate and compare the adult neural stem cells (NSCs) derived from the subventricular zone (SVZ), adipose tissue (AD) and bone marrow (BM) in SD rat in terms of their morphologies, their potential of neural differentiation and their ability to secrete neurotrophins (NTs).Methods Tissues from the suventricular zone, adipose tissue and bone marrow in the same SD rat were chosen and induced in vitro into SVZ-NSCs, AD-NSCs and BM-NSCs, respectively. The abilities of proliferation and differentiation of these 3 NSCs were compared. Immunocytochemistry and Western blotting were employed to detect the expressions of surface markers of neurons and neuroglia, including nestin,βtubulin, galactocerebroside C (GalC) and glial fibrillary acidic protein (GFAP). The secretions of BDNF and NGF were detected by ELIS A. Results No obvious differences of morphology between SVZ-NSCs and both BM-NSCs and AD-NSCs were found (P>0.05). The proliferation ability of SVZ-NSCs was stronger than that of BM-NSCs and AD-NSCs. The percentage of nestin-positive cells in the SVZ-NSCs was significantly higher than that in the BM-NSCs or AD-NSCs (P<0.05). No obvious differences in the expressions of βtubulin, GFAP, and GalC among the 3 groups were found (P>0.05).The secretions of BDNF and NGF in all the 3 groups could be noted; those in the SVZ-NSCs was significantly higher than those in the BM-NSCs and AD-NSCs (P<0.05); those in the AD-NSCs was slightly higher than those in the BM-NSCs. Conclusions SVZ-NSCs, AD-NSCs and BM-NSCs show similar morphological and phenotypic characteristics; however, SVZ-NSCs present more powerful proliferation, differentiation and secretion abilities than AD-NSCs and BM-NSCs. Considering about such problems as immuno-repulsion, ethic and the origins, AD-NSCs appear to be the better choice than BSVZ-NSCs and M-NSCs.

Objective To investigate the effect of collagen scaffold loaded with collagen-binding domain neurotrophin-3 (CBD-NT3) on the extension of cellular processes of dorsal root ganglions (DRGs), and explore the significance of this kind of combinatorial strategies in the spinal cord injury repair. Methods The tail tendons of SD neonatal rats were performed the removal of cellular components to prepare the collagen scaffold; HE staining was employed to evaluate whether the cells were completely removed from the collagen scaffold. The collagen scaffold was loaded with CBD-NT3,and then, they were co-cultured with primary DRGs for 1, 3 and 5 d, respectively. NT3 and PBS were also co-cultured with primary DRGs for 1, 3 and 5 d, respectively, as controls. Cells on the scaffold were stained by fluorescein diacetate (FDA) for morphology observation and the lengths and angles of the processes in each group were also quantitatively analyzed. Scanning electron microcopy (SEM) was employed to observe the topography of scaffold and the ultrastructure of DRGs 3 d after the co-culture.Results HE staining indicated that the cellular components in the scaffold were removed completely.The length of processes elongation in CBD-NT3 treatment group was significantly longer than that in the controls 3 d after the co-culture (P＜0.05). The 95% confidence interval of the angle between the line which the process emerged from the cell soma to the growing tip of the process and the long axis of fiber was 18.8-20.7 degrees. The results of SEM showed that cells could rely on the topography of the scaffold to anchor and grow. Conclusion The combinatorial strategies of collagen scaffold with CBD-NT3 can play a double function for oriented guiding and inducing extension of cellular processes effectively,which may provide a better therapeutic approach for spinal cord injury repair.

Objective To study the effect of the EGF-L functional region containing human TN-R protein in prokaryocytes and anti-EGF-L serum on the cortical neuronsin vitro in rats. Methods The anti-EGF-L serum was obtained from the rabbits immunized with the protein of EGF-L, then combine with TN-R coat petri dish to prepare various culture substrate.The effect on adhere,migrate and neurite outgrowth of neuron by differ culture substrate was observed. Results High concentration of antiserum against protein of EGF-Lwas successfully obtained from the rabbits,and it has no effect on the neuron when coated on petri dish as culture substrate with PLL in vitro,but can add the adhesive to the neuron,partly neutralize the inhibitory on the neuron neurite of TN-R.The antiserum can also allow the neuron with its neurite to migrate to the area coated with TN-R. Conclusion The anti-EGF-L serum can weaken the inhibitory on the neuron of TN-R in vitro,the function in vivo need further research,

To observe the effects of phenylallyl compounds on prostaglandin E2 (PGE2) release in mouse cerebral microvascular endothelial cells (bEnd. 3) stimulated by IL-1beta, and to analyze their structure-activity relationship. Different concentrations of phenylallyl compounds were added separately, and the content of PGE2 induced by IL-1beta in the culture media was measured by ELISA assay. The 50% inhibitory concentration (IC50) of PGE2 was calculated. Studies showed that phenylallyl compounds could affect the PGE2 release differently in bEnd. 3 cells induced by IL-1beta. Close relationships were shown between the inhibitory activities and the location and number of the substituent groups. In conclusion, phenylallyl compounds exhibited inhibitory activities at different extent on PGE2 release in bEnd. 3 cells stimulated by IL-1beta and presented certain structure-activity relationship.
Acrolein ; analogs & derivatives ; isolation & purification ; pharmacology ; Animals ; Brain ; blood supply ; Cells, Cultured ; Cinnamates ; isolation & purification ; pharmacology ; Dinoprostone ; antagonists & inhibitors ; secretion ; Drugs, Chinese Herbal ; chemistry ; Endothelial Cells ; cytology ; metabolism ; Inhibitory Concentration 50 ; Interleukin-1beta ; pharmacology ; Mice ; Microvessels ; cytology ; Propanols ; isolation & purification ; pharmacology ; Structure-Activity Relationship

OBJECTIVETo investigate the influences of Shensu Yin to RAW 264.7 on the expression of TLR3, TLR4 and the factors of the downstream in RAW 264. 7 cells.

METHODRAW 264.7 cell line was stimulated with Lipopolysaccharide and POLY I: C, respectively, and treated with the drug serum of Shensuyin simultaneously. 24 hours later, collected the supernatant and measured the inflammatory factors TNF-alpha and IFN-beta, extracted mRNA and measured the expression of TLR3, TLR4 and other correlated indexes of the downstream, analyzed and evaluated Shensu Yin's substance basis of pharmacodynamic actions.

RESULTShensu Yin drug serum depressed the expression of TLR4, MyD88, TRAF-6, TRAM and TRIF mRNA, as a result, it decreased the amount of TNF-alpha and IFN-beta.

CONCLUSIONDepressing the expression of TLR3, MyD88, TRAM and TRIF mRNA may be the elementary basis of Shensu Yin to play heat-clearing and detoxicating effect.

Adaptor Proteins, Vesicular Transport ; genetics ; Animals ; Cell Line ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Interferon-beta ; secretion ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; drug effects ; metabolism ; Male ; Mice ; Myeloid Differentiation Factor 88 ; genetics ; Plants, Medicinal ; chemistry ; Poly I-C ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin ; genetics ; Signal Transduction ; drug effects ; Toll-Like Receptor 3 ; genetics ; Toll-Like Receptor 4 ; genetics ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo explore the pathological mechanisms of Guizhi Decoction () syndrome and the therapeutic molecular mechanisms of the Guizhi Decoction, Mahuang Decoction (), Sangju Decoction ( ) and Yinqiao Powder (), as well as the potentially biological basis that Guizhi Decoction is most effective only for the patients with Guizhi Decoction syndrome in clinical practice.

METHODSWe first got serum samples from the patients suffering from both upper respiratory tract infection and Guizhi Decoction syndrome identified by the doctors of Chinese medicine (CM) in the clinic. Four formulas with therapeutic actions of pungent warmth or pungent coolness for superficial syndromes were chosen and four kinds of rat serum samples each containing one of the above-mentioned herbal formulas were collected, then the effects of Guizhi Decoction syndromes' patient serum as well as the effects of sera containing the formulas after being stimulated by the patient serum samples on both the mRNA expression of certain toll-like receptor (TLR) subtypes and the release of some inflammatory cytokines in RAW264.7 cells were tested and analyzed in vitro.

RESULTSThe expression of TLR-3, TLR-4 and TLR-9 mRNA among the 9 tested TLR subforms were up-regulated in the macrophages stimulated by the sera from untreated upper respiratory infection patients with the Guizhi Decoction syndrome (symptomcomplex). The products such as interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α and interferon (IFN)-β from stimulated macrophages through TLR signaling pathways were also increased correspondingly. Interestingly, the changes induced by the Guizhi Decoction syndrome patients' sera were masked significantly after the macrophages were incubated with the sera from donors treated with Guizhi Decoction. Similarly, the three other exterior-releasing formulas were all effective in reversing the up-regulated changes of certain TLR subforms to different degrees, but both the number of targeted TLRs and efficacy of them seemed to be inferior to that of Guizhi Decoction.

CONCLUSIONEvidence from these experiments might contribute to the scientific explanation of both the pharmacological mechanisms of Guizhi Decoction and also the CM theory that Guizhi Decoction is specifically prescribed for the treatment of Guizhi Decoction syndrome (The gearing formula to the symptom-complex).

Animals ; Cell Survival ; drug effects ; genetics ; Cytokines ; secretion ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; drug effects ; Healthy Volunteers ; Humans ; Inflammation Mediators ; metabolism ; Inhibitory Concentration 50 ; Macrophages ; drug effects ; metabolism ; Male ; Mice ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Toll-Like Receptors ; genetics ; metabolism