OBJECTIVE: To establish the UPLC fingerprint of Gardeniae Fructus for providing reference for effective quality control. METHODS: The samples were analyzed on a Waters ACQUITY UPLC BEH C18(2.1 mm × 50 mm, 1.7 μm) column maintained at 30℃ and eluted with methanol and water containing 0.02% phosphoric acid as mobile phases in a linear gradient mode. The flow rate was set at 0.4 mL·min-1 and the injection volume was 0.2 μL. The detection wavelengths were set at 238 nm (0-5.4 min) and 440 nm (5.4-10 min). RESULTS: The fingerprints of 16 batches of samples were analyzed with similarity evaluation, principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). The common mode of the fingerprint was established with 24 common peaks, and four of them were identified, which were genipin 1-gentiobioside (4), gardenoside (6), crocin I (15), and crocin II (17). The similarity degrees of the 16 batches of samples were between 0.986 0 and 0.995 0.The samples were divided into two groups, ZheJiang (S1-S6) and other areas (S7-S16), analyzed by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). Eleven significantly different components were found, including genipin1-gentiobioside, gardenoside, crocin I, crocin II and some others. CONCLUSION: The establishment of UPLC fingerprint of Gardeniae Fructus from different producing areas and the application of chemical pattern recognition can provide more comprehensive references for the quality control and evaluation of genuine Gardeniae Fructus from Zhejiang Province.

Objective To investigate the effects of aspirin(aspi) on rat cardiac fibroblasts (CFs) proliferation induced by aldosterone(ald) and the underlying molecular mechanisms .Methods Primary CFs from 1-3 day neonatal rats were digested by 0.08%trypsin and then purified by differential adhesion .The rats were divided into four groups:control group, DMEM medium ( free calf serum ) , ald group [ DMEM medium ( free calf serum ) +ald 1 ×10 -8 mol/L ] , aspi group [DMEM medium (free calf serum)+ald 1 ×10 -8 mol/L+aspi 1.11 ×10 -6 mol/L] and spiro group [DMEM medium (free calf serum)+ald 1 ×10 -8 mol/L +spiro 1 ×10 -6 mol/L].The morphology of CFs was assayed by HE staining methods .MTT Methods were used to measure cell proliferation .Western blotting was used to determine protein expression of TGF-β-Smad 2,3,4.Results HE Staining results showed that compared with the control group , ald activated cell proliferation and increased the cell division phase significantly (P<0.01).Compared with ald group, aspi+ald as well as spiro+ald could reduce cell division significantly ( P<0 .05 ) .MTT assay showed that compared with control group , ald could significantly improve the metabolism of MTT in CF (P <0.01).Compared with ald group, aspi +ald as well as spiro+ald could reduce the metabolism of MTT (P<0.01).Western blotting revealed that the expression levels of TGF-β-Smad 2, 3, 4 in CF were significantly increased by the stimulation of ald but were significantly reduced in aspi +ald and spiro+ald groups compared with ald group (P<0.01).Conclusion Aspi can inhibit the proliferation of CFs induced by ald,possibly by down-regulating the expression of Smad 2, Smad3 and Smad4.

BACKGROUND:Skul repair materials cannot only restore the normal shape of the skul , but also play an important role in brain functional recovery. OBJECTIVE:To summarize the research status of polyetheretherketone (PEEK), titanium al oy and tissue engineering technique in cranioplasty and the prospect of three-dimensional (3D) printing technology. METHODS:Literatures related to skul repair materials were retrieved in databases of CNKI and PubMed published from 1995 to 2016, using the keywords of“bone regeneration material in calvarial, 3d printing bone scaffold”in Chinese and English, respectively. RESULTS AND CONCLUSION:Although titanium and PEEK have been used in clinic, titanium holds conductivity, thermal conductivity, while PEEK that may be displaced or lost is not involved in osseointegration. Tissue engineering technology participates in the skul tissue reconstruction, achieving satisfactory repair outcomes, but the problems of scaffold selection and preparation, seed cel obtainment, and growth factor release need to be overcomed. 3D printing technology can print personalized shape, fit the defect precisely, but the raw materials should have good biocompatibility and biomechanical property. Combination of tissue engineering technology with 3D printing technology shows a broad prospect in cranioplasty.

Drugs are exogenous compounds for human bodies, and will be metabolized by many enzymes after administration. CYP450 enzyme, as a major metabolic enzyme, is an important phase I drug metabolizing enzyme. In human bodies, about 75% of drug metabolism is conducted by CYP450 enzymes, and CYP450 enzymes is the key factor for drug interactions between traditional Chinese medicine( TCM) -TCM, TCM-medicine and other drug combination. In order to make clear the interaction between metabolic enzymes and TCM metabolism, we generally chose the enzymatic activity as an evaluation index. That is to say, the enhancement or reduction of CYP450 enzyme activity was used to infer the inducing or inhibitory effect of active ingredients and extracts of traditional Chinese medicine on enzymes. At present, the common method for measuring metabolic enzyme activity is Cocktail probe drugs, and it is the key to select the suitable probe substrates. This is of great significance for study drug's absorption, distribution, metabolism and excretion (ADME) process in organisms. The study focuses on the interaction between TCMs, active ingredients, herbal extracts, cocktail probe substrates as well as CYP450 enzymes, in order to guide future studies.
Animals ; Cytochrome P-450 Enzyme Inhibitors ; metabolism ; pharmacology ; Cytochrome P-450 Enzyme System ; chemistry ; metabolism ; Drugs, Chinese Herbal ; metabolism ; pharmacology ; Enzyme Activation ; drug effects ; Enzyme Activators ; metabolism ; pharmacology ; Humans

To compare the differences of the active ingredient contents and the sulfur dioxide residue in Astragali Radix before and after sulfur fumigation and provide a basis for establishing an alternative processing method. Astragali Radix, harvested at the same time in Longxi Gansu, were processed with different methods. high performance liquid chromatography (HPLC) was used to determine the contents of the active ingredients in Astragali Radix and the revised method of the pharmacopoeia of China in 2011 was applied to determine the sulfur dioxide residue. The results show that the three-fold sulfur-fumigation group has the highest level of astragaloside IV and the dried sulfur-fumigation group with 10% water has the lowest level; the content of calycosin-7-O-β-D-glucoside is the highest in naturally dried group and the lowest in the group of sulfur fumigating for 3 times; the sulfur dioxide residue of all sulfur-fumigation groups exceeds certain limit significantly and the group of sulfur fumigating for 3 times reaches the highest level.
Astragalus Plant ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; adverse effects ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods

[Objective] To estimate the effects of ginsenoside Rb1 on melanogenesis in human melanocytes and underlying mechanisms.[Methods] Epidermal melanocytes were obtained from circumcision specimens of children,and subjected to primary culture.After 2 to 5 passages,the melanocytes were treated with different concentrations of ginsenoside Rb1,dimethyl sulfoxide (DMSO,vehicle control),forskolin at 10 μmol/L(positive control) or remained untreated (blank control).After additional culture for 72 hours,methyl thiazolyl tetrazolium (MTT) assay and NaOH lysis method were used to evaluate cell viability and melanin content in melanocytes respectively,spectrophotometer to determine dopa oxidase activity of tyrosinase,Western blot to quantify the protein level of tyrosinase,microphthalmia-associated transcription factor (MITF),phosphorylated and total cAMP response element binding protein (p-CREB and t-CREB) in melanocytes.[Results] After treatment with ginsenoside Rbl of 25,50 and 100 μmol/L for 72 hours,the melanocytes experienced no significant changes in viability (P ＞ 0.05 ),but a significant dose-dependent increase in melanin content (112.4％± 5.7％,155.7％ + 6.3％,217.2％ ± 11.7％ vs.100％,P＜ 0.05 or 0.01) and tyrosinase activity(117.9％ ± 5.7％,158.2％ ± 9.6％,182.6％ ± 10.0％ vs.100％,P＜ 0.05 or 0.01 ) compared with the vehicle control melanocytes.The protein expressions of tyrosinase,MITF and p-CREB were statistically higher in melanocytes treated with ginsenoside Rb1 of 100 μmol/L for 72 hours than in the vehicle control melanocytes (225.4％ ± 12.8％ vs.100％ ± 7.9％,313.5％ ± 16.7％ vs.100％ ± 9.8％,322.5％ ± 21.1％ vs.100％ ± 9.1％,all P＜ 0.01).The increase in MITF protein expression was inapparent in melanocytes at 8 hours after the treatment with ginsenoside Rb1 of 100 μmol/L,but statistically significant at 24 hours compared with the melanocytes at baseline (P＜ 0.01).The pretreatment with H-89 (a 8elective inhibitor of PKA) at 10 μmol/L,significantly suppressed the ginsenoside Rb1 (100 μmol/L for 72 hours) -induced phosphorylation of CREB,increase in MITF,tyrosinase expression,as well as tyrosinase activity and melanin content in melanocytes (all P ＜ 0.01 ).[Conclusion]s Ginsenoside Rb1could enhance the melanogenesis and tyrosinase activity in normal human melanocytes.The PKA/CREB/MITF/ tyrosinase signaling pathway may contribute to the pro-melanogenic effect of ginsenoside Rb1.

Objective To observe the efficacy of underwater partial body-weight-supported treadmill training in repairing spinal cord injury (SCI) and its relationship with spinal nerve plasticity. Methods A total of 40 Sprague-Dawley rats were randomly divided into five groups: a sham model group, a model control group, an underwater training group, a partial body-weight-supported treadmill training (PBWSTT) group and an underwater PBWSTT group. A rat model of SCI was induced by contusion of the T10 segment with a Multicenter Animal Spinal Cord Injury Study (MASCIS) impactor. One week post-operation, different rehabilitation strategies, such as free exercise in water, BWSTT and underwater PBWSTT, were administered to the rats in the underwater training groups for 8 weeks.Those in the sham model group and model control group were given no training. The Basso, Beattie and Bresnahan (BBB) locomotor rating scale and a climbing test were used to evaluate the recovery of hindlimb locomotor function.The expression of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) in the spinal cords was detected with immunohistochemical methods. Results Nine weeks post-operation, hindlimb locomotor function had improved significantly more in the underwater PBWSTT group than in underwater training group or the BWSTT group.The expression of BDNF in the 3 training groups was significantly higher than in the model control group, though there was no significant difference among the 3 training groups. The expression of NT-3 in the underwater PBWSTT group increased more significantly than in the BWSTT group, however there was no significant difference between the underwater PBWSTT group and the underwater training group. Conclusion Underwater PBWSTT can promote the recovery of hindlimb locomotor function in rats after SCI, probably through increasing the expression of BDNF and NT-3 and thus promoting neural plasticity in the spinal cord.

Objective:To analyze the volatile components in compound Shenqiyizhi powder. Methods: The technique of head-space solid phase micro extraction( HS-SPME) with gas chromatography-mass spectrometry ( GC-MS) was applied to study the vola-tile components in the compound powder. The compound essential oil was analyzed by steam distillation( SD) with GC-MS. Results:A total of 117 kinds of volatile components were detected, and among them, 70 kinds of chemical compositions were identified from the compound powder, accounting for 94. 46% of the total volatile components. The component with the highest content wasβ-himachalene followed by ylangene (15. 56%) and β-asarone (7. 36%). Totally 66 kinds of volatile components were detected from the compound essential oil by SD-GC-MS, and among them, 43 kinds were identified, accounting for 77. 41% of the total volatile components. The results indicated that the main volatile components were β-asarone (16. 73%), eugenol methyl ether (6. 74%) and ylangene (5. 81%). Conclusion:The main volatile components in Shenqiyizhi powders areβ-cedrene, ylangene,β-asarone,α-asarone and el-emicin etc. The research can provide scientific basis for the analysis of volatile components.

Objective:To optimize the forming process of Luomai granules. Methods: With the qualified granule yield and the moisture absorption as the indicators, the excipients were screened, and through the study of granulation effect, the proportion of excip-ients was optimized. With the normalized value of qualified granule yield, solubility, hygroscopicity and sensory evaluation as the eval-uation indices, and excipients times, ethanol concentration and amount of citric acid as the main influencing factors with five levels for each, the forming process of Luomai granules was optimized by central combination design-response surface methodology. Results:The option formula for Luomai granules was the follows:the excipients amount was 5-fold of the extract, 85% ethanol was used as the wet-ting agent, and citric acid content was 0. 3%. Conclusion:Optimized by the central combination design-response surface method, Lu-omai granules has such advantages as high qualified yield, low hygroscopicity, good solubility, soft taste and appropriate dosage, sug-gesting the optimized preparation process of Luomai granules is reasonable and feasible.

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley