This study established a rapid UPLC-TQ-MS/MS method for determination of eight active ingredients in Lilium lancifolium. The contents range of regaloside E, F, C and B are as follows: 0.604 0×10⁻¹-18.62×10⁻¹, 0.680 0×10⁻²-44.75×10⁻², 0.700 0×10⁻³-29.65×10⁻¹, 0.170 0×10⁻¹-4.724 mg•g⁻¹; the contents of chlorogenic acid, caffeic acid, protocatechualdehyde and ferulic acid, within the range of 6.827×10⁻³-16.07×10⁻³, 0.011 1×10⁻³-79.71×10⁻³, 0.593 7×10⁻³-2.962×10⁻³, 2.606×10⁻²-45.89×10⁻² mg•g⁻¹, respectively. According to PCA (principal components analysis) plotting, 35 batches can be divided into two categories, namely Anhui Huoshan and Hunan Longshan. The main different elements between these two categories are caffeic acid and ferulic acid according to the VIP (variable importance in the projection) points figure. Based on comprehensive principal component values, there are eight batches of L. lancifolium from Huoshan among the comprehensive ranking of ten. The UPLC-TQ-MS method for simultaneous analysis of eight active ingredients is accurate, efficient and convenient. This result can provide scientific basis for quality control of L. lancifolium.

OBJECTIVEThis study investigated the effect of clock gene PER1 on the expression levels of other clock genes in clock gene networks in oral squamous cell carcinoma cells.

METHODSWe used RNA interference mediated by short hairpin RNAs (shRNAs) to effectively knock down PER1 in SCC15 human oral squamous cell carcinoma cells. Flow cytometry was used to detect the degree of proliferation and apoptosis of the cells after PER1 knockdown, and quantitative real-time PCR was used to detect the mRNA expression levels of the clock genes CLOCK, BMAL1, PER1, PER2, PER3, DEC1, DEC2, CRY1, CRY2, TIM, CKIE, RORA, NPAS2, and REV-ERBA.

RESULTSThe proliferation index of SCC15 cells increased significantly while the apoptotic index decreased significantly after PER1 knockdown (P<0.05). The mRNA expression levels of PER1, PER2, DEC1, DEC2, CRY1, CRY2, and NPAS2 markedly decreased (P<0.05) while those of PER3, TIM, RORA, and REV-ERBA markedly increased (P<0.05). By contrast, no obvious changes were observed in the mRNA expression levels of CLOCK, BMAL1, and CKIE (P>0.05).

CONCLUSIONSThe clock gene PER1 can regulate the expression levels of other clock genes in the clock gene networks; these genes include PER2, DEC1, DEC2, CRY1, CRY2, NPAS2, PER3, TIM, RORA, and REV-ERBA. PER1 gene thus plays an important role in the regulation of clock gene networks.
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Apoptosis ; Carcinoma, Squamous Cell ; Gene Regulatory Networks ; Head and Neck Neoplasms ; Humans ; Mouth Neoplasms ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

To evaluate the promoting blood circulation and removing blood stasis effects of Danshen-Honghua(DH) herb pair with different preparations (alcohol, 50% alcohol and water) on blood rheology and coagulation functions in acute blood stasis rats, and optimize the best preparation method of DH based on principal component analysis(PCA), hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods. Ice water bath and subcutaneous injection of adrenaline were both used to establish the acute blood stasis rat model. Then the blood stasis rats were administrated intragastrically with DH (alcohol, 50% alcohol and water) extracts. The whole blood viscosity(WBV), plasma viscosity(PV), erythrocyte sedimentation rate(ESR) and haematocrit(HCT) were tested to observe the effects of DH herb pair with different preparations and doses on hemorheology of blood stasis rats; the activated partial thromboplastin time(APTT), thrombin time(TT), prothrombin time(PT), and plasma fibrinogen(FIB) were tested to observe the effects of DH herb pair with different preparations on blood coagulation function and platelet aggregation of blood stasis rats. Then PCA, hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods were all used to comprehensively evaluate the total promoting blood circulation and removing blood stasis effects of DH herb pair with different preparations. The hemorheological indexes and coagulation parameters of model group had significant differences with normal blank group. As compared with the model group, the DH herb pair with different preparations at low, middle and high doses could improve the blood hemorheology indexes and coagulation parameters in acute blood stasis rats with dose-effect relation. Based on the PCA, hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods, the high dose group of 50% alcohol extract had the best effect of promoting blood circulation and removing blood stasis. Under the same dose but different preparations, 50% alcohol DH could obviously improve the hemorheology and blood coagulation function in acute blood stasis rats. These results suggested that DH herb pair with different preparations could obviously ameliorate the abnormality of hemorheology and blood coagulation function in acute blood stasis rats, and the optimized preparation of DH herb pair on promoting blood effects was 50% alcohol extract, providing scientific basis for more effective application of the DH herb pair in modern clinic medicine.

To investigate the effects of Liu Tea extracts(LTE) on proliferation, apoptosis and drug sensitivity of drug-resistant gastric cancer cell BGC823/5-FU. MTT assay was used to analyze effect of LTE on cell growth and sensitivity chemotherapeutic drugs, and synergistic effect of the combination of LTE with 5-FU on BGC823/5-FU cells. Combination index (CI) was calculated by CompuSyn. Cell apoptosis was measured by flow cytometry (FCM). Protein expressions of P-gp, Bcl-2, Bax and Caspase-3 (17KD) were detected by Western blot at different concentrations of LTE in BGC823/5-FU cells (100, 200, 400 mg•L⁻¹). The results showed that LTE had an inhibitory effect on growth of BGC823/5-FU cell in a dose-time-dependent manner and significantly reduced IC₅₀ of 5-FU, CDDP, PTX and ADM to BGC823/5-FU cells(P<0.05), indicating it could reverse tolerance of drug resistant cells to chemotherapeutic drugs, with reversion multiples of 2.35, 1.68, 1.96, 0.52. The combination of LTE with 5-FU had positive synergistic effect on the BGC-823 cell line. FCM assay suggested that LTE could induce BGC823/5-FU apoptosis. The apoptosis rate was up to 46.2% when the cells were treated with 800 mg•L⁻¹ LTE after 24 h(P<0.01). According to the protein detection results, with the increase in concentration of LTE, the protein expression of Bcl-2 was gradually decreased(P<0.01), the expression of Bax and Caspase-3 were extremely increased(P<0.01), with statistical significance in difference(P<0.01) but no difference in the expression of P-gp between experiment group and control group. LTE can inhibit the growth of drug-resistant human gastric cancer cell BGC823/5-FU and reverse its chemotherapeutic tolerance to some extent. Inhibition of antiapoptotic proteins, activation of proapoptotic proteins and induction of apoptosis of resistant cells may be its main mechanisms.

This study intends to explore the potential resource-orientedutilization value of the flower of Sophora flavescents by analyzing alkaloids and flavonoids in the flower of S. flavescens from Shanxi province. This study established a rapid UPLC-TQ-MS/MS method that is used for determination of seven alkaloids and seven flavonoids in the flower of S.flavescens. The different florescences all have the seven detected alkaloids such as cytisine, oxy-matrine, oxy-sophocarpine, sophoridine, N-methylcytisine, matrine, sophocarpine.The total contents of detected alkaloids are as follows: flower buds 1.47%, primal flowers 1.34%, full bloomed flowers 1.17%, faded flowers 1.01%. The top three contents of alkaloids are N-methylcytisine , oxy-sophocarpine and oxymatrine, accounting for about 83% of the total amount of detected alkaloids. All the samples in different florescences have the seven detected flavonoids such as rutin, luteolin, quercetin, isoquercitrin, trifolirhizin, kurarinone, and kushenol I. The total contents of detected alkaloids are as follows: flower buds 495.2 μg•g⁻¹, primal flowers 313.7 μg•g⁻¹, faded flowers 224.2 μg•g⁻¹, full bloomed flowers 193.0 μg•g⁻¹. The content of luteolinis relatively higher than other detected flavonoids, accounting for about 89%-94% of the total amount of detected flavonoids. The results indicated that the flower of S.flavescens could be an important material resource to obtain the resourceful alkaloids. This result can provide scientific basis for resource-oriented utilization and industrial development of the flower of S. flavescens.

Xin-Sheng-Hua granule (XSHG) is a popular remedy commonly used in clinic for the treatment of lochiostasis after delivery. To comparatively investigate the roles of herb pairs containing Angelicae Sinensis Radix (Danggui) upon the formula by evaluating the blood coagulation and hemorheology function in acute blood stasis rats, acute blood stasis rat model was established by ice water bath and subcutaneous injection of adrenaline. And the blood stasis mice were administrated intragastrically with different samples of the formula minus herb pairs containing Danggui and the whole formula (XSHG, SHD, DY, DC, DT, DH, DJ and DZ). The whole blood viscosity (WBV), plasma viscosity (PV), erythrocyte sedimentation rate (ESR) and haematocrit (HCT) were applied to evaluate the effects of the formula minus herb pairs containing Danggui on hemorheology of blood stasis rats. The thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT), and plasma fibrinogen (FIB) were used to observe the effects of the formula minus herb pairs containing Danggui on blood coagulation function of blood stasis rats. Additionally, the maximum aggregation induced by adenosine diphosphate (ADP) was tested to observe the effect of different samples on platelet aggregation index of blood stasis rats.Afterwards, multi-attribute comprehensive index methods and principal component analysis were both applied to comprehensively assess the total effects of the formula minus herb pairs containing Danggui on promoting blood circulation and dissipating blood stasis. Compared with normal group, the hemorheological parameters and coagulation indexes of model group had statistical differences (P<0.01). Compared with model group, different samples (XSHG, SHD, DY, DC, DT, DH, DJ and DZ) could improve the blood hemorheology indexes, coagulation parameters and platelet aggregation in acute blood stasis rats. According to multi-attribute comprehensive index methods and the principal component analysis, the effects of promoting blood circulation by removing blood stasis became poor when excluding herb pairs containing Danggui from the formula, the sample DY and DC had the weakest effect of activating blood circulation and dissipating blood stasis, and the effect of sample DY was slightly poorer than DC. The orders of contribution of herb pairs containing Danggui on the formula were Danggui-Yimucao>Danggui-Chuanxiong>Danggui-Honghua>Danggui-Zhigancao>Danggui-Taoren>Danggui-Jiangtan. In conclusion, various herb pairs containing Danggui played different roles on the effects of improving the abnormality of hemorheology and coagulation function. And the herb pairs Danggui-Yimucao were particularly important for the formula, which was consistent with the characteristics of XSHG and the traditional effect of Yimucao. Moreover, it could lay foundation to further reveal the compatibility mechanism of XSHG.

To investigate the stability and the conversion rules of cantharidin and cantharidic acid contained in the Mylabris aqueous solution under different conditions. The contents of cantharidin and cantharidic acid under different conditions (pH, temperature and light) were determined by HPLC-TQ-MS. The results showed that the content of cantharidin was gradually decreased with the rising of pH value, while on the contrary, the content of cantharidic acid was increased gradually; after Mylabris aqueous solution with different pH values were placed at 25, 40 ℃ and 25 ℃ respectively for lighting for 90 days, the samples were collected for analysis. The results showed the contents of cantharidin and cantharidic acid were changed greatly in the first 10 days, mainly including the decrease of cantharidic acid and increase of cantharidin when the solution was acidic, and the increase of cantharidic acid and decrease of cantharidin when the solution was alkaline. After that, the contents of the above two components basically remained stable. This study revealed that pH was the main factor to affect the contents of cantharidin and cantharidic acid, and they could be converted into each other with the changes of pH value. High temperature and light can accelerate the speed of achieving such balance. The study can provide certain reference for the quality control of the medicines using the Mylabris as raw material.

OBJECTIVETo observe analgesic and sedative effect of acupuncture combined with medicine (ACM) on patients undergiong cardiac surgery.

METHODSA total of 50 patients with cardiac surgery from January 2012 to October 2014 were randomly assigned to the conventional analgesia group (group A) and the ACM analgesia group (group B), 25 in each group. Patients in group A were subjected to analgesia and sedation by injecting dexmedetomidine, while patients in group B were subjected to analgesia and sedation by electro-acupuncture [EA, Shenting (GV24); Yintang (EX-HN3)] combined with injection of dexmedetomidine. Morphine hydrochloride injection was performed when analgesia and sedation effect was ineffective in the two groups. The indicators of patients at different time points in the two groups were observed, such as static and dynamic VAS scores, SAS scores, mean arterial pressure (MAP), heart rate (HR), oxygen saturation (SpO2). The injection dosage of dexmedetomidine and morphine hydrochloride, analgesia satisfaction rate, sedation satisfaction rate, the incidences of adverse reactions during treatment such as bradycardia and low blood pressure, mechanical ventilation time, ICU time, and hospitalization expense were observed and recorded in the two groups.

RESULTSThere was no statistical difference in static and dynamic VAS scores, SAS score, MAP, HR and SpO2 between the two groups at different time points (P > 0.05). The injection dosage of dexmedetomidine and morphine hydrochloride was significantly reduced in group B than in group A (P < 0.05). The analgesia satisfaction rate of patients in group B was much higher than that in group A (P < 0.05). The incidence of bradycardia also obviously decreased more in group B than in group A (P < 0.05). There was no statistical difference in patients' sedation satisfaction rate, incidences of low blood pressure, delirium, vomiting; mechanical ventilation time, ICU time, or hospitalization expense between the two groups (P > 0.05).

CONCLUSIONThe analgesia method of ACM could reduce the dosage of traditional analgesic drugs and the occurrence of partial adverse reactions.

Acupuncture Analgesia ; Analgesia ; methods ; Analgesics ; therapeutic use ; Cardiac Surgical Procedures ; Dexmedetomidine ; therapeutic use ; Electroacupuncture ; Heart Rate ; Humans ; Hypnotics and Sedatives ; therapeutic use ; Morphine ; therapeutic use ; Pain ; prevention & control ; Pain Management ; methods ; Respiration, Artificial

San'ao Decoction (SD) and its analogous formulas derived in the following generations are common used prescriptions for treating pulmonary diseases with principal symptoms such as cough and asthma. They are usually compatible with Chinese herbs for facilitating Fei, dispelling wind, resolving phlegm and fluid retention. Material bases in these formulas are mainly derived from Chinese drugs, but dissolution contents of active components are changed and new components are produced after compatibility. By multilevel effect evaluation, these analogous formulas all have commonness in ventilating Fei and superiorities of evidence-based derivation. The effect pathway of commonness was involved in cell structure protection, anti-inflammation, antioxidant, and immunoregulation.
Asthma ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; Medicine, Chinese Traditional

OBJECTIVETo study the pharmacodynamic interaction of nourishing and tonifying blood effects of the herb pair consisting of Angelica sinensis and Ligusticum chuanxiong by response surface method.

METHODSThe blood deficiency rat model was induced by injecting N-acetylphenylhydrazine and cyclophosphamide. The effects of Angelica sinensis and Ligusticum chuanxiong in different proportions (0:1, 1:5, 2:5, 2:3, 1:1, 3:2, 5:2, 5:1, and 1:0) and at different concentrations on the peripheral blood index and the organ indices were observed. Then all indices were integrated to the total nourishing effect value by comprehensive index method. The interaction was analyzed by response surface method. The model parameters were estimated with nonlinear regression. The three-dimensional response surfaces were constructed with Matlab Software.

RESULTSIn the response surface, most compatibility of Angelica sinensis and Ligusticum chuanxiong showed synergistic action, some showed addition action, and few of them showed obvious antagonist action. The proportion of Angelica sinensis and Ligusticum chuanxiong from 4:1 to 2:1 and the dose of Angelica sinensis and Ligusticum chuanxiong from low to high showed addition action, while the other proportions showed obvious addition action at low dose and synergistic action at high dose.

CONCLUSIONSThe research results could provide scientific evidence for reasonable application of Angelica sinensis and Ligusticum chuanxiong in clinics of Chinese medicine. The quantitative analysis on drug interactions of herbal compatibility by response surface method could provide reference for relative studies.

Angelica sinensis ; chemistry ; Animals ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; Female ; Ligusticum ; chemistry ; Mice ; Mice, Inbred ICR