3'-Methoxydaidzein exerts analgesic activity by inhibiting voltage-gated sodium channels.

Isoflavones are widely consumed by people around the world in the form of soy products, dietary supplements and drugs. Many isoflavones or related crude extracts have been reported to exert pain-relief activities, but the mechanism remains unclear. Voltage-gated sodium channels (VGSCs) play important roles in excitability of pain sensing neurons and many of them are important nociceptors. Here, we report that several isoflavones including 3'-methoxydaidzein (3MOD), genistein (GEN) and daidzein (DAI) show abilities to block VGSCs and thus to attenuate chemicals and heat induced acute pain or chronic constriction injury (CCI) induced pain hypersensitivity in mice. Especially, 3MOD shows strong analgesic potential without inducing addiction through inhibiting subtypes NaV1.7, NaV1.8 and NaV1.3 with the IC50 of 181 ± 14, 397 ± 26, and 505 ± 46 nmol·L-1, respectively, providing a promising compound or parent structure for the treatment of pain pathologies. This study reveals a pain-alleviating mechanism of dietary isoflavones and may provide a convenient avenue to alleviate pain.

[Research progress of pesticide residues in traditional Chinese medicines].

The residue of the pesticides is closely related to the quality and safety of traditional Chinese medicines(TCMs) and has attracted widespread attention at home and abroad.This article analyzed the current status of pesticide residues in TCMs by summarizing the research results of recent years.At the same time,the methods for the detection of pesticide residues in TCMs were summarized,and the domestic and foreign pesticide residue limit standards for TCMs were compared,intending to provide reference and basis for the detection and control of pesticide residues in TCMs.

Structural characterization of a pectin-type polysaccharide from Curcuma kwangsiensis and its effects on reversing MDSC-mediated T cell suppression.

Myeloid-derived suppressor cells (MDSCs) accumulate in tumor-bearing hosts and play a major role in tumor-induced immunosuppression. The potent modulatory effects of polysaccharides on the innate and adaptive immune system stimulate antitumor responses. In this study, a polysaccharide with an apparent molecular weight of 14.0 kD was isolated from Curcuma kwangsiensis and designated as CKAP-2. The polysaccharide was characterized through high-performance gel permeation chromatography, chemical derivative analyses, GC-MS, FT-IR, and NMR. Results revealed that CKAP-2 is a highly methyl-esterified pectin-type polysaccharide. It is predominantly composed of a homogalacturonan region and small amounts of type-I rhamonogalacturonan regions. Its degree of methyl-esterification is approximately 62.4%. The effect of CKAP-2 on MDSC-medicated immunosuppression was primarily tested. CKAP-2 recovered the MSC2-supressed proliferation of CD4+ and CD8+ T-cells. This finding suggested that CKAP-2 can reverse MDSC-mediated T-cell suppression and that CKAP-2 can be potentially applied in antitumor therapy.

[Optimization of extraction technology from Paeoniae Radix Alba using response surface methodology].

To ensure the stability of chemistry components and the convenience of operation, ultrasound method was chosen to study in this investigation. As the total common peaks area in chromatograms was set to be evaluation index, the influence on the technology caused by extraction time, ethanol concentration and liquid-to-solid ratio was studied by using single factor methodology, and the extraction technology of Paeoniae Radix Alba was optimized by using response surface methodology. The results showed that the extracting results were most affected by ethanol concentration; liquid-to-solid ratio came the second and extraction time thirdly. The optimum ultrasonic-assisted extraction conditions were as follow: the ultrasonic extraction time was 20.06 min, the ethanol concentration in solvent was 72.04%, and the liquid-to-solid ratio was 53.38 mL · g(-1), the predicted value of total common peaks area was 2.1608 x 10(8). Under the extraction conditions after optimization, the total common peaks area was 2.1422 x 10(8), and the relative deviation between the measured and predicted value was 0.86%, so the optimized extraction technology for Paeoniae Radix Alba is suitable and feasible. Besides, for the purpose of extracting more sufficiently and completely, the optimized extraction technology had more advantages than the extraction method recorded in the monogragh of Paeoniae Radix Alba in Chinese Pharmacopoeia, which will come true the assessment and utilization comprehensively.

[Study on chemical components distribution in Paeoniae Radix Alba and its processing methods].

After studying the discipline of chemical components distribution in peony root, it was found that the content distribution of albiflorin, paeoniflorin, benzoylpaeoniflorin and paeonol was inhomogeneous in different parts of peony root, as same as in different roots belonging to a same plant. Therefore, in order to minimize the effects of different qualities of materials on experimental result, the content changing percentages of four chemical components was chosen as indicators to study processing method of Paeoniae Radix Alba. The processing technique of Paeoniae Radix Alba was put peony roots in the boiled water for at most ten minutes, and cut them into 2-3 mm slices after peeling the root bark, finally dry them in a oven under 55 degrees C, which was similar to the traditional processing method used in Bozhou. And the peeling, boiling and drying methods would affect the contents of albiflorin, paeoniflorin, benzoylpaeoniflorin and paeonol on several levels.

[Protective effects of notoginsenoside R1 against amyloid-ß (1-42) induced mitochondrial apopototic death in SH-SY5Y cells].

OBJECTIVE: To investigate the effects and underlying mechanism of notoginsenoside R1 on amyloid-ß (1-42) (Aß(1-42)) induced mitochondrial apoptotic death in SH-SY5Y cells. METHOD: Cell viability was assayed by MTT, apoptotic rates were analyzed with PI/Annexin V flow cytometry, Bax and Bcl-2 expression were detected with Western blotting, enzymatic activity of caspase-3, caspase-8 and caspase-9 were measured by ELISA assay. RESULT: The 6.25-100 nmol x L(-1) of notoginsenoside R1 attenuate Aß(1-42) induced apoptotic death of SH-SY5Y in dose dependent manner. The ratio of Bcl-2/Bax was elevated in SH-SY5Y with notoginsenoside R1 treatment. Caspase-3 and caspase-9 were activated with notoginsenoside R1 treatment while caspase-8 was not affected. CONCLUSION: Notoginsenoside R1 could protect SH-SY5Y cells from Aß(1-42) induced apoptosis via mitochondria related apoptotic pathway.

[Determination of chemical components of Paeoniae Radix Alba decoction pieces and its quality evaluation].

OBJECTIVE: An UPLC method was developed to evaluate the quality of Paeoniae Radix by simultaneously determining four components, paeoniflorin, albiflorin, benzoylpaeoniflorin and paeonol in Paeoniae Radix Alba decoction pieces. METHOD: The UPLC chromatographic column was ACQUITY UPLC® HSS T3. The mobile phase was acetonitrile-0.05% phosphoric acid water with gradient elution. The column temperature was 30 °C and detection wavelength was 230 nm with a flow rate of 0.4 mL · min(-1). A linear model was obtained through principal component analysis (PCA), and PCA scores were used to evaluate the quality of Radix Paeoniae Alba decoction pieces comprehensively. RESULT: Paeoniflorin, albiflorin, benzoylpaeoniflorin and paeonol could be well separated from other components, and the results of specificity, precision, repeatability, linearity, recovery rate and stability reached the standards, respectively. The content of paeoniflorin in 9 batches of Paeoniae Radix Alba decoction pieces was below the standard given by Chinese Pharmacopoeia (2010 edition). Using the comprehensive scoring method with principal component analysis, the results showed that the samples from Zhejiang province have better quality than those from Anhui and Shandong provinces. CONCLUSION: The method established in this study can effectively determine the content of paeoniflorin, albiflorin, benzoylpaeoniflorin and paeonol, which could be used for quality control of Paeoniae Radix Alba.

[Advance in studies on TRPV1 and analgesic effect of traditional Chinese medicines].

Transient receptor potential vanilloid 1 (TRPV1) is a non-selective positive ion channel that is mainly expressed in sensory neurons and a member of transient receptor potential (TRP) family. The receptor could be activated by mechanical irritation, chemical irritation or endogenous ligand to mediate pains and cause injury to body functions. Traditional Chinese medicine believes that the mechanism of pain is that "stagnation leads to pain". Specifically, both of the contracture and tautness caused by cold and the blood stasis could result in blood impassability and pain. Most of traditional Chinese medicines for clearing heat and removing toxicity have the anti-inflammatory effect, while those for warming interior, and promoting blood circulation to remove blood stasis have the effect in smoothening blood vessels. Therefore, either with the anti-inflammatory effect or the effect in smoothening blood vessels, traditional Chinese medicines for clearing heat and removing toxicity, warming interior, and promoting blood circulation have the analgesic effect In this paper, the authors summarize the analgesic effect of the above three traditional Chinese medicines, with TRPV1 as the target.

Pharmacokinetics and brain distribution differences of baicalin in rat underlying the effect of Panax notoginsenosides after intravenous administration.

Baicalin (BA) is the most well-known flavonoid present in Radix Scutellariae. The aim of this study was to explore whether the pharmacokinetic behavior of BA in rat brain can be affected by Panax notoginsenosides (PNS), and to assess the possible mechanism for the observed effects. Specific HPLC and HPLC/MS/MS methods were developed and validated for the determination of BA in the rat plasma and brain using carbamazepine as an internal standard. BA was found to enter rat brain quickly after a single intravenous dose. When co-administered with PNS, clearance (CL) of BA from rat plasma decreased by 50.00%, while the area under the curve AUC0-t and AUC0-∞ increased 94.69% and 87.68%, respectively. On the other hand, some pharmacokinetic parameters of BA in rat brain had obvious differences after PNS was administered, such as an increase in Tmax from 5 min to 15 min, an increase in AUC0-t and AUC0-∞ by 42.75% and 29.39%, respectively, as well as a decrease in CL by 27.95%. Together, these results indicate that PNS can decrease the elimination rate of BA from rat plasma, promote the penetration of BA into rat brain, increase the concentration and slow down the elimination of BA from rat brain. The data provide important information that compatibility with PNS can promote the consequent effects of BA for the treatment of encephalopathy.

[Protective effect of jiedu tongluo injection on cerebral edema in rats with lesion of cerebral ischemia/reperfusion].

OBJECTIVE: To investigate the protective effects of Jiedu Tongluo injection on cerebral edema induced by focal lesion of cerebral ischemia/reperfusion, the hydrous content of brain and the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin and MMP-9 in rats. METHOD: The model of brain middle cerebral artery ischemia/reperfusion was established by the thread approach. After 24 hours of reperfusion, cerebral edema formation was determined by the hydrous content of brain. The permeability of blood brain barrier was evaluated based on the leakage of Evans blue. Enzyme-linked immunoadsordent assay (ELISA)was used to examine the expression of ICAM-1, VCAM-1, E-selectin. The expression of MMP-9 was measured by immunohistochemistry. RESULT: JDTL, in the dose of 2 mL x kg(-1) and 4 mL x kg(-1), relieved cerebral edema (P < 0.05, P < 0.01), reduced the expressions of ICAM-1, VCAM-land E-selectin and decreased MMP-9 activity (P < 0. 05, P < 0.01) in model rats. CONCLUSION: Jiedu Tongluo injection has a protective effect on rat brain from cerebral edema induced by the injury of focal cerebral ischemia/reperfusion. The mechanism is related to that Jiedu Tongluo injection can reduce the expressions of ICAM-1, VCAM-1 and E-selectin and inhibit of MMP-9 activation in rat brain.

[Advance in studies of Panax notoginseng saponins on pharmacological mechanism of nervous system disease].

The pharmacological mechaisms of Panax notoginseng saponins on nervous system diseases (Alzheimer's disease, Parkinson's disease, ischermic cerebral apoplexy and depressive disorder) , including panax notoginseng saponins, protoparaxotriol saponins, panasadiol saponins, ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re and notoginsenoside R1 were summarized to analyze the study hotspots and potential advantages (such as estrogen-like effect) of notoginsenoside's pharmacological actions, provide reference for further pharmacological studies and new ideas for clinical treatment of nervous system diseases and drug studies and development.

[30% intralipid shortens the duration of anesthesia induced by etomidate in rats].

OBJECTIVE: To examine whether 30% intralipid could shorten the duration of anesthesia induced by etomidate and its underlying mechanism. METHODS: Male Sprague-Dawley rats were anesthetized by etomidate (2.85 mg/kg) through caudal vein. After loss of righting reflex, 40 rats were randomly divided into four groups, each being administered intravenously with 2 mL 30% intralipid, 2 mL normal saline, 0.5 mL 30% intralipid, and 0.5 mL normal saline, respectively. The durations from loss of righting reflex to revival of righting reflex and to normal activities were recorded. In the study in vitro, the mixture containing 30% intralipid (3.03%, 1.52%, 0.76%, 0.37% and 0.00% respectively), rat plasma and 0.066 mg/mL etomidate was vortex mixed and centrifuged. The concentration of etomidate in aqueous phase was determined by high-performance liquid chromatographic (HPLC) method. RESULTS: Compared with those of normal saline at a dose of 2 mL, 30% intralipid significantly shortened the duration from loss of righting reflex to revival of righting reflex [(493 +/- 27) s vs. (755 +/- 64) s, P < 0.01) and from loss of righting reflex to normal activities [(643 +/- 36) s vs. (1104 +/- 114) s, P < 0.01] of the rats. Intralipid reduced the concentration of etomidate in aqueous phase in vitro. CONCLUSION: Intralipid shortens the duration of anesthesia induced by etomidate in rats, possibly through extractive effect of intralipid on etomidate.

Inhibition of advanced glycation end product formation by Pu-erh tea ameliorates progression of experimental diabetic nephropathy.

Accumulation of advanced glycation end products (AGEs) has been implicated in the development of diabetic nephropathy. We investigated the effects of Pu-erh tea on AGE accumulation associated with diabetic nephropathy. Although it did not affect blood glucose levels and insulin sensitivy, Pu-erh tea treatment for 8 weeks attenuated the increases in urinary albumin, serum creatinine, and mesangial matrix in db/db mice. We found that Pu-erh tea prevented diabetes-induced accumulation of AGEs and led to a decreased level of receptor for AGE expression in glomeruli. Both production and clearance of carbonyl compounds, the main precursor of AGE formation, were probably attenuated by Pu-erh tea in vivo independent of glyoxalase I expression. In vitro, HPLC assay demonstrated Pu-erh tea could trap methylglyoxal in a dose-dependent manner. Our study raises the possibility that inhibition of AGE formation by carbonyl trapping is a promising approach to prevent or arrest the progression of diabetic complications.

The interaction between emulsified isoflurane and lidocaine is synergism in intravenous regional anesthesia in rats.

BACKGROUND: Our previous studies have demonstrated that emulsified isoflurane (EI) produced epidural anesthesia and blockade of nerve conduction. We designed this study to observe whether EI could produce an anesthetic effect in IV regional anesthesia (IVRA) and to investigate the underlying interaction between EI and lidocaine when they were combined in IVRA. METHODS: IVRA was evaluated using tail-flick and tail-clamping tests in a rat model. In experiment 1, Sprague-Dawley rats were assigned to 4 groups (n = 10 per group), receiving 0.5 mL of 8%EI, 0.5% lidocaine, 30% Intralipid, or normal saline to observe whether EI could produce an anesthetic effect in IVRA. In experiment 2, for tail IVRA, EC(50) (median effective concentration) of EI alone and EC(50) of lidocaine alone, as well as EC(50) of lidocaine with the addition of Intralipid (0.0% EI) or with the addition of EI at different concentrations (0.4%, 0.8%, and 1.6%) were determined using an up-and-down method. Isobolographic analysis was used to evaluate the interaction between EI and lidocaine. RESULTS: For experiment 1, successful IVRA was observed in 8 of 10 rats with 8% EI, 10 of 10 rats with 0.5% lidocaine, and 0 of 10 rats with 30% Intralipid or normal saline. The anesthetic effect was not different in onset time (1.5 ± 0.9 vs 1.0 ± 0.0 minutes, P = 0.104) or recovery time (15 ± 9 vs 18 ± 12 minutes, P = 0.394) between 8% EI and 0.5% lidocaine. For experiment 2, EC(50) of EI was 4.467% ± 0.375% and EC(50) of lidocaine was 0.183% ± 0.072%; EC(50) of lidocaine was 0.173% ± 0.036% with the addition of Intralipid, and 0.064% ± 0.008%, 0.035% ± 0.005%, and 0.028% ± 0.006% with the addition of 0.4%, 0.8%, and 1.6% EI, respectively. With the addition of EI, the requirement for lidocaine was reduced in a synergistic manner. CONCLUSIONS: EI produced IVRA, and a synergistic interaction was found between EI and lidocaine for IVRA in a rat tail model.

Possible protection by notoginsenoside R1 against glutamate neurotoxicity mediated by N-methyl-D-aspartate receptors composed of an NR1/NR2B subunit assembly.

Notoginsenoside R1 (NTR1) is the main active ingredient in Panax notoginseng, a herbal medicine widely used in Asia for years. The purpose of this study was to investigate pharmacological properties of NTR1 on neurotoxicity of glutamate (Glu) in primary cultured mouse cortical neurons along with its possible mechanism of action. We found that NTR1 significantly protected neurons from the loss of cellular viability caused by brief exposure to 10 microM Glu for 1 hr in a dose-dependent manner at concentrations from 0.1 to 10 microM, without affecting the viability alone. NTR1 significantly inhibited the increased number of cells positive to propidium iodide (PI) staining, increase of intracellular free Ca(2+) ions, overproduction of intracellular reactive oxygen species, and depolarization of mitochondrial membrane potential in cultured neurons exposed to Glu, in addition to blocking decreased Bcl-2 and increased Bax expression levels. We further evaluated the target site at which NTR1 protects neurons from Glu toxicity by using the acquired expression strategy of N-methyl-D-aspartate (NMDA) receptor subunits in human embryonic kidney 293 cells. We found that 10 microM NTR1 protected NR1/NR2B subunit expressing cells from cell death by 100 microM NMDA, but not cells expressing NR1/NR2A subunits, when determined by PI staining. These results suggest that NTR1 may preferentially protect neurons from Glu excitotoxicity mediated by NMDA receptor composed of an NR1/NR2B subunit assembly in the brain.

[Regulatory effect of anthraquinone derivatives from rhubarb on aquaporin 4 expression in colon of rats and in LoVo cell line].

OBJECTIVE: To investigate the cathartic effect of total anthraquinone (AQ) from rhubarb on SD rats and its regulatory effect on aquaporin 4 (AQP4) expression in rat colon and in vitro cultured LoVo cell line. METHODS: Twenty-four SD rats were randomly divided into the normal control group treated with distilled water, and the two AQ groups administered with AQ suspension in cathartic and high dose (AQcd and AQhd) respectively via gastrogavage for 5 days. Water content in colonic stool was detected and the expression of AQP4 in rat's proximal colon was measured using Western blot and RT-PCR. LoVo cells cultured in vitro were used in the experimental study. The AQP4 protein and mRNA expressions in the cells were detected by Western blot and semiquantitative RT-PCR after they were cultured for 24 h with RPMI-1640 medium containing rhein/emodin in different concentrations, and those cultured with RPMI-1640 containing 20 mg/L rhein/emodin for different time points. RESULTS: After treatment, the stool water content in the AQcd and AQhd groups was higher than that in the control group and the AQP4 expression in rats treated with AQ decreased in a dose-dependent manner. The study showed that rhein/emodin could significantly down-regulate the protein and mRNA expressions of AQP4 in cultured LoVo cells, with the effectiveness related with dose and acting time. CONCLUSION: At the same time of playing cathartic action, total AQ of rhubarb can effectively down-regulate the expression of AQP4 in rat's proximal colon; rhein/emodin can suppress the AQP4 expression in LoVo cells in vitro. One mechanism of cathartic effect of rhubarb AQ is possibly its down-regulation on AQP4 expression.

[Preliminary analysis of ginseng industry in Wisconsin].

OBJECTIVE: To study the case of Wisconsin as the top ginseng state in United States which has come through four developing steps: beginning, stagnating, flourishing and now, downturn. METHOD: The current situation of the ginseng industry in Wisconsin was briefly introduced, the federal and state management on ginseng cultivation and export, the organization of Ginseng Board of Wisconsin and their marketing style based on the field investigation and data collected from USDA and Wisconsin state. RESULT: The advantages and disadvantages of Wisconsin ginseng industry were analyzed in order to provide some suggestions for Chinese medicine industry. CONCLUSION: Chinese ginseng industry should learn the organization system from Wisconsin.

[Effect of total anthraquinone in rheum on aquaporin 2 expression in rat distal colon].

OBJECTIVE: To investigate effect of total anthraquinone in rheum on aquaporin 2 expression in rat distal colon. METHOD: SD rats were randomly divided into control group, low dose group, middle dose group and high dose group. Gavaged to control group, and treated group were administered saline and total anthraquinone in rheum with dosage of 0.14, 2.5, 4.5 g x kg(-1) x d(-1), respectively. All rats were put sacrificed after 5 days and stool in full length colon was gently collected to detect water content stool. Distal colon was removed to detect AQP2 expression with immunohistochemistry, western blot and RT-PCR. RESULT: No diarrhea was found in low dose group and control group, there were not significant difference water content of stool and AQP2 expression between low dose group and control group. However, soft feces and loose stools occurred in diarrheic dose group, loose stools and watery stool appeared in high dose group. Stool water content increased in diarrheic dose group and High dose group, expression of AQP2 decreased evidently in these two groups (P < 0.01). CONCLUSION: Total anthraquinone in rheum can reduce the transcription and translation of AQP2 in rats' distal colon, increase fecal water content, which probably is one of the mechanisms of diarrhea caused by total anthraquinone in rheum.

[Regulative effects of aquaporin 4 expression by rhein in rhubarb to intestinal epithelial cell line LoVo].

OBJECTIVE: To investigate the effects of rhein on regulating aquaporin4 (AQP4) to LoVo cells cultured with RPMI-1640 medium containing rhein. METHODS: LoVo cells were cultured with RPMI-1640 medium containing different concentration rhein for 24 hours and were cultured with RPMI-1640 containing rhein (20 mg/L) for different time. Four groups were assigned as LoVo cells were cultured respectively with RPMI-1640 medium containing different concentration rhein (40, 20, 10 mg/L and control group), while six groups were assigned as LoVo cells were cultured with RPMI-1640 medium containing rhein (20 mg/L) for different time (3, 6, 12, 24, 48 h and control group). The location of AQP4 protein in LoVo cells was definited by immuocytochemistry dying. Western blotting and semiquantitative RT-PCR were adopted to detect the relative expression of AQP4 protein and mRNA. RESULTS: AQP4 was located mainly in plasma membrane of LoVo cells while partly in cytoplasm. The relative expression of AQP4 protein and mRNA decreased with the increasing of rhein concentration; there was no significant difference of the relative expression of AQP4 in 10 mg/L group compared with that in control group, but it decreased significantly in 40 and 20 mg/L groups. The relative expression of AQP4 in 3 and 6 h groups was lower than that in control group but there was no statistical significance, however that in 12, 24, 48 h groups was lower significantly compared with that in control group. CONCLUSION: Rhein can inhibit the genetic transcription and the translation of AQP4 gene in LoVo cells, which demonstrates that the change of AQP4 expression regulated by rhein may be related to the cathartic effect of rhubarb.