[Optimal processing technology of Zhangbang vinegar-processed Olibanum with multi-indicator-response surface methodology and anticoagulant effect evaluation].

This study first optimized the processing technology for Zhangbang vinegar-processed Olibanum and investigated its in vitro anticoagulant activity. A multi-index-response surface methodology was used, with yield, powder yield, and the relative percentage of the content of six non-volatile components [11-keto-boswellic acid(KBA), 3-acetyl-11-keto-boswellic acid(AKBA), ß-elemonic acid, α-boswellic acid(α-BA), ß-boswellic acid(ß-BA), and α-acetyl-boswellic acid(α-BA)] and three volatile components(octyl acetate, incensole, and incensole acetate) as evaluation indicators. Analytical hierarchy process(AHP) combined with coefficient of variation method was used to calculate the weight of each indicator and calculate the comprehensive score(OD). Furthermore, response surface methodology was used to investigate the effects of frying temperature(A), burning time(B), rice vinegar dosage(C), and steaming time(D) on the processing technology of vinegar-processed Olibanum. Vinegar-steamed Olibanum was prepared according to the optimal processing technology for in vitro anticoagulant experiments. The results showed that the weights of octyl acetate, incensole, incensole acetate, KBA, AKBA, ß-elemonic acid, α-BA, ß-BA, α-ABA, yield, and powder yield were 0.358 2, 0.104 5, 0.146 4, 0.032 9, 0.123 7, 0.044 4, 0.022 1, 0.042 2, 0.110 1, 0.012 2, and 0.0032, respectively. The optimal processing technology for Zhangbang vinegar-processed Olibanum was as follows. Olibanum(50 g) with a particle size of 1-5 mm was continuously stir-fried at a low heat of 150-180 ℃ until in a gel-like state, ignited for burning for 15 s, sprayed with 7.5 g of rice vinegar(15%), and steamed for 3 min without fire. Subsequently, the cover was removed, and the product was continuously stir-fried at 150-180 ℃ until in a soft lump shape, removed, cooled, and crushed. The results of the in vitro anticoagulant experiments showed that compared with the blank group, both Olibanum and vinegar-processed Olibanum significantly prolonged the activated partial thromboplastin time(APTT), thrombin time(TT), and prothrombin time(PT) of rat platelet-poor plasma(PPP), and the effect of vinegar-processed Olibanum was significantly better than that of Olibanum(P<0.05). The optimized processing technology for Zhangbang vinegar-processed Olibanum is stable, feasible, and beneficial for the further development and utilization of Olibanum slices. At the same time, using the content of volatile and non-volatile components, yield, and powder yield as indicators, and verifying through pharmacological experiments, the obtained results are more reasonable and credible, and have positive guiding significance for the clinical application of characteristic processed Olibanum products.

Tracing analgesic constituents from crude and vinegar-processed resin of Boswellia carterii by integrating ultra-performance liquid chromatography tandem mass spectrometry-based determination, analgesic evaluation in mice, and gray relationship analysis.

The analgesic effect of the resin of Boswellia carterii (BC) is well known; however, the constituents that contribute to the analgesic effect remain elusive. The current study integrates ultrasonic-assisted extraction, quantitative determination, analgesic evaluation in rats, and gray relationship analysis for tracing analgesic constituents from the resin of BC. First, a robust and precise ultra-performance liquid chromatography tandem mass spectrometry approach with multiple reaction monitoring mode was developed for the simultaneous quantification of seven major constituents in crude and vinegar-processed resin of BC. Glycyrrhetinic acid was chosen as the internal standard. The approach showed good linearity. The intra- and inter-day precisions of each constituent were within 3.0%. The recoveries of each constituent were in the range of 96.4-102.7%. The approach was then applied to determine the seven constituents in 10 batches of crude and vinegar-processed resin of BC. Second, the analgesic effects of crude and vinegar-processed resin of BC were assessed in mice. Third, chemometrics methods, gray relationship analysis, and partial least squares regression were employed for determining the relationship between the contents of seven constituents and their analgesic effects. 11-Keto-ß-boswellic acid, 3-acetyl-ß-boswellic acid, 3-acetyl-α-boswellic acid, 3-acetyl-11-keto-ß-boswellic acid, and ß-sitosterol were identified as the key analgesic constituents of BC.

[Toxicity comparison of raw and vinegar-processed Bupleuri Radix based on ~1H-NMR metabolomics].

This study compared the toxicity of raw Bupleuri Radix(BR) and vinegar-processed Bupleuri Radix(VPBR) based on proton nuclear magnetic resonance(~1H-NMR), and explored the mechanism of toxicity. Thirty-two male Sprague-Dawley(SD) rats were randomly divided into four groups: a control group(distilled water), a raw BR group(15 g·kg~(-1)·d~(-1)), a rice VPBR(R-VPBR) group(15 g·kg~(-1)·d~(-1)), and a shanxi VPBR(S-VPBR) group(15 g·kg~(-1)·d~(-1)). After administration for 30 d, pathological sections were treated and observed, and biochemical indexes related to liver and renal function were determined. The serum, liver, and kidney of rats were collected and analyzed by ~1H-NMR. The principal component analysis(PCA) and orthogonal partial least squares-discrimination analysis(OPLS-DA) were performed. The results showed that, as compared with the control group, alanine aminotransferase(ALT), aspartate aminotransferase(AST), and alkaline phosphatase(ALP) in the raw BR group were increased significantly, while ALT and ALP in the R-VPBR and S-VPBR groups were significantly decreased(P<0.05), which indicated that BR showed certain hepatotoxicity, and vinegar processing reduced its hepatotoxicity. No significant difference of blood urea nitrogen(BUN) and creatinine(CREA), the biochemical indexes related to renal function, was observed in the control group and administration groups, indicating that BR had less effect on the renal function. The results of multivariate statistical analysis showed that the biomarkers of BR affecting liver metabolism were methionine, glutamine, and glutamic acid, and affecting kidney metabolism were taurine, ornithine, and inosine. These biomarkers were mainly involved in amino acid metabolism, energy metabolism, lipid metabolism, and taurine metabolism. VPBR alleviated the effect on the biomarkers, and S-VPBR had smaller effect than R-VPBR. Combining the results of biochemical indexes and metabolomics analysis, both raw BR and VPBR showed toxic effect on rats, whereas vinegar processing reduced its toxicity. S-VPBR has smaller effect on kidney and liver metabolism than R-VPBR, which indicates that the vinegar used for processing has certain effect on the toxicity of BR.

[Mechanism and material basis of Sparganii Rhizoma and vinegar-processed Sparganii Rhizoma in treatment of hyperlipidemia based on network pharmacology].

The present study explored the effective components, functional targets, and mechanism of Sparganii Rhizoma(vinegar-processed Sparganii Rhizoma) in the treatment of hyperlipidemia based on network pharmacology and experimental verification. In the network pharmacology, the screening of active components, target prediction, and pathway enrichment analysis of Sparganii Rhizoma were carried out, followed by the comparison with targets and pathways related to hyperlipidemia. In the experimental verification, the hyperlipidemia model in rats was induced to detect hemorheological parameters and coagulation function. The liver index was observed by HE staining, and PCR technology was used to verify the results of the network pharmacological analysis. Compared with the model group, the Sparganii Rhizoma and vinegar-processed Sparganii Rhizoma groups showed decreased liver index(P<0.05), reduced liver lipid deposition, dwindled serum low-density lipoprotein cholesterol(LDL-c) level(P<0.05), diminished blood viscosity, and increased prothrombin time(PT), thrombin time(TT), and activated partial thrombin time(APTT)(P<0.05). As revealed by the PCR assay, Sparganii Rhizoma could affect LDL-c and high-density lipoprotein cholesterol(HDL-c) levels and reduce the inhibitory effect of cholesterol ester transporter by regulating the expression of Apol2, Apof, and Stab2, thereby treating hyperlipidemia. Vinegar-processed Sparganii Rhizoma could enhance triglyceride metabolism and cholesterol reversal by regulating the expression of Hmgcr, Hmgcs2, Abca1, Abcg1, Cyp7 b1, and Stab2. Compared with the Sparganii Rhizoma, the vinegar-processed one was potent in treating hyperlipidemia. The active components of Sparganii Rhizoma in the treatment of hyperlipidemia may be L-alpha-palmitin,(1S,2S)-1,2-bis(2-furyl)ethane-1,2-diol, cis-zimtsaeure, o-acetyl-p-cresol, sanleng, and 9-hexadecenoic acid. Based on the network pharmacology and experimental verification, this study preliminarily explored the potential active components and possible mechanism of Sparganii Rhizoma in the treatment of hyperlipidemia, which is expected to provide a certain basis for in-depth research on active components, mechanism, and clinical application.

A biochemometrics strategy combining quantitative determination, bioactivity evaluation and relationship analysis for identification of analgesic alkaloids of raw and vinegar-processed Corydalis turtschaninovii.

A biochemometrics strategy combining quantitative determination, bioactivity evaluation, and relationship analysis was proposed for identification of analgesic components of herbs. First, a robust liquid chromatography tandem mass spectrometry method was developed for simultaneous determination of nine major alkaloids in crude and vinegar-processed Corydalis turtschaninovii. Nine alkaloids were separated on a BEH C18 column with a mobile phase consisting of acetonitrile and water spiked with 0.1% formic acid and then detected by multiple reactions monitoring in the positive ion mode. Nitidine chloride was employed as the internal standard. The method displayed good linearity and the precisions of intra-day and inter-day were all within 3.0%. The recovery rates of each alkaloid ranged from 97.1 to 102.9%. The method was successfully applied for quantitative analysis of nine alkaloids in ten batches of crude and vinegar-processed Corydalis turtschaninovii. Second, the analgesic effects of crude and vinegar-processed Corydalis turtschaninovii were evaluated in mice. Third, principle component analysis, canonical correlation analysis, and partial least squares regression were used to analysis the relationship between the contents of nine major alkaloids and the analgesic effect of different crude and vinegar-processed samples. Tetrahydropalmatine, coptisine, and dehydrocorydaline have a close positive correlation with the analgesic effect.

[Identification of curcumin content in raw and vinegar-processed rhizomes of Curcuma kwangsiensis based on electronic nose combined with back propagation neural network].

This study aimed to establish a rapid and accurate method for identification of raw and vinegar-processed rhizomes of Curcuma kwangsiensis, in order to predict the content of curcumin compounds for scientific evaluation. A complete set of bionics recognition mode was adopted. The digital odor signal of raw and vinegar-processed rhizomes of Curcuma kwangsiensis were obtained by e-nose, and analyzed by back propagation(BP) neural network algorithm, with the accuracy, the sensitivity and specificity in discriminant model, correlation coefficient as well as the mean square error in regression model as the evaluation indexes. The experimental results showed that the three indexes of the e-nose signal discrimination model established by the neural network algorithm were 100% in training set, correction set and prediction set, which were obviously better than the traditional decision tree, naive bayes, support vector machine, K nearest neighbor and boost classification, and could accurately differentiate the raw and vinegar products. Correlation coefficient and mean square error of the regression model in prediction set were 0.974 8 and 0.117 5 respectively, and could well predict curcumin compounds content in Curcuma kwangsiensis, and demonstrate the superiority of the simulation biometrics model in the analysis of traditional Chinese medicine. By BP neural network algorithm, e-nose odor fingerprint could quickly, conveniently and accurately realize the discrimination and regression, which suggested that more bionics information acquisition and identification patterns could be combined in the field of traditional Chinese medicine, so as to provide ideas and methods for the rapid evaluation and stan-dardization of the quality of traditional Chinese medicine.

The Effects of Vinegar Processing on the Changes in the Physical Properties of Frankincense Related to the Absorption of the Main Boswellic Acids.

Boswellic acids (BAs), as the main components of frankincense, exhibit notable anti-inflammatory properties. However, their pharmaceutical development has been severely limited by their poor oral bioavailability. Traditional Chinese medicinal processing, called Pao Zhi, is believed to improve bioavailability, yet the mechanism is still completely unclear. Previous research suggested that the bioavailability of a drug can be influenced by physical properties. This paper was designed to investigate the physical properties of frankincense and processed frankincense, including the surface morphology, particle size, polydispersity index (PDI), zeta potential (ZP), specific surface area, porosity, and viscosity. The differences in the intestinal absorption characteristics and equilibrium solubilities between frankincense and processed frankincense were determined by an ultra-high-performance liquid chromatography coupled with a triple quadrupole electrospray tandem mass spectrometry (UHPLC-TQ-MS) analysis method. The results showed that vinegar processing can alter the surface morphology, decrease the particle size and PDI, raise the absolute values of the ZP, specific surface area and porosity, and drop the viscosity of frankincense. Meanwhile, the rates of absorption and dissolution of the main BAs were increased after the processing of frankincense. The present study proves that the physical properties were changed after processing, in which case the bioavailability of frankincense was enhanced.

[Effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma on immune hepatic fibrosis, proliferation of HSC-T6 and expression of α-SMA and Procollagen I].

To compare the effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizomaon immune hepatic fibrosis, proliferation of HSC-T6, and expressions of α-SMA and Procollagen I. The immunological liver fibrosis model was prepared through intraperitoneal injection with porcine serum 0.5 mL in each rat, twice a week, for 14 weeks. Expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN, HA and HYP, MDA in liver tissues were observed after administration of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma (0.95, 1.90 g•kg⁻¹). The pathological changes in liver tissues were observed by HE staining. Masson staining and Sirius red staining were used to observe the expression of collagen in rat liver. HSC-T6 was cultured, and the proliferation of HSC-T6 was determined by MTT assay at different concentrations in 12, 24, 36, 48 h. The expressions of α-SMA and Procollagen I were detected by Real-time PCR. The results showed that expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN and HA in Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma groups (0.95, 1.90 g•kg⁻¹) were significantly lower than model group; in terms of effect, vinegar-processed Curcumae Rhizoma group was superior to Curcumae Rhizoma group. Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma containing serum could inhibit the proliferation of HSC-T6 in a dose-effect and time-effect manner. Expressions of α-SMA and Procollagen I in HSC-T6 were decreased after 24 h, especially in 20% vinegar-processed Curcumae Rhizoma containing serum group (P<0.01). Both Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma could reduce immune hepatic fibrosis to varying extent. Their anti-hepatic fibrosis mechanism may be correlated with inhibition of the proliferation of HSC-T6, and reduction of the formation of extracellular matrix and promotion of its degradation.

Discovery and verification of antidepressant active ingredients of raw and vinegar-processed Bupleurum marginatum var. Stenophyllum based on plant metabolomics and serum pharmacology.

The dried root of Bupleurum marginatum var. stenophyllum (H. Wolff) R.H. Shan & Y. Li (BM), which has been used as a Bupleuri radix in Guizhou Province and is listed in the 2003 edition of the Guizhou Quality Standard for Traditional Chinese Medicines and Ethnic Materia Medica, is effective at dispersing the liver and relieving depression and often used in the form of raw or vinegar-processed product (VBM). However, the potential depression-relieving components of BM are unclear. The aim of this study was to determine the potential antidepressant constituents of BM and investigate the effect of vinegar processing on these components. The antidepressant effect and mechanism of BM and VBM were investigated in depressed mice and BV2 cells, respectively. The pharmacodynamic constituents were screened through serum pharmacochemistry, which combined the results of metabolomics analysis of BM and VBM, high-performance liquid chromatography (HPLC) content determination, and verification of the antidepressant effect and mechanism of differential components of SSb2 to clarify the connotation of vinegar processing. Our results demonstrated that BM can exert a significant antidepressant effect by inhibiting microglia polarization and that this effect was enhanced after vinegar processing. Thirty-eight components were identified in the BM, 13 of which were blood-absorbable, mainly saponins, and defined as potential antidepressant components of the BM. The contents of 17 components-6 of which were absorbed into the blood-changed considerably after processing. It was finally determined that vinegar processing can enhance the antidepressant effect of BM by increasing the contents of SSb1 and SSb2. SSb2 exerts this effect via the samemechanism as BM. In conclusion, in this study we clarified the antidepressant effects and potential active components of BM and examined the mechanism of vinegar processing. These findings lay a foundation for the future research on the antidepressant effects of BM as well as for the complete development and application of BM's ethnomedicinal resources.

Variations of the fungal microbiome in Corydalis Rhizoma with different collection areas, processing methods, and storage conditions.

Corydalis Rhizoma (CR, Yanhusuo in Chinese) has been widely used as an analgesic in herbal medicine and functional food. Cases of fungal and mycotoxin contamination in CR have been reported. In this study, the composition and diversity of fungal microbiome in CR samples from four herbal markets and two processing methods were investigated by DNA metabarcoding. Variations of the fungal microbiome in CR during cold and conventional storage were monitored. Results showed that Aspergillus was the dominant genus and saprotroph was the dominant trophic mode. Six potential toxigenic fungi, namely, Aspergillus fumigatus, Aspergillus ostianus, Aspergillus terreus, Penicillium citrinum, Penicillium oxalicum, and Trichothecium roseum, were detected. Differences in fungal composition and diversity among various groups based on collection areas and processing methods were also observed. Moreover, the relative abundance of dominant genera in CR samples stored at different temperatures was significantly different and changed with storage time. This study is the first to reveal the influence of collection areas, processing methods, and storage conditions on the fungal microbiome in CR, which was expected to provide a basis for control strategies of fungal contamination in the industrial chain of CR.

Network pharmacology and experimental verification reveal the mechanism of Hedysari Radix and Curcumae Rhizoma with the optimal compatibility ratio against colitis-associated colorectal cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: The herb pair Astragali Radix (AR) and Curcumae Rhizoma (vinegar-processed, VPCR), derived from the traditional Chinese medicine (TCM) text 'Yixuezhongzhongcanxilu', have long been used to treat gastrointestinal diseases, notably colitis-associated colorectal cancer (CAC). Hedysari Radix (HR), belonging to the same Leguminosae family as AR but from a different genus, is traditionally used as a substitute for AR when paired with VPCR in the treatment of CAC. However, the optimal compatibility ratio for HR-VPCR against CAC and the underlying mechanisms remain unclear. AIM OF THE STUDY: To investigate the optimal compatibility ratio and underlying mechanisms of HR-VPCR against CAC using a combination of comparative pharmacodynamics, network pharmacology, and experimental verification. MATERIALS AND METHODS: The efficacy of different compatibility ratios of HR-VPCR against CAC was evaluated using various indicators, including the body weight, colon length, tumor count, survival rate, disease activity index (DAI) score, Haemotoxylin and Eosin (H&E) pathological sections, inflammation cytokines (IL-1ß, IL-6, IL-10, TNF-α), tumor markers (K-Ras, p53), and intestinal permeability proteins (claudin-1, E-cadherin, mucin-2). Then, the optimal compatibility ratio of HR-VPCR against CAC was determined based on the fuzzy matter-element analysis by integrating the above indicators. After high-performance liquid chromatography (HPLC) analysis for the optimal compatibility ratio of HR-VPCR, potential active components of HR-VPCR were identified by TCMSP and the previous bibliographies. Swiss Targets and GeneCards were adopted to predict the targets of the active components and the targets of CAC, respectively. Then, the common targets of HR-VPCR against CAC were obtained by Venn analysis. PPI networks were constructed in STRING. GO and KEGG enrichments were visualized by the David database. Finally, the predicted pathway was experimentally validated via Western blot. RESULTS: Various compatibility ratios of HR-VPCR demonstrated notable therapeutic effects to some extent, evidenced by improvements in body weight, colon length, tumor count, pathological symptoms (DAI score), colon and organ indexes, survival rate, and modulation of inflammation factors (IL-1ß, IL-6, IL-10, TNF-α), as well as tumor markers (K-Ras, p53), and down-regulation of intestinal permeability proteins (claudin-1, E-cadherin, mucin-2) in CAC mice. Among these ratios, the ratio 4:1 represents the optimal compatibility ratio by the fuzzy matter-element analysis. Thirty active components of HR-VPCR were carefully selected, targeting 553 specific genes. Simultaneously, 2022 targets associated with CAC were identified. 88 common targets were identified after generating a Venn plot. Following PPI network analysis, 29 core targets were established, with AKT1 ranking highest among them. Further analysis via GO and KEGG enrichment identified the PI3K-AKT signaling pathway as a potential mechanism. Experimental validation confirmed that HR-VPCR intervention effectively reversed the activated PI3K-AKT signaling pathway. CONCLUSIONS: The optimal compatibility ratio for the HR-VPCR herb pair in alleviating CAC is 4:1. HR-VPCR exerts its effects by alleviating intestinal inflammation, improving intestinal permeability, and regulating the PI3K-AKT signaling pathway.

Frankincense vinegar-processing improves the absorption of boswellic acids by regulating bile acid metabolism.

BACKGROUND: Boswellic acids in Olibanum (known as frankincense) are potent anti-inflammatory properties in treating ulcerative colitis (UC), but its low bioavailability limited drug development. Evidence accumulated that vinegar processing of frankincense exerts positive effects on improving absorption of compositions. The underlying mechanism is unknown. In recent decades, spectacular growth and multidisciplinary integration of metabolic application were witnessed. The relationship between drug absorption and curative effect has been more or less established. However, it remains a knowledge gap in the field between drug absorption and endocrine metabolism. PURPOSE: To investigate the enhancement mechanism of vinegar processing in the absorption of boswellic acids via the aspect of bile acid metabolism. METHODS: The effects of raw frankincense (RF) and processed frankincense (PF) were compared by the UC model of rats. The plasma concentration of boswellic acids and the hepatic and colonic bile acids contents were quantified by UPLC-TQ-MS. The levels of mRNA and protein associated with bile acid metabolism were also compared. RESULTS: The results showed that PF exhibited re-markable mitigating effects on UC with the elevated plasma level of boswellic acid and upregulated expression of the absorption-related protein multidrug resistance-associated protein 2 (MRP2) and organic anion transporting polypeptide 1B3 (OATP1B3) in the liver and colon. It improved colonic lithocholic acid (LCA), which promoted the expression of bile acid nuclear receptors constitutive androstane receptor (CAR) and pregnane X receptor (PXR), resulting in the upregulation of MRP2 and OATP1B3. CONCLUSION: This paper revealed the mechanisms behind the absorption promotion effects of processing. Bile acids metabolism exhibits potential status in pharmaceutical development. The results shed light on the interdisciplinary collaboration between the metabolism and drug absorption fields.

Raw and vinegar processed Curcuma wenyujin regulates hepatic fibrosis via bloking TGF-ß/Smad signaling pathways and up-regulation of MMP-2/TIMP-1 ratio.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma wenyujin Y.H. (CW), a variety of Curumae Rhizoma, which documented in China Pharmacopeia, has long been used as plant medicine for its traditional effect on promoting Qi, activating blood stagnation and expelling blood stasis. Nowadays, it is often used in clinic for extraordinary effect on liver diseases. It is worthy to be noted that CW processed with vinegar has been applied in clinic for 1500 years which started in the northern and southern dynasties. AIM OF STUDY: Liver fibrosis is a worldwide clinical issue. It is worth developing a multi-target and multicellular approach which is high efficiency and low side effects for the treatment of hepatic fibrosis. The anti-hepatic fibrosis molecular mechanisms of CW and vinegar Curcuma wenyujin (VCW) need to be explored and elucidated. Furthermore, the study aimed to discuss the efficiency and mechanism differences between CW and VCW in hepatic fibrosis. METHODS AND RESULTS: Biochemical assays and histopathology were adopted to evaluate the anti-hepatic fibrosis effect of CW and VCW. The TGF-ß/Smad signaling involving TGF-ß1, TGF-ßRⅠ, TGF-ßRⅡ and Smad2, Smad3, Smad7 in fibrosis is examined, which is a critical step towards the evaluation of anti-hepatic fibrosis agents. Meanwhile, the MMP/TIMP balance is a potential therapy target by modulating extracellular matrix, which is also examined. Both CW and VCW inhibit the activation and proliferation of hepatic stellate cells and induce apoptosis via blocking TGF-ß/Smad signaling pathways. Additionally, the level of MMP-2/TIMP-1 regulated significantly, which suggest CW and VCW participate in the degradation process, and maintain the formation and production of extracellular matrix. CONCLUSION: Raw and vinegar processed Curcuma wenyujin regulates hepatic fibrosis via bloking TGF-ß/Smad signaling pathways and up-regulation of MMP-2/TIMP-1 ratio. And VCW has more exhibition than CW.

Establishment of a rapid and sensitive UPLC-MS/MS method for pharmacokinetic determination of nine alkaloids of crude and processed Corydalis turtschaninovii Besser aqueous extracts in rat plasma.

Corydalis turtschaninovii (CT) is a traditional Chinese medicine which is known to have analgesic effects, and is under investigation for the management of chronic pain. Our study aims to establish a UPLC-MS/MS method for pharmacokinetic determination of nine bioactive alkaloids of raw and processed CT in rat plasma. Nitidine chloride was selected as internal standard. After protein precipitation with methanol, the plasma samples were separated on a reversed phased column with a mobile phase of acetonitrile and water (including 0.1% formic acid). The MRM parameters were optimized as follows: m/z 354.0 → 188.1 for protopine, m/z 321.0 → 293.1 for coptisine, m/z 371.1 → 189.1 for allocryptopine, m/z 357.1 → 193.3 for tetrahydropalmatine, m/z 324.1 → 176.1 for tetrahydrocoptisine, m/z 340.1 → 176.2 for tetrahydroberberine, m/z 368.1 → 289.1 for corynoline, m/z 370.5 → 192.1 for corydaline, m/z 367.2 → 351.2 for dehydrocorydaline, and m/z 406.0 → 300.3 for the IS. The linearity, accuracy, precision, stability, recovery and matrix effect of the method were well validated. This method was successfully employed for a pharmacokinetic study of raw and vinegar-processed CT in rats. The absorption of the nine alkaloids was accomplished in an hour. The double peak phenomenon of the nine alkaloids may be ascribed to enterohepatic recirculation. Compared with the raw group, AUC0→t and Cmax of the nine alkaloids were significantly elevated in the vinegar-processed group. Our findings suggest that vinegar-processing could increase the bioavailability of the nine alkaloids of CT in rats. The pharmacokinetic information obtained will provide basis for application of processed CT in future clinical therapy.

Herbal Textual Research on Olibanum in Famous Classical Formulas / 中国实验方剂学杂志

By consulting the ancient and moderm literature， this paper makes a textual research on the name， origin， quality evaluation， harvesting and processing of Olibanum， so as to provide a basis for the development of the famous classical formulas containing this medicinal material. According to the herbal textual research， the results showed that Olibanum was first described as a medicinal material by the name of Xunluxiang in Mingyi Bielu（《名医别录》）， until Ruxiang had been used as the correct name since Bencao Shiyi（《本草拾遗》） in Tang dynasty. The main origin was Boswellia carterii from Burseraceae family. The mainly producing areas in ancient description were ancient India and Arabia， while the modern producing areas are Somalia， Ethiopia and the southern Arabian Peninsula. The medicinal part of Olibanum in ancient and modern times is the resin exuded from the bark， which has been mainly harvested in spring and summer. It is concluded that the better Olibanum has light yellow， granular， translucent， no impurities such as sand and bark， sticky powder and aromatic smell. There were many processing methods in ancient times， including cleansing（water flying， removing impurities）， grinding（wine grinding， rush grinding）， frying（stir-frying， rush frying， wine frying）， degreasing， vinegar processing， decoction. In modern times， the main processing methods are simplified to cleansing， stir-frying and vinegar processing. Nowadays， the commonly used specifications include raw， fried and vinegar-processed products. Among the three specifications， raw products is the Olibanum after cleansing， fried products is a kind of Olibanum processed by frying method， vinegar-processed products is the processed products of pure frankincense mixed with vinegar. Based on the research results， it is recommended to select the resin exuded from the bark of B. carterii for the famous classical formulas such as Juanbitang containing Olibanum， processing method should be carried out in accordance with the processing requirements of the formulas， otherwise used the raw products if the formulas without clear processing requirements.

Optimal processing technology of Zhangbang vinegar-processed Olibanum with multi-indicator-response surface methodology and anticoagulant effect evaluation / 中国中药杂志

This study first optimized the processing technology for Zhangbang vinegar-processed Olibanum and investigated its in vitro anticoagulant activity. A multi-index-response surface methodology was used, with yield, powder yield, and the relative percentage of the content of six non-volatile components [11-keto-boswellic acid(KBA), 3-acetyl-11-keto-boswellic acid(AKBA), β-elemonic acid, α-boswellic acid(α-BA), β-boswellic acid(β-BA), and α-acetyl-boswellic acid(α-BA)] and three volatile components(octyl acetate, incensole, and incensole acetate) as evaluation indicators. Analytical hierarchy process(AHP) combined with coefficient of variation method was used to calculate the weight of each indicator and calculate the comprehensive score(OD). Furthermore, response surface methodology was used to investigate the effects of frying temperature(A), burning time(B), rice vinegar dosage(C), and steaming time(D) on the processing technology of vinegar-processed Olibanum. Vinegar-steamed Olibanum was prepared according to the optimal processing technology for in vitro anticoagulant experiments. The results showed that the weights of octyl acetate, incensole, incensole acetate, KBA, AKBA, β-elemonic acid, α-BA, β-BA, α-ABA, yield, and powder yield were 0.358 2, 0.104 5, 0.146 4, 0.032 9, 0.123 7, 0.044 4, 0.022 1, 0.042 2, 0.110 1, 0.012 2, and 0.0032, respectively. The optimal processing technology for Zhangbang vinegar-processed Olibanum was as follows. Olibanum(50 g) with a particle size of 1-5 mm was continuously stir-fried at a low heat of 150-180 ℃ until in a gel-like state, ignited for burning for 15 s, sprayed with 7.5 g of rice vinegar(15%), and steamed for 3 min without fire. Subsequently, the cover was removed, and the product was continuously stir-fried at 150-180 ℃ until in a soft lump shape, removed, cooled, and crushed. The results of the in vitro anticoagulant experiments showed that compared with the blank group, both Olibanum and vinegar-processed Olibanum significantly prolonged the activated partial thromboplastin time(APTT), thrombin time(TT), and prothrombin time(PT) of rat platelet-poor plasma(PPP), and the effect of vinegar-processed Olibanum was significantly better than that of Olibanum(P<0.05). The optimized processing technology for Zhangbang vinegar-processed Olibanum is stable, feasible, and beneficial for the further development and utilization of Olibanum slices. At the same time, using the content of volatile and non-volatile components, yield, and powder yield as indicators, and verifying through pharmacological experiments, the obtained results are more reasonable and credible, and have positive guiding significance for the clinical application of characteristic processed Olibanum products.

Exploring Synergistic Mechanism of Vinegar-processed Olibanum in Treatment of Ulcerative Colitis Based on Bile Acid-regulated "Gut-liver" Crosstalk / 中国实验方剂学杂志

ObjectiveTo investigate the synergistic mechanism of vinegar-processed Olibanum on ulcerative colitis（UC） via the bile acids regulating "gut-liver" crosstalk. MethodRats were randomly divided into normal group， model group， Olibanum group and vinegar-processed Olibanum group. UC model of rats was induced by intracolonic instillation of 2，4，6-trinitrobenzenesulfonic acid（TNBS）. Ultra high performance liquid chromatography-triple quadrupole-mass spectrometry（UPLC-QQQ-MS） was used to perform the qualitative analysis of 30 bile acids in the colon of rats. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to detect changes in the expression of farnesoid X receptor（FXR）， fibroblast growth factor 15（FGF15） and FGF receptor 4（FGFR4） in "gut-liver" crosstalk at mRNA and protein levels. And with the help of HcoEpiC cell model intervened by conjugated bile acids， simulating the UC state， and according to the different modes of intervention， they were divided into the blank group， conjugated bile acid group， Olibanum group， vinegar-processed Olibanum group and 3-O-acetyl-9，11-dehydro-β-boswellic acid（ADHBA） group. The effect of Olibanum before and after processing with vinegar and the main differential component ADHBA on the mRNA expression of FXR and FGF19 were explored by Real-time PCR. ResultCompared with the normal group， the levels of conjugated bile acids in the model group increased significantly（P<0.01）， and the mRNA and protein expressions of "gut-liver" crosstalk factors FXR， FGF15 and FGFR4 decreased significantly（P<0.05， P<0.01）. Compared with the model group， the content of conjugated bile acids in the Olibanum group and vinegar-processed Olibanum group was significantly decreased（P<0.01）， the mRNA and protein expressions of FXR， FGF15 and FGFR4 were significantly elevated（P<0.05， P<0.01）， and vinegar-processed Olibanum exhibited superior effects than Olibanum. In cellular experiments， a significant decrease in mRNA expression of FXR and FGF19 was observed in the conjugated bile acid group when compared with the blank group（P<0.01）. Compared with the conjugated bile acid group， the mRNA expressions of FXR and FGF19 were significantly higher in the Olibanum， vinegar-processed Olibanum and ADHBA groups（P<0.05， P<0.01）， and the effect of vinegar-processed Olibanum was more favorable. ConclusionVinegar-processed Olibanum may enhance the ameliorating effect on UC by enhancing the down-regulation of conjugated bile acids in the colon and the up-regulation of FXR-FGF15/19-FGFR4 "gut-liver" crosstalk pathway， and ADHBA may be the main material basis for the synergy.

Establishment of Characteristic Pattern of Vinegar-processed Schisandra chinensis Formula Granules and Content Determination of 5 Components / 中国药房

OBJECTIVE：To establ ish characteristic pattern of vinegar-processed product of Schisandra chinensis formula granules from different habitats ，and to determine the contents of 5 components. METHODS ：HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C 18 column with mobile phase consisted of acetonitrile-water （gradient elution）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃，and detection wavelength was set at 220 nm. The sample size was 10 µL. Using schisandrin as reference ，HPLC characteristic pattern of 19 batches of vinegar-processed S. chinensis formula granules was drawn. The similarity evaluation was performed with Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition），and common peaks were confirmed. The contents of schisandrin ，schisandrol， angeloylgomisin H ，schizandrin and deoxyschizandrin were determined by same method. RESULTS ：There were 8 common peaks in 19 batches of vinegar-processed S. chinensis formula granules ，and the similarities were all above 0.996；five of them were identified as schisandrin ，schisandrol，angeloylgomisin H ，schizandrin and deoxyschizandrin ，respectively. The linear range of schisandrin，schisandrol，angeloylgomisin H ，schizandrin and deoxyschizandrin were 0.030-0.380，0.016-0.195，0.009-0.115， 0.006-0.078 and 0.011-0.138 μg（r＞0.999），respectively. RSDs of precision ，stability（24 h）and reproducibility tests were all lower than 2％. Average recoveries were 99.84％，99.54％，99.28％，100.03％，100.27％（RSD＜1.4％，n＝9）. Average contents of five components in 19 batches of samples were in the range of 0.15％-0.36％，0.02％-0.16％，0.02％-0.06％，0.02％-0.08％ and 0.08％-0.17％，respectively；among them ，total contents of five components in sample S 18 and S 19 from Hebei province were relatively high ，while those were relatively low in sample S 16 and S 17（RSD＝42％）；RSD of total content in other samples （S1-S15）was 12％，and was lower than that of Hebei province ；total content of five components were higher in sample from Jilin province. CONCLUSIONS ： Established characteristic pattern and method for the content determination are specific and reproducible，and can be used for the quality evaluation of vinegar-processed S. chinensis formula granules. The total content fluctuation of vinegar-processed product of S. chinensis formula granules from Liaoning ，Jilin and Heilongjiang provinces is lower than that in Hebei province ，and the quality of vinegar-processed Δ 基金项目：国家中药标准化项目（No.ZYB2H-Y-GD-13） ＊主管中药师 ，硕士 。研究方向 ：中药质量标准 。E-mail： S. chinensis formula granules from Jilin province is the best. lzz332@126.com

Anti-herpes Activity of Vinegar-processed Daphne genkwa Flos Via Enhancement of Natural Killer Cell Activity.

Herpes simplex virus (HSV) is a common causative agent of genital ulceration and can lead to subsequent neurological disease in some cases. Here, using a genital infection model, we tested the efficacy of vinegar-processed flos of Daphne genkwa (vp-genkwa) to modulate vaginal inflammation caused by HSV-1 infection. Our data revealed that treatment with optimal doses of vp-genkwa after, but not before, HSV-1 infection provided enhanced resistance against HSV-1 infection, as corroborated by reduced mortality and clinical signs. Consistent with these results, treatment with vp-genkwa after HSV-1 infection reduced viral replication in the vaginal tract. Furthermore, somewhat intriguingly, treatment of vp-genkwa after HSV-1 infection increased the frequency and absolute number of CD3(-)NK1.1(+)NKp46(+) natural killer (NK) cells producing interferon (IFN)-γ and granyzme B, which indicates that vp-genkwa treatment induces the activation of NK cells. Supportively, secreted IFN-γ was detected at an increased level in vaginal lavages of mice treated with vp-genkwa after HSV-1 infection. These results indicate that enhanced resistance to HSV-1 infection by treatment with vp-genkwa is associated with NK cell activation. Therefore, our data provide a valuable insight into the use of vp-genkwa to control clinical severity in HSV infection through NK cell activation.

Functional restoration of exhausted CD4(+) and CD8(+) T cells in chronic viral infection by vinegar-processed flos of Daphne genkwa.

T-cell exhaustion has become an important issue in chronic infection because exhausted antigen-specific T cells show impaired abilities to eradicate persistently infected pathogens and produce effector cytokines, such as IFN-γ and TNF-α. Thus, strategies to either restore endogenous exhausted T cell responses or provide functional T cells are needed for therapeutics of chronic infection. Despite promising developments using antibodies and cell immunotherapy, there have been no reported attempts to restore exhausted T cells using treatment with materials derived from natural resources. Here, using a mouse model of chronic infection with lymphocytic choriomeningitis virus (LCMV), we found that vinegar-processed flowers (flos) of Daphne genkwa (vp-genkwa), which was composed mainly of four index components, restored exhausted CD4(+) and CD8(+) T cells significantly, as corroborated by evidence that vp-genkwa treatment enhanced functional LCMV-specific CD4(+) and CD8(+) T cells, both quantitatively and qualitatively. Furthermore, pretreatment with vp-genkwa prevented the generation of exhausted LCMV-specific CD8(+) T cells. Such restorations of exhausted LCMV-specific CD4(+) and CD8(+) T cells by vp-genkwa were closely associated with reduced viral burden in sera and tissues. More interestingly, vp-genkwa treatment induced down-regulation of negative molecules, such as PD-1 and Tim-3, in exhausted CD4(+) and CD8(+) T cells with more apparent down-regulation of Tim-3, suggesting that Tim-3 molecule may be a major target in restoring exhausted T cell responses. Collectively, these results provide valuable new insights into the use of vp-genkwa to develop a therapeutic strategy for chronic human diseases, such as hepatitis B and C virus, human immunodeficiency virus, and cancers.