Effects of the adjunctive therapy of Sun's abdominal acupuncture on acute ischemic stroke and inflammatory factors: a randomized controlled trial. / å­™æ°è ¹é’ˆè¾ åŠ©æ²»ç–—æ€¥æ€§è„‘æ¢—æ­»åŠå¯¹ç‚Žæ€§å› å­çš„å½±å“ï¼šéšæœºå¯¹ç §è¯•éªŒ.

OBJECTIVES: To compare the clinical effect on acute ischemic stroke (AIS) between the combined treatment of Sun's abdominal acupuncture combined with the routine acupuncture and the simple routine acupuncture, and explore the influences on inflammatory factors i.e. interleukin (IL)-1ß and IL-10. METHODS: Eighty patients with AIS were randomly divided into an observation group (40 cases, 1 case dropped out) and a control group (40 cases, 1 case dropped out). The routine regimens of western medicine were administered in the two groups. In addition, the routine scalp acupuncture and the body acupuncture were used in the patients of the control group. The scalp acupuncture stimulation region and acupoints included the anterior parietal-temporal oblique line, Hegu (LI 4), Chize (LU 5), Shousanli (LI 10), etc. of affected side. In the observation group, on the base of the intervention of the control group, electroacupuncture was applied to "fouth abdominal area" of Sun's abdominal acupuncture, with the continuous wave and the frequency of 5 Hz. In the two groups, the intervention was given twice per day, once in every morning and afternoon separately, with the needles retained for 40 min in each intervention. The interventions were delivered for 6 days a week, lasting 3 weeks. The scores of Fugl-Meyer assessment scale (FMA), Berg balance scale (BBS) and the modified Barthel index (MBI), and the levels of IL-1ß and IL-10 in the serum were observed before and after treatment in the two groups; the effect and safety of interventions were compared between the two groups. RESULTS: After treatment, the scores of FMA, BBS and MBI increased in comparison with those before treatment in the two groups (P<0.01), and these scores in the observation group were higher than those in the control group (P<0.05, P<0.01). After treatment, in the two groups, the levels of IL-1ß in the serum were reduced in comparison with those before treatment (P<0.01), and the IL-1ß level in the observation group was lower than that in the control group (P<0.05); the levels of IL-10 in the serum were elevated in comparison with those before treatment in the two groups (P<0.01) and the IL-10 level in the observation group was higher than that in the control group (P<0.05). The total effective rate was 92.3% (36/39) in the observation group, which was superior to that in the control group (84.6% [33/39], P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups (P>0.05). CONCLUSIONS: Sun's abdominal acupuncture combined with the routine acupuncture can ameliorate the motor impairment, adjust the balance dysfunction and improve the activities of daily living in the patients with AIS. The therapeutic effect of this combined regimen is better than that of the routine acupuncture, which may be associated with the regulation of the inflammatory factors after cerebral infarction.

[Component identification and analysis in vivo of Sanhan Huashi formula].

Sanhan Huashi formula(SHF) is the intermediate of a newly approved traditional Chinese medicine(TCM) Sanhan Huashi Granules for the treatment of COVID-19 infection. The chemical composition of SHF is complex since it contains 20 single herbal medicines. In this study, UHPLC-Orbitrap Exploris 240 was used to identify the chemical components in SHF and in rat plasma, lung and feces after oral administration of SHF, and heat map was plotted for characterizing the distribution of the chemical components. Chromatographic separation was conducted on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 µm) using 0.1% formic acid(A)-acetonitrile(B) as mobile phases in a gradient elution. Electrospray ionization(ESI) source was used to acquire data in positive and negative mode. By reference to quasi-molecular ions and MS/MS fragment ions and in combination with MS spectra of reference substances and compound information in literature reports, 80 components were identified in SHF, including 14 flavonoids, 13 coumarins, 5 lignans, 12 amino-compounds, 6 terpenes and 30 other compounds; 40 chemical components were identified in rat plasma, 27 in lung and 56 in feces. Component identification and characterization of SHF in vitro and in vivo lay foundations for disclosure of its pharmacodynamic substances and elucidation of the scientific connotation.

Construction of a Meta-Evidence Prototype Database of Traditional Chinese Medicine Splenogastric Diseases and Its Application in an Automatic Meta-Analysis System.

Background: Traditional Chinese medicine splenogastric diseases (TCMSDs) are equivalent to digestive system diseases in modern medicine. The forms of clinical evidence of TCMSDs include clinical trials, such as randomized controlled trials (RCTs) and systematic reviews (SRs). SRs mainly rely on manual operations and have the shortcomings of time consumption and low efficiency; therefore, they cannot meet the needs of rapid clinical decision-making. It is urgent to establish a new and smart form of a database to support the progress of SRs. Methods: We searched and screened all TCMSD RCT reports, in both Chinese and English, and extracted them into meta-evidence through predesigned structural Microsoft Excel tables. All meta-evidence was imported into an online clinical meta-evidence collection and management system after data quality checking. The meta-evidence database of traditional Chinese medicine (TCM) splenogastric disease (MED-TCMSD) was then tested as a backend of an automatic meta-analysis system. Results: A total of 405 cases of TCMSD RCTs were processed into meta-evidence. The most common diseases were stomach stuffiness disease, epigastralgia, and chronic atrophic gastritis. Banxiaxiexin decoction and its modifications were the most used interventions. More than half of the cases employed TCM in conjunction with regular therapeutics. The top reported outcomes included clinical effects, adverse events, and TCM syndromes. The MED-TCMSD worked well as a part of the automatic meta-analysis system. Conclusions: We developed and tested a new form of clinical evidence, meta-evidence, for automatic SR and fast evidence-based decision-making. As an example of the MED, the MED-TCMSD can improve the production and updating efficiency of the evidence of TCMSDs. The methods of constructing the MED-TCMSD can be further applied to the development of MEDs of other diseases.

Comparison of triterpene compounds of four botanical parts from Poria cocos (Schw.) wolf using simultaneous qualitative and quantitative method and metabolomics approach.

Poria cocos (Schw.) Wolf (PCW) is a widely used traditional Chinese medicine and dietary supplement. Its four parts including Poriae Cutis (PC), Rubra Poria (RP), White Poria (WP) and Poria cum Radix Pini (PRP) have different pharmacological effects and clinical applications. It is necessary to establish analytical platforms for differentiating the chemical composition of four botanical parts for the rational utilization. We established a simultaneous qualitative and quantitative method based on UHPLC-MS combined metabolomics approach to give an explanation of the distribution of triterpene compounds in four parts. Eight triterpene compounds were determined absolutely and a total of 51 triterpene compounds were tentatively identified in PCW. PC and PRP showed a quite clear discrimination by the principal component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) and twelve differential compounds were found. Four compounds including poricoic acid D, 16α-hydroxydehydrotrametenolic acid, 3-epidehydrotumulosic acid, 25-hydroxypolyporenic acid C were speculated to be related to diuretic effects.

Metabolites study on 5-O-methylvisammioside in vivo and in vitro by ultra high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry.

5-O-Methylvisammioside is one of major chromones of Radix Saposhnikoviae possessing definite pharmacological activities, but there are few reports with respect to the metabolism of 5-O-methylvisammioside. In this work, metabolites in vivo were explored in male Sprague-Dawley rats and in vitro investigated on rat intestinal bacteria incubation model and were identified by using ultra high performance liquid chromatography/quadrupole time-of-flight mass spectrometry. An online data acquisition method based on a multiple mass defect filter and dynamic background subtraction was developed to trace all probable metabolites. As a result, 26 metabolites in vivo (including 18, 15, 10, and 10 in rat urine, faece, bile, and blood) and 7 metabolites in vitro were characterized, respectively. Additionally, the main metabolic pathways in vivo and in vitro, including deglycosylation, deglycosylation + demethylation, deglycosylation + oxidation, N-acetylation, and sulfate conjugation, were summarized by calculating the relative content of each metabolite. The obtained results significantly enriched our knowledge about 5-O-methylvisammioside metabolism and will lead to a better understanding of its safety and efficacy.

Structural characterization and screening of chemical markers of flavonoids in Lysimachiae Herba and Desmodii Styracifolii Herba by ultra high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry based metabolomics approach.

In traditional Chinese medicine, Lysimachiae Herba (LH) and Desmodii Styracifolii Herba (DSH) have been widely used for the treatment of calculi, but there is a certain focus in clinical application. Flavonoids as their pharmacologically active substances were focusly studied to make clear of their chemical compositions and reveal the similarities and differences between LH and DHS by analysis of characteristic marker components at the molecular level. An ultra high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) approach based on metabolite profiling was established. The high-resolution data was acquired through data dependent acquisition (DDA) mode. Based on the targeted and untargeted analytical strategies, a total of 113 compounds were identified, of which 80 compounds existed in LH and 61 in DSH. Then multivariate statistical analysis was applied to further find the characteristic marker components, and a total number of 21 variables were screened as the valuable variables for discrimination. By matching with identified flavonoids, these 21 variables were corresponding to 15 flavonoids (including 6 from LH and 9 from DSH) which were firstly identified as the marker compounds. These results indicated that the UPLC-QTOF-MS/MS method with analysis strategy was a powerful tool for rapidly identification and screening of marker compounds of flavonoids between LH and DSH, and the 15 screened marker compounds provide a chemical basis for the further researches on the mechanisms of LH and DSH in the treatment of cholelithiasis and nephrolithiasis respectively.

Determination and Quantitative Comparison of Nucleosides in two Cordyceps by HPLC-ESI-MS-MS.

A hybrid quadruple linear ion trap liquid chromatography-tandem mass spectrometry (LC-MS-MS) analytical method has been developed for simultaneous determination of 13 nucleosides from Cordyceps cicadae and Cordyceps sinensis for assessing whether C. cicadae can become a substitute for C. sinensis, based on that C. cicadae has in common with C. sinensis for treating chronic diseases. Among the 13 compounds, three compounds including cytidine, hypoxanthine and 2'-deoxyguanosine were firstly identified and quantified in C. cicadae. Ideal separation was achieved in one single LC-MS-MS run of 12 min by optimized chromatographic conditions. The identification and quantification analysis of target compounds were performed in electrospray ionization tandem mass spectrometry. The limit of detection and quantity for 13 compounds were <42.0 and 84.2 ng/mL, respectively. The determination results of 12 batches of C. sinensis and 20 batches of C. cicadae were then analyzed and classified by multivariate statistical analysis. C. cicadae contained a relatively higher level of three nucleosides (cytidine, uracil and 2'-deoxyuridine) than those in C. sinensis. The results of this study provided the theoretical basis for the substitution of C. sinensis with C. cicadae. The optimized method could be used for the quality control and investigate bioactive compound variation of Cordyceps.

Simultaneous detection of four flavonoids and two alkaloids in rat plasma by LC-MS/MS and its application to a comparative study of the pharmacokinetics between Abri Herba and Abri mollis Herba extract after oral administration.

Abri Herba and Abri mollis Herba both were important members of the Leguminosae family in southwestern China. Abri mollis Herba was often used as Abri Herba due to their proximity, but there are few studies on pharmacokinetics to compare their main identical active compositions. A sensitive and selective high-performance liquid chromatography with tandem mass spectrometry method in the positive/negative electrospray ionization switching mode was developed and validated for the simultaneous analysis of four flavonoids and two alkaloids in rat plasma. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of methanol and 0.5% acetic acid. The detection of the target compounds was conducted in multiple-reaction monitoring mode with a hybrid triple quadrupole linear ion trap mass spectrometer equipped with positive/negative ion-switching electrospray ion source. The differences in pharmacokinetics were discovered, which indicated that the substitution between them is some degree of irrationality. The validated method was successfully applied to pharmacokinetic study of the six components in male rat plasma after oral administration of Abri Herba and Abri mollis Herba extract and the results in the study would provide a useful guide for the clinical application of Abri Herba with those in Abri mollis Herba.

[Simultaneous determination of 8 constituents of Desmodii Styracifolii Herba in rat bile with HPLC-MS/MS and their excretion pharmacokinetics].

To establish a HPLC-MS/MS quantitative method for analysis of 8 constituents in rat bile. The method was applied in the biliary excretion study after oral administration of Desmodii Styracifolii Herba extract. The rat bile samples were collected during 0-1, 1-2, 2-4, 4-6, 6-8, 8-12, 12-24 h. Diamonsil C18column (4.6 mm×150 mm, 5 µm) was adopted and eluted with methanol and 0.01% acetic acid in a gradient mode. The flow rate was 0.8 mL•min⁻¹, and the column temperature was set at 40 ℃. The detection was carried out by a triple quadrupole linearion trap mass spectrometer in the negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. The calibration curves showed a good linearity, with correlation coefficients (r≥0.991 5) for all of the analytes within the concentration range. The intra-day and inter-day precisions (RSD) were both less than 15%, and the accuracies (RE) ranged from -15% to 15%. The method had a good stability, and was suitable for the content determination of 8 constituents in rat bile. The extraction recoveries of the 8 analytes were more than 63.2%, and the RSD of IS-normalized matrix factors were no more than 15%, which met the requirements for analysis. According to the biliary excretion study, the bile excretion rates of the 8 analytes were relatively low, with great difference among the individuals. The results showed that the established method was simple, selective and specific, thus could be applied in the biliary excretion study of the 8 analytes in rat bile.

Simultaneous determination of nine coumarins in rat plasma by HPLC-MS/MS for pharmacokinetics studies following oral administration of Fraxini Cortex extract.

A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for the simultaneous determination of nine coumarins including aesculin, aesculetin, fraxin, fraxetin, scopoletin, isoscopoletin, 6-hydroxy-7,8-dimethoxy coumarin, 8-hydroxy-6,7-dimethoxy coumarin and umbelliferone in rat plasma using nodakenin as the internal standard (IS). The plasma samples were pretreated by a one-step direct protein precipitation with methanol. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of methanol and water (containing 0.05% acetic acid). All analytes and IS were quantitated through electrospray ionization in negative ion multiple reaction monitoring (MRM) mode. This method was fully validated in terms of the sensitivity, specificity, accuracy, precision (intra- and inter-day), matrix effect, recovery as well as the stability of the analyte under various conditions, and the results satisfied the requirements of biological sample measurement. The validated method was successfully applied to pharmacokinetic study of the nine coumarins in rat plasma after oral administration of Fraxini Cortex aqueous extract, among which the pharmacokinetics of four coumarins including fraxetin, isoscopoletin, 6-hydroxy-7,8-dimethoxy coumarin and 8-hydroxy-6,7-dimethoxy coumarin were studied for the first time.

Comparison of pharmacokinetic profiles of Terminalia phenolics after intragastric administration of the aqueous extracts of the fruit of Terminalia chebula and a Mongolian compound medicine-Gurigumu-7.

ETHNOPHARMACOLOGICAL RELEVANCE: The dried fruit of Terminalia chebula (fructus chebulae) is an important Traditional Medicine used for intestinal and hepatic detoxification. Gurigumu-7 which is made of fructus chebulae and 6 other traditional medicines is one of the most frequently used compound Mongolian and Tibet medicines for liver diseases. Terminalia phenolics are considered as the bioactive constituents of fructus chebulae and consequently of Gurigumu-7. AIM OF THE STUDY: To compare the pharmacokinetic profiles of Terminalia phenolics after intragastric administration of the aqueous extracts of fructus chebulae and Gurigumu-7 and to evaluate the possible influence of intestinal bacterial metabolism on these pharmacokinetic profiles. MATERIALS AND METHODS: An ultra performance liquid chromatography with triple quadrupole mass spectrometry method was established and validated for simultaneously determining the pharmacokinetic profiles of seven Terminalia phenolics after intragastric administration of pure compounds, fructus chebulae extract, and Gurigumu-7 extract. In vitro rat fecal lysates experiments were carried out to explore the metabolic discrepancy between fructus chebulae and Gurigumu-7. RESULTS: Seven Terminalia phenolics were detected in rat plasma after intragastric administration of the aqueous extracts of fructus chebulae and Gurigumu-7. Administration of Gurigumu-7 could promote the absorption and increase the Cmax and AUC values of these phenolic constituents compared to fructus chebulae administration. The fecal lysates studies showed that the Terminalia phenolics in Gurigumu-7 were less rapidly bio-transformed than those in fructus chebulae. This may be a contributing factor to the pharmacokinetic discrepancy between the phenolics in fructus chebulae and Gurigumu-7. CONCLUSION: Administration of Gurigumu-7 could increase the absorption of Terminalia phenolics through slowing down the intestinal bacteria metabolism. These results provide, in part, an in vivo rationale for the formulation of the traditional Mongolia / Tibet medicine, Gurigumu-7.

Simultaneous Detection of Flavonoids, Phenolic Acids and Alkaloids in Abri Herba and Abri Mollis Herba using Liquid Chromatography Tandem Mass Spectrometry.

INTRODUCTION: Abri Herba has remarkable properties, such as cleanup heat detoxification, dampness and activating blood circulation to dissipate blood stasis; as a result, it has been applied to treat acute or chronic hepatitis and mastitis. Abri mollis Herba is often used as Abri Herba. Hierarchical cluster analysis (HCA) was applied to compare the similarities and differences of the chemical compositions in the two types of medicinal materials. OBJECTIVE: To establish a high-performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS) method for the simultaneous analysis of 15 flavonoids, two phenolic acids and three alkaloids in Abri Herba and Abri mollis Herba. METHODOLOGY: The chromatographic separation was performed on a C18 column with a mobile phase of methanol (A), acetonitrile (B) and 0.5‰ acetic acid in water (C) using gradient elution. The detection of the target compounds was performed in multiple-reaction monitoring (MRM) mode using a hybrid quadrupole linear ion trap mass spectrometer equipped with positive/negative ion-switching electrospray ionisation (ESI) source. RESULTS: The developed method is reliable, sensitive and specific. In addition, the method has been successfully applied to differentiate 15 batches of Abri Herba and 27 batches of Abri mollis Herba stems. Furthermore, a comparison of the contents among stems, roots and leaves from the same strain in seven batches of Abri mollis Herba and four batches of Abri Herba has also been performed. CONCLUSION: HPLC-MS/MS method is sensitive and selective and can be suitable for the reliable quality control of Abri mollis Herba and Abri Herba.

[Effect of Preconditioning of Catgut Implantation at Acupoints on Changes of Behavior and EEG in Epileptic Rats].

OBJECTIVE: To observe the effect of preconditioning of catgut implantation on changes of behavior and EEG of rats with epilepsy. METHODS: Forty male Wistar rats were randomly divided into control, model, catgut implantation, medication groups (10 rats/group). The catgut implantation was performed at "Yintang" (EX 2) to "Baihui" (GV 20) and "Changqiang" (GV 1) on day 1, 4 and 7. The epileptic model was made by intraperitoneal injection of penicillin sodium on the last day of the treatment. In the medication group, intragastric administration of valproic acid sodium was conducted once a day for consecutive 7 days. Behavior measurement and EEG recording were carried out shortly after the penicillin administration. RESULTS: Compared with the model group, the seizure latency was notably increased in the catgut implantation group and the medication group (P < 0.01), without significant difference between the last two groups (P > 0.05). The degree of the seizure attack was less severe in the catgut implantation group and the medication group (P < 0.01), without significant difference between the two groups (P > 0.05). Similar to valproic acid sodium, catgut implantation significantly reduced the amplitude and accelerated the frequency of EEG (P < 0.01). CONCLUSION: Catgut implantation can delay the seizure attack latency, alleviate the severity, regulate the amplitude and frequency of the epileptiform discharges, giving rise to evident antiepileptic effect.

Simultaneous quantification of 25 active constituents in the total flavonoids extract from Herba Desmodii Styracifolii by high-performance liquid chromatography with electrospray ionization tandem mass spectrometry.

A sensitive and selective high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry method has been developed and validated for the simultaneous determination of 25 active constituents, including 21 flavonoids and four phenolic acids in the total flavonoids extract from Herba Desmodii Styracifolii for the first time. Among the 25 compounds, seven compounds including caffeic acid, acacetin, genistein, genistin, diosmetin, diosmin and hesperidin were identified and quantified for the first time in Herba Desmodii Styracifolii. Chromatographic separation was accomplished on a ZORBAX SB-C18 (250 mm×4.6 mm, 5.0 µm) column using gradient elution of methanol and 0.1‰ acetic acid v/v at a flow rate of 1.0 mL/min. The identification and quantification of the analytes were achieved using negative electrospray ionization mass spectrometry in multiple-reaction monitoring mode. The method was fully validated in terms of limits of detection and quantification, linearity, precision and accuracy. The results indicated that the developed method is simple, rapid, specific and reliable. Furthermore, the developed method was successfully applied to quantify the 25 active components in six batches of total flavonoids extract from Herba Desmodii Styracifolii.

Symptom clustering in chronic gastritis based on spectral clustering.

OBJECTIVE: Apply spectral clustering to analyze the patterns of symptoms in patients with chronic gastritis (CG). METHODS: Based on 919 CG subjects, we applied mutual information feature selection to choose the positively correlated symptoms with each pattern. Then, we used the Shi and Malik spectral clustering algorithm to select the top 20 correlated symptoms. RESULTS: We ascertained the results of six patterns. There were three categories for the pattern of accumulation of damp heat in the spleen-stomach (0.00332). There were six categories for the pattern of dampness obstructing the spleen-stomach (0.02466). There were two categories for the pattern of spleen-stomach Qi deficiency (0.013 89). There were three categories for the pattern of spleen-stomach deficiency cold (0.009 15). There were five categories for the pattern of liver-Qi stagnation (0.01910). There were four categories for the pattern of stagnant heat in the liver-stomach (0.00585). CONCLUSION: Most of the spectral clustering results of the symptoms of CG patterns were in accordance with clinical experience and Traditional Chinese Medicine theory. Most categories suggested the nature and/or location of the disease.

Simultaneous determination of four flavonoids and one phenolic acid in rat plasma by LC-MS/MS and its application to a pharmacokinetic study after oral administration of the Herba Desmodii Styracifolii extract.

A sensitive and selective liquid chromatography and tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of four flavonoids (schaftoside, isovitexin, luteolin, and apigenin) and one phenolic acid (ferulic acid) in rat plasma using sulfamethoxazole as the internal standard (IS). The separation was performed using a Diamonsil C18 column, which was eluted with methanol (A) and 0.1‰ acetic acid (B). The gradient condition was as follows: 0-5min, 40-60% A; 5-6min, 60-95% A; and 6-10min, maintained at 95% A. The analytes were detected using a hybrid quadrupole linear ion trap mass spectrometer that was equipped with an electrospray ionization source in the negative ion and multiple-reaction monitoring modes. A full validation of the method was performed. The linearity of the analytical response was good, with correlation coefficients greater than 0.9925 for all of the compounds within the concentration range. The lower limits of quantitation (LLOQ) of schaftoside, isovitexin, luteolin, apigenin, and ferulic acid in rat plasma were 1.66, 0.84, 3.69, 1.70, and 3.91ng/mL, respectively. The intra-day and inter-day precisions of the investigated components exhibited an RSD within 13.20%, and the accuracy (RE%) ranged from -8.47% to 10.90%. The results indicated that the developed method is sufficiently reliable for the pharmacokinetic study of schaftoside, isovitexin, apigenin, luteolin, and ferulic acid in rats following oral administration of the Herba Desmodii Styracifolii extract.

Uniqueness of the Gossypium mustelinum genome revealed by GISH and 45S rDNA FISH.

Gossypium mustelinum ((AD)4 ) is one of five disomic species in Gossypium. Three 45S ribosomal DNA (rDNA) loci were detected in (AD)4 with 45S rDNA as probe, and three pairs of brighter signals were detected with genomic DNA (gDNA) of Gossypium D genome species as probes. The size and the location of these brighter signals were the same as those detected with 45S rDNA as probe, and were named GISH-NOR. One of them was super-major, which accounted for the fact that about one-half of its chromosome at metaphase was located at chromosome 3, and other two were minor and located at chromosomes 5 and 9, respectively. All GISH-NORs were located in A sub-genome chromosomes, separate from the other four allopolyploid cotton species. GISH-NOR were detected with D genome species as probe, but not A. The greatly abnormal sizes and sites of (AD)4 NORs or GISH-NORs indicate a possible mechanism for 45S rDNA diversification following (AD)4 speciation. Comparisons of GISH intensities and GISH-NOR production with gDNA probes between A and D genomes show that the better relationship of (AD)4 is with A genome. The shortest two chromosomes of A sub-genome of G. mustelinum were shorter than the longest chromosome of D sub-genome chromosomes. Therefore, the longest 13 chromosomes of tetraploid cotton being classified as A sub-genome, while the shorter 13 chromosomes being classified as D sub-genome in traditional cytogenetic and karyotype analyses may not be entirely correct.

Simultaneous detection of flavonoids and phenolic acids in Herba Lysimachiae and Herba Desmodii Styracifolii using liquid chromatography tandem mass spectrometry.

A sensitive liquid chromatography and tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous analysis of 15 flavonoids and 3 phenolic acids in Herba Lysimachiae and Herba Desmodii Styracifolii. Separation was performed using a Diamonsil C(18) column, which was eluted with methanol (A) and 0.1‰ acetic acid (B). The gradient condition was as follows: 0-34 min, 20-34% A; 34-38 min, 34-95% A; maintained 95% A for the next 4 min. Analytes were detected using a hybrid quadrupole linear ion trap mass spectrometer that was equipped with an electrospray ionisation source in the negative ion and multiple-reaction monitoring modes. A full validation of the method was performed, including the linearity, precision, and accuracy, as well as limits of detection and quantification. The results indicated that the developed method was simple, sensitive and reliable. Furthermore, the method was successfully applied to differentiate 16 batches of Herba Lysimachiae and 21 batches of Herba Desmodii Styracifolii.

Simultaneous determination of seven components in Qibaomeiran pill by HPLC-DAD.

A high-performance liquid chromatography coupled with photo diode array detection method is developed for simultaneous determination of seven active components in Qibaomeiran pill, including rutin, 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glycoside, ferulic acid, psoralen, isopsoralen, emodin, and physcion. The analysis is performed on a C(18) column using a mobile phase composed of A (0.1% acetic acid) and B (acetonitrile) with linear gradient elution. Four wavelengths at 245, 290, 320, and 350 nm were chosen as the monitoring wavelength to determine seven active components, respectively. All the compounds show good linearity (r > 0.999). The developed method is fully validated in respect to precision, repeatability, and accuracy. The proposed method is successfully applied to quantify the seven active components in different Qibaomeiran pill samples. The results indicate that the developed assay could be considered as a suitable quality control method for the Qibaomeiran pill.

Determination of piceid in rat plasma and tissues by high-performance liquid chromatographic method with UV detection.

A rapid, sensitive and selective HPLC method was developed and validated for determination of piceid in rat plasma and tissues. The drug was isolated from plasma and tissues by a simple protein precipitation procedure. Chromatographic separation was performed on a C(18) column with acetonitrile-water (26:74, v/v) as mobile phase. The method was successfully applied to the pharmacokinetics and tissue distribution research after oral administration of a 50 mg/kg dose of piceid to healthy male Wistar rats. The pharmacokinetic parameters showed that piceid was quickly absorbed, distributed and eliminated within 4 h after oral administration. The tissue distribution results showed that, at 10 min, the concentrations of piceid in most tissues reached peak level except in heart and testis. The highest level of piceid was found in stomach, then in small intestine, spleen, lung, brain, testis, liver, kidney and heart. The amount of piceid in testis and heart reached the peak level at 30 min. At 120 min, the amount of piceid in all tissues decreased to a low percentage of the initial concentration. Piceid was absorbed throughout the gastrointestinal tract with considerable absorption taking place in the stomach and small intestine. There was no long-term accumulation of piceid in rat tissues.