Lactobacillus rhamnosus GG induced protective effect on allergic airway inflammation is associated with gut microbiota.

Great interest has been taken in the use of beneficial bacteria for allergic diseases recently, but the underlying mechanisms through which probiotics induces immune regulation or immune tolerance are poorly understood. We aimed to explore whether Lactobacillus rhamnosus GG (LGG)-induced beneficial effect relates to the change of microbiota. LGG was administered orally to mouse model of ovalbumin (OVA)-induced allergic airway inflammation. Our findings manifested that LGG-treatment contributes to protect against OVA-induced allergic airway inflammation by expanding mesenteric CD103+DCs and accumulating mucosal Tregs. Moreover, protective effect induced by LGG is associated with gut microbiota instead of lung flora. Collectively, our findings indicate that LGG induced protective effect is associated with gut microbiota and provide a new evidence of probiotic application in the allergic airway inflammation.

Pharmacokinetic study of methylnaltrexone after single and multiple subcutaneous administrations in healthy Chinese subjects.

1. Pharmacokinetics of methylnaltrexone (MNTX) were evaluated after subcutaneous administrations (s.c.) in healthy Chinese subjects. 2. In a cross-over single dose study, 12 subjects were given 0.075, 0.15, and 0.3 mg/kg of MNTX bromide injection. In a multiple doses study, another 12 subjects subcutaneously received 0.15 mg/kg of MNTX bromide injection every 48 h, in total five administrations. The concentrations of MNTX in plasma were quantified by LC-MS/MS. 3. After single s.c. administrations of 0.075, 0.15, and 0.3 mg/kg of MNTX bromide, Cmax values of MNTX were 93.5 ± 28.6, 191 ± 37, and 364 ± 54 ng/mL, respectively, and AUC0-∞ were 88.8 ± 8.8, 181 ± 16, and 357 ± 34 ng⋅h/mL, respectively. The t1/2 of MNTX was about 7.7 h. After multiple doses administration, the Cmax, Cav, AUCss, and MRT0-∞ values were 191 ± 50, 3.79 ± 0.40 ng/mL, 182 ± 19 ng⋅h/mL, and 3.56 ± 1.17 h, respectively. 4. Methylnaltrexone bromide displayed dose-proportional pharmacokinetics in the dose range of 0.075-0.3 mg/kg. After multiple doses administration, t1/2 was slightly prolonged, with the cumulative factor of 1.02. This study provides a pharmacokinetic reference after a single dose and multiple doses of MNTX bromide in Chinese subjects.

An LC-MS/MS method for simultaneous determination of cefprozil diastereomers in human plasma and its application for the bioequivalence study of two cefprozil tablets in healthy Chinese volunteers.

A rapid and sensitive liquid chromatography-tandem mass spectrometric method was developed for the first time and validated for the determination of cefprozil diastereomers in human plasma. The plasma samples were prepared by protein precipitation using acetonitrile. Detection was performed using an electronic spray ion source in the negative ion mode, operating in the multiple reaction monitoring of the transitions m/z 388.0 to m/z 205.0 for cefprozil diastereomers and m/z 346.1 to m/z 268.1 for cephalexin (the internal standard). The calibration curves of cis-cefprozil and trans-cefprozil were linear in the ranges 0.125-16.0 µg/mL and 0.0403-1.72 µg/mL, respectively. The lower limits of quantification of cis- and trans-cefprozil were 0.125 and 0.0403 µg/mL in human plasma, respectively. The intra- and inter-day precisions of cis- and trans-cefprozil were all <9.7%, and the accuracy ranged from 99.2 to 104.7% and from 100.6 to 102.2%, respectively. The validated method was successfully applied to a bioequivalence study of two cefprozil formulations in 24 healthy Chinese volunteers. The two cefprozil tablets were bioequivalent by measurement of cis-, trans- and total cefprozil. We suggest that the bioequivalence of cefprozil formulations can be evaluated only using cis-cefprozil as the analyte in future studies.

In Vivo Metabolite Profiling of a Purified Ellagitannin Isolated from Polygonum capitatum in Rats.

Ellagitannin is a common compound in food and herbs, but there are few detailed studies on the metabolism of purified ellagitannins. FR429 is a purified ellagitannin with antitumor potential, which is from Polygonum capitatum Buch.-Ham.ex D. Don. The present study was designed to investigate the metabolic profiles of FR429 in rats in vivo. Using liquid chromatography coupled to ion trap time-of-flight mass spectrometry (LC/MS(n)-IT-TOF), total eight metabolites were found in rat bile and urine after intravenous administration of FR429, but could not be detected in plasma. These metabolites were ellagic acid, mono-methylated FR429, ellagic acid methyl ether glucuronide, ellagic acid methyl ether diglucuronide, ellagic acid dimethyl ether glucuronide, and ellagic acid dimethyl ether diglucuronide. It was concluded that methylation and subsequent glucuronidation were the major metabolic pathways of FR429 in rats in vivo. This is the first report on the in vivo metabolism of the purified ellagitannin in rats.

Identification of Haploid Maize Kernel Using NIR Spectroscopy in Reflectance and Transmittance Modes: A Comparative Study.

The spectra measurements mode that suitable for haploid maize kernel identification was explored using MicroNIR-1700 series of miniature near infrared spectrometer by JDSU company. Based on Near Infrared Spectroscopy (NIRS) qualitative analysis techniques, we conducted a comparative study using reflectance and transmittance spectra to identify haploid maize kernels. Partial least squares-discriminant analysis (PLS-OLDA) was used to compress the pretreated spectral data, and then the identification models were built based on Support Vector Machine (SVM). The measured data were recorded in reflectance and transmittance modes and the recognition correct rates were calculated. For measurements taken in reflectance mode, the average recognition rate was less than 60% regardless of embryo side positions. In transmittance mode, however, the average recognition rate reached 93.2%. The experiment results show that diffuse reflection spectrum could only obtain corn grain surface information, so embryo side positions severely affect haploid maize kernel identification effect when reflectance measurements mode have been employed, but they have far less impact on transmittance mode. The near infrared diffuse transmittance spectra analyzes non-uniform samples can achieve the analysis of optical path depth information accumulation, all information of the sample interior can be obtained, so transmittance spectra could identify haploid maize effectively and be desensitized to kernel positions. NIRS qualitative analysis techniques with features of rapid, nondestructive could identify the haploid and Micro-NIR spectrometer scan fast and cost less, which have utility for automatically selecting haploid maize kernels from hybrid kernels.

Biotransformation and in vitro metabolic profile of bioactive extracts from a traditional Miao-nationality herbal medicine, Polygonum capitatum.

Polygonum capitatum Buch.-Ham.ex D. Don, a traditional Miao-nationality herbal medicine, has been widely used in the treatment of various urologic disorders. Recent pharmacological studies demonstrated that a pure compound, FR429, isolated from the ethanol extracts of P. capitatum could selectively inhibit the growth of four hepatocellular carcinoma (HCC) cell lines in a dose-dependent manner. Thus, P. capitatum probably exhibits potential antitumor activity. However, there is very little information on the metabolism of substances present in P. capitatum extracts. In this study, gallic acid, quercetrin, ethanol extracts and ethyl acetate fraction of ethnolic extract (EtOAc fraction) of P. capitatum were cultured anaerobically with rat intestinal bacteria. A highly sensitive and selective liquid chromatography electrospray ionization-ion trap-time of fight mass spectrometry (LC/MSn-IT-TOF) technique was employed to identify and characterize the resulting metabolites. A total of 22 metabolites (M1-M22), including tannins, phenolic acids and flavonoids, were detected and characterized. The overall results demonstrated that the intestinal bacteria played an important role in the metabolism of P. capitatum, and the main metabolic pathways were hydrolysis, reduction and oxidation reactions. Our results provided a basis for the estimation of the metabolic transformation of P. capitatum in vivo.

The feasibility and clinical significance of lateral approach thyroidectomy.

BACKGROUND: By comparing the three lateral approaches to thyroidectomy, the feasibility and clinical effects were analyzed, and the advantages of the lateral approach were summarized. METHODS: From January 2022 to January 2023, 52 patients with thyroid cancer admitted to our department were selected and subjected to Lateral approach for thyroidectomy. Among them, 31 patients underwent thyroidectomy via the supraclavicular approach, 13 patients underwent endoscopic thyroidectomy via the subclavicular approach, and 8 patients underwent endoscopic thyroidectomy via the axillary approach. The basic conditions, surgical conditions, complications, postoperative pain scores and postoperative satisfaction of patients in the three approach surgery groups were recorded and analyzed. RESULTS: There were no significant differences among the three approach groups in terms of patient characteristics, number of central lymph node dissections, intraoperative blood loss, postoperative drainage volume, duration of drainage tube placement, length of hospital stay, postoperative pain, satisfaction, and complications. However, the operation time was longest in the subclavicular approach group, followed by the axillary approach group, and shortest in the supraclavicular approach group. The total hospitalization cost was highest in the axillary approach group, followed by the subclavicular approach group, and lowest in the supraclavicular approach group. CONCLUSION: The lateral approach for thyroidectomy is deemed a safe and effective method. The three different approach paths gradually increase in length, allowing for the accumulation of anatomical experience. This approach has a shorter learning curve for clinical doctors and is a favorable choice for patients seeking aesthetic benefits.

[Effects of echinacoside on extracellular acetylcholine and choline levels of hippocampus and striatum of cerebral ischemia rats].

The aim of this study is to investigate the effect of echinacoside (ECH) on cholinergic neurotransmitter extracellular of hippocampus and striatum and its possible mechanisms of neuro-protective effect against vascular dementia rats. In this study brain microdialysis technique combined with HPLC-IMER-ECD (high-performance liquid chromatography-immobilized enzyme reactor-electrochemical detector) was used. The bilateral common carotid arteries occluded in two times operation at 72 h interval for vascular dementia model rats were used and the successful vascular dementia model rats were examined by Morris water maze. The content of acetylcholine (ACh) and choline (Ch) of microdialysate extracellular of hippocampus and striatum was determined by HPLC-IMER-ECD and the AChE activity in the hippocampus was measured. The results showed that the success rate of vascular dementia model was 83.08% after six weeks; the results also showed that echinacoside and galantamine could increase the content of ACh and reduce the content of Ch extracellular of hippocampus and striatum significantly and the AChE activity increased significantly compared with that of the model group. The results suggested that echinacoside could promote the recovery of cholinergic neurotransmitter levels in vascular dementia rats' brain, which may be one of the mechanisms of neuro-protection.

Molecular characterization of hexokinase (HK) in Haemaphysalis longicornis and evaluation of HK protein- and DNA-based vaccines against adult ticks.

BACKGROUND: Haemaphysalis longicornis is an obligate hematophagous ectoparasite, which transmits various pathogens to humans, livestock and wild animals. Hexokinase (HK) is a key regulatory enzyme of the glycolytic pathway in the organisms. However, little is known about hexokinase and its functions in ticks. RESULTS: The open reading frame of the H. longicornis HK (HlHK) gene was 1425 bp and encoded a protein of 474 amino acids, containing conserved domains for glucose, glucose 6-phosphate, and adenosine triphosphate. The expression of HlHK gene was detected at different developmental stages and in different tissues of unfed female ticks. Enzyme-linked immunosorbent assay revealed that both HK protein- and DNA-based vaccines increased the antibody levels of the immunized animals. A vaccination trail on rabbits against H. longicornis infestation indicated that the rHlHK protein and HlHK DNA vaccines reduced the number of attached female ticks by 9% and 12%, egg mass weight by 36% and 34%, and egg hatching rate by 41% and 17%, respectively. Overall, protein vaccination conferred 65.6% protection against adult female ticks, whereas the DNA vaccine conferred 51.8% protection. CONCLUSION: This is the first report of the molecular characterization of the HK protein and sequencing of the HK gene from H. longicornis. Positive results from vaccination trials on rabbits of the recombinant HK protein and HK DNA suggest that these novel anti-tick vaccines potentially can be used as viable tick control tools for the management of the Asian longhorned tick. Additionally, inhibition of glucose metabolism may be a new strategy for tick control. © 2023 Society of Chemical Industry.

Angiosarcoma of the breast with hypofibrinogenemia: A rare case report and review of the literature.

Background: Breast angiosarcoma is a rare malignant tumor, accounting for approximately 0.04% of all breast malignancies. Angiosarcoma of the breast with hypofibrinogenemia is even rarer and has not been described in man. Breast angiosarcoma is associated with high metastatic potential and poor prognosis, and there is no specific manifestation in imaging. At present, surgery is considered to be the only effective treatment. There is no unified standard for postoperative adjuvant radiotherapy and chemotherapy. Case Presentation: A 30-year-old female patient underwent left breast mass resection under local anesthesia on May 22, 2014. Postoperative pathology showed a vasogenic tumor. On November 10, 2017, she visited us again due to left breast swelling and pain during lactation, and underwent breast mass puncture. She was diagnosed with breast hematoma and fibrinogen reduction. On November 14, 2017, mastectomy was performed under tracheal intubation and general anesthesia, and the fibrinogen gradually returned to normal after surgery. Pathological examination showed a hemangiosarcoma with hematoma formation in the left breast. According to the pathological findings and after comprehensive evaluation, the patient underwent modified radical mastectomy for left breast cancer and right axillary sentinel lymph node biopsy on December 5, 2017. The patient died on January 28, 2018 due to rupture and hemorrhage of liver cancer and hemorrhagic shock. Conclusion: Breast angiosarcoma with hypofibrinogenemia is a rare and highly aggressive malignancy. Clinicians should be familiar with its clinicopathological features and diagnostic criteria. Multidisciplinary approach is recommended to benefit the patients.

Achieving Partial Nitrification-Anammox Process Dependent on Microalgal-Bacterial Consortia in a Photosequencing Batch Reactor.

Partial nitrification coupled with anammox (PN/A) process is an energy-efficient approach for nitrogen removal from low C/N wastewater. In this study, PN/A was achieved with optimal oxygen supply from a green microalga, Chlorella sorokiniana. The PN process was first initiated within 35 days, and the following algae-intensified PN then reached the steady state within the next 32 days. The dissolved oxygen (DO) concentration was gradually maintained at 0.6 mg L-1 via adjusting the photoperiod to 6-h light/18-h dark cycles, when the accumulation ratio of NO2 --N and the removal ratio of NH4 +-N were both more than 90%. The nitrogen removal capability of anammox was acclimated via elevating the individual effluent NH4 +-N and NO2 --N levels from 100 to 200, to 300 mg L-1. After acclimation, the removal rates of NH4 +-N and total nitrogen (TN) reached more than 70 and 80%, respectively, and almost all the NO2 --N was removed. Then, the algae-intensified PN/A, algammox biofilm system, was successfully started up. When the NH4 +-N level increased from 100 to 300 mg L-1, the TN removal varied between 78 and 82%. In the photosequencing bioreactor, C. sorokiniana, ammonia-oxidizing bacteria (AOB), and anammox coexisted with an illumination of 200 µmol m-2 s-1 and a 6-h light/18-h dark cycles. The DO levels ranged between 0.4 and 0.5 mg L-1. In addition, the microbial community analysis by Illumina MiSeq sequencing showed that the dominant functional bacteria in the algae-intensified PN/A reactors included Nitrosomonas (AOB) and Candidatus Brocadia (anammox), while Nitrospira and Nitrobacter (nitrite oxidizing bacteria), together with Denitratisoma (denitrifier) were largely inhibited. Further studies are required to optimize the microalgal-bacterial consortia system to achieve superior nitrogen removal rates under controllable conditions.

Differences in Glycerolipid Response of Chlamydomonas reinhardtii Starchless Mutant to High Light and Nitrogen Deprivation Stress Under Three Carbon Supply Regimes.

Carbon source serves as a crucial factor for microalgal lipid biosynthesis. The supplied exogenous inorganic or organic carbon affects lipid accumulation in microalgae under stress conditions. However, the impacts of different carbon availability on glycerolipid metabolism, triacylglycerol (TAG) metabolism in particular, still remain elusive in microalgae. Chlamydomonas starchless mutant BAFJ5 has emerged as a model system to study TAG metabolism, due to its property of hyper-accumulating TAG. In this study, the glycerolipidomic response of the starchless BAFJ5 to high light and nitrogen-deprived (HL-N) stress was deciphered in detail to distinguish glycerolipid metabolism under three carbon supply regimes. The results revealed that the autotrophically and mixotrophically grown BAFJ5 cells aerated with air containing 2% CO2 presented similar changes in growth, photosynthetic activity, biochemical components, and glycerolipid metabolism under HL-N conditions. But the mixotrophically grown BAFJ5 aerated with air containing 0.04% CO2 exhibited more superior accumulation in TAG, which was esterified with a significantly higher proportion of C18:1n9 and prominently the lower proportions of polyunsaturated fatty acids. In addition, these cells increased the relative levels of C18:2n6 in the membrane lipids, i.e., monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), in priority, and decreased that of C18:3n3 and C18:4n3 in the betaine lipid, N,N,N-trimethylhomoserine diacylglycerol (DGTS), subsequently, to adapt to the HL-N stress conditions, compared to the cells under the other two conditions. Thus, it was suggested that C. reinhardtii starchless mutant appeared to present distinct metabolism for TAG biosynthesis involving membrane lipid remodeling under distinct carbon supply regimes. This study provides insights into how the different carbon supply regimes affect lipid metabolism in Chlamydomonas starchless cells, which will benefit the optimized production of storage lipids in microalgae.

Chinese herbal medicines for mild cognitive impairment: A protocol for meta-analysis and trial sequential analysis.

BACKGROUND: Mild cognitive impairment (MCI), as a common neurodegenerative aging disease representing an intermediate stage between normal cognitive functioning and dementia, poses an excessive burden on health care. The clinical benefit of Chinese herbal medicines (CHMs) for MCI remains inconclusive. This study is aimed at evaluating the efficacy and acceptability of CHMs through meta-analysis and trial sequential analysis (TSA). METHODS: We applied extensive strategies on preliminary literature screening to identify relevant randomized controlled trials which meticulously compare any of CHMs interventions with placebo groups as monotherapy for MCI. The primary outcome of this study is the change of global cognitive function, and the secondary outcomes include assessments of activities of daily living, mood, and adverse events. Data synthesis, risk of bias assessment, sensitivity and subgroup analyses, and TSA will be conducted with application of Review Manager, Stata, and TSA software. The quality of the evidence will be evaluated using the Grading of Recommendations Assessment, Development and Evaluation instrument. INPLASY registration number: INPLASY202190006 (https://inplasy.com/inplasy-2021-9-0006/). RESULTS: This study will confirm the clinical efficacy and safety of CHMs when used in the treatment of patients with MCI. CONCLUSION: This study will provide reliable evidence and references for the selection of CHMs in therapy and future clinical research of MCI.

Antimicrobial peptides: bridging innate and adaptive immunity in the pathogenesis of psoriasis.

ABSTRACT: Antimicrobial peptides (AMPs) are small molecules produced by a myriad of cells and play important roles not only in protecting against infections and sustaining skin barrier homeostasis but also in contributing to immune dysregulation under pathological conditions. Recently, increasing evidence has indicated that AMPs, including cathelicidin (LL-37), human ß-defensins, S100 proteins, lipocalin 2, and RNase 7, are highly expressed in psoriatic skin lesions. These peptides broadly regulate immunity by interacting with various immune cells and linking innate and adaptive immune responses during the progression of psoriasis. In this review, we summarize the recent findings regarding AMPs in the pathogenesis of psoriasis with a main focus on their immunomodulatory abilities.

Evaluation of the toxicity of iron-ion irradiation in murine bone marrow dendritic cells via increasing the expression of indoleamine 2,3-dioxygenase 1.

High linear energy transfer radiation is known to deposit higher energy in tissues and cause greater toxicity compared to low-LET irradiation. Local immunosuppression is frequently observed after irradiation (IR). Dendritic cells (DCs) play important roles in the initiation and maintenance of the immune response. The dysfunction of DCs contributes to tumor evasion and growth. However, molecular mechanisms underlying the establishment of immune tolerance induced by heavy ion IR through this DC population are poorly understood. Therefore, here we report our findings on the dysfunction of bone marrow-derived dendritic cells (BMDCs) induced by 1 Gy iron ion radiation and promotions of expressions of JNK1/2/3, indoleamine 2,3-dioxygenase 1 (IDO1), p-ERK1/2 and p38/MAPK; and decrease of IDO2, MHC class II, CD40, CD80 expressions and IFN-γ and TNF-α secretion after total-body IR in mice. JNK+IDO1+ BMDCs showed up-expression of p-ERK1/2 and p-p38/MAPK, reduced expression of MHC class II and CD80, and were not able to effectively stimulate allogeneic spleen T cells. The inhibition of IDO1 expressions could partly restore the function of BMDCs. In all, our study shows that elevated JNK and IDO1 expression induced by Fe ion IR could result in dysfunction of BMDCs via p-p38/MAPK and p-ERK1/2 signal pathway, and it may represent a new mechanism in radiation-induced immune tolerance.

Neuroprotection of Cilostazol against ischemia/reperfusion-induced cognitive deficits through inhibiting JNK3/caspase-3 by enhancing Akt1.

Cilostazol(CTL) is a phosphodiesterase inhibitor, which has been widely used as anti-platelet agent. It also has preventive effects on various central nervous system (CNS) diseases, including ischemic stroke, Parkinson's disease and Alzheimer disease. However, the molecular mechanism underlying the protective effects of CTL is still unclear, and whether CTL can prevent I/R induced cognitive deficit has not been reported. Transient global brain ischemia was induced by 4-vessel occlusion in adult male Sprague-Dawley rats. The open field tasks and Morris water maze were used to assess the effect of CTL on anxiety-like behavioral and cognitive impairment after I/R. Western blotting were performed to examine the expression of related proteins, and HE-staining was used to detect the percentage of neuronal death in the hippocampal CA1 region. Here we found that CTL significantly improved cognitive deficits and the behavior of rats in Morris water maze and open field tasks (P<0.05). HE staining results showed that CTL could significantly protect CA1 neurons against cerebral I/R (P<0.05). Additionally, Akt1 phosphorylation levels were evidently up-regulated (P<0.05), while the activation of JNK3, which is an important contributor to I/R-induced neuron apoptosis, was reduced by CTL after I/R (P<0.05), and caspase-3 levels were also decreased by CTL treatment. Furthermore, all of CTL's protective effects were reversed by LY294002, which is a PI3K/Akt1 inhibitor. Taken together, our results suggest that CTL could protect hippocampal neurons and ameliorate the impairment of learning/memory abilities and locomotor/ exploratory activities in ischemic stroke via a PI3K-Akt1/JNK3/caspase-3 dependent mechanism.

Metabolite profiling analysis of FR429, an ellagitannin purified from Polygonum capitatum, in rat and human liver microsomes, cytosol and rat primary hepatocytes in vitro.

FR429, an ellagitannin (a type of polyphenol), is isolated and purified from Polygonum capitatum Buch.-Ham.ex D. Don which is the original herbal medicine of the "Re-Lin-Qing" formula used clinically to treat urinary tract infection in China. FR429 has been investigated for its antitumor potential in tumor-bearing nude mice in vivo, but its in vitro anti-tumor effect in hepatoma cell lines was low. Thus, it was of our interest to investigate its metabolism pathways for supporting its in vivo antitumor potential. The metabolic profiles of FR429 were studied in vitro by liquid chromatography coupled to ion trap time-of-flight mass spectrometry. Total eight metabolites were identified in rat and human liver microsomes, cytosol, and rat primary hepatocytes in vitro. Ellagic acid, a reported anti-angiogenic agent, was one of the main metabolites in these biological matrices. Methylated metabolites catalyzed by catechol-O-methyl transferase (COMT) were observed mainly in the in vitro incubation with rat liver cytosol, which was verified by using a COMT specific inhibitor entacapone and supported by molecular docking analysis. Methylated and sulfated metabolites were also found in rat primary hepatocytes in a time-dependent manner. In conclusion, the in vitro metabolism pathways of FR429 were hydrolysis, methylation and sulfation. The anti-tumor effects of its major metabolites should be further studied.

In vitro identification of cytochrome P450 isoforms responsible for the metabolism of 1-hydroxyl-2,3,5-trimethoxy-xanthone purified from Halenia elliptica D. Don.

1-Hydroxyl-2,3,5-trimethoxyxanthone (HM-1) is one of the main constituents extracted from Halenia elliptica D. Don, which is a traditionally used Tibetan medicinal plant. The aim of this study was to illustrate the proposed metabolic pathways of HM-1 and identify which cytochrome P450 (CYP450) isoforms involved in its metabolism by using pooled human liver microsomes (HLMs) and recombinant CYP450 isoforms with selective chemical inhibitors. Metabolites were identified by high performance liquid chromatography coupled to ion trap time-of-flight mass spectrometry (LCMS(n)-ESI-IT-TOF) and nuclear magnetic resonance spectroscopy (hydrogen-1 NMR and carbon-13 NMR). Three metabolites (M1-M3) were identified, which demonstrated that demethylation and hydroxylation were the major Phase I metabolic reactions for HM-1 in HLMs. The structure of another metabolite (M4) was still unclear. The enzymatic kinetics of M1 (K(m)=23.19±14.20 µM) and M2 (Km=32.06±17.09 µM) exhibited substrate inhibition; whereas, the formation of M3 (K(m)=5.73±0.70 µM) and M4 (K(m)=16.43±5.12 µM) displayed Michaelis-Menten kinetics. The intrinsic clearance (V(max)/K(m)) of M3 was highest among these metabolites, suggesting that M3 was the major metabolite of HM-1. Moreover, CYP3A4 and CYP2C8 were the primary CYP450 isoform responsible for the metabolism of HM-1. CYP1A2, CYP2A6, CYP2B6, CYP2C9 and CYP2C19 were also involved in HM-1 metabolism, especially in the formation of M3. This study finally provides evidence of substrate inhibition and metabolism-based drug-drug interaction for the medicinal preparations containing HM-1 used in clinic.

Excretion of berberine and its metabolites in oral administration in rats.

Berberine (BBR) has been confirmed to show extensive bioactivities for the treatments of diabetes and hypercholesterolemia in clinic. However, there are few pharmacokinetic studies to elucidate the excretions of BBR and its metabolites. Our research studied the excretions of BBR and its metabolites in rats after oral administration (200 mg/kg). Metabolites in bile, urine, and feces were detected by liquid chromatography coupled to ion trap time-of-flight mass spectrometry; meanwhile, a validated liquid chromatography coupled with tandem mass spectrometry method was developed for their quantifications. Sixteen metabolites, including 10 Phase I and six Phase II metabolites were identified and clarified after dosing in vivo. Total recovered rate of BBR was 22.83% (19.07% of prototype and 3.76% of its metabolites) with 9.2 × 10(-6) % in bile (24 h), 0.0939% in urine (48 h), and 22.74% in feces (48 h), respectively. 83% of BBR was excreted as thalifendine (M1) from bile, whereas thalifendine (M1) and berberrubine (M2) were the major metabolites occupying 78% of urine excretion. Most of BBR and its metabolites were found in feces containing 84% of prototype. In summary, we provided excretion profiles of BBR and its metabolites after oral administration in rats in vivo.