Optimization of a laparoscopic procedure for advanced intrahepatic cholangiocarcinoma based on the concept of "waiting time": a preliminary report.

INTRODUCTION: Clinicians increasingly perform laparoscopic surgery for intrahepatic cholangiocarcinoma (ICC). However, this surgery can be difficult in patients with advanced-stage ICC because of the complicated procedures and difficulty in achieving high-quality results. We compared the effects of a three-step optimized procedure with a traditional procedure for patients with advanced-stage ICC. METHODS: Forty-two patients with advanced-stage ICC who received optimized laparoscopic hemihepatectomy with lymph node dissection (LND, optimized group) and 84 propensity score-matched patients who received traditional laparoscopic hemihepatectomy plus LND (traditional group) were analyzed. Surgical quality, disease-free survival (DFS), and overall survival (OS) were compared. RESULTS: The optimized group had a lower surgical bleeding score (P = 0.038) and a higher surgeon satisfaction score (P = 0.001). Blood loss during hepatectomy was less in the optimized group (190 vs. 295 mL, P < 0.001). The optimized group had more harvested LNs (12.0 vs. 8.0, P < 0.001) and more positive LNs (8.0 vs. 5.0, P < 0.001), and a similar rate of adequate LND (88.1% vs. 77.4%, P = 0.149). The optimized group had longer median DFS (9.0 vs. 7.0 months, P = 0.018) and median OS (15.0 vs. 13.0 months, P = 0.046). In addition, the optimized group also had a shorter total operation time (P = 0.001), shorter liver resection time (P = 0.001), shorter LND time (P < 0.001), shorter hospital stay (P < 0.001), and lower incidence of total morbidities (14.3% vs. 36.9%, P = 0.009). CONCLUSIONS: Our optimization of a three-step laparoscopic procedure for advanced ICC was feasible, improved the quality of liver resection and LND, prolonged survival, and led to better intraoperative and postoperative outcomes.

Comparison of predictive value of risk scores for gastrointestinal bleeding in antiplatelet therapy.

Gastrointestinal bleeding (GIB) is the most common serious bleeding complication of antiplatelet therapy. The bleeding risk score (BRS) of GIB may help to determine the risk of bleeding, and provides a reference for the formulation of antiplatelet therapy regimen in clinical practice, but we found that no specific risk scores are available in East Asian patients. This study analyzed patients who were administered antiplatelet therapy from May 2015 to December 2018 in two medical centers. Patient's baseline data were obtained. We assessed four BRSs (New Score, RIETE Score, Cuschieri Score, de Groot Score) and compared them using the area under the receiver operating characteristic curve (AUC). The 4,052 patients enrolled in this study had an average age of 69.6 ± 10.8 years, and 65.9% of them were male. Among the 4,052 patients included, 171 patients experienced GIB within 6 months of follow-up. In the study population, the AUCs for the New, RIETE, Cuschieri, and de Groot scores were 0.673 (95% confidence interval (CI) 0.616-0.729, P < .001), 0.742 (95% CI 0.690-0.794, P < .001), 0.598 (95% CI 0.537-0.659, P = .002), and 0.875 (95% CI 0.839-0.912, P < .001), respectively. After validation, the de Groot Score has better performance. Among the four scores, the de Groot Score might be more suitable for helping Chinese clinicians to predict the risk of GIB in patients taking antiplatelet drugs, and reduce GIB events.

Comparison of clinical performance of four gastrointestinal bleeding risk scores in Chinese patients with atrial fibrillation receiving oral anticoagulants.

Gastrointestinal bleeding is the most common bleeding complication during anticoagulant therapy. A reliable bleeding risk score can help the clinician assess risk of bleeding in individual patients and select the anticoagulant regimen. This study retrospectively analyzed the data of patients with atrial fibrillation who received anticoagulant therapy from July 2015 to December 2018 at two centers-the Fujian Medical University Union Hospital and Fuzhou Second Hospital Affiliated to Xiamen University. Demographic data, clinical findings, and laboratory results were collected from the hospital records. Patients were followed up for 6 months. The performance of four bleeding risk scores (New Score, RIETE Score, Cuschieri et al. Score, de Groot et al. Score) for prediction of gastrointestinal bleeding was assessed using the area under the curve. A total of 3462 patients (mean age, 66.3 ± 11.5 years; 59.6% males; 1055 direct oral anticoagulants users and 2407 warfarin users) were followed up for 6 months. While 99/3462 (2.9%) patients had gastrointestinal bleeding. The area under the curves for the New, RIETE, Cuschieri et al., de Groot et al. scores were 0.652 (95% CI 0.576-0.728), 0.862 (95% CI 0.809-0.914), 0.606 (95% CI 0.527-0.685), and 0.873 (95% CI 0.816-0.929), respectively. Among the four BRSs evaluated, the RIETE score and the de Groot et al. score appear to have the good predictive value, while the NEW score and the Cuschieri et al. score did not sufficiently predict gastrointestinal bleeding risk within the study Chinese population.

Improvement of analgesic efficacy for total hip arthroplasty by a modified ultrasound-guided supra-inguinal fascia iliaca compartment block.

BACKGROUND: Fascia iliaca compartment block (FICB) is an anterior approach to the lumbar plexus block and provides the effective adjunctive analgesia for total hip arthroplasty (THA). METHODS: As a case series study, 28 patients (≥ 65 years old) with THA were received a modified in-plane ultrasound-guided supra-inguinal (S-FICB) as an analgesic adjunct to evaluate the analgesic effectiveness and the local anesthetic diffusion with magnetic resonance imaging (MRI). A combination of propofol and sufentanil was administered to conduct target-controlled infusion. RESULTS: The pain scores were 1 (0-4), 2 (1-5), 3 (1-6) and 3 (1-6) at 4, 8, 12, and 24 h. The cumulative opioids were 8 (8-12), 18 (16-32), 28 (24-54) and 66 (48-104) mg of i.v. morphine equivalents at 4, 8, 12, and 24 h. The patient-controlled analgesia (PCA) times were 0 (0-1), 1 (0-2), 2 (0-5) and 5 (3-8) at 4, 8, 12, and 24 h. In lateral, anterior and medial part of thigh, the sensory blockade in 28 patients was 23 (82 %), 21 (75 %) and 19 (68 %) at 5 min; 28 (100 %) at 10 and 20 min. Motor blockade of femoral nerve (FN) and obturator nerve (ON) was present in 13 (46 %) and 3 (11 %) patients at 5 min, 24 (86 %) and 9 (32 %) at 10 min, 26 (93 %) and 11 (39 %) at 20 min. Injectate permeated to the FN and extended superiorly over the surface of iliac muscle (IM) and pectineus muscle (PM) in all patients. CONCLUSIONS: The modified S-FICB has provided an effective postoperative analgesic adjunct after THA with the satisfactory blockade of femoral (FN), obturator (ON) and sciatic (SN) nerves, especially for ON, when compared with the existing techniques.

MicroRNA-107 regulates anesthesia-induced neural injury in embryonic stem cell derived neurons.

Ketamine, though widely used in pediatric anesthesia, may induce cortical neurotoxicity in young patients. This study focused on an in vitro model of rat brain embryonic stem cell (ESC)-derived neurons to investigate the effects of microRNA-107 (miR-107) on ketamine-induced neural injury. Rat brain ESCs were proliferated in vitro and differentiated toward neuronal fate. Ketamine induced neural injury in ESC-derived neurons was inspected by TUNEL and neurite growth assays. Ketamine-induce aberrant miR-107 expression was examined by qRT-PCR. MiR-107 was downregulated in ESCs through lentiviral transduction. Its effect on ketamine-induced neural injury in ESC-derived neurons was then examined. Potential downstream target of miR-107, brain derived neurotrophin factor (BDNF), was inspected by dual-luciferase reporter assay and qRT-PCR. BDNF was knocked down, through siRNA transfection, in NSCs to investigate its functional involvement in miR-107 mediated neural protection in ketamine-injured NSC-derived neurons. Ketamine induced apoptosis, neurite degeneration, and upregulated miR-107 in NSC-derived neurons. Lentivirus-mediated miR-107 downregulation attenuated ketamine-induced neural injury. BDNF was proven to be directly and inversely regulated by miR-107 in NSC-derived neurons. SiRNA-mediated BDNF inhibition reversed the protective effect of miR-107 downregulation on ketamine injury in NSC-derived neurons. MiR-107 / BDNF was demonstrated to be an important epigenetic signaling pathway in regulating ketamine-induced neural injury in cortical neurons. © 2018 The Authors. IUBMB Life published by Wiley Periodicals,Inc. on behalf of International Union of Biochemistry and Molecular Biology., 71(1):20-27, 2019.

Newly discovered COLQ gene mutation and its clinical features in patients with acetyl cholinesterase deficiency.

To investigate the relationship between acetyl cholinesterase associated collagen gene (COLQ) mutation in patients with acetyl cholinesterase deficiency and its clinical characteristics. Serum and red blood cell acetyl cholinesterase from patients with acetyl cholinesterase deficiency (n=6) and normal controls (n=20) were measured by butyryl thiocholine substrate. COLQ gene variations were detected by sequencing. And the cholinesterase (ChE) genotypes were measured by dibucaine inhibition in vitro. The distributions of ChE surrounded the blood vessels and nerve fibers in lung or pancreas tissues were detected by immunohistochemical staining and indirect immunofluorescence. Serum lactic acid, ammonia and other clinical data were analyzed. Serum ChE in patients with acetyl cholinesterase deficiency were only 1/50 to 1/1000 fold of normal controls. Comparing to controls, dibucaine inhibition values of patients were significantly lower, while there were no differences in red blood cells acetyl cholinesterase. Serum lactic acid and ammonia in patients were significantly higher than controls. Inser 1281-1282 GC of COLQ gene was found in 2 patients, while IVS 6 + 21 T > A, IVS 6 + 30 G > T, IVS 6 + 34 T > C and IVS66 + 12 inser T mutations were found in the other 4 patients, respectively. In addition, the patients with COLQ gene mutation were resistant to regular doses of anesthetics. COLQ gene mutation may be an important reason for the lack of serum ChE in patients with acetyl cholinesterase deficiency.

Effect of parthanatos on ropivacaine-induced damage in SH-SY5Y cells.

Ropivacaine is one of the most common but toxic local anaesthetics, and the mechanisms underlying its neurotoxicity are still largely unknown. This study was conducted to prepare a ropivacaine-induced neuronal injury model and research the effects of ropivacaine on PARP-1 activation and nicotinamide adenine dinucleotide (NAD)+ depletion. The cell death and apoptosis of ropivacaine-induced SH-SY5Y cells were detected with flow cytometry. The lactate dehydrogenase cycling reaction measured the NAD+ level, and western blots were used to analyze the expression levels of PARP-1 and apoptosis-inducing factor (AIF) after ropivacaine treatments with different concentrations and durations. A PARP-1 inhibitor (PJ-34) was used to confirm the relationship between PARP-1 activation and NAD+ depletion. Hoechst 33258 nuclear staining and a mitochondrial membrane potential (Δψm) assay were used to detect the role of exogenous NAD+ in ropivacaine-induced neuronal injury. Ropivacaine-induced SH-SY5Y cell death and apoptosis, PARP-1 activation, and AIF increase as well as intracellular NAD+ depletion occurred in a time- and concentration-dependent manner (P<.05). PARP-1 activation led to NAD+ depletion (P<.05). Exogenous NAD+ impaired ropivacaine-induced nuclear injury (P<.05). Ropivacaine treatment induced PARP-1 activation and NAD+ depletion (P<.05). Parthanatos (PARP-1-dependent cell death) was definitely involved in ropivacaine-induced neuronal injury, and exogenous NAD+ may be a novel therapeutic method for parthanatos-dependent neuronal injury.

Determination of Tomato's SSC and TS Based on Diffuse Transmittance Spectroscopy.

In order to meet the demands for rapid and safe nondestructive testing of fruit and vegetable quality，tomato detection system with a special circular light source was built based on the visible / near infrared diffuse transmission principle. Taking soluble solids content (SSC) and total sugar (TS) as the internal quality index, the prediction of 58 tomato samples was carried out by using this system. First, we collected the spectral data of four points for each tomato. Second, Savitzky-Golay smooth(SG-Smooth), standard normal variable transformation(SNV), multiplication scattering correction(MSC), first derivative (FD) and other methods were used to process the original spectral curve before the partial least squares regression(PLSR) model was established. Finally, we validated the established model. The results show that the correlation coefficient (r) of calibration and prediction of the SSC prediction model -are 0.995 6 and 0.976 0 when using 10 point SG-smooth, and the root mean square error of calibration and prediction are 0.052 4% and 0.082 3%. The partial least square regression (PLSR)model, with respect to the first derivative (FD) spectra, provides better prediction performance for total sugar of tomato, with correlation coefficient (r) of calibration of 0.969 1 and 0.972 9, and prediction, root mean standard error of 0.423 8% and 0.454 9%. In the experimental verification of the prediction model, the relationship of SSC between predicted and true value is 0.985 5, root mean square error is 0.066 3°Brix, the relationship of TS between predicted and true value is 0.944 9 while root mean square error is 0.571 5%. The results show that the content of soluble solids and total sugar in tomato can be realized by using visible / near infrared diffuse reflectance spectroscopy. It provides a real-time, nondestructive and rapid detection method for the evaluation of the internal quality of tomato, and provides a theoretical basis for its online grading.

[A Point Raman Scanning Method for Rapidly Detection of Spinach Quality and Safety Parameters].

According to actual market demand for nondestructive detection of vegetables quality and safety, combined with the heterogeneity of quality and safety parameters such as pesticide residues on leaf vegetables surface and to realize the automatic point scanning for the whole leaf vegetables samples, a suction device based on laboratory (self-designed) Raman spectroscopy hardware and a GUI application software based on the LabVIEW development platform were developed. This system can test the Raman spectroscopy of the whole spinach including the automatic collection, display and storage of the Raman signal of all the scanned points by set up different scan step. A new method to remove the Raman spectrum background was proposed based on data replacement with linear equation at the range of threshold spectrum on both sides of the effective peaks according to the characteristics of spinach original spectra. Its principle is to determine the starting position of linear fitting by judging whether there is trough on both sides of the crest, and then to generate and replace the original spectra data in peak position through the linear fitting equation. Spinach samples were used for the experiment showed that the chlorophyll content and distribution of chlorpyrifos pesticide residue on each scanning point can be obtained after scanning. Therefore, the point scanning Raman system could improve detection accuracy of the quality and safety parameters for the non-uniform samples effectively.

Textbook outcomes of hepatocellular carcinoma patients with sarcopenia: A multicenter analysis.

BACKGROUND: The impact of sarcopenia on textbook outcome (TO) after hepatectomy in hepatocellular carcinoma (HCC) patients remains unclear. This study aimed to investigate the association between sarcopenia and TO, to clarify its long and short-term prognostic value, and to develop a nomogram model based on sarcopenia and TO for survival prediction. METHODS: Patients who underwent HCC resection between January 2012 and March 2017 in three large hospitals in Fujian were retrospectively recruited and divided into sarcopenia and non-sarcopenia groups based on skeletal muscle index (SMI) values. TO was defined as no 30-day morality, no 30-day readmission, negative margins, no prolonged hospital stay, and no major complications. Multivariate regression was used to screen for clinical factors associated with TO. Nomograms of overall survival (OS) and recurrence-free survival (RFS) after hepatectomy for HCC were developed. RESULTS: A total of 1172 patients were included in the study. The TO rates were 28.74% (121/421 patients) in the sarcopenia group and 43.4% (326/751 patients) in the non-sarcopenia group. The results showed that sarcopenia was an independent predictor of TO (p < 0.001), TO was an independent predictor of perioperative treatment-related sarcopenia (PTRS)(p = 0.002), and TO was an independent predictor of OS and RFS (p < 0.001). Nomogram models based on sarcopenia and TO were generated and accurately predicted OS and RFS at 1, 3, and 5 years. CONCLUSION: Both sarcopenia and TO are independent predictors of OS and RFS after HCC resection. Sarcopenia was an independent predictor of TO. Sarcopenia influenced long-term survival by affecting short-term postoperative outcomes.

Impact of preoperative carbohydrate loading on gastric volume in patients with type 2 diabetes.

BACKGROUND: Enhanced recovery after surgery advocates that consuming carbohydrates two hours before anesthesia is beneficial to the patient's recovery. Patients with diabetes are prone to delayed gastric emptying. Different guidelines for preoperative carbohydrate consumption in patients with diabetes remain controversial due to concerns about the risk of regurgitation, aspiration and hyperglycemia. Ultrasonic gastric volume (GV) assessment and blood glucose monitoring can comprehensively evaluate the safety and feasibility of preoperative carbohydrate intake in type 2 diabetes (T2D) patients. AIM: To evaluate the impact of preoperative carbohydrate loading on GV before anesthesia induction in T2D patients. METHODS: Patients with T2D receiving surgery under general anesthesia from December 2019 to December 2020 were included. A total of 78 patients were randomly allocated to 4 groups receiving 0, 100, 200, or 300 mL of carbohydrate loading 2 h before anesthesia induction. Gastric volume per unit weight (GV/W), Perlas grade, changes in blood glucose level, and risk of reflux and aspiration were evaluated before anesthesia induction. RESULTS: No significant difference was found in GV/W among the groups before anesthesia induction (P > 0.05). The number of patients with Perlas grade II and GV/W > 1.5 mL/kg did not differ among the groups (P > 0.05). Blood glucose level increased by > 2 mmol/L in patients receiving 300 mL carbohydrate drink, which was significantly higher than that in groups 1 and 2 (P < 0.05). CONCLUSION: Preoperative carbohydrate loading < 300 mL 2 h before induction of anesthesia in patients with T2D did not affect GV or increase the risk of reflux and aspiration. Blood glucose levels did not change significantly with preoperative carbohydrate loading of < 200 mL. However, 300 mL carbohydrate loading may increase blood glucose levels in patients with T2D before induction of anesthesia.

Comparing the minimum local anesthetic dose of ropivacaine in real-time ultrasound-guided spinal anesthesia and traditional landmark-guided spinal anesthesia: a randomized controlled trial of knee surgery patients.

BACKGROUND: Through previous studies and clinical practice, we have found that real-time ultrasound-guided (UG) spinal anesthesia (SA) and traditional landmark-guided (LG) SA each require a different minimum local anesthetic dose (MLAD) of ropivacaine. For this study, we used Dixon's up-and-down sequential method to analyze and compare the MLAD of different ropivacaine concentrations required for the UG and LG SA methods. METHODS: A total of 120 patients undergoing knee surgery were consecutively recruited and randomly divided into four groups (30 patients per group). These groups were categorized as follows: Group I: high ropivacaine ultrasound-guided (HRUG), Group II: low ropivacaine ultrasound-guided (LRUG), Group III: high ropivacaine landmark-guided (HRLG), and Group IV: low ropivacaine landmark-guided (LRLG). SA was established by a bolus administration of up-and-down doses of 0.75% or 0.5% plain ropivacaine. Initial doses of 16, 18, 12, and 14 mg were administered to groups I-IV, and after that, increased or decreased by 1.5 mg according to dose effectiveness. Upon identifying the intervertebral puncture level, a lumbar X-ray was performed with metal markers, and actual radiographic findings were identified and compared to the initial markings. RESULTS: For UG groups, the MLAD in the LRUG group was significantly higher than in the HRUG group [20.192 mg (95% CI, 19.256-21.174) versus 17.176 mg (95% CI, 16.276-18.124), respectively; P<0.001]. For LG groups, the MLAD in the LRLG group was significantly higher than in the HLRG group [14.478 mg (95% CI, 13.364-15.500) versus 13.201 mg (95% CI, 11.959-14.571), respectively; P=0.047]. When comparing both high ropivacaine groups (HRGs: I/III) to the low ropivacaine groups (LRGs: II/IV), we found that both UG subgroups (I/II) had a significantly higher MLAD than LG subgroups (III/IV) (P<0.001). US identified L4-5 in up to 90% of cases. Comparatively, palpation was successful in only 33.3% of patients. The rates of cephalad localization by US and palpation were 6.67% vs. 66.67%, respectively (P=0.002). CONCLUSIONS: We found a higher MLAD of ropivacaine was required for UG SA at the L4-5 level due to the method providing a more accurate (less cephalad) localization than traditional LG SA. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2000033158.

Botrychium schaffneri Underw. extract acts via DIABLO to induce apoptosis and inhibit proliferation of non-small cell lung carcinoma in vitro and in vivo.

BACKGROUND: Botrychium schaffneri Underw. has been popularly consumed since ancient times as a traditional medicine in China to treat whooping cough, bronchial asthma, and febrile convulsive twitch disease. This led us to investigate whether Botrychium schaffneri Underw. extract (BSE) may be effective against lung cancer, especially non-small cell lung carcinoma (NSCLC). METHODS: In this study, we extracted the ethanolic root extract of the grass, Botrychium schaffneri Underw. In vitro study, the change of NCI-H1299 cell proliferation was observed with CCK8 and MTT assays. Cell apoptosis was assessed using a kit based on staining with FITC-conjugated annexin V. In vivo study, we establish a stable animal model of NSCLC in nude mice, tumor volume and weight was measured twice a week. We conduct gene microarray screened for differentially expressed genes (DEGs), between NCI-H1299 cells treated by BSE or not. Then the DEGs were functionally annotated and path enriched. RESULTS: It was revealed that BSE significantly suppressed NSCLC cell proliferation (IC50 134 µg/mL) and induced apoptosis. It also slowed tumor growth without affecting body weight, and a dose of 25 g/kg led to significantly smaller tumors than in control animals (13.85±3.36 vs. 23.40±6.05, P=0.044). Apoptosis-related protein direct IAP Binding protein with low PI (DIABLO) expression was up-regulated by BSE, and DIABLO knockdown significantly attenuated the anti-tumor effects of the extract. CONCLUSIONS: In conclusion, BSE reduces the viability of NSCLC cells and promotes apoptosis, and these effects may be mediated by DIABLO.

L1, ß-catenin, and E-cadherin expression in patients with colorectal cancer: correlation with clinicopathologic features and its prognostic significance.

OBJECTIVE: Currently, there are no clinically used/routine biomarkers that accurately predict whether colorectal cancer (CRC) patients will or will not respond to adjuvant chemotherapy. The aim of this study was to investigate L1, ß-catenin, and E-cadherin expression in patients with CRC and their relationship to tumor progression, and to identify patients who will respond to chemotherapy. METHODS: A total of 142 patients who underwent surgical treatment for CRC were chosen retrospectively. The samples from these patients were analyzed by immunohistochemistry. SPSS-14 program package was used for statistical calculation. RESULTS: Expression of L1, ß-catenin, and E-cadherin were found to be strongly associated with invasion and metastasis of CRC. Cox multivariate analysis results indicated that L1 expression and stage of Dukes could be considered as the independent prognostic factors for survival. Furthermore, our study found that the 5-year survival rate was the significantly associated with the expression of L1, ß-catenin (normal and ectopic expression), and E-cadherin for Dukes' stage B (P < 0.01) patients. However, no such result was found for Dukes' stage A (P > 0.05) and C (P > 0.05) patients. CONCLUSION: Our study provided reference for identifying patients who need adjuvant chemotherapy. L1, ß-catenin, and E-cadherin could be considered as biomarkers to predict whether CRC patients will or will not respond to adjuvant chemotherapy.

L1CAM is involved in lymph node metastasis via ERK1/2 signaling in colorectal cancer.

L1-cell adhesion molecule (L1CAM, L1) belongs to the immunoglobulin superfamily and was originally found to play a role in nerve cells. Recently, the expression and prognostic value of L1 has been established in several cancers, including colorectal cancer (CRC). However, its association with lymph node metastasis in CRC and the mechanisms underlying its effects remain unclear. In this study, we evaluated the L1 transcript levels in CRC (n=12) and normal intestinal tissues (n=15) by qRT-PCR. Western blotting was used to evaluate L1 and pERK1/2 expression levels. Immunohistochemistry was performed to evaluate the relationship between L1 and pERK1/2 in CRC tissues with different levels of differentiation. The mRNA expression levels in CRC tissues were significantly higher compared to normal intestinal tissues. Western blotting demonstrated that both L1 and pERK1/2 levels were higher in CRC than in normal tissues. Immunohistochemistry confirmed that L1 and pERK1/2 levels in adenomas with lymph node metastasis were significantly higher than in poorly and well-differentiated adenomas, indicating that L1 and pERK1/2 levels correlated with CRC lymph node metastasis. In conclusion, L1 and pERK1/2 were significantly up-regulated in CRC tissues and lymph node metastasis may occur via the L1CAM-mediated ERK pathway in CRC.

Determination of asiatic acid in beagle dog plasma after oral administration of Centella asiatica extract by precolumn derivatization RP-HPLC.

A novel precolumn derivatization reversed-phase high-performance liquid chromatography (RP-HPLC) method with UV-vis detection for the quantitative determination of total concentration of asiatic acid (AA) in beagle dog plasma is described. AA was extracted with n-hexane-dichloromethane-2-propanol (20:10:1, v/v/v) from plasma, which had been hydrolyzed by acid and derivatized with p-Toluidine. Chromatographic separation was achieved on a C(18) column using gradient elution in a water-methanol system. Detection was set at UV wavelength of 248nm. A calibration curve ranging from 0.01 to 1.5microg/mL was shown to be linear, and the lower limit of quantification (LLOQ) was 0.01microg/mL. The intra- and inter-day precisions which were determined by three different concentrations (0.05, 0.2 and 0.8microg/mL) ranged from 4.4% to 13.1% and 4.6% to 14.2%, respectively. Mean extraction recoveries were no less than 65% for AA and ursolic acid (IS). Plasma samples containing asiatic acid were stable for 30 days at -20 degrees C. The method was successfully applied to a pharmacokinetic study in beagle dogs after oral administration of Centella asiatica extract, and the main pharmacokinetic parameters obtained were: T(1/2), 4.29h; T(max), 2.70h; C(max), 0.74microg/mL; AUC(0-t) and AUC(0-infinity), 3.74 and 3.82microgh/mL, respectively.