Neomangiferin, a natural C-glucosyl xanthone, has recently received a great deal of attention due to its multiple biological activities. In this study, a rapid and sensitive ultra-high performance liquid chroma-tography tandem mass spectrometry (UHPLC–MS/MS) method for the quantification of neomangiferin in rat plasma was developed. Using chloramphenicol as an internal standard (IS), plasma samples were subjected to a direct protein precipitation process using methanol (containing 0.05% formic acid). Quan-tification was performed by multiple reactions monitoring (MRM) method, with the transitions of the parent ions to the product ions of m/z 583.1-330.9 for NG and m/z 321.1-151.9 for IS. The assay was shown to be linear over the range of 0.2–400 ng/mL, with a lower limit of quantification of 0.2 ng/mL. Mean recovery of neomangiferin in plasma was in the range of 97.76%–101.94%. Relative standard deviations (RSDs) of intra-day and inter-day precision were both o 10%. The accuracy of the method ranged from 94.20%to 108.72%. This method was successfully applied to pharmacokinetic study of neomangiferin after intravenous (2 mg/kg) and intragastric (10 mg/kg) administration for the first time. The oral absolute bioavailability of neomangiferin was estimated to be 0.53%7 0.08%with an elimination half-life (t1/2) value of 2.74 7 0.92 h, indicating its poor absorption and/or strong metabolism in vivo.

Objective To investigate the clinical value of double contrast-enhanced ultrasonography (DCUS) for the diagnosis of gastric cancer.Methods A retrospective analysis was performed to review the DCUS data of 26 patients which were diagnosed as gastric cancer by pathology in the third affiliated hospital of Kunming Medical University from October 2014 to July 2015.The analysis results were compared with postoperative pathology to get accuracy rates.Results The located accuracy,qualitative accuracy and accuracy at T phase of DUCS was 100％ (26/26),100％ (26/26) and 88.5％ (23/26),respectively.Compared with color doppler flow imaging (CDFI),the qualitative accuracy of DUCS was much higher (P ＜ 0.05).The accuracy of DUCS at T phase was higher than that of CDFI with no statistical significance (P ＞ 0.05) Conclusion DUCS has an important application value in the diagnosis of gastric cancer.

OBJECTIVE:To study the bioequiavailability of two sertraline hydrochloride formulations in healthy volunteers.METHODS:A randomized, crossover study of 22 healthy volunteers receiving single oral dose of 50mg sertraline hydrochloride solution(test preparation) or sertraline hydrochloride tablets(comparator preparation) was conducted and the blood concentrations determined by LC/MS/MS.RESULTS: The pharmacokinetic parameters for the test sertraline and the comparator sertraline were as follows:t1/2 were (27.3?5.2)h and (26.3?3.0)h,respectively;Cmax were(9.56?2.49)?g?L-1 and(9.43?2.91)?g?L-1,respectively;tmax were(5.18?1.47)h and(6.00?1.07)h,respectively;AUC0～108 were(329?112)?g?h?L-1 and (297?111)?g?h?L-1,respectively;AUC0～∞ were(354?127)?g?h?L-1 and (316?122)?g?h?L-1,respectively.There were no significant differences in main pharmacokinetics parameters between the 2 preparations,except in tmax from analysis of variance and one-side & two-sides t tests.The relative bioavailability of the test sertraline was(115.5?26.7)%.CONCLUSION:These two sertraline hydrochloride formulations are bioequivalent.

Ischemia-reperfusion injury (IRI) is a common pathophysiological phenomenon, secondary to multiple pathological processes, such as organ transplantation, acute kidney injury and myocardial infarction. IRI may significantly aggravate the severity of diseases and increase the fatality of patients. Aseptic inflammation is one of the critical mechanisms of IRI. Damage-associated molecular pattern (DAMP) is a pivotal substance, which mediates aseptic inflammation. After released into extracellular space, it could effectively activate the immune system, and initiate and maintain the inflammatory responses by binding with pattern recognition receptor (PRR). Neutrophil extracellular trap (NET) is a DNA-based network structure released by neutrophils during the process of inflammatory responses, which contains histones and multiple granular proteins. Recent studies have demonstrated that DAMP and NET may aggravate IRI via aseptic inflammation. In this article, relevant studies of DAMP, NET and their relationship in IRI were reviewed, which was of great significance for understanding the pathophysiological mechanism of IRI and studying the corresponding prevention and treatment strategies.

Objective:To preliminarily explore the effects of tumor treating fields (TTF) arrays on the dose distribution in the treatment of Glioblastoma (GBM) using combined radiotherapy and concurrent TTF.Methods:EDR2 and MatriXX plate ionization chamber were employed to measure the absorbed doses of tissues at different depths (< 1 mm, 3 mm, 5 mm, 1 cm, 1.5 cm, 3 cm, 5 cm, 10 cm, and 15 cm) in the case that TTF arrays and latex-free foam were attached and not attached on the surface. Then the absorbed doses were calculated, compared, and analyzed. For the volumetric arc therapy (VMAT) of 10 GBM patients, deep dose verification was performed using the Sun Nuclear ArcCheck 3D dose verification system and the D99%, Dmean, and D1% of tumors and OARs were assessed. Results:The surface dose increased by 173% in the case that TTF arrays and latex-free foam were attached to the surface compared with the case of the surface with nothing attached. The surface dose increased by 61.7% due to the attachment of low-density latex-free foam. The dose deviation gradually decreased with an increase in the depth and stabilized (about 4%) at a depth of greater than 1.5 cm. As indicated by the VMAT verification result, the D99%, Dmean, and D1% of PTV and CTV decreased by 1.1%-1.2% and the Dmean and D1% of OARs (i.e., brainstem, pituitary gland, optic chiasma, optic nerve, eyeball, and eye crystal) decreased by 0.7%-1.5% in the case that TTF array and latex-free foam were attached on the surface compared with the case the surface with nothing attached. Conclusions:The combined radiotherapy and concurrent TTF in the GBM treatment will lead to a slight reduction of the absorbed doses of targets and OARs but a significant increase in the absorbed doses of the scalp. Therefore, it is recommended that the scalp doses should be reduced as far as possible in the design of the radiation treatment plan to reduce the adverse reactions on the scalp of GBM patients.

BACKGROUND:Intervertebral disc degeneration is an important cause of low back pain.At present,there are many modeling methods for disc degeneration in China and abroad,but there is not a model for low back pain due to disc degeneration. OBJECTIVE:To compare the effect of mechanical puncture combined with tumor necrosis factor α and complete Freund's adjuvant with a conventional disc mechanical puncture alone. METHODS:A total of 18 male adult Sprague-Dawley rats were randomly divided into 3 groups,with 6 animals in each group.No treatment was given in the blank group.Animal models of intervertebral disc degeneration were made in the L4-5 segments of rats in the control using conventional mechanical puncture.In the experimental group,on the basis of mechanical puncture,tumor necrosis factor α+complete Freund's adjuvant was injected into the L4-5 intervertebral discs using a microinjector to establish a model of disc degeneration induced by mechanical puncture combined with inflammatory factors.Four weeks after surgery,the pain threshold of rats was measured by the hot plate method for assessing the perception of heat injury in rats with intervertebral disc degeneration.MRI examination was performed to observe the disc degeneration in each group.ELISA was used to detect the levels of serum tumor necrosis factor α,interleukin 1β,interleukin 6 and prostaglandin E2.Hematoxylin-eosin and Safranin O-fast green staining were used to observe the morphological changes of the disc. RESULTS AND CONCLUSION:In terms of pain,the behavioral pain threshold of the experimental group was continuously decreased,and the levels of serum inflammatory factors were significantly higher compared with the control group.In terms of morphology,the MRI results showed that the L4-5 nucleus pulposus signal completely disappeared in the experimental group.Histopathological results showed that in the control group,the nucleus pulposus was intact,more notochord cells were visible,and some fiber rings were ruptured,while in the experimental group,there are fewer notochord cells and the structure of the nucleus pulposus and fibrous ring is disturbed,with the boundary disappearing.To conclude,mechanical puncture combined with tumor necrosis factor alpha and complete Freund's adjuvant can successfully establish a discogenic low back pain model in rats.This operation is simple and economical to achieve obvious disc degeneration and low back pain,with greatly shortened molding cycle.This model can be used as a reference for studying discogenic low back pain models.

Objective:To observe the effect of Yunpi-Xiefei-Huatan Decoction on airway mucus hypersecretion of asthmatic rats and its regulation on epidermal growth factor receptor (EGFR) / mucin 5AC (MUC5AC) signal pathway. Methods:Seventy SD rats were randomly divided into normal group, model group, high dose group, medium dose group, low dose group, western medicine group and combined group, with 10 rats in each group. Except the normal group, the other groups were sensitized with 1 ml ovalbumin and aluminum hydroxide mixture to establish the asthma rat model. On the 16th day of the experiment, the high, medium and low dose groups were given Yunpi-Xiefei-Huatan Decoction of 40, 20, 10 g/kg, respectively, the western medicine group was given carboxymethylstein tablets of 150 mg/kg, and the combined group was given Yunpi-Xiefei-Huatan Decoction of 20 g/kg and carboxymethylstein tablets of 150 mg/kg, once a day, for 4 weeks. The levels of tumor necrosis factor (TNF-α) and interleukin-13 (IL-13) in serum of rats were detected by Enzyme-linked Immunosorbent Assay (ELISA), the total number and classification of leukocytes in BALF were observed by Wright Giemsa staining, the pathological changes of lung tissue were observed by glycogen staining (PAS). The protein expression of epidermal growth factor receptor (EGFR) and MUC5AC (MUC5AC) were detected by Western blotting, and the mRNA expression of EGFR and MUC5AC was detected by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Results:Compared with the model group, the level of IL-13 and TNF-α in the high, medium and low dose groups of traditional Chinese medicine, western medicine group and combined group was significantly decreased ( P<0.05). The levels of WBC, eosinophils and neutrophils in rat alveolar lavage fluid were significantly decreased ( P<0.05). The expression of EGFR (0.466 ± 0.023, 0.354 ± 0.047, 0.667 ± 0.066, 0.553 ± 0.065, 0.290 ± 0.033 vs. 0.782 ± 0.047) and MUC5AC (0.424 ± 0.022, 0.313 ± 0.033, 0.603 ± 0.051, 0.495 ± 0.041, 0.243 ± 0.024 vs. 0.806 ± 0.090) significantly decreased ( P<0.05), the m RNA expression of EGFR (2.302 ± 0.321, 2.549 ± 0.623, 3.084 ± 0.453, 2.585 ± 0.314, 1.810 ± 0.379 vs. 4.101 ± 0.567), MUC5AC (3.243 ± 0.742, 3.283 ± 1.064, 4.419 ± 0.572, 3.817 ± 0.637, 2.469 ± 0.424 vs. 5.840 ± 0.661) in the high, medium and low dose groups, western medicine group and combined group was significantly decreased ( P<0.05). Conclusion:Yunpi-Xiefei-Huatan Decoction ccould inhibit asthma, and its mechanism mightbe related to the EGFR/MUC5AC signaling pathway.

Objective To investigate the effect of MMP-8 on cornea. Methods Fifteen C57BL/6J healthy mice were selected. The right eyes corneal stroma was injected by 10μL MMP-8 as the experimental group and the left eyes were injected by same amount of normal saline as the control group. At 0,4,8 h, the two-photon microscope second harmonic generation imaging technology was used to scan mice corneal stroma layer by layer in vivo. The obtained images were performed the 3D reconstruction by Imaris software and the signal intensity of the images were calculated. At 4,8 h, the corneal opacity degree was evaluated under slit lamp. At 8 h,mice were killed and corneas were collected to determine the hydroxyproline concentration. Results The cornea stromal fiber signal strengthes at 0 h in the experimental group and control group were (89.7±11.2) and (85.3±7.0),which at 4 h were (14.5±3.4) and (46.6±14. 0) respectively,which at 8 h were (11.0±4.6) and (34.6±12.5) respectively. The cornea stromal signal strength at 4,8 h in the experiemental group was significantly decreased compared with that in the control group (P<0.05) ;the cornea at 4 ,8 h in the experimental group was significantly turbid than that in the control group (P<0.05);the cornea hydroxyproline concentrations detected at 8h in the experiemental group and control group were (0.433±0. 090) μg/mg and (0. 590±0. 133) μg/mg respectively,the experimental group was significantly lower than the control group (F=7. 193,P=0. 014). Conclusion MMP-8 has obvious degradation and destroy effect on mice corneal stroma collagen,which leads to the decrease of corneal opacity.

OBJECTIVE To study the pharmacokinetic characters of Baihe -zhimu decoction （BZD） and its single herbs . METHODS A liquid chromatography -tandem mass spectrometry method was developed to simultaneously determine the blood concentrations of 7 effective components （neomangiferin，mangiferin，regaloside A ，regaloside Ⅰ，timosaponin B Ⅱ，timosaponin E and timosaponin A Ⅲ）in rats after oral administration of BZD extract ，single herb extract of Lilium brownii and Anemarrhena asphodeloides. The pharmacokinetic parameters were calculated . RESULTS The linear range of 7 effective components as neomangiferin were 1-1 000，1-1 000，0.1-100，0.4-400，1-1 000，0.1-100 and 0.5-500 ng/mL（all r＞0.998），respectively. The accuracy of the method ranged from 87.50% to 115.00%，and the RSDs of intra -day and inter -day precision were 0.62%-14.35%. RSDs of matrix factor were 2.68%-14.03%，and deviation of stability were within ±15%. Compared with L. brownii and A. asphodeloides，AUC0－24h，AUC0－∞ and cmax of 7 effective components in BZD were increased significantly （P＜0.05），while CL z/Fof 6 effective components （except timosaponin B Ⅱ）decreased significantly （P＜0.05）. CONCLUSIONS BZD can increase the absorption of effective ingredients in rats ，slow down their elimination and prolong their retention time ，indicating pharmacokinetic behaviors of effective components in BZD possess more advantages over those of single herbs .

OBJECTIVE： To establish the method for simultaneous determination of clopidogrel （CLP）， its intermediate metabolite （2-O-CLP）， inactive metabolite （CLPCA） and active metabolite （CLPTM） in human plasma. METHODS： Totally 90 patients diagnosed as stroke were selected from the First Affiliated Hospital of Army Medical University. They were given one CLP tablet （75 mg/tablet） orally on an empty stomach in the morning. Blood samples were collected 2 h after taking the tablet. CLPTM- D was formed by derivation of CLPTM with 2-bromo-3’-methoxyacetophenone and extracted by precipitation of acetonitrile protein together with the other three substances to be measured. LC-MS/MS method was adopted. The determination was performed on Agilent poroshell 120 EC-C18 column with mobile phase consisted of acetonitrile （0.1％ formic acid） and water （0.1％ formic acid） （90 ∶ 10， V/V）. The quantitation analysis was performed using multiple reaction monitoring at the specific ion transitions of m/z 308.1→198.1 （CLPCA）， 322.3→212.0 （CLP）， 338.3→155.0 （2-O-CLP）， 504.4→354.1 （CLPTM-D） and 264.0→154.1 （ticlopidine， internal standard）， respectively. RESULTS： The retention time of CLPCA， CLP， 2-O-CLP， CLPTM-D and internal standard were 2.01， 3.32， 2.83， 2.68， 1.87 min， respectively. The linear range of CLPCA， CLP， 2-O-CLP and CLPTM-D were 100-10 000， 0.2-20， 0.3-30， 0.5-50 ng/mL （all r≥0.999 5）. The intra-day and inter-day RSD were all less than 9.5％ （n＝5）. Accuracy ranged from 93.5％-98.9％ （n＝5）， and extraction recovery was from 85.4％ to 95.9％ （n＝5）. The matrix effect ranged from 2.7％-6.2％ （n＝5）. In stability tests （storing at －80 ℃ for 3 months， 3 freeze-thaw cycles， storing at 4 ℃ for 8 h）， RE of CLP， CLPCA and CLPTM-D were all lower than 10.0％ （n＝5）. CONCLUSIONS： Established LC-MS/MS method has the advantages of high specificity， accuracy and reliability， and can be used to detect the concentration of CLP and its three metabolites in human plasma.