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OBJECTIVE To explore the improvement effect and potential mechanism of Baicao fuyanqing suppository on bacterial vaginitis （BV） in rats. METHODS The female SD rats were randomly divided into normal group， model group， metronidazole group （positive control， 0.03 g/kg）， Baicao fuyanqing suppository low-dose， medium-dose and high-dose groups （0.18， 0.36， 0.72 g/kg）， with 8 rats in each group. Except for the normal group， the rats in other groups were injected subcutaneously with 0.2 g of Estradiol benzoate injection+20 μL of Escherichia coli suspension （2×108～3×108 cfu/mL） through the vaginal opening to establish the BV rat model. Administration groups were given relevant medicine vaginally， while the normal group and the model group were given normal saline， once a day， for 6 consecutive days. Twenty-four hours after the last administration， the vaginal appearance score and vaginal pH were measured for each group of rats. The levels of cytokines [interleukin-1β （IL-1β）， IL-2， IL-13， immunoglobulin A （IgA）] in vaginal lavage fluid were determined. The morphology of the uterus and accessories， and pathological changes in the vaginal tissue were observed. The expressions of Toll-like receptor 2 （TLR2）， TLR4 and nuclear factor-κB （NF-κB） in vaginal tissues were determined. RESULTS Compared with the normal group， the uterus edema， the irregular shape of uterus and accessories， the vaginal mucosa hyperplasia， and the massive desquamation of epithelial cells were observed in the model group， complicating with massive infiltration of inflammatory cells； vaginal opening redness and swelling score and secretion score， vaginal pH， the levels of proinflammatory cytokine IL-1β and IL-2， the protein expressions of TLR2， TLR4 and NF- κB were all increased or up-regulated， while the levels of IgA and anti-inflammatory cytokine IL-13 decreased significantly （P＜0.05 or P＜0.01）. Compared with the model group， varying cn degrees of improvement in uterine and accessories， and vaginal tissue lesions in rats were observed in administration groups， and most of the quantitative indicators mentioned above showed significant improvement （P＜0.05 or P＜0.01）. CONCLUSIONS Baicao fuyanqing suppository has a certain improvement effect on inflammatory symptoms in BV rats， and its mechanism may be related to the inhibition of the TLR/NF-κB signaling pathway.
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OBJECTIVE To analyze quality maker （Q-marker） of Ka nggongyan soft capsule （KSC）. METHODS The fingerprints of 20 batches of KSC were established by ultra high performance liquid chromatography （UPLC）method. Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）were used to evaluate the similarity and confirm common peaks. The contents of norisoboldine ，leonurine hydrochloride ，forsythoside B ，acteoside，poliumoside and isoacteoside were determined by the same UPLC method. Targets and pathways related to KSC in the treatment of cervicitis were screened and analyzed by network pharmacology and molecular docking method to construct a “component-target-pathway”network，and analyze its potential Q-marker. RESULTS Twelve common peaks were identified in the fingerprints of 20 batches of KSC ，and the similarity was greater than 0.99. Six common peaks were identified ，including norisoboldine ，leonurine hydrochloride ，forsythoside B，acteoside，poliumoside and isoacteoside. The contents of the above 6 components were 1.336-1.774，0.093-0.143，4.970-5.888， 0.505-0.623，5.206-6.226 and 0.785-0.895 mg/g，respectively. By network pharmacology analysis ，14 key targets and 94 pathways were obtained ，and their binding energies to the core targets （protein kinase B 1，tumor necrosis factor ）were all less than －6.4 kJ/cal. CONCLUSIONS Six components such as norisoboldine and leonurine hydrochloride are potential Q-marker of KSC.
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OBJECTIVE： Compare the contents of free ellagic acid and total ellagic acid in fruits， roots and levels of Rosa roxburghii， and to evaluate the in vitro anti-oxidant activity of ethanol extract of three medicinal parts of R. roxburghii. METHODS： Free ellagic acid and total ellagic acid were obtained from different medicinal parts of R. roxburghii by ultrasonic extraction and acid hydrolysis， respectively. The contents of them were determined by UPLC. Using half-clearance value （IC50） as anti-oxidant evaluation index， free radical scavenging test of 1，1-diphenyl-2-trinitrophenylhydrazine （DPPH） and 2，2′ -binitro-bis（3-ethylbenzothiazoline-6-sulfonic acid） （ABTS） were conducted for ethanol extract of different medicinal part， using vitamin C （VC） as positive control. RESULTS： There were significant differences in the content of free ellagic acid and total ellagic acid in different medicinal parts of R. roxburghii. The content level of free ellagic acid was in descending order： R. roxburghii leaves （38.49 mg/g）＞R. roxburghii fruits （20.59 mg/g）＞R. roxburghii roots （11.35 mg/g）； the content level of total ellagic acid was in descending order： R. roxburghii leaves （197.08 mg/g） ＞ R. roxburghii fruits （49.36 mg/g） ＞ R. roxburghii roots （21.86 mg/g）. The contents of total ellagic acid in the fruits and roots of R. roxburghii were twice as much as that of free ellagic acid in corresponding parts； the contnets of total ellagic acid in the leaves of R. roxburghii were five times higher than that of free ellagic acid in corresponding parts. In the DPPH free radical scavenging test and ABTS free radical scavenging test， the order of antioxidant activity was VC ＞ R. roxburghii leaves＞R. roxburghii fruits＞R. roxburghii roots. There was no statistical significance in the effects of R. roxburghii leaves [IC50 to DPPH free radical and ABTS free radical were （4.57±0.70）、（115.99±2.21） μg/mL] and R. roxburghii fruits [IC50 to DPPH free radical and ABTS free radical were （5.12±0.24）、（127.61±3.31） μg/mL]， compared with the effects of VC [IC50 to DPPH free radical and ABTS free radical were （4.47±0.38）、（121.42±2.65） μg/mL] （P＞0.05）. CONCLUSIONS： Among fruits， roots and leaves of R. roxburghii， the content of free （total） ellagic acid is the highest in the R. roxburghii leaves and in vitro anti-oxidant activity of its ethanol extract is the strongest.
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Objective@#To investigate the impact of ultrasonic assisted rapid processing technique combined with the environment friendly reagent (which can be utilized in fixing,dehydrating and clearing) on processing tumor biopsy specimens and the subsequent target detection.@*Methods@#Postoperative tissue samples of 56 cases of breast cancer, colorectal cancer, lung cancer, stomach cancer, liver mass, bladder mass, uterus mass were obtained at the National Cancer Center, Cancer Hospital, Chinese Academy of Medical Sciences from February to April, 2017. Three specimens ranging in size from 1 to 3 mm were collected from each sample, and were separated into control group (traditional tissue-processing method); experiment group 1 (3.7% neutral buffered formaldehyde fixation, composite environment friendly reagent and ultrasonic assisted rapid processing) and experimental group 2 (composite environment friendly reagent direct fixation, higher temperature and longer time for tissue processing). Two pathologists blinded to the experimental groups scored totally the nuclear, cytoplasmic, and membrane staining of 43 cases of immunohistochemistry (IHC), four HER2 fluorescence in situ hybridization (FISH), 20 extracted DNA quality and four EGFR gene mutation detection in lung adenocarcinoma; the results were compared with the control group.@*Results@#There was no difference in the IHC staining, HER2 FISH, the DNA quality, and EGFR genetic results between experimental group 1 and control group. For experiment group 2, comparing results of IHC staining, HER2 FISH and the quality of DNA, there was no obvious difference from control group and experiment group 1, but might show an increase in the background of IHC staining. The difference between the treatment temperature and time in the experimental group 2 did not affect the results of the gene mutation detection.@*Conclusions@#Environment freindly reagent and ultrasonic assisted rapid processing equipment could be used for rapid processing and diagnosis for tumor biopsies. Using complex environment-friendly reagents supplement fixation, higher treatment temperature and longer treatment time do not significantly affect the IHC, FISH and molecular detection accuracy.