Combretum indicum can kill insects and benefit the spleen and stomach, which is the most important medicine to treat children's diseases. The classics of materia medica, calendar edition of Chinese Pharmacopoeia, local processing standards and related literature were reviewed to sort out the processing history of Combretum indicum, compare the ancient medicinal parts and processing methods, and summarize the inclusion in national and local standards. The history of the evolution of Combretum indicum was summarized in order to provide ideas for rational drug use in clinical and standard improvement.

Objective To investigate the rules of Traditional Chinese Medicine in the treatment of perimenopausal syndrome (PMS) and provide a theoretical basis for the clinical treatment of PMS. Methods The literature related to PMS were collected from China Knowledge Network (CNKI), Wanfang database and Weipu database in the past 20 years, the herbal compound prescriptions for the treatment of PMS were screened and a relevant database were established and analyzed by SPSS. Results The relevant literatures contains 184 Chinese medicine prescriptions/proprietary Chinese medicines, 122 flavors of traditional Chinese medicines, and the drug categories were mainly tonic drugs, antipyretic drugs, astringent drugs, and tranquilizers. The core single-flavor Chinese medicines were Baishao(Radix Paeoniae Alba), Shudihuang(Rehmannia glutinosa ), Danggui(Radix Angelicae Sinensis), Fuling (Indian Buead). The property and flavor covered sweet, bitter, cold, etc.; and the channel tropism belonged to the liver, kidneys, heart, lungs, spleen and meridians. The cluster analysis of high-frequency Traditional Chinese Medicine obtained two main combinations. Conclusion TCM treatment of PMS focused on replenishing qi, soothing the liver, nourishing the kidneys, nourishing blood and calming the heart, and then according to clinical compatibility with drugs such as soothing the nerves, clearing heat and removing dampness; most of its clinical treatment were Xiaoyaosan, Liuwei Dihuang pills, and Zhibo Rehmanniae decoction and other prescriptions which were added and subtracted.

Objective To establish and improve the quality standard of ginkgo semen decoction pieces. Methods The morphological character for 29 batches of ginkgo gemen and 12 batches of stir-fried ginkgo gemen were observed, and the moisture contents were assayed using the method in Chinese Pharmacopoeia 2015 edition, the first supplement. Results The character of ginkgo gemen and stir-fried ginkgo gemen were consistent in different batches. The moisture content of ginkgo gemen was 8.8% to 12.2%, with an average of 10.5%. The moisture content of stir-fried ginkgo gemen was from 5.4% to 12.3%, with an average of 8.9%. Considering that ginkgo semen decoction pieces are stored for a long time, they are prone to the attack of mildew and insects, and the moisture limit is set to be no more than 10.0% in ginkgo gemen and stir-fried ginkgo. Conclusion Compared with the Chinese Pharmacopoeia 2015 edition, the first supplement, the character identification for ginkgo semen decoction pieces was added and the quality standards were improved. The moisture content of ginkgo semen decoction pieces needs to be strictly controlled under 10.0% to prevent mildew and insects.

Objective To establish the quality standard of Gansu granules (GGs) and effectively control the quality of GGs. Methods TLC identification method of Gansu granules was improved by using pinocembrin-7-O-β-D-glucoside as the indicator. In HPLC analysis, Agilent ZorbaxSB-C₁₈ (4.6 mm×250 mm, 5 μ m) was used as a chromatographic column, and acetonitrile-0.5% formic acid solution was used as the mobile phase. The detection wavelength of 280 nm was used to analyze the content of pinocembrin-7-O-β-D-glucoside in GGs. Results The separation of characteristic spots was good and clear when the established TLC method was used to identify Gansu granules. Quantitative analysis showed that the concentration of pinocembrin-7-O-β-D-glucoside has a good linear relationship with the peak area in the range of 2.4-240 µg/ml (r= 0.9999). The average sample recovery rate of pinocembrin-7-O-β-D-glucoside is 100.76% with RSD value of 0.66%. The accuracy was good. Conclusion The TLC identification method and HPLC content determination method established in this experiment have strong specificity and good reproducibility, and can be used as an improved standard for the quality control of Gansu granules.

Objective To establish an HPLC method for simultaneous assay of macrocyclic polyphenols from Penthorum chinense Pursh, pinocembrin-7-O-[4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucose (PHG), pinocembrin-7-O-[3''-O-galloyl-4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucose (PGHG) and pinocembrin dihydrochalcone-7-O-[3''-O-galloyl-4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucoside or thonningianin A (THA), and optimize the extraction process. Methods The total extraction rate of PHG, PGHG, THA was used as an investigated index to analyze the extracts from Penthorum chinense Pursh. Orthogonal design was applied to evaluate solvent amount, extraction time, solvent concentration and extraction times as the influencing factors for the optimal extraction process of macrocyclic polyphenols from Penthorum chinense Pursh. Results When this content assay method was adopted, there were good linear relationships for PHG, PGHG, THA in the linear range. The recoveries were between 100.90% to 102.04% with the RSDs below 1.5%. The optimal extraction process was involved in cutting Penthorum chinense Pursh into 3-5 cm, adding 10 times 80% ethanol aqueous solution by volume and refluxing 2 hours twice. The extraction rate of macrocyclic polyphenols was above 90% with this process. Conclusion This assay method is accurate, stable, and repeatable. The optimized extraction process is stable and feasible for further development and utilization.

Objective To provide the experimental basis for the subsequent genetic diversity research through establishing and optimizing the inter-simple sequence repeat PCR (ISSR-PCR) reaction system of Gnaphalium affine. Methods The single-factor experimental method and full experimental method were used to optimize the ISSR-PCR reaction system of Gnaphalium affine. Under the optimal system, after screening primers and corresponding annealing temperatures, the systematic feasibility was verified. Results The optimal ISSR-PCR reaction system was consisted of 10 μl Premix Taq DNA polymerase, 0.3 μmol/L primer, 10 ng DNA template, and sterilized water added to 20 μl. Finally, 10 primers were screened from 100 universal primers, and verification results indicated the system had high stability, good reproducibility, and the selected primers had good polymorphism. Conclusion The ISSR-PCR amplification system of Gnaphalium affine was established for the first time and the primers with appropriate annealing temperatures were filtered out, which provided a reference for the subsequent genetic diversity research of Gnaphalium affine.

The effective components are the material basis for efficacy of Traditional Chinese Medicine (TCM), also the core of its quality control. This paper focused on two key scientific issues that how do the effective components form?, How can we improve the effective components?, combining with examples of our group's research, to dis-cuss the research thoughts, technical methods for TCM quality-design. This paper provided a methodological refer-ence for the study of TCM quality-control.

Objective To evaluate the performance of modified Hodge test on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Methods Fortynine Enterobacteriaceae isolates with decreased susceptibility to carbapenems ( MIC of imipenem, meropenem or ertapenem was ≥ 2 μg/ml ) were collected from 16 teaching hospitals from 2004 to 2008. MICs of imipenem, meropenem and etapenem were determined by agar dilution method. Carbapenemases were detected by modified Hodge test. Carbepenemase-causing positive results and AmpCs-causing positive results were differentiated by phenyl boronic acid and oxacillin. Beta-lactamases encoding genes including blaNDM-1were detected by PCR and sequencing. Results Thirty-six of 49 isolates were non-susceptible to imipenem (MIC >4 μg/ml), 31 were non-susceptible to meropenem (MIC > 4 μg/ml) and 47 were non-susceptible to ertapenem (MIC > 2 μg/ml). Twenty-three isolates showed positive modified Hodge test result, including 9 weak-positive results and 14 strong-positive results. Through PCR detection and sequencing, 2 out of 9 isolates showing weak-positive results carried blaKPC-2 and other 7 did not carry any carbapenemase genes but AmpCs/ESBLs genes. Among the 14 isolates showing strong-positive results, 4 carried blaKPC-2, 8 carried blaIMP-4 and 2 caried blaIMP-8. All 26 isolates with negative modified Hodge test result didn't carry any carbapenemase genes. No isolate carried blaNDM-1. Carbapenemases genes PCR detection was regarded as a gold standard, and the sensitivity, specificity, positive predictive value and negative predictive value of modified hodge test was 100%, 79%, 70% and 100% on the detection of carbapenemases among Enterobacteriaceae with decreased susceptibility to carbapenems. Conclusions Modified Hodge test revealed great sensitivity but showed a few false positive results. True and false positive results can be effectively differentiated by phynel boronic acid and oxacillin.

Objective To evaluate the influences of susceptibility interpretation of Escherichia coli,Klebsiella pneumonia and Proteus mirabilis in China mainland according to the old and new ceftazidime,cefotaxime and ceftriaxone breakpoints in CLSI M100-S20 and CLSI M100-S19. Methods First, We analyzed the antibacterial susceptibility results of the three bacteria by agar dilution method in the SEANIR surveillance item, which were collected from 15 national hospitals between the year of 2005 and 2007 and excluded the AmpC enzyme positive isolates according to the PGR-DNA sequencing method and/or the antibacterial susceptibility phenotype. ESBL phenotype was confirmed by the CLSI phenotypic confirmatory test. Antibacterial susceptibility of the total 2733 Escherichia coli, Klebsiella pneumonia, Proteus mirabilis isolates was retrospectively analyzed by WHONET 5. 4 software according to the breakpoints of the CLSI M100-S19 (S19) and CLSI M100-S20 (S20). Second, 207 isolates of Peking Union Medical College Hospital with the results of both agar dilution method and disk diffusion method were performed by recurrent analysis. Then we observed the inter-method agreement through the scatter diagram according to the breakpoints of S19 and S20. Results First, as to the ESBL positive Escherichia coli, Klebsiella pneumonia and Proteus mirabili.s, the resistant rate of cefotaxime increased from 65.2% , 55.5%, 14. 6% under S19 (64 μg/ml) to 99. 7%, 96. 2% , 93. 8% under S20 (4 μg/ml). The susceptibility rates decreased from 6. 0%, 11.5%, 33.3% under S19 (8 μg/ml) to 0%, 0. 2%, 0% under S20 ( 1 μg/ml). Ceftriaxone had the same trend as cefotaxime. Though ceftazidime was more active than cefotaxime and ceftriaxone, as to the ESBL positive Escherichia coli and Klebsiella pneumonia, the resistant rates slightly increased from 30. 3%,43. 2% under S19 (32 μg/ml) to42.0%, 56. 0% under S20 (16 μg/ml). The susceptibility rates slightly decreased from 58. 1%, 44. 1% under S19 (8 μg/ml) to 44. 7%, 28.0% under S20 (4 μg/ml). Second,as to the ESBL negative Escherichia coli, Klebsiella pneumonia and Proteus mirabilis, all the susceptibility rates of ceftazidime, cefotaxime and ceftriaxone were between 99. 2%-100. 0%, the resistant rate were between 0%-0. 4%. Third, the S20 MIC breakpoints had a good correspondence with the ESBL phenotype.Fourth, according to the recurrent analysis of MIC testing and disk dilution method, r value was 0. 67,0. 79, 0. 77 for ceftazidime, cefotaxime and ceftriaxone, respectively, and all P value were under 0. 01. The intermethod rates of S19 and S20 were both acceptable. Conclusions If the cefotaxime and ceftriaxone S20 new breakpoints were used, the concordance of antibacterial susceptibility results and ESBL phenotype would increase greatly. The clinician could select proper antibiotics according to the antibacterial susceptibility results and clinical symptoms. It is no longer necessary to edit results for cephalosporins, aztreonam, or penicillins from susceptible to resistant. However, until laboratories implement the new interpretive criteria,ESBL testing should be performed as described in Supplemental Table 2A-S1. The relationship between the new breakpoints of ceftazidime and clinical outcomes need to be further evaluated.

Objective To observe the effects of the chronic adventitia injury on the vasoconstriction of the rat carotid artery. Methods A non-occlusive silicone collar was positioned around the right carotid artery of rats. Blood flow and vascular reactivity to 5-HT were examined, and both carotids were harvested for morphometry at day 3,7 and 14 after operation. Results In the early stage of advenfitia injury induced by positioning a silicone collar around file rat carotid artery, there appeared the characteristic histological changes of chronic vasospasm in collared artery, such as the reduction of the luminal area for （12.15±2.29）% at day 3 after operation （P =0.003 ） and （45.17±3.84）% at day 7 （P 〈 0.001 ）] ,corrugation of the internal elastic lamina,medial thickening up to [ （23.04±5.96）% at day 3 （P =0.009）, （61.65±10.32）% for day 7 （P 〈 0.001 ）] ,the reduction of the blood flow and the increase of vascular reactivity to 5-HT when compared to con- tralateral arteries. Two weeks after collar placement, the vascular wall remodeling was observed in injured artery, such as the medial thickening for [（31.52±4.56） %,P =0.012] and a diffuse intimal hyperplasia,the reduction of the lunfinal area [（37.17±4.57）% （P 〈 0.001）] and the carotid artery blood flow. The average neointima area was （0.19±0.05） rom2 in collared arteries. The vascular reactivity to 5-HT came back to the normal level. Conclusions Collar-induced advenfitia injury caused the enhancement of vascular contractility and the neointima formation. The change of vascular contractility appeared before the formation of neointima.