OBJECTIVE:To establish a RP-HPLC method for the determination of glycyrrhizic acid in sulfonamideglycyrrhiza mixture METHODS:Lichrosorb RPC18 analytical column was used The mobile phase was composed of methanol-(CH)4NH4OH(65∶35) The pH of the mobile phase was adjusted to 6 5 with 10% phosphoric acid Detection wavelength was 254nm RESULTS:Linearity was good in the range of 0 5～50?g/ml(r=0 9 996);the average recovery was 99 75%,RSD=1 87% CONCLUSION:The method is simple and convenient

Objective To screen the effective silencing targets of P21 gene at the cellular level in rhesus monkey . Methods To detect the expression of P21 gene in COS-7 cells ( derived from the kidney of African green monkey , Cerco-pithecus aethiops).Four small hairpin RNA (shRNA) sequences targeting rhesus monkey P21 gene were designed and in-serted into lentivirus-based gene silencing constructs FUGW-TDT.The vectors were transfected into COS-7 cells respective-ly.The suppression of P21 mRNA was detected by real-time PCR, and the expression of P21 protein was detected by West-ern blot assay .Results Four gene-silencing sequences were screened that lied in 541-561 bp, 542-562 bp, 215-239 bp, and 624-648 bp of the rhesus monkey P21 mRNA.Their silencing rate was (91.82 ±3.21)%, (82.47 ±2.48)%, (81.31 ±2.69 )% and ( 87.35 ±4.59 )%, and the protein expression was ( 11.97 ±0.70 )%, ( 20.22 ±0.65 )%, ( 23.21 ± 0.63)%and (14.42 ±0.86)%, respectively.Conclusions Four effective silencing target sequences are screened at cel-lular level , which can be used in gene silencing research of rhesus monkeys .

Objective To decrease the p53 gene expression at cellular and animal levels in marmoset using RNA interference technique.Methods The shRNA interference sequences were designed and inserted into the adeno-associated virus vector plasmid after bioinformatics analysis.The plasmids were transfected into African green monkey kidney cos-7 cells.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western bolt.The adeno-associated virus-8 was injected through the hind leg vein.The changes of p53 protein expression in the liver tissue was detected by Western blot and immunohistochemistry.Results We screened two RNA interference effective arget sequences.The expression of p53 mRNA was suppressed ( 82.7 ±8.1 )% and ( 80.7 ± 7.5)%, respectively (P<0.05), and the expression of p53 protein was decreased (77.3 ±11.5)% and (73.7 ± 10.7)%, respectively (P<0.05).The two marmosets after virus infection showed that there were virus distributions in the liver, testes, and neck detected by in vivo fluorescence imaging.The expression of p53 in the marmoset liver was detected by western blot, immunohistochemistry analysis showing no obvious changes.Conclusions In the present study, the decrease of P53 gene expression at cellular level is achieved, however, the liver P53 protein in the marmoset liver is not significantly changes.Further optimization of the way of infection is needed in the future.

Objective To investigate the correlation between carotid intraplaque neovascularization and acute ischemic cerebrovascular disease. Methods The patients examined with contrast-enhanced ultrasound were enroled and divided into either a symptomatic group or an asymptomatic group according to their cerebral ischemic symptoms. The patients were also divided into a low-echo group, an equal-echo group, and an heterogeneous echo group according to the plaque echo characteristics on conventional ultrasound. The carotid intraplaque neovascularization was evaluated with contrast-enhanced ultrasound. Multivariate logistic regression analysis was used to identify the correlation between carotid intraplaque neovascularization and acute ischemic cerebrovascular disease. Results A total of 73 patients with acute ischemic cerebrovascular disease were enroled, 32 patients in the symptoms group (19 ischemic stroke, 13 transient ischemic attack), 41 patients in the asymptomatic group. Plaque echo characteristics: low-echo 15, equal-echo 41, and heterogeneous echo 17. The proportions of the patients with plaque enhancement (84. 4% vs. 61. 0% ; χ2 = 4. 802, P = 0. 028) and enhanced intensity (21. 78 ± 8. 50 dB vs. 15. 93 ± 8. 82 dB; t = 2. 440, P = 0. 018) in the symptomatic group were significantly higher than those in the asymptomatic group. The proportions of the patients with enhanced plaque in the low-echo, equal-echo and heterogeneous echo group were 93. 3% , 58. 5% , and 82. 4% , respectively (χ2 = 7. 826, P = 0. 020 ). The low-echo group and heterogeneous echo group were significantly higher than the equal-echo group (al P < 0. 05), but there was no significant difference between the low-echo group and the heterogeneous echo group (P > 0. 05). The intraplaque enhanced intensities in the low-echo group, equal-echo group, and heterogeneous echo group were 22. 62 ± 9. 33 dB, 14. 38 ± 8. 02 dB, and 18. 15 ± 9. 64 dB, respectively (F = 3. 877, P = 0. 027). The low-echo group was significantly higher than the equal-echo group (P = 0. 024 ). Multivariate logistic regression analysis showed that carotid intraplaque neovascularization (odds ratio 3. 456, 95% confidence interval 1. 103 - 10. 828; P = 0. 033) was independently associated with acute ischemic cerebrovascular disease. Conclusions Carotid intraplaque neovascularization is closely associated with acute ischemic cerebrovascular disease.

Objective In order to establish a rhesus monkey model of p53 gene silencing, firstly we screened and determined the effective silencing targets of p53 gene at the cellular level in rhesus monkey.Methods The expression of p53 gene was detected in COS-7 cells ( derived from the kidney of the African Green Monkey, Cercopithecus aethiops).Three small hairpin RNA ( shRNA) sequences targeting rhesus monkey p53 gene were designed, analysed by bioinformatics, and inserted into lentivirus-based gene silencing constructs FUGW-TDT.The plasmids of p53-RNAi and control vector were transfected into the COS-7 cells, respectively.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western blot assay.Results p53 gene expression was detected in COS-7 cells.Bioinformatics analysis showed that three gene-silencing sequences were screened which lied in the open reading frame ( ORF) region and targeted 238 -258bp, 681 -701bp, 169 -189bp of the rhesus monkey p53 mRNA.At 48 hrs after transfection of the three silencing constructs, p53 mRNA was suppressed by(87.17 ±4.03)%, ( 72.62 ±4.11)% and(76.22 ±0.98 )%, and p53 protein was suppressed by ( 84.44 ±2.18 )%, ( 71.04 ±1.18)% and ( 74.17 ±0.95 )%, respectively. Conclusions We obtained three effective target sequences showing high efficiency in p53silencing, which can be used in further studies on gene silencing in rhesus monkey.

Entinostat plus exemestane in hormone receptor-positive (HR+) advanced breast cancer (ABC) previously showed encouraging outcomes. This multicenter phase 3 trial evaluated the efficacy and safety of entinostat plus exemestane in Chinese patients with HR + ABC that relapsed/progressed after ≥1 endocrine therapy. Patients were randomized (2:1) to oral exemestane 25 mg/day plus entinostat (n = 235) or placebo (n = 119) 5 mg/week in 28-day cycles. The primary endpoint was the independent radiographic committee (IRC)-assessed progression-free survival (PFS). The median age was 52 (range, 28-75) years and 222 (62.7%) patients were postmenopausal. CDK4/6 inhibitors and fulvestrant were previously used in 23 (6.5%) and 92 (26.0%) patients, respectively. The baseline characteristics were comparable between the entinostat and placebo groups. The median PFS was 6.32 (95% CI, 5.30-9.11) and 3.72 (95% CI, 1.91-5.49) months in the entinostat and placebo groups (HR, 0.76; 95% CI, 0.58-0.98; P = 0.046), respectively. Grade ≥3 adverse events (AEs) occurred in 154 (65.5%) patients in the entinostat group versus 23 (19.3%) in the placebo group, and the most common grade ≥3 treatment-related AEs were neutropenia [103 (43.8%)], thrombocytopenia [20 (8.5%)], and leucopenia [15 (6.4%)]. Entinostat plus exemestane significantly improved PFS compared with exemestane, with generally manageable toxicities in HR + ABC (ClinicalTrials.gov #NCT03538171).