Objective:To optimize and establish the experimental methods for the determination of 2,4-dihydroxybenzophenone (BP-1) in mouse brain. Methods:BP-1 was determined by high performance liquid chromatography (HPLC) and separated by Waters Symmetry? C18 (4. 6 mm × 250 mm, 5 μm) using isocratic elution, and the sample preparation conditions were optimized by orthogonal experiment design. The mobile phase was methanol-water (volume ratio 3∶1) containing 3% (volume fraction) ace-tic acid (pH 3. 40) at a flow rate of 1. 0 mL/min, and ultraviolet (UV) detection wavelength was set at 290 nm. Retention time was used for qualitative analysis and internal standard method for quantitative analysis. Results: Under the optimized experimental conditions, the calibration curve was linear with a correlation coefficient of 0. 999 8 over the concentration range of 0. 2-10. 0 mg/L. The recoveries of BP-1 were between 96. 8% and 104. 5%. The intra-day and inter-day precision of BP-1 were 3. 5% -5. 7% and 4. 5% -6. 4%, respectively. The extraction recoveries of BP-1 at three concentrations (0. 5, 2. 0, 8. 0 mg/L) in the mouse brain were 90. 5%, 89. 5%, and 97. 7%, and the matrix effect of BP-1 at these three concentrations were 102. 9%, 102. 7%, and 90. 9%, respectively. Conclusion:The method is simple, ac-curate, and suitable for determination of the contents of BP-1 in mouse brain.

OBJECTIVE To study the nephrotoxicity induced by bakuchiol alone and bakuchiol combined with psoralen and to explore its mechanism. METHODS The cytotoxicities of bakuchiol and bakuchiol combined with psoralen were investigated using human renal tubular epithelial cell lines (HK-2), in presence or absence of hepatic S9 mixture. The HK-2 cells were exposed to culture medium alone (blank control), 0.5% DMSO (vehicle control), aristolochic acid Ⅰ (AAⅠ;positive control), psoralen 5 μmol·L~(-1) group, bakuchiol 5,10,20,30 and 40 μmol·L~(-1) groups, and bakuchiol+psoralen (20+5), (30+5) and (40+5)μmol·L~(-1) groups, respectively. The cell viabilities were examined by MTT assay; cell membrane injuries were examined by detecting lactate dehydrogenase (LDH) release rate; and the morphological changes in HK-2 cells were observed with contrast microscope. The rate of cell apoptosis was detected by AnnexinⅤ/PI staining, and cell cycle was detected by PI staining with flow cytometry. RESULTS No cytotoxicity was found in psoralen 5 μmol·L~(-1) group. The HK-2 cell viabilities were significantly reduced after 4, 24, 48 and 72 h of exposure to either bakuchiol 20, 30 and 40 μmol·L~(-1)groups or bakuchiol+psoralen (20+5), (30+5) and (40+5)μmol·L~(-1) groups in a time- and concentration-dependent manner. The IC_(50) values of bakuchiol were (26.4±4.8), (21.8±0.6) and (24.1±0.8)μmol·L~(-1) for 24, 48 and 72 h exposure, respectively. The cytotoxicity of bakuchiol was significantly decreased in presence of hepatic S9 mixture. The LDH release rate of HK-2 cell increased significantly after 24 h of exposure to bakuchiol 20,30 and 40 μmol·L~(-1) or bakuchiol+psoralen groups. With the concentration and time increasing, the HK-2 cells became more and more contracted and rounded. In bakuchiol 40 μmol·L~(-1) or bakuchiol+psoralen (20+5), (30+5) and (40+5)μmol·L~(-1) groups, HK-2 cells showed apoptotic characters. In bakuchiol or bakuchiol+psoralen groups, apoptotic cells significantly increased and cells in G2 phase markedly decreased. CONCLUSION Bakuchiol has a significant cytotoxicity in HK-2 cells, and combined with psoralen can not decrease its toxicity. The cytotoxicity of bakuchiol is significantly reduced in the presence of hepatic S9 mixture. The possible mechanisms of the renal cytorotoxicity of bakuchiol are as follows: ① direct damage to the cell membrane; ② inducing cell apoptosis; ③ inhibiting intracellular DNA synthesis and block cell mitosis and proliferation.

Objective:To investigate the possible reversal effects of 1 ,3-diphenyl-1 ,3-propanedione (DPPD)for cocaine-induced content changes of neurotransmitters of brain in mice.Methods:In this study,36 healthy ICR male mice were randomly divided into control group,cocaine group,three DPPD pretreatment groups (200,400,and 800 mg/kg)and DPPD alone group (800 mg/kg).The mice in control group were administered intragastrically with 1 % Tween 80 for 3 d,and the mice in cocaine group were administered intragastrically with 1 % Tween 80 for 2 d before cocaine was injected subcutaneously on the 3rd day.The mice in the three DPPD pretreatment groups were administered intragastrically (DPPD 200,400,and 800 mg/kg)for 3 d before cocaine was injected subcutaneously 30 min after the administration on the 3rd day.The mice in DPPD alone group were administered intragastrically with DPPD at dose of 800 mg/kg for 3 d.The mice were sacrificed 20 minutes after cocaine injection.The contents of dopamine (DA)and 5-hydroxytryptamin (5-HT)in the mice brain were determined by high performance liquid chromatography (HPLC)-fluorescence detector,the contents of glutamic acid (Glu) and γ-aminobutyric acid (GABA)in the mice brain were determined by HPLC-ultraviolet detector,and the neurotransmitter levels were compared between the groups.Results:The results showed that as com-pared with the control group,DA and GABA contents in cocaine group increased significantly (P <0.01 and P <0.05),while Glu content decreased (P <0.05).As compared with cocaine group,the DA levels in the three DPPD pretreatment groups (200,400,and 800 mg/kg)all decreased significantly (P <0.01 ).In DPPD 200 mg/kg pre-administration group,GABA content decreased (P <0.05),and the contents of the four kinds of neurotransmitters had no statistical differences with those of the control group.Conclusion:DPPD may have potential reversal effects of the content changes of neurotransmitters in mice brain induced by cocaine at a lower dose.

One influenza H3N2 virus, A/swine/Shandong/3/2005 (Sw/SD/3/2005), was isolated from pigs with respiratory disease on a farm in eastern China. Genetic analysis revealed that Sw/SD/3/2005 was a triple-reassortant virus with a PB2 gene from human-like HIN1, NS from classical swine H1NI, and the remaining genes from human-like H3N2 virus. These findings further support the concept that swine can serve as reservoir or mixing vessels of influenza virus strains and maintain genetic and antigenic stability of viruses. Furthermore, we have successfully established a reverse genetics system based on eight plasmids and rescued Sw/SD/3/2005 through cell transfection. HI tests and RT-PCR confirmed that the rescued virus maintained the biological properties of the wild type Sw/SD/3/2005. The successful establishment of the reverse genetics system of Sw/SD/3/2005 will enable us to conduct extensive studies of the molecular evolution of H3N2 influenza viruses in swine.

Objective:To explore the clinical observation of Xiaozhang Plaster for external use in the treatment of postprandial discomfort syndrome of qi stagnation functional dyspepsia.Methods:Randomized controlled trial. From January 2019 to January 2022, 128 patients with postprandial discomfort syndrome of functional dyspepsia of qi stagnation type in our hospital were selected and divided into two groups by random number table, with 64 cases in each group. Both groups were orally treated with mosapride citrate tablets. On this basis, the treatment group was combined with external use of Xiaozhang Plaster, and the control group was combined with placebo patch. Both groups were treated for 2 weeks. Traditional Chinese Medicine syndrome score, symptom severity and attack frequency score were performed before and after treatment. The quality of life of the patients was assessed with SF-36 Short-form health survey-36 (SF-36), anxiety and depression were assessed with Hospital Anxiety and Depression Scale (HADS), and the growth hormone releasing peptide (Ghrelin), calcitonin gene-related peptide (CGRP) The levels of gastrin 17 (G-17), pepsinogen I (PG I) and pepsinogen Ⅱ (PG Ⅱ) levels were observed by ELISA, and clinical efficacy was evaluated.Results:The total effective rate was 96.88% (62/64) in the treatment group and 81.25% (52/64) in the control group. The difference between the two groups was statistically significant ( χ2=8.02, P<0.05). After treatment, the score and total score of epigastric distension, epigastric distension and pain, fullness of both flanks, loss of appetite in the treatment group were significantly lower than those in the control group ( t=38.43, 32.39, 38.43, 32.87, 33.74, P<0.01), the score of symptom severity and attack frequency were significantly lower than those in the control group ( t=61.42, 33.46, P<0.01), and the score of SF-36 was significantly higher than that in the control group ( t=8.26, P<0.01); The score of HADS was significantly lower than that of the control group ( t=38.06, P<0.01). After treatment, the levels of CGRP and G-17 in the treatment group were significantly lower than those in the control group ( t values were 6.22, 29.51, P<0.01), the level of Ghrelin, and the levels of PG Ⅰ, PG Ⅱ were significantly higher than those in the control group ( t=23.85, 13.26, 19.53, P<0.01). Conclusion:On the basis of basic drug treatment, the application of Xiaozhang Plaster for external use to intervene patients with postprandial discomfort syndrome of qi stagnation functional dyspepsia can improve the quality of life, gastrointestinal hormones, anxiety and depression symptoms, and the effect is remarkable.

Objective To investigate the perioperative risk factors associated with prognoses of patients with acute ischemic stroke accepted mechanical thrombectomy under general anesthesia.Methods The clinical data of 108 patients with acute ischemic stroke,admitted to and accepted mechanical thrombectomy under general anesthesia in our hospital from January l,2016 to October 31,2018,were collected.According to modified Rankin scale (mRS) scores 90 d after surgery,patients were divided into good prognosis group (mRS scores ≤2) and poor prognosis group (mRS scores ≥3).Univariate analysis was used to compare the general data (age,gender,body mass index,and underlying diseases) and perioperative conditions (immediate heart rate,systolic and diastolic blood pressures immediately after admission,operative time,and anesthesia time) between the two groups of patients.Multivariate Logistic regression analysis was used to identify the perioperative risk factors influencing the prognoses of patients with acute ischemic stroke accepted mechanical thrombectomy.Results Among the 108 patients,65 had good prognosis and 43 had poor prognosis.Univariate analysis showed that there was no significant difference in general data between the two groups (P＞0.05),but there were significant differences in heart rate immediately after admission,National Institutes of Health Stroke Scale (NIHSS) scores immediately after admission and 3 d after operation,maximum hemoglobin and blood glucose values from immediately after admission to 3rd d of operation,and thrombolysis in myocardial infarction (TIMI) blood flow classification (P＜0.05).Multivariate Logistic regression analysis showed that heartrate immediately after admission (OR=1.035,95％CI:1.002-1.067,P=0.037) and NIHSS scores 3 d after operation (OR=1.153,95％CI:1.016-1.272,P=0.030) were the perioperative risk factors influencing the prognoses of patients with acute ischemic stroke accepted mechanical thrombectomy.Conclusion For patients with acute ischemic stroke who have rapid heart rate immediately after admission and high NIHSS scores 3 d after mechanical thrombectomy,possibility of poor prognosis should be noticed.

The choice of females to accept or reject male courtship is a critical decision for animal reproduction. Serotonin (5-hydroxytryptamine; 5-HT) has been found to regulate sexual behavior in many species, but it is unclear how 5-HT and its receptors function to regulate different aspects of sexual behavior. Here we used Drosophila melanogaster as the model animal to investigate how 5-HT and its receptors modulate female sexual receptivity. We found that knockout of tryptophan hydroxylase (Trh), which is involved in the biosynthesis of 5-HT, severely reduced virgin female receptivity without affecting post-mating behaviors. We identified a subset of sexually dimorphic Trh neurons that co-expressed fruitless (fru), in which the activity was correlated with sexual receptivity in females. We also found that 5-HT_1A and 5-HT₇ receptors regulate virgin female receptivity. Our findings demonstrate how 5-HT functions in sexually dimorphic neurons to promote virgin female receptivity through two of its receptors.
Animals ; Male ; Female ; Drosophila/physiology* ; Drosophila melanogaster/physiology* ; Serotonin ; Drosophila Proteins/physiology* ; Sexual Behavior, Animal/physiology* ; Transcription Factors ; Nerve Tissue Proteins