Objective To investigate the hepatoprotective effect of Propolis alcohol-extract (PAE) against acetaminophen (APAP)-induced acute hepatic damage in mice and to explore the possible mechanism.Methods Sixty-three C57BL/6 MT(-/-)mice were equally randomized into 9 groups.The normal control group and the model group received gastric gavage of normal saline (20 mL?kg-1),four PAE groups were given PAE in the dose of 12.5,25,50 and 100 mg?kg-1 respectively,alcohol +APAP group and alcohol control group received 20 %alcohol 20 mL?kg-1 and PAE control group was given PAE in the dose of 100 mg?kg-1 qd,for 4 consective days.Thirty miniutes after last administration,the mice in the normal control group,PAE control group and alcohol control group received saline 10 mL?kg-1,and the mice in other groups received APAP 380 mg?kg-1 to induce acute hepatic injury.The activities of serum alanine aminotransferase (ALT),aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were determined,and the liver tissues were collected for histopathological assessment by HE staining under light microscope.The ratio of glutathione (GSH) and oxidized glutathione (GSSG),and the content of GSH,GSSG and malondialdehyde (MDA) in liver homogenate were also measured.Results Compared with the model group,PAE could markedly decrease serum ALT,AST and LDH activity,reduce the MDA level in liver homogenate,and increase hepatic GSH content and the ratio of GSH/GSSG in the liver homogenate.The hepatic histopathological changes in liver were also significantly ameliorated.In PAE control group,GSH content and the ratio of GSH/GSSG were also increased.However,the above indexes remained unchanged in alcohol control group.Conclusion The propolis alcohol-extract can prevent the liver from PAP-induced acute hepatic injury.

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.

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.

The interferon-γ testing and the comparative cervical skin test were compared with the simple cervical hypersensitivity test routinely used in our country in herds infected with Mycobacterium bovis, in which 167 infected cattle from 5 herds were tested with interferon-γ testing and the comparative cervical test (CCT) simultaneously and other 106 cattle were tested with other hypersensitivity tests used as control for comparison. In these 167 cattle tested with both tests, 89 cattle were proved to be positive both in interferon-γ testing and CCT using the bovine PPD of Netherland with a coincidence rate of 92.7% (89/96). In 106 cattle the coincidence rate of positive reactions detected by γ-interferon test and CCT was 93.41% (78/83.5), that detected by the routine skin of foreign country was 62.26%(66/106); and that between the hypersensitivity test of China and foreign country was 92.19% (59/64). It is obvious that the interferon-γ testing and CCT seem to have higher coincidence rate and the specificity of the skin tests used in our country was rather low. Consequently, the use of the comparative cervical skin test should be considered to replace the routinely used shin tests.

OBJECTIVE To study the protective effect of Orychophragmus violaceus(OV)seed against acute hepatotoxicity induced by the traditional Chinese Medicine Cortex Dictamni in mice. METHODS Twenty-five mice were randomly divided into 5 groups:control group,Cortex Dictamni group(70 g · kg-1)and OV seed groups(36,54 and 72 g · kg-1). OV Seed groups were orally adminis?tered with the aqueous extract of OV seed for 4 consecutive days while the other groups were ig given water. On the 4th day,Cortex Dictamni group and OV seed groups were ig given the aqueous extract of Cortex Dictamni,and normal control group was ig given water. Twenty-four hours later,all the mice had their blood and liver samples taken after anesthesia. The serum chemical parameters were measured, including glutamic oxaloacetic transaminase (GOT),glutamate pyruvate transaminase (GPT) and lactate dehydrogenase(LDH),as well as malondialdehyde(MDA),glutathione(GSH)and oxidized glutathione(GSSG)levels in the liver. GSH/GSSG ratio was calculated. Histopathologic changes in the liver were observed and the area was calculated after HE staining. RESULTS Compared with normal control group,Cortex Dictamni(70 g · kg-1)increased the serum GOT,GPT and LDH levels by 500, 140 and 40 fold(P<0.01). OV seed reduced serum GOT,GPT and LDH levels increased by Cortex Dictamni(P<0.05,P<0.01),by as much as 62%,75% and 99% for GPT,70%,82% and 98% for GOT,and 55%,75%and 96%for LDH,respectively. The level of MDA and the ratio of GSH/GSSG in Cortex Dictamni group were 1.39 ± 0.58 and(3.53 ± 1.27)μmol · g-1,a 10-fold increase and 40%decline compared with normal control group(P<0.01). OV seed of 72 g·kg-1 lowered the level of MDA by 22%(P<0.05),and OV seed(36,54 and 72 g · kg-1)increased the GSH/GSSG ratio by 47%,42%and 54%(P<0.05). Histopathologic results showed that OV seed alleviated the liver damage induced by Cortex Dictamni from(64.1±8.5)%to(37.5±7.1)%and (20.0±0.8)%(P<0.01). CONCLUSION OV seed can effectively protect mice from the acute hepatotoxicity induced by Cortex Dictamni.

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.

OBJECTIVETo study the immunotoxicity induced by 9,10-dimethyl-1,2-benzathrancene (DMBA) in metallothionein gene-knocked-out mice [MT(-/-)] as compared with that in wild-type mice [(MT(+/+)].

METHODSFemale mice were treated with 25 mg/kg and 50 mg/kg of DMBA i.p., respectively and immunized with sheep red blood cells (SRBC) i.v. on the following day and rechallenged by injection of SRBC via footpad s.c. on the fourth day post-immunization. Humoral and cell-mediated immune function was assessed by the number of spleen IgM antibody plaque formation cells (PFC) to SRBC and cell-mediated delayed-type hypersensitivity (DTH) measured by footpad swelling thickness.

RESULTSAfter treatment with 25 mg/kg DMBA, a decrease in weight of their spleen and thymus and PFC/spleen were observed in MT(-/-) mice, while only decrease in thymus weight of MT(+/+) mice. The humoral function was suppressed by 72% in MT(-/-) mice. No obvious change in cell-mediated immune function was observed both in MT(-/-) and MT(+/+) mice. Both humoral and cell-mediated immune function were suppressed more severe (91%) in MT(-/-) mice treated with 50 mg/kg DMBA than those treated with 25 mg/kg DMBA (72%). DTH was not altered by DMBA in MT(+/+) mice. The weight of their spleen and thymus decreased and humoral immune function suppressed in MT(+/+) mice, but these changes were significantly less severe. No obvious suppression of cell-mediated immune function was observed in MT(+/+) mice.

CONCLUSIONTheir humoral and cell-mediated immune function was more susceptible to being suppressed by DMBA in MT(-/-) mice, indicating that MT could protect their immune function from damage caused by DMBA.

9,10-Dimethyl-1,2-benzanthracene ; toxicity ; Animals ; Immunity ; drug effects ; Metallothionein ; physiology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Organ Size ; drug effects

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.