Objective:To evaluate the role of nuclear factor erythroid 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in the improvement in postoperative cognitive function by electroacupuncture preconditioning in aged rats.Methods:Eighty clean-grade healthy male Sprague-Dawley rats, aged 24 months, weighing 550-600 g, were divided into 4 groups ( n=20 each) using a random number table method: control group (C group), POCD group, electroacupuncture+ POCD group (EA group), and Nrf2 inhibitor ML385+ electroacupuncture+ POCD group (I group). A rat POCD model was prepared by performing exploratory laparotomy under 3% sevoflurane anesthesia. Nrf2 inhibitor ML385 15 mg/kg was intraperitoneally injected at 30 min before surgery in group I, while the equal volume of normal saline was given instead in the remaining groups. Electroacupuncture stimulation of the Zusanli, Hegu and Neiguan acupoints (disperse-dense waves, current intensity 1 mA, frequency 2/15 Hz, 30 min) was performed at 8: 00 a.m. every day for 5 days prior to model preparation in EA group and I group. An open field test was conducted at 3 days after operation to measure the autonomous motor function, and Morris water maze test was conducted after the end of the open field test at 3-7 days after operation to evaluate the cognitive function. After the Morris water maze test, the rats were sacrificed under anesthesia and hippocampal tissues were taken for determination of the level of reactive oxygen species (ROS, by flow cytometry) and expression of Nrf2 and HO-1 (by Western blot) and for examination of the mitochondrial ultrastructure of hippocampal neurons (with a transmission electron microscope). Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was decreased, the hippocampal ROS levels were increased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and mitochondria were swollen and mitochondrial cristae were broken in the hippocampal CA1 neurons in group POCD. Compared with group POCD, the escape latency was significantly shortened, the number of crossing the original platform quadrant was increased, the hippocampal ROS levels were decreased, the expression of Nrf2 and HO-1 was up-regulated ( P<0.05), and the swelling of mitochondria and rupture of cristae structure were improved in the hippocampal CA1 neurons in group EA. Compared with group EA, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was decreased, the hippocampal ROS levels were increased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and the swelling of mitochondria and rupture of cristae structure were aggravated in the hippocampal CA1 neurons in group I. Conclusions:Activation of the hippocampal Nrf2/HO-1 signaling pathway is involved in improvement in postoperative cognitive function by electroacupuncture preconditioning in aged rats.

OBJECTIVE To investigate the changes in high performance liquid chromatography （HPLC） fingerprint spectra and nucleoside components between Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and its processed product Succus bambusae pinella preparata， providing a reference for the quality evaluation of the latter. METHODS HPLC fingerprint was established for 10 batches of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and its processed product Succus bambusae pinella preparata following the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 Edition）. Hierarchical cluster analysis （HCA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS- DA） were conducted on their common peaks. The contents of four nucleoside components， hypoxanthine， uridine， adenine， and guanosine， in both Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and Succus bambusae pinella preparata were determined. RESULTS The similarity between the fingerprints of the 10 batches of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine， Succus bambusae pinella preparata， and their corresponding reference fingerprints ranged from 0.851 to 0.990. A total of 10 common peaks were obtained for both samples， and 4 components were identified as hypoxanthine， uridine， adenine， and guanosine. The results of HCA， PCA and OPLS-DA showed that the samples of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and Succus bambusae pinella preparata were clustered into separate categories， with OPLS-DA selecting 4 differential components between them， ranked by variable importance projection values as peak 8， peak 1， peak 6 （adenine） and peak 10. The content determination results showed that the average contents of hypoxanthine， uridine， adenine and guanosine in Succus bambusae pinella preparata declined by 15.90%， 12.00%， 26.04% and 22.18% compared to Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine， respectively， with statistically significant differences in the contents of hypoxanthine， adenine and guanosine （P＜0.05 or P＜0.01）. CONCLUSIONS The established fingerprint and content determination methods are simple to operate and have good repeatability， which are suitable for qualitative and quantitative analysis of Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and Succus bambusae pinella preparata. The average contents of the four nucleoside components decreased after the processing of Succus bambusae pinella preparata.

Objective:To evaluate the role of nuclear factor erythroid 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in the improvement in postoperative cognitive function by electroacupuncture preconditioning in aged rats.Methods:Eighty clean-grade healthy male Sprague-Dawley rats, aged 24 months, weighing 550-600 g, were divided into 4 groups ( n=20 each) using a random number table method: control group (C group), POCD group, electroacupuncture+ POCD group (EA group), and Nrf2 inhibitor ML385+ electroacupuncture+ POCD group (I group). A rat POCD model was prepared by performing exploratory laparotomy under 3% sevoflurane anesthesia. Nrf2 inhibitor ML385 15 mg/kg was intraperitoneally injected at 30 min before surgery in group I, while the equal volume of normal saline was given instead in the remaining groups. Electroacupuncture stimulation of the Zusanli, Hegu and Neiguan acupoints (disperse-dense waves, current intensity 1 mA, frequency 2/15 Hz, 30 min) was performed at 8: 00 a.m. every day for 5 days prior to model preparation in EA group and I group. An open field test was conducted at 3 days after operation to measure the autonomous motor function, and Morris water maze test was conducted after the end of the open field test at 3-7 days after operation to evaluate the cognitive function. After the Morris water maze test, the rats were sacrificed under anesthesia and hippocampal tissues were taken for determination of the level of reactive oxygen species (ROS, by flow cytometry) and expression of Nrf2 and HO-1 (by Western blot) and for examination of the mitochondrial ultrastructure of hippocampal neurons (with a transmission electron microscope). Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was decreased, the hippocampal ROS levels were increased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and mitochondria were swollen and mitochondrial cristae were broken in the hippocampal CA1 neurons in group POCD. Compared with group POCD, the escape latency was significantly shortened, the number of crossing the original platform quadrant was increased, the hippocampal ROS levels were decreased, the expression of Nrf2 and HO-1 was up-regulated ( P<0.05), and the swelling of mitochondria and rupture of cristae structure were improved in the hippocampal CA1 neurons in group EA. Compared with group EA, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was decreased, the hippocampal ROS levels were increased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and the swelling of mitochondria and rupture of cristae structure were aggravated in the hippocampal CA1 neurons in group I. Conclusions:Activation of the hippocampal Nrf2/HO-1 signaling pathway is involved in improvement in postoperative cognitive function by electroacupuncture preconditioning in aged rats.

Objective:To explore the molecular mechanism of the effect of the histone methylase zeste gene enhancer homolog 2 (EZH2) on the proliferation and apoptosis of human hypertrophic cardiomyocytes AC16.Methods:The AC16 hypertrophic cardiomyocyte model was constructed by adding angiotensin Ⅱ to the AC16 cell culture medium. The cells were divided into four groups, including the blank control group, the angiotensin Ⅱ group, the empty vector + angiotensin Ⅱ group, and the EZH2 overexpression + angiotensin Ⅱ group. The expression levels of EZH2 and brain natriuretic peptide ( BNP) genes were measured using fluorescent quantitative PCR. The EZH2, trimethylation of lysine at position 27 of histone H3 (H3K27me3), and BNP proteins expression were detected by Western Blot. The MTS method was used to detect the proliferation of AC16 cell. The Annexin V-FITC/PI double staining method was used to detect the apoptosis of AC16 cell. Results:Compared with the blank control group, the expression levels of EZH2 and H3K27me3 in the angiotensin Ⅱ group were decreased, the expression level of BNP was increased, cell proliferation was decreased, and apoptosis was increased (all P < 0.001). Compared with the empty vector + angiotensin Ⅱ group, the expression levels of EZH2 and H3K27me3 in the EZH2 overexpression + angiotensin Ⅱ group were increased, the expression level of BNP was decreased, the cell proliferation level was increased, and the apoptosis level was decreased (all P < 0.001). There was no significant difference between the angiotensin Ⅱ group and the empty vector + angiotensin Ⅱ group (all P > 0.05). Conclusions:Histone methylase EZH2 has an effect on the proliferation and apoptosis of AC16 cell, providing a reference for the treatment of myocardial hypertrophy and revealing the exact pathogenesis of myocardial hypertrophy.

ObjectiveTo determine the pathogens and drug resistance in patients with bacterial diarrhea in Lishui City of Zhejiang Province from 2015 to 2019， and provide scientific evidence for prevention and treatment of bacterial diarrhea. MethodFecal specimens were collected from patients with diarrhea in the People’s Hospital of Lishui City from 2015 to 2019. Bacteria were identified by time-of-flight mass spectrometer and serum agglutination reaction. Drug sensitivity in the suspected bacteria was identified by VITEK 2 Compact system. ResultsA total of 2 937 fecal samples were tested from 2015 to 2019， of which 191 were positive for bacteria. The prevalence was 6.65% in male and 6.32% in female. It was highest in the age group 21‒30 years old， followed by the group 51‒60 years old. Summer was the season with the highest prevalence of bacteria. Furthermore，the bacterial species included salmonella （3.98%）， Vibrio parahaemolyticus （1.43%）， aeromonas （0.48%）， shigella （0.37%） and other bacteria （3.66%）. Salmonella had high resistance to cefuroxime and amikacin. Vibrio parahaemolyticus and shigella had high resistance to ampicillin. Aeromonas had high resistance to ampicillin and ampicillin/sulbactam. ConclusionPrevalence of bacteria differs by gender， age and seasons in patients with bacterial diarrhea in Lishui from 2015 to 2019. Salmonella is the principal pathogen in bacterial diarrhea. Additionally， multiple drug resistance is commonly identified. Therefore， it warrants strengthening the pathogenic surveillance on bacteria and drug resistance in bacterial diarrhea.

Objective:To investigate the effects of doxorubicin (Dox)-loaded tumor-derived extracellular vehicles (EVs) on cell proliferation and apoptosis of human hepatocellular carcinoma.Methods:The extracellular vesicles loaded with Adriamycin (EVs-Dox) were prepared by the method of directly co-incubation. The morphology of EVs-Dox was detected by transmission electron microphotometer. The diameter of EVs-Dox was determined by dynamic light scattering (DLS). Western blotting was utilized to detect the expression of CD63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was calculated by tandem mass spectrometry (LC-MS/MS). The drug release experiment in vitro was utilized to simulate the drug release of drug-loaded vesicles in vivo. PKH67-labeled EVs-Dox was showed cellular uptake. After treatment with EVs-Dox, MTS assay and flow cytometry assay were conducted to investigate the effects of EVs-Dox on cell proliferation and apoptosis of PLC/PRF/5.Results:The EVs-Dox showed an elliptical double-layer membrane structure of different sizes under transmission electron microscope. The diameter of EVs-Dox was (115.9±5.2) nm.Western blotting data showed high expression of CD 63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was 0.77%. The in vitro release experiment showed that the drug-loaded vesicles could release the drug slowly. PKH67-labeled EVs-Dox showed that carcinoma cells can uptake EVs-Dox within 16h. MTS assay showed that the cell viability rate of (54.9±3.2) % was significantly lower than that of in the Dox group [(77.7±5.4)%, P<0.05]. EVs-Dox inhibited hepatocellular carcinoma proliferation. Flow cytometry assay showed that the apoptosis rate of EVs-Dox (47.9±7.0) % was higher than that in the Dox group [(38.0±1.5)%, P<0.05]. Conclusion:EVs-Dox inhibits cell proliferation and accelerates apoptosis of hepatocellular carcinoma cells.

Objective:To evaluate the drug-drug interactions and the tolerability of combined medication between yimitasvir phosphate capsules with sofosbuvir tablets, omeprazole magnesium enteric-coated tablets, and rosuvastatin calcium tablets in healthy volunteers.Methods:A randomized, open, and continuous administration design was used in trial 1 (yimitasvir phosphate capsules with sofosbuvir tablets). 28 subjects were randomly divided into two groups. A non-randomized, open design was used in trial 2 (yimitasvir phosphate capsules with omeprazole magnesium enteric-coated tablets), and included 42 subjects divided into three groups. The open design method was used in trial 3 (yimitasvir phosphate capsules with rosuvastatin calcium tablets), and included 14 subjects. The plasma concentrations of yimitasvir phosphate, sofosbuvir and their main metabolites GS-331007, omeprazole and rosuvastatin were validated by a liquid chromatography/tandem mass spectrometry (LC-MS/MS). The pharmacokinetic parameters were calculated by Phoenix winNonlin software.Results:(1) in trial 1, after single and co-administration, the 90% CI of sofosbuvir C max and AUC 0-tau geometric mean ratio (GMR) were 152.0% (118.0% ~ 197.0%) and 230.0% (184.0% ~ 287.0%), with an increase of 52.0% and 130.0% compared to single dose of sofosbuvir, respectively. The 90% CI of GS-331007 C max GMR was 74.0% (67.5% ~ 81.2%) and reduced by 26% compared to single dose of sofosbuvir. (2) in trial 2, the 90% CI of C max GMR after yimitasvir single or co-administration at the same time, with a 4-hours interval, or with a 12- hours interval were 68.9% (44.5% ~ 106.7%) , 64.0% (43.8% ~ 93.6%) and 56.4%(38.9% ~ 81.9%), and the 90% CI of AUC 0-t GMR were 68.6% (46.5% ~ 101.2%), 68.3% (47.6% ~ 98.0%) and 60.5% (41.8% ~ 87.5%), respectively. Compared with single dose of yimitasvir, the C max and AUC 0-t were decreased by 31.1% and 31.4%, 36.0% and 31.7%, 43.6% and 39.5%, respectively. (3) In trial 3, after single and co-administration, the 90% CI of rosuvastatin C max and AUC 0-72 GMR were 172.4% (153.6% ~ 193.5%) and 158.0% (144.3% ~ 172.9%), respectively, with an increase of 74.9% and 60.5% compared to single dose of rosuvastatin. There were no serious adverse events and adverse events leading to withdrawal from the trial. Conclusion:Yimitasvir phosphate capsules have drug-drug interactions with sofosbuvir tablets, omeprazole magnesium enteric-coated tablets, and rosuvastatin calcium tablets.

OBJECTIVE： To provide reference for the evaluation of the correlation between drugs and adverse drug reaction （ADR） and the implementation of medication therapy management （MTM）. METHODS： Clinical information of a elderly patient with chronic disease （hypertension and coronary heart disease） whose suffered from leukocyte and platelet counts reduction and abnormal liver biochemical examination after taking candesartan were analyzed retrospectively in outpatient department of Tianjin Third Central Hospital. MTM pharmacists analyzed the correlation of candesartan with ADR using Naranjo evaluation scale method. The reasons for abnormal liver biochemical examination were analyzed by Naranjo evaluation scale method combined with Roussel Uclaf causality analysis method （called RUCAM method for short）. The medication reconciliation was conducted according to the results， and pharmacists cooperated with doctors to set individualized medication regimen and follow-up. RESULTS： By Naranjo evaluation scale method， analysis results showed that candesartan was “probably related” to ADR. By RUCAM method， analysis results showed that candesartan was “probably related” to liver biochemical abnormalities. MTM pharmacists suggested that candesartan should be stopped in time and the patient’s medication should be adjusted. The physician and the patient adopted the pharmacist’s advice. After 38 days of drug withdrawal， the patient’s ADR symptoms disappeared， and leukocyte count， platelet count and liver biochemical examination were normal. After adjustment of medication， the patient was followed up for 6 months with normal blood pressure. CONCLUSIONS： Naranjo evaluation scale method and RUCAM are simple and feasible in evaluating the correlation of drugs with ADR and hepatotoxicity. The two methods are consistent in evaluating the correlation between drugs and hepatotoxicity. Naranjo scale method and RUCAM method can be combined to analyze the correlation between drugs and ADR with abnormal liver biochemical examination.

ObjectiveTo examine the characteristic of criminal parenting styles and personality.Methods 379 prisoners took part in by using EMBU and PDQ-4 +.Results ( 1 ) There exist significant difference of parenting styles as parental emotional warmth and understanding,rejection and deny,punishment and strictness among the different criminals type (F =5.70,3.45,3.61,4.66,4.13,2.79,P ＜ 0.01 ).(2) There exist significant difference of total PDQ-4 + scores and the subscales for different type of personality disorder except obsessive-compulsive and schizotypal personality disorder among different criminals type (F =7.67,4.06,14.02,6.94,4.20,4.19,4.01,7.52,7.21,7.21,9.11 ; P ＜ 0.01 or P ＜ 0.05 ). ( 3 ) There were negative correlations between the score in the subscales for different type of personality disorder except obsessive-compulsive personality disorder and the factor of parental emotional warmth and understanding ( r =- 0.12 ～ - 0.32 ; P ＜ 0.01 or P ＜ 0.05 ).And there were positive correlations between the score in the all subscales for different type of personality disorder and the factors of parental rejection and deny,punishment and strictness ( r=0.11 ～ 0.42 ; P＜ 0.01 or P＜ 0.05).(4)Paternal punishment and strictness,maternal emotional warmth and understanding,maternal partiality make a certain prediction about characteristic of criminal personality.ConclusionThere exist different characteristic of criminal parenting styles and personality among different criminal type,and some parenting styles make a certain prediction about characteristic of criminal personality characteristics.

Objective To compare the treatment effect of different doses methylprednisolone(MP)combined with monosialotetrahexosyl ganglioside(GM-1)on experimental spinal cord injury.Methods Based on the duplication of compressed spinal cord animal model,36 rats were divided into three groups randomly:the first group with high dose of MP,the second with moderate dose of MP and GM-1,the third with low dose of MP and GM-1.The motor evoked potential(MEP)and the grades of BBB were detected on the 7th day and 14th day after injury.Results The peak latencies and the amplitudes of MEP and the BBB grades changed significantly after injury in the three groups.After treatment by different administration,the recovery of the two combined groups was more obvious than that of the high dose MP group,and the effect with the low dose of MP and GM-1 group was most significant.Conclusion The treatment effect of low dose MP combined with GM-1 is better than the other two therapies on experimental spinal cord injury.