Objective:To simultaneously determine six components ( paeoniflorin, rosmarinic acid, salvianolic acid B, ligustilide, clyptotanshinone and tanshinone ⅡA ) in refined coronary tablets by UHPLC-DAD. Methods: A Dikma Endevaorsil C18 column (2. 1 mm × 100 mm,1. 8 μm) was used to perform the determination, which was maintained at 30℃ during the analysis. The mobile phase was composed of acetonitrile and 0. 05% phosphoric acid at a flow rate of 0. 3 ml·min-1 with gradient elution. The detection wavelength was respectively set at 230,270,288 and 321 nm. Results:Paeoniflorin, rosmarinic acid, salvianolic acid B, ligustilide, clyptotanshinone and tanshinone ⅡA showed good linearity within the range of 0. 001 0-0. 010 2 μg ( r = 0. 999 8 ), 0. 005 7-0.056 9 μg(r =1.000 0), 0.005 3-0.052 7 μg(r = 1.000 0),0.002 1-0.020 6 μg(r = 1.000 0),0.001 1-0.011 2 μg(r =1. 000 0) and 0. 001 4-0. 014 4 μg(r=0. 999 8),respectively. The average recovery was 98. 78%(RSD=0. 50%), 97. 99%(RSD=0. 76%),98. 44%(RSD=0. 85%),99. 12%(RSD=0. 66%), 98. 82%(RSD=0. 81%) and 97. 80%(RSD=0. 80%), respec-tively. Conclusion:The method is simple, rapid and accurate, which can be used for the quality control of refined coronary tablets.

Objective:To determine the content of lead, cadmium, arsenic, mercury and copper in Xiaohuoluo pills produced by 34 enterprises. Methods:Using microwave digestion, the contents of lead, cadmium and arsenic were determined by graphite furnace atomic absorption spectrometry;the content of mercury was determined by hydride generation atomic absorption spectrophotometry;the content of copper was determined by air-acetylene flame atomic absorption. Results: Lead, cadmium, arsenic and mercury in Xiao-huoluo pills from 34 different manufacturers were beyond the limits in various degrees. Conclusion: The method is simple and accu-rate, and can be used in the heavy metal determination of Xiaohuoluo pills.

Objective To investigate the effect of high frequency stimulation of bilateral nucleus accumbens on relapse behaviors in morphine-dependent rats.Methods Twenty adult SD rats were employed in the experiment.Through stereotactic operation,outer electrode cannula was implanted into rats' bilateral nucleus accumbens.After 5 days of rest,the morphine-dependent rat model with conditioned place preference (CCP) was established through intraperitoneal morphine injection (10 mg/kg).After acquisition of CPP,normal saline was replaced with morphine for CPP extinction training.CPP test was used to exam the effect of extinction.The rats,after being randomly divided into experimental group (morphine+DBS) and control group (morphine+sham DBS),were electrically stimulated using modified DBS circuits.Rats in the experimental group were given high frequency electrical stimulation while the control group was sham stimulation.After consecutive stimulation for 7 days,rats in the two groups were given small dose of morphine (3 mg/kg)to trigger relapse.Results (1) The CPP score increased after the establishment of rat models compared with pre-establishment of the rat models((616.2±74.7) s vs (353.9±84.3) s,P＜0.01).(2) The CPP score after extinction training decreased compared with pre-conditioned CPP score ((456.4± 148.8) s vs (353.9±84.3) s),P=0.0847) and had no statistical difference compared with post-conditioned CPP score ((456.4±148.8) s vs (616.2± 74.7) s,P=0.0219).(3) When the relapse was induced by small doses of morphine within 24h after the last stimulation,the CPP score of the experimental group decreased compared with the CPP score of control group ((330.1 ±212.6) s vs (684.2±230.2)s,P=0.0029),and the relapse was restrained.Conclusion High frequency DBS of bilateral nucleus accumbens can attenuate relapse behavior in rats

Objective To investigate the influence of deep brain stimulation (DBS) at high frequency to the bilateral nucleus accumbens on morphine-induced conditioned place preference (CPP) and relapse behaviors during extinction phase in rats. Methods Twenty adult SD rats were employed in the experiment. Through stereotactic operation, outer electrode cannula was implanted into rats' bilateral nucleus accumbens. After 5 days of rest, the morphine-dependent rat model with CPP was established through intraperitoneal morphine injection (10 mg/kg). The rats, after being randomly divided into experimental group (morphine+DBS) and control group (morphine+sham DBS), were electrically stimulated using DBS circuits. Rats in the experimental group were given high frequency electrical stimulations while the control group was given sham stimulation. The CPP score of the two groups was recorded the day after stimulation until successful extinction and then the extinction time was compared between the two groups. After successful extinction the rats were given small dose of morphine to trigger relapse within 24 hours, and the CPP score was recorded and compared between the two groups.Results Compared with the control group (six days), the experimental group (26 days) had a longer extinction time. After relapse, the retention time within the drug-paired chamber of the experimental group was (357.01±192.72) s, obviously shorter than that of the control group ((704.91±181.35) s;t=2.370, P=0.034 6). Conclusion High frequency DBS to rats' bilateral nucleus accumbens can prolong extinction time but inhibit relapse behavior.

OBJECTIVE:To establish a method for the determination of related substances in bifonazole raw material. METH-ODS:HPLC method was performed on the column of Kromasil C18 with mobile phase of methanol-0.02 mol/L phosphoric acid(ad-justed pH to 7.5 with triethylamine)(70:30,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 258 nm,volume injection was 20 μl,and the column temperature was 40 ℃. RESULTS:The linear range was 0.05-0.25 μg/ml for bifonazole(r=0.9996), 0.05-0.25 μg/ml for impurity A(bifonol)(r=0.9997)and 0.05-0.25 μg/ml for impurity B(4-C isomer)(r=0.9995);the detec-tion limits were 8.2 ng/ml,7.5 ng/ml and 8.4 ng/ml,and the quantification limits were 27.1 ng/ml,24.7 ng/ml and 27.8 ng/ml,re-spectively;RSDs of precision and reproducibility tests were lower than 1%;recovery of impurity B was 95.13%-101.29%(RSD=1.89%,n=9);both impurity A and impurity B were were detected in the 3 batches of samples. CONCLUSIONS:The method is accurate,sensitive and reproducible,and can be used for the determination of the related substances in bifonazole raw material.

Objective To clone,express and characterize a tegument protein gene of Schistosoma japonicum(Sj29),and investigate the immune protection of the recombinant protein against S.japonicum in mice.Methods The gene coding for Sj29 protein was amplified by PCR,and the sequence was analyzed by bioinformatics tools.Partial fragment of Sj29 gene was subcloned into the prokaryotic expression vector pET28c(+).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced the recombinant with IPTG.The recombinant protein(rSj29) was purified by His-binding-resin affinity chromatography and characterized by Western blotting.Three groups each with 10 BALB/c mice were immunized subcutaneously three times(two weeks interval) respectively with 100 ?l recombinant rSj29(0.1 mg/ml),adjuvant or PBS.At the 15th day after the final inoculation,each mouse was challenged by 40 ?2 cercariae of S.japonicum.At the 53th day after infection,the mice were sacrificed to obtain the number of adult worms,number of eggs in liver and feces.Serum samples were collected at pre-immunization and certain time after immuniza-tion,and were analyzed for IgG by ELISA.The localization of rSj29 in worms of different developmental stages was demonstrated by immunofluorescent technique.mRNA expression level of Sj29 gene in worms of different developmental stages and three groups after infection was detected by quantitative real-time PCR.Results A 576 bp Sj29 gene fragment was obtained.The recombinant protein rSj29 with Mr 22 900 was expressed in the form of inclusion body.The recombinant rSj29 can be recognized by sera of mice immunized with rSj29 and sera of infected mice.The number of adult worms(15.4?5.9),number of hepatic eggs(40 143.3?2 995.9) and number of fecal eggs(3 803.9?110.9) in re-combinant protein group were significantly higher than those of PBS control group(20?3.4,49 318.1?6 648.3,5 238.1? 303.5,respectively)(P

Objective:To investigate the occurrence of carotid artery abnormalities in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients and the related risk factors affecting the occurrence of carotid artery abnormalities.Methods:A total of 169 regular follow-up outpatients with HIV/AIDS from April 2003 to December 2019 in Peking Union Medical College Hospital, whose carotid artery ultrasound examination were performed from July 2015 to December 2019 were included. The patients were divided into young (≤44 years old), middle-aged and elderly (≥45 years old) according to their ages, and the physical examination data of the included patients were collected.The statistical methods were logistic regression analysis and single sample t test. Results:Among the 169 HIV/AIDS patients, 40(23.7%) had abnormal carotid artery and 129(76.3%) had no abnormal carotid artery. Middle-aged and elderly people (odds ratio ( OR)=3.85, 95%confidence interval (95% CI) 1.54-9.65, P<0.01), hypertension ( OR=6.24, 95% CI 1.95-20.00, P<0.01), hyperlipidemia ( OR=2.44, 95% CI 1.00-5.93, P<0.05), and elevated human leucocyte antigen (HLA)-DR + CD8 + /CD8 + ( OR=1.03, 95% CI 1.01-1.06, P<0.05) were the risk factors for carotid artery abnormality. The common carotid artery inner medium film thickness (IMT) of patients in HIV/AIDS group Ⅰ (20 to 30 years old), group Ⅱ (31 to 40 years old), group Ⅲ (41 to 50 years old) were (0.061 0±0.001 2), (0.062 9±0.001 4) and (0.065 6±0.002 6) cm, respectively, which were thicker than the control groups ((0.051±0.003), (0.056±0.004) and (0.063±0.002) cm, respectively). The differences were all statistically significant ( t=5.119, 4.775 and 1.739, respectively, all P<0.05). The common carotid artery IMT of patients in HIV/AIDS group A (30 to 44 years old) and group B (45 to 59 years old) were (0.062 6±0.001 1) and (0.072 3±0.003 4) cm, respectively, which were thicker than the control groups ((0.052±0.011) and (0.064±0.015) cm, respectively), the differences were both statistically significant ( t=9.520 and 3.012, respectively, both P<0.01). Conclusion:Younger HIV-positive people have a higher probability of abnormal carotid arteries and may be at greater risk of cardiovascular disease than HIV-negative people of the same age, suggesting that HIV-positive people, especially young people, should be examined early with ultrasound of the neck arteries to detect abnormalities and intervene as soon as possible.

Objective To report the effects of surgical treatment of old die-punch fractures of the distal radius.Methods A retrospective study of case series was made of the 21 old die-punch fractures of the distal radius which had been surgically treated from January 2012 to January 2017 at Department of Orthopaedics,Shandong Provincial Hospital.There were 13 males and 8 females,aged from 34 to 63 years (average,46.9 years).According to the preoperative definite diagnoses by the X-ray and CT images,there were 14 metacarpal compression fractures and 7 dorsal compression fractures.The palmar approach was used in 13 cases,the dorsal approach in 7 and the combined approach in one.Autogenous iliac bone grafting was performed for all the cases.Their preoperative and postoperative Cooney scoring and visual analogue scale (VAS) of the wrist,and postoperative complications as well,were observed and recorded.Results This cohort was followed up for 8 to 14 months (average,9.2 months).All their postoperative wounds healed by the first intention.Their Cooney functional scores of the wrist were,respectively,58.5 ± 4.2 and 84.0 ± 3.1 points preoperatively and at the final follow-up,and their corresponding VAS scores 7.6 ± 0.9 and 3.4 ± 1.3 points,showing significant differences between preoperation and the final follow-up (P ＜ 0.05).Carpal arthritis was observed in one case.Conclusion For old die-punch fractures of the distal radius,surgical treatment can obviously reduce pain,improve function of the wrist and enhance the quality of life of the patients.

Objective To explore the application value of external fixator in obsolete hip dislocation.Methods Retrospective analysis of 9 cases of obsolete hip dislocation treated with external fixator from January 2010 to January 2017.There were 6 males and 3 females,with an average of 32.0±8.3 years old (range 19-47).The time from injury to operation was 3-9 months,with an average of 6.2± 1.9 months.The patient's limb shortening length,traction time,traction length,operation time,surgical bleeding volume,preoperative and last follow-up VAS score,Epstein score,D'Aubigne functional score were recorded.Results The length of the limb shortened 5-11cm,with an average of 6.7±2.0 cm.The preoperative traction time ranged 20-45 d,with an average of 26.9±8.2 d.The length of traction ranged 5-12 cm,with an average of 7.1±2.2 cm.The second reconstruction operation time was 2.0-4.0 hours,with an average of 2.8±0.7 h.The bleeding volume was 800-2 000ml,with an average of 1 216.7±422.8 ml.The Epstein score at the last follow-up was excellent in 4 cases,good in 4 cases and improved in 1 case.The preoperative VAS score and the final follow-up VAS score were 0.9±0.8 points and 4.6±0.9 points respectively.There was a significant difference between preoperative and postoperative VAS scores (t=8.981,P=0.000).The D'Aubigne functional scores of preoperative and final follow-up were 5.2±1.0 points and 13.1±2.0 points respectively and the difference was statistically significant (t=-12.044,P=0.000).Conclusion The external fixation in the treatment of obsolete dislocation of the hip is helpful and accepted in patients with femoral head dislocation.It is easy to carry and tract for patients and convenient for functional exercise.The external fixation makes operation simple and flexible;the traction force is strong and the effect of the femoral head reduction is obvious,the operation method is simplified and the quality of the reduction is improved.

Objectives:To investigate clinical characteristics, outcomes and antimicrobial resistance of community-acquired Pseudomonas aeruginosa (CAPA) infections in Chinese pediatric patients. Methods:This retrospective study was conducted at 6 tertiary hospitals in China during January 2016 to December 2018. The clinical and microbiological data of CAPA infected hospitalized children in Hainan and in other regions were collected and compared, and the antimicrobial resistance patterns, clinical characteristics and antibiotic therapy were analyzed. Between different groups were compared using the Chi-square test and Mann-Whitney U test. Results:Among 91 patients, 63 cases were males, 28 cases were females, and 74 cases were from Hainan province, 17 cases were from other regians. The age of consultation was 22.5 (5.4, 44.0) months. Twenty-four cases (26%) had underlying diseases. Fever (79 cases (87%)) and cough (64 cases (70%)) were common initial symptoms. Other concomitant symptoms included wheezing 8 cases (9%), diarrhea 3 cases (3%) and vomiting 4 cases (4%). Twenty-eight cases (31%) had organ infections, including pneumonia 22 cases (24%), skin infection 5 cases (5%), meningitis, intra-abdominal infection and upper urinary tract infection each 1 case (1%). The resistance rate of CAPA isolates to cefepime (4% (4/90)), amikacin (1% (1/90)), ciprofloxacin (2% (2/90)) and levofloxacin (1% (1/89)) was low, and to ceftazidime, piperacillin, piperacillin-azobactam, carbapenem was 12% (11/90), 3/16, 18% (10/56) and 6% (5/90), respectively. Antimicrobial combination therapy accounted for 52% (47/91) of empirical therapy and 59% (52/88) of definite therapy. Two cases (2%) were hopeless discharged, and 3 cases (3%) died during hospitalization. The worse prognosis of CAPA infection is significantly different among children in other regions and in Hainan (4/17 vs. 1% (1/74), χ2=9.74, P<0.05). Conclusions:The invasive CAPA-infection has regional difference in incidence and prognosis in China. Clinical symptoms and signs are non-specific. CAPA strains isolated from pediatric patients display low level of resistance to most of the common antipseudomonal antibiotics. The proportion of poor prognostic outcome is lower in Hainan than in other regions.