OBJECTIVE To establish the fingerprint and multi-component content determination method of Crataegus pinnatifida leaves from different producing areas， and to evaluate the quality of C. pinnatifida leaves and screen the differential markers. METHODS Seventy-eight batches of C. pinnatifida leaves were collected from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province. High-performance liquid chromatography （HPLC） and Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints （2012 edition） were used to draw the fingerprints and conduct similarity evaluation. Grey correlation analysis， cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed by using SPSS 19.0， MetaboAnalyst 5.0 and SIMCA 14.1 software. The differential markers affecting the quality of C. pinnatifida leaves were screened with variable importance in the projection （VIP） value greater than 1 and the error line not exceeding the origin as the criterion. Using vitexin rhamnoside as an internal reference， the contents of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin in 78 batches of C. pinnatifida leaves were determined by the same HPLC combined with quantitative analysis of multi- components by single-marker （QAMS）， and the results were compared with external standard method. RESULTS Eight common peaks were calibrated in the fingerprints for 78 batches of C. pinnatifida leaves from 4 producing areas. Five known components were identified， including chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， vitexin rhamnoside （peak 4）， hypericin （peak 7） and isoquercetin （peak 8）； their similarities ranged from 0.871 to 0.998. Average relative correlations of samples from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province were 0.538， 0.528， 0.462 and 0.435， respectively. CA and PCA showed that the samples from Chengde of Hebei Province and Huludao of Liaoning Province were roughly classified into one category， while the samples from Linyi of Shandong Province and Yuncheng of Shanxi Province were roughly classified into one category； VIP values of peak 1， 2， 3 and 5 were all greater than 1. By QAMS， the relative correction factors of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin were 0.401， 0.993， 1.670 and 1.615 （RSD＜2%）. Compared with external standard method， except for isoquercetin in the two batches of samples （S39 and S41）， there was no significant difference in the content of each component in other batches of samples （the relative deviations≤ 5%）. CONCLUSIONS The established fingerprint and QAMS method are simple to operate and can be used to evaluate the quality of C. pinnatifida leaves. The sample from Chengde of Hebei Province is relatively good in quality. Chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， and the corresponding components of peaks 2 and 5 may be differential markers affecting the quality of C. pinnatifida leaves.

OBJECTIVE To determi ne the contents of total fla vonoids i n Scutellaria barbata standard decoction ，evaluate in vitro antioxidant activity ，establish the fingerprint and conduct chemical pattern recognition analysis. METHODS The contents of total flavonoids in S. barbata standard decoction （calculated by scutellarein ）were determined by ultraviet-visible spectrophotometry. In vitro antioxidant activity of S. barbata standard decoction was investigated by free radical scavenging tests of 1，1-diphenyl- 2-trinitrophenylhydrazine（DPPH）and 2，2′-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid ）ammonium salt （ABTS）；HPLC method was adopted. Using scutellarin as reference ，the fingerprints of 16 batches of S. barbata standard decoction were drawn and evaluated by Similarity Evaluation System of TCM Chromatogram Fingerprint （2004 A edition ），and the common peaks were determined；Pearson correlation analysis was carried out by using SPSS 24.0 software to screen substances with in vitro antioxidant activity. Taking them as variables ，cluster analysis and principal component analysis were carried out by using SPSS 24.0 and SIMCA 14.1 software. RESULTS The linear range of total flavonoids were 2.106-21.06 μg/mL（R2＝0.999 3）；RSDs of precision ， reproducibility and stability tests （120 min）were all lower than 2％；the recovery was 100.62％（RSD＝0.55％，n＝6）；the contents of total flavonoids were 0.634-1.053 mg/mL. Median inhibitory concentration （IC50） of DPPH radical scavenging experiment ranged 1.120-3.602 mg/mL，and IC 50 of ABTS radical scavenging e xperiment range d 0.684-1.327 mg/mL. The results of correlation analysis showed that the content of total flavonoids Δ 基金项目 ：河北省高校省级重点学科建设项目 （No.冀教 in S. barbata standard decoction was negatively correlated 高〔2013〕4号）；承德医学院自然科学研究计划项目（No.201824） ＊讲师，硕士。研究方向：中药质量控制 。电话：0314-2291186。 with the IC 50 of DPPH free radical and ABTS free radical E-mail：duyilongww@sina.com scavenging experiment ，and the correlation coefficients were # 通信作者 ：教授，硕士。研究方向 ：中药质量控制 。电话： －0.976 and －0.940 respectively（P＜0.01）. There were 18 0314-2291186。E-mail：phf2301@163.com common peaks in the fin gerprints of 16 batches of S. barbata 中国药房 2022年第33卷第4期 China Pharmacy 2022Vol. 33 No. 4 ·425· standard decoction ；the s imilarities were 0.964-0.997. A total of 4 common peaks were identified ，such as scutellarin （peak 8）， scutellarein（peak 14），luteolin（peak 15），apigenin（peak 17）.In the HPLC fingerprints of S. barbata standard decoction ，the peak areas of peak 3-4，8-9，12-15 and 17 were significantly negatively correlated with the IC 50 of DPPH free radical and ABTS free radical scavenging experiment （P＜0.05）. The results of cluster analysis showed that 16 batches of S. barbata standard decoction could be clustered into two categories ，of which S 2，S7-S8 and S 14-S16 were clustered into one category ，S1，S3-S6 and S 9-S13 were clustered into one category. By principal component analysis ，16 batches of S. barbata standard decoction were divided into two categories ，of which S 2，S4，S7 and S 14-S16 were clustered into one category ，and S 1，S3，S5-S6 and S 8-S13 were clustered into one. The comprehensive scores were high in the samples of S 4，S13，S15. CONCLUSIONS Established HPLC fingerprint and chemical pattern recognition analysis method can be used to evaluate the quality of S. barbata standard decoction ； peak 3-4，8-9，12-15 and 17 and total flavonoids are the potential material basis for S. barbata standard decoction to scavenge DPPH free radical and ABTS free radical.

OBJE CTIVE To establish a method for quality evaluation of Xin ’an capsule by combining fingerprint ， multi-component quantitative analysis and chemical pattern recognition analysis. METHODS High performance liquid chromatography（HPLC）combined with Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 edition）were used to establish the fingerprints of 24 batches of Xin ’an capsules and evaluate the similarity. The common peaks were determined. The contents of glucosylvitexin ，rhamnosylvitexin，vitexin，hyperoside and isoquercetin in Xin ’an capsules were determined by the same HPLC method. Taking the common peak area of fingerprint as the variable ，MetaboAnalyst 5.0 tool was used to draw the cluster analysis （CA）heat map. SIMCA 14.1 software was used to perform principle component analysis （PCA）and partial least squares-discriminant analysis （PLS-DA）. RESULTS Twelve common peaks were identified with the similarity greater than 0.97. Six common peaks were identified as chlorogenic acid ，glucosylvitexin，rhamnosylvitexin，vitexin，hyperoside and isoquercetin.The linear range of glucosylvitexin ，rhamnosylvitexin，vitexin， hyperoside and isoquercetin were 2.36-151.35，9.15-585.20， 1.20-76.50， 0.68-43.20， 0.44-27.90 µg/mL（all r＞0.999）.RSDs of precision ，repeatability and stability （24 h）tests were 163.com all less than 2.00％ . The average recoveries were 95.80％（RSD＝0.96％ ，n＝6），102.10％ （RSD＝0.93％ ，n＝6）， 103.26％（RSD＝1.28％，n＝6），103.89％（RSD＝0.73％，n＝6） and 102.09％（RSD＝1.79％，n＝6），respectively. The contents of the five components were 0.988 8-1.559 1，4.336 6-11.220 1， 0.065 1-0.830 5，0.043 8-0.692 5 and 0.023 2-0.427 2 mg/grain，respectively. The results of CA and PCA showed that 24 batches of samples could be divided into three categories ，i.e. S 1-S15，S16-S18 and S 19-S24. PLS-DA showed that variable importance in projection values of the corresponding component of peak 6 and glucosylvitexin （peak 7），rhamnosylvitexin（peak 8），hyperoside （peak 10） and isoquercetin （peak 11） were greater than 1. CONCLUSIONS The established HPLC fingerprint and multi-component quantitative method are simple and feasible. Combined with chemical pattern recognition analysis ，it can be used for the quality control of Xin ’an capsules. Glucosylvitexin ，rhamnosylvitexin and other components may be differentital markers affecting the quality of each batch of samples.

Background: and Purpose Despite administration of evidence-based therapies, residual risk of stroke recurrence persists. This study aimed to evaluate the residual risk of recurrent stroke in acute ischemic stroke or transient ischemic attack (TIA) with adherence to guideline-based secondary stroke prevention and identify the risk factors of the residual risk. Methods: Patients with acute ischemic stroke or TIA within 7 hours were enrolled from 169 hospitals in Third China National Stroke Registry (CNSR-III) in China. Adherence to guideline-based secondary stroke prevention was defined as persistently receiving all of the five secondary prevention medications (antithrombotic, antidiabetic and antihypertensive agents, statin and anticoagulants) during hospitalization, at discharge, at 3, 6, and 12 months if eligible. The primary outcome was a new stroke at 12 months. Results: Among 9,022 included patients (median age 63.0 years and 31.7% female), 3,146 (34.9%) were identified as adherence to guideline-based secondary prevention. Of all, 864 (9.6%) patients had recurrent stroke at 12 months, and the residual risk in patients with adherence to guidelinebased secondary prevention was 8.3%. Compared with those without adherence, patients with adherence to guideline-based secondary prevention had lower rate of recurrent stroke (hazard ratio, 0.85; 95% confidence interval, 0.74 to 0.99; P=0.04) at 12 months. Female, history of stroke, interleukin-6 ≥5.63 ng/L, and relevant intracranial artery stenosis were independent risk factors of the residual risk. Conclusions There was still a substantial residual risk of 12-month recurrent stroke even in patients with persistent adherence to guideline-based secondary stroke prevention. Future research should focus on efforts to reduce the residual risk.

Objective　To investigate the clinical efficacy of anterior cervical surgery by comparative analyzing hand dysfunction using brief Michigan hand questionnaire(Brief MHQ)in Hirayama disease patients.Methods　From Aug 2011 to Dec 2016,27 patients of hirayama disease who underwent surgery were enrolled in this study.The study group consisted of 27 men.The mean follow-up period was 41.1 months.The levels of surgery included 18 cases of C4-C7,6 cases of C3-C6,2 cases of C4-C6 and 1 case of C5-T1.Brief MHQ were evaluated for the 27 patients.According to the Wilcoxon analysis,the unchanged domains were analyzed with the multifactor Logistic regression analysis by preoperative duration of symptoms,age of onset,and number of affected extremities.Dynamic flexion-extension lateral X-rays were performed at baseline and at final follow-up.Results　No failure of internal fixation was detected on dynamic flexion-extension lateral X-rays.Five domains of preoperative Brief MHQ had lower scores,including Function,Satisfaction,Aesthetics,Activities of daily living,Work domain.With the exception of Aesthetics and Pain domain,all the other four domains showed significant improvement after surgery.The total score was 38.44±5.83 at base-line and 43.19±4.47 at follow-up.The score of Function was 5.19±1.36 at baseline 6.37±1.15 at follow-up;The score of Satisfaction was 5.56±1.22 at baseline 6.60±1.05 at follow-up;The score of Activities of daily living was 6.33±1.84 at baseline 7.60±1.47 at follow-up;The score of Work was 6.85±1.75 at baseline 7.67±1.33 at follow-up.The risk factors of postoperative outcomes reported in the literature included duration of disease at the time of surgery,age of onset,and extremity involvement.According to the Logistic regression,pre-operation duration was the risk factor for Aesthetics domain and the cut-off time was 1.75 years.Conclusion　Four domains of Brief MHQ score were improved significantly after anterior surgery for patients with hirayama disease.Brief MHQ was useful to evaluate the hand dysfunction and clinical efficacy in patients with hirayama disease.

Objective　To investigate the relationship of motor unit number estimation(MUNE)by multiple point stimulation with the outcome of surgical treatment patients with Hirayama disease(HD).Methods　A total of 36 consecutive patients including unilateral in 26 cases and bilateral in 10 cases with Hirayama disease treated by anterior cervical discectomy decompression and fusion in Peking University Third Hospital from October 2007 to May 2015 were reviewed retrospectively.There were 35 males and 1 female,aged from 16-26 years(average,19.2 years).A total of 46 hands were enrolled.Odom criteria was used to evaluate the subjective outcome of surgical treatment.Multiple point stimulating technique was used to estimate the motor unit number of abductor pollicis brevis and abductor digitiminimi preoperatively and at the time of pre-operation and the latest follow-up.Hands were divided into two groups based on Odom criteria(Group A with excellent and good;Group B with fair and poor).The difference between the two groups were examined by t text.Results　A total of 46 hands with complete clinical and electrophysiology data were followed up for 12-96 months(average,28.2 months).The outcome at the final follow-up according to Odom criteria was:Excellent in 8 cases,Good in 18 cases,Fair in 20 cases and no Poor case.MUNE of abductor pollicis brevis increased significantly after surgery from 139.6±68.4 to 188.2±60.4(t=-5.86,P<0.001).MUNE of abductor digitiminimi increased significantly after surgery from 75.0±66.3 to 104.2±80.4 significantly(t=-3.86,P<0.001).For two groups in age,follow-up period,preoperative MUNE of abductor pollicis brevis,and preoperative abductor digitiminimi,there was no significant difference.The illness course of Group A was 24.0±11.3 months,which was significantly shorter than Group B 34.9±21.2 months(t=-4.452,P<0.01).Group A had more increased MUNE of abductor pollicis brevis 65.6±64.1 compared with Group B 26.7±34.7(t=2.446,P<0.05)and Group A had more increased MUNE of abductor digitiminimi 42.6±59.3 compared with Group B 11.8±32.4 after surgery(t=2.088,P<0.05).Conclusion　MUNE by multiple point stimulating technique could be used to evaluate the neurological function of Hirayama disease and the outcome of surgical treatment quantitatively.

Objective To explore the effect of bladder muscle flaps for long segment defect of ureter middle-lower segment and reconstruction method in laparoscopic surgery. Methods Clinical data of 3 patients with long segment defect of ureter middle-lower segment, all of whom underwent laparoscopic surgery from May 2014 to April 2016 was retrospectively evaluated. There were 1 male and 2 females, in 2 cases with history of ureteroscopy holmium laser lithotripsy in ureter middle-lower segment, in 2 cases with history of repeated ESWL. Preoperative urinary tract ultrasound, CT and intravenous urography imaging showed severe hydronephrosis, ureter middle-upper segment severe hydroureter, ureter middle-lower segment severe stricture. Results Operations were successful in 3 cases. After reconstruction bladder muscle flaps average length of is 9.6 cm, The average operation time of 180 min, The average length of hospital stay for 10 d, Postoperative eighth weeks extracted the double J tube and used ureteroscopy showed anastomotic unobstructed, it may smooth Through 8.5 F ureteroscopy, and no infection and urinary leakage occurred, Follow-up ranged from 3 to 18 months. 3 cases hydronephrosis and hydroureter significantly reduce, ureter unobstructed, no narrow in ureter and muscle flap of tube joint, serum creatinine valueswere in normal range. Conclusions The bladder muscle flaps for the treatment of long segment defect of ureter middle-lower segment in laparoscopic surgery was a safe and effective therapy, but it must be accomplished by seasoned doctors.

Prolonged P-wave duration has been observed in diabetes. However, the underlying mechanisms remain unclear. The aim of this study was to elucidate the possible mechanisms. A rat model of type 2 diabetes mellitus (T2DM) was used. P-wave durations were obtained using surface electrocardiography and sizes of the left atrium were determined using echocardiography. Cardiac inward rectifier K+ currents (I(k1)), Na+ currents (I(Na)), and action potentials were recorded from isolated left atrial myocytes using patch clamp techniques. Left atrial tissue specimens were analyzed for total connexin-40 (Cx40) and connexin-43 (Cx43) expression levels on western-blots. Specimens were also analyzed for Cx40 and Cx43 distribution and interstitial fibrosis by immunofluorescent and Masson trichrome staining, respectively. The mean P-wave duration was longer in T2DM rats than in controls; however, the mean left atrial sizes of each group of rats were similar. The densities of I(k1) and I(Na) were unchanged in T2DM rats compared to controls. The action potential duration was longer in T2DM rats, but there was no significant difference in resting membrane potential or action potential amplitude compared to controls. The expression level of Cx40 protein was significantly lower, but Cx43 was unaltered in T2DM rats. However, immunofluorescent labeling of Cx43 showed a significantly enhanced lateralization. Staining showed interstitial fibrosis was greater in T2DM atrial tissue. Prolonged P-wave duration is not dependent on the left atrial size in rats with T2DM. Dysregulation of Cx40 and Cx43 protein expression, as well as fibrosis, might partly account for the prolongation of P-wave duration in T2DM.
Action Potentials ; Animals ; Blotting, Western ; Connexin 43/metabolism ; Connexins/metabolism ; Diabetes Mellitus, Type 2/*physiopathology ; Disease Models, Animal ; Echocardiography ; Electrocardiography ; Fibrosis/pathology ; Heart Atria/*diagnostic imaging/physiopathology ; In Vitro Techniques ; Male ; Membrane Potentials ; Microscopy, Fluorescence ; Patch-Clamp Techniques ; Potassium Channels/metabolism ; Rats ; Rats, Wistar

Objective To construct the genetic engineered cell line which can continuously secrete human tumor necrosis factor ( hTNFα) in Chinese Hamster Ovary cells( CHO cells) ,and observe the change of its protein secretion function.Methods Constructed plas-mid which carries hTNFαgene expression through vector GV141.Selected stable transfection cell lines by G418 and transfection with lipo-fectamine.Identified its gene expression with Real-Time PCR,and identified its protein secretion by ELISA.Results GV141-hTNFαexpres-sion vectors were constructed successfully which were proved by sequence alignment.Real-Time PCR proved that it contained hTNFαgene in hTNFα/CHO cell line.ELIAS identification results showed that the cell lines can continuously secrete hTNFαwithin a certain cell propaga-tion.Conclusion The hTNFα/CHO cell line can continuously secrete human tumor necrosis factor within a certain cell propagation.

OBJECTIVE:To establish a method for the simultaneous determination of chlorogenic acid,vitexin glucoside,vi-texin rhamnoside,vitexin,rutin and hyperoside in Crataegus pinnatifida. METHODS:With reference peak of vitexin glucoside, HPLC was conducted to calculate the relative correction factor(RCF)of chlorogenic acid,vitexin glucoside,vitexin rhamnoside, vitexin,rutin and hyperoside,then the contents of above-mentioned 5 components in C. pinnatifida were calculated. The column was Agilent ZORBAX SB C18 with mobile phase of 0.1% formic acid-acetonitrile-tetrahydrofuran (gradient elution) at a flow rate of 1.0 ml/min,the detection wavelength was 350 nm,column temperature was 30 ℃,and the injection volume was 10 μl. RE-SULTS:The linear range was 12.50-400.0 μg for chlorogenic acid(r=0.999 8),25.00-800.0 μg for vitexin glucoside(r=0.999 9), 31.25-1 000.0 μg for vitexin rhamnoside(r=0.999 9),6.470-260.0 μg for vitexin(r=0.999 9),2.50-80.0 μg for rutin(r=0.999 8) and 9.375-300.0 μg for hyperoside(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%;re-coveries were 99.2%-103.9%(RSD=1.6%,n=6),97.9%-100.8%(RSD=1.2%,n=6),99.2%-100.8%(RSD=0.5%,n=6), 97.3%-101.3%(RSD=1.5%,n=6),98.0%-103.0%(RSD=1.9%,n=6)and 95.5%-101.5%(RSD=2.2%,n=6). RCFs of vitex-in glucoside with chlorogenic acid,vitexin rhamnoside,vitexin,rutin and hyperoside were 1.119,1.009,0.706,1.063 and 0.830, respectively. CONCLUSIONS:The method is simple with good precision,stability and reproducibility,and it can be sued for the simultaneous determination of 6 components in C. pinnatifida.