OBJECTIVE To establish the fingerprint of Pinghuo tea standard decoction and a method for determination of multi-component to clarify the transfer relationship of quantities and quality from pieces and standard decoction. METHODS Fifteen batches of Pinghuo tea standard decoction were prepared and the extract rate was determined； the fingerprint of the preparation was established by using high-performance liquid chromatography（HPLC）； the similarity evaluation and the determination of common peaks were performed， and chemometric analysis was performed； the same method was used to determine the content of indicator components and the transfer rate was calculated. The chromatographic column was Venusil C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution）； the column temperature was 30 ℃， and the detection wavelengths were 238 nm （0-37 min， 85-102 min） and 330 nm （37-85 min） at a flow rate of 1.0 mL/min with an injection volume of 10 μL. RESULTS The similarity of HPLC fingerprints for 15 batches of Pinghuo tea standard decoction was not lower than 0.968. A total of 24 common peaks were calibrated and 9 peaks were recognized， which were as follows neochlorogenic acid （peak 3）， chlorogenic acid （peak 6）， geniposide （peak 9）， glycyrrhizin （peak 10）， galuteolin （peak 11）， isochlorogenic acid A （peak 14）， luteolin （peak 21）， kaempferol （peak 23） and glycyrrhizic acid （peak 24）. Cluster analysis， principal component analysis and orthogonal partial least squares discriminant analysis showed consistent results， all of which could classify the 15 batches of samples into three categories. The linear range of indicator components in 15 batches of Pinghuo tea standard decoction， such as geniposide， luteolin， isochlorogenic acid A， glycyrrhizin， and glycyrrhizic acid， were 0.020 580-0.411 600， 0.001 617-0.080 850， 0.006 076-0.607 600， 0.005 125-0.071 740， and 0.017 288-0.432 200 mg/mL， respectively； RSDs of precision， repeatability， stability and recovery rate tests were all not higher than 4% （n＝6）. The mass fractions ranged 3.227 9-10.002 2， 0.297 4-0.554 6， 3.350 1-6.159 6， 0.720 6-1.073 3， 2.003 1-3.030 1 mg/g； transfer rates from the pieces and standard decoction were 19.762 8%-35.840 5%， 12.123 3%-21.254 0%， 46.097 2%-82.869 4%， 58.708 8%-91.629 6%， 39.114 3%-63.710 6%. The transfer rates of the extract from 15 batches of Pinghuo tea standard decoction ranged from 61.15%-84.68%. CONCLUSIONS Established HPLC fingerprint and content determination methods in this study are simple and accurate， which can provide reference for the quantitative value transfer study， quality control， clinical application and the development of subsequent formulations of Pinghuo tea standard decoction.

BACKGROUND:Ischemic postconditioning is one of the effective ways to reduce ischemia-reperfusion injury and has been more and more widely used in clinical practice in recent years,but its specific molecular mechanism has yet to be studied. OBJECTIVE:To investigate the role and mechanism of piRNA-005854 in the aging cardiomyocytes caused by hypoxic postconditioning. METHODS:In vitro,cardiomyocytes were administered 8 mg/mL D-galactose for 9 days to induce their aging.β-Galactosidase staining was used to observe the aging of cardiomyocytes.Senescent cells were treated with hypoxia/reoxygenation and hypoxic postconditioning.ELISA was utilized to detect changes in myocardial injury markers creatine kinase isoenzyme MB and lactate dehydrogenase levels.Western blot assay was applied to detect the expression changes of autophagy-related proteins LC3II,p62,ULK1 and phosphorylated ULK1 in aging cardiomyocytes.qRT-PCR was employed to determine the expression level of piRNA-005854.piRNA-005854 inhibitor and piRNA-005854 mimics were transferred into aging cardiomyocytes and followed with hypoxic postconditioning.Western blot assay was used to examine the expression of LC3II,p62,ULK1 and phosphorylated ULK1. RESULTS AND CONCLUSION:(1)D-galactose induced obvious senescence of cardiomyocytes 9 days later.(2)Compared with the normoxia group,creatine kinase isoenzyme MB and lactate dehydrogenase levels increased in the hypoxia/reoxygenation group(P<0.01);LC3 II/I expression was increased;p62 expression was decreased;ULK1 phosphorylation level was increased,and piRNA-005854 expression was increased(P<0.01).(3)Compared with the hypoxia/reoxygenation group,creatine kinase isoenzyme MB and lactate dehydrogenase levels significantly reduced in the hypoxic postconditioning group(P<0.01);LC3 II/I expression significantly decreased(P<0.05);p62 expression increased(P<0.01);ULK1 phosphorylation level decreased(P<0.05),and piRNA-005854 expression decreased(P<0.01).(4)After transfection of piRNA-005854 inhibitor,LC3II/I expression was decreased(P<0.01);the expression of p62 was increased significantly(P<0.05);the phosphorylation level of ULK1 was decreased significantly(P<0.01).After transfection of piRNA-005854 mimics,LC3II/I expression was increased significantly;the expression of p62 was decreased,and the phosphorylation level of ULK1 was increased significantly(P<0.01).(5)The results show that piRNA-005854-mediated reduction of ULK1-dependent autophagy level is a possible mechanism that hypoxic postconditioning exerts its protective effect on aging cardiomyocytes.

Objective:To analyze the feasibility and clinical efficacy of reperfusion-expanding-thrombectomy-stenting (RETS) technique in the endovascular treatment of acute carotid artery tandem lesion.Methods:The general clinical data of 88 patients with carotid artery tandem lesion who received emergency endovascular treatment from January 2018 to December 2020 in Department of Neurology, Linyi People′s Hospital were reviewed, the Modified Rankin Scale (mRS) was used as the evaluation standard for the prognosis of patients at 90 days after endovascular treatment, and the clinical data were analyzed, including the recanalization (modified thrombolysis in cerebral infarction ≥2b), perioperative complications and 90-day prognosis, and good prognosis was defined as a mRS score of 0-2.Results:A total of 88 patients with tandem carotid artery disease were included,48 of whom were treated with RETS technique, 40 were treated with anterograde approach. Compared with antegrade recanalization, RETS technique had significant differences in the time from puncture to recanalization [(72.06±17.29) min vs (98.88±26.09) min, t=-5.56, P<0.001] and the primary recanalization rate [35/48(73.0%) vs 21/40(52.5%),χ2=3.93 ,P=0.047], with statistically significant difference. There was no significant difference in clinical prognosis and surgical complications between the two methods (all P>0.05). Conclusions:RETS technique can shorten the operation time and increase the primary recanalization rate. RETS technique is safe and feasible for the treatment of carotid tandem lesions.

Objective: To compare the efficacies of the two techniques of "micro-hemostasis and micro-cutting" with straight bipolar electrocoagulation forceps and traditional clamp-ligation for hemostasia in thyroid surgery. Methods: A total of 228 patients who underwent surgical treatment for thyroid neoplasms in our hospital between January 2015 and December 2018 were retrospectively analyzed, including 50 males and 178 females, aged 23-68 years old. Of those, 150 cases as electric knife group received traditional thyroid surgery between January 2015 and December 2018 and 78 cases as bipolar electrocoagulation group received thyroid surgery by using the technique of bipolar electrocoagulation with meticulous anatomy between January 2018 and December 2018. The total operation time, single operation time, intraoperative hemorrhage, postoperative drainage volume on the first day, postoperative hoarseness and hypocalcemia were compared between the two groups. SPSS 16.0 was used to analyze the data. Results: The total operation time and intraoperative hemorrhage in the bipolar electrocoagulation group were significantly lower than those in the electric knife group ((59.33±18.29)min vs (77.21±25.39)min, (14.83±9.22)ml vs (36.86±11.80)ml, all P<0.01). The single operation time of the bipolar electrocoagulation group was shorter than that of the electric knife group((10.25±6.16) min vs (20.34±7.24)min, (16.25±7.15)min vs (35.68±8.25)min, (12.12±5.25)min vs (20.68±7.26)min, t value was 3.948,16.262,8.238, all P<0.01).There was no significant difference between the two groups in postoperative drainage volume on the first day (P>0.05) and the incidence of postoperative hoarseness (P>0.05), while the incidence of hypocalcemia in the bipolar electrocoagulation group(10.26%) was lower than that in the electric knife group(21.33%,χ²=4.353, P<0.05). Conclusions The fine dissection for thyroid operation can be achieved by using straight bipolar electrocoagulation tweezers. The use of "micro-hemostasis" and "micro-cutting" technique with bipolar electrocoagulation tweezers can greatly reduce intraoperative bleeding, operation time and postoperative complication.

GGGGCC repeat expansions in the C9ORF72 gene are the most common cause of amyotrophic lateral sclerosis and frontotemporal dementia (c9ALS/FTD). It has been reported that hexanucleotide repeat expansions in C9ORF72 produce five dipeptide repeat (DPR) proteins by an unconventional repeat-associated non-ATG (RAN) translation. Within the five DPR proteins, poly-PR and poly-GR that contain arginine are more toxic than the other DPRs (poly-GA, poly-GP, and poly-PA). Here, we demonstrated that poly-PR peptides transferred into cells by endocytosis in a clathrin-dependent manner, leading to endoplasmic reticulum stress and cell death. In SH-SY5Y cells and primary cortical neurons, poly-PR activated JUN amino-terminal kinase (JNK) and increased the levels of p53 and Bax. The uptake of poly-PR peptides by cells was significantly inhibited by knockdown of clathrin or by chlorpromazine, an inhibitor that blocks clathrin-mediated endocytosis. Inhibition of clathrin-dependent endocytosis by chlorpromazine significantly blocked the transfer of poly-PR peptides into cells, and attenuated poly-PR-induced JNK activation and cell death. Our data revealed that the uptake of poly-PR undergoes clathrin-dependent endocytosis and blockade of this process prevents the toxic effects of synthetic poly-PR peptides.

Objective To explore the choice of handling methods for bilateral internal ring during the transumbilical single-site laparoscopic orchiopexy for cryptorchidism with bilateral processus vaginalis unclosed in children,and evaluate the clinical effect.Methods Retrospective analysis was conducted for the clinical data of 102 children with cryptorchidism and bilateral processus vaginalis unclosed who were hospitalized at Children's Hospital Affiliated to Capital Institute of Pediatrics from January 2011 to January 2016.They were divided into the observation group (55 cases) and the control group(47 cases).In the observation group,the internal rings of the affected side were destroyed and stitched with a needle between the edge of arcuate of musculus trasversus abdominis and fascia trans versalis of posterior peritoneum.If the diameter of opposite internal ring was less than 0.5 cm,only a circle was destroyed.Otherwise,a circle was destroyed and sutured with a needle.In the control group,the processing methods for orchiopexy and affected side internal ring were same as the observation group.Purse string suture was done for opposite internal rings of all cases in the control group.The parameters of operative duration,intraoperative blood loss,postoperative hospital stay,postoperative complications were compared between 2 groups.Results All operations were successful in both groups,spermatic cords were reserved and testicles were in scrotum of all cases.Operative duration was significantly shorter in the observation group than that in the control group [(42.02 ± 3.21) min vs.(48.43 ± 4.18) min,t =-8.739,P ＜ 0.01].The differences in intraoperative blood loss,postoperative hospital stay and postoperative complications between 2 groups were not statistically significant[(4.38 ± 1.42) mL vs.(4.80 ± 1.37) mL,t =-1.533,P ＞0.05;(2.87 ±0.64) dvs.(2.98 ±0.61) d,t =-0.853,P ＞0.05;1.8％ (1/55 cases) vs.2.1％(1/47 cases),x2 =0.013,P ＞ 0.05].During a mean follow-up of 30(12-72) months,there was no case of testicular ascent or atrophy,or hernia,or hydrocele.Conclusions The transumbilical single-site 3-port laparoscopic orchiopexy for cryptorchidism has stable efficacy.The improved method for bilateral internal ring is simple and has satisfactory effect,which is worthy of clinical promotion.

Objective To evaluate the clinical effect of treatment for high intra-abdominal cryptorchidism of children with transumbilical single-site and multichannel laparoscopic single stage Fowler-Stephens (F-S) orchiopexy.Methods The case records of the intra-abdominal cryptorchidism of children who had undergone transumbilical single-site laparoscopic single stage F-S orchiopexy were reviewed retrospectively in Children's Hospital Affiliated to Capital Institute of Pediatrics between January 2011 and January 2017,were assigned as the observation group,whose age ranged from 1 to 8 years and average age was 18 months,with 22 unilateral and 8 bilateral,38 testis in total.A total of 31 children with intra-abdominal cryptorchidism who had undergone laparoscopic two stage F-S orchiopexy were assigned as the control group,whose age was from 11 months to 9 years and average age was 20 months,with 23 unilateral and 8 bilateral,39 testis in total.Postoperative follow-ups were conducted with the evaluation index included the testical position,with or without atrophy.The procedure effect and postoperative complications were observed,and the difference between two operation methods was evaluated.Results Operations in all cases were successful in both groups without intraoperative complication.A total of 38 testis were operated with single stage F-S orchiopexy in the observation group,and 39 testis were operated with two stage F-S orchiopexy in the control group.Postoperative complications included scrotum wound infection one case and scrotum hematoma in one case in the observation group,and abdominal wall emphysema in one case,intestinal obstruction in one case as well as umbilicus infection 1 case in control group.Follow-ups ranged from 6 months to 6 years,median 24 months.All testicals were within the scrotum,and each group had 1 case of testical atrophy.The difference of postoperative complication and effect between two groups had no statistical significance(x2 =0.184,0.107,all P ＞ 0.05).Conclusions Outcomes between single stage and two stage F-S orchiopexy are similar.The transumbilical single-site laparoscopic F-S orchiopexy not only has the satisfactory effect,but also saves some patients from reoperation and secondary anaesthesia,but doctors must be aware of the indications and contraindications of this procedure should be brought to attention.

OBJECTIVE:To investigate the effect of polyamide on the separation on phenolic acids components by nanofiltra-tion process. METHODS:Phenolic acids components were selected as targets,extracted from the Salvia miltiorrhiza,Lonicera ja-ponica or intermediate preparations (tea polyphenols) with water,and filtered with membranes with different pore sizes. The con-tent changes of components were analyzed by HPLC. RESULTS:Danshensu,protocatechuic aldehyde and caffeic acid could better transmit nanofiltration membranes with 3 different pore sizes;with the decrease of pore size,the rejection rates of neochlorogenic acid,chlorogenic acid,rosmarinci acid,ECG and EGCG grew in different levels. But salvianolic acid B was almost retained by the membrane with low molecular weight except 600-800 Da. CONCLUSIONS:Molecular weight importantly influenced nanofiltra-tion separation on phenolic acid components;nanofiltration has a good prospect in enrichment of phenolic acids components.

Objective To investigate the effects of mindfulness cognitive behavioral training on self -perception burden and self-efficacy of patients with shoulder-hand syndrome ( SHS ) after brain infarction . Methods A total of 65 cases with SHS were randomly divided by coin-tossing into two groups:control group (n=32) and experimental group (n =33).Patients in control group received routine rehabilitation , while patients in experimental group were given mindfulness cognitive behavioral training for 6 weeks consecutively based on routine rehabilitation;both groups of patients were evaluated using Barthel index (BI), self-perceived burden scale ( SPB ) , general self-efficacy scale ( GSES ) , mindful attention awareness scale ( MAAS ) and quality of life short-form 36(SF-36) before and after 6 weeks of intervention.Results There was no significant difference in the scores of BI,SPB,GSES,MAAS and SF-36 between two groups before the intervention (P>0.05).After the intervention, scores of BI,GSES and MAAS in experimental group were (72.4 ±11.6), (7.3 ±1.3),(79.9 ±11.8) respectively,which all higher than that of control group significantly (t=3.38, 6.50,5.41;P<0.05), while the score of SPBS was (38.2 ±5.7) lower than that of control group (47.3 ± 6.6) significantly (t=5.72;P<0.05); The scores of SF-36 for each dimension of experimental group were higher than that of control group significantly (P<0.05).Furthermore, MAAS score was correlated positively with scores of general health , vitality, social function , emotional function , mental health in SF-36 with significant difference ( P<0.05 ) .Conclusions The cognitive behavioral training based on mindfulness can weaken the self-perceived burden and strengthen self-efficacy of patients with SHS after brain infarction greatly , so as to improve the life quality of patients .