ObjectiveTo establish and preliminarily verify a digital polymerase chain reaction(dPCR)method for the detection of genome titer of adeno-associated virus(AAV)gene therapy products.MethodsThrough dPCR detection of the same batch of AAV gene therapy products,the primers,probes,sample treatment methods and concentrations of primer and probe were optimized. The specificity,linearity,relative accuracy and repeatability of the optimized method were verified,and the limit of quantitation(LOQ)and detection range were determined.ResultsNFS-05 vector titer 5.0 crop primer probe was selected,the final primer concentration was 800 nmol/L,the final probe concentration was 400 nmol/L,and the sample pretreatment method was digestion by Proteinase K. The established method had good specificity. The correlation coefficient(R2)of the standard curve was greater than 0. 999 in the range of 3. 3 × 10~5-1. 3 × 10~8vg/mL. The relative accuracy was between 70% and 130%,the CV of repeatability was not more than 15%,and the LOQ was 3. 3 × 10~5vg/mL.ConclusionThe established dPCR method has good specificity,linearity,accuracy and repeatability,and can be applied to the determination of the genome titer of NFS-05 recombinant adeno-associated virus(rAAV)stock solution products. 关键词(KeyWords)：数字PCR;腺相关病毒;基因组滴度;基因治疗 Digital polymerase chain reaction(dPCR);Adeno-associated virus(AAV);Genome titer;Gene therapy 基金项目(Foundation): 北京市科技计划课题资助（Z221100007922015） 作者(Author): 安怡方,毕华,刘家池,魏云欢,赵琴,于雷,史新昌,裴德宁,李响,周勇,秦玺,梁成罡 AN Yifang,BI Hua,LIU Jiachi ,WEI Yunhuan ,ZHAO Qin ,YU Lei,SHI Xinchang,PEI Dening,LI Xiang,ZHOU Yong,QIN Xi,LIANG Chenggang

OBJECTIVE:To explore the effects of different doses of sufentanil on hemodynamics and postoperative stress re-sponse of children with congenital heart disease in cardiac surgery anesthesia. METHODS:40 children underwent ventricular septal defect repair in our hospital were divided into group A,B,C and D with 10 cases in each group according to the dose of sufent-anil. Groups A,B,C and D were given intravenous injection of sufentanil(0.6,0.8,1.0,1.2 μg/kg)respectively during anesthe-sia induction. Heart rate(HR)and systolic blood pressure(SBP)of 4 groups were recorded before induction(T1),after induction (T2),intubation(T3),2 min after intubation(T4),5 min after intubation(T5),and 10 min(T6)after intubation. The contents of acetylcholine(ACH),cortisol,glucagon,IL-6 and lactic acid were recorded when getting into the operating room(t1),after intu-bation(t2),after spliting the sternum(t3),before rewarming(t4),10 minutes after the shutdown the CPB(t5)and 24 h after oper-ation(t6). RESULTS:There was no significant differences in HR and SBP between 4 groups at T1(P>0.05);but there was statis-tically significant difference between 4 groups at T2-T6(P<0.05). There was no statistical significant difference in ACH,glucagon, IL-6 and lactic acid between 4 groups at T1 (P>0.05);but there was statistically significant difference between 4 groups at T2-T6 (P<0.05). There was no statistically significant difference in cortisol between 4 groups at T1-T4(P>0.05);but there was statisti-cally significant difference between 4 groups at T5-T6(P<0.05). CONCLUSIONS:During pediatric cardiac surgery anesthesia,the concentration of sufentanil kept in 1.0μg/kg is the most suitable,and has small effect on hemodynamics and postoperative stress re-sponse.

A series of etiological investigations were made on muscular metabolism in low Se children and rats, including (1) low Se environment and low Se state in children, (2) moasurement of some biochemical indexes concerning muscular metabolism (selenium, creatine, ereatinine, phosphate, CPK, etc.), (3) Se supplemented experiment on children in low Se state, (4) experimental study on Se deficiency and Se supplemented dietary in rats, (5) patho-histological findings on the muscular tissues of children with KBD, and (6) effect of Se deficiency on muscular metabolism in children. The results show that there may be a causal relationship between Se deficiency in environment and muscular metabolism di- sturbance in low Se children. A hyphothesis on the disturbance of energy metabolism in mitochondria caused by Se deficiency is put forward that Se deficiency may blunt creatinephosphate synthesis in mitochondria and cause lesions in the mitochondrial structure, which was different from the antioxidant fiction. There is a marginal deficiency in Se for children in this paper, i.e. to take more than 20 ?tg Se/day is necessary. We suggest that muscular atrophy in low Se children be precisely designated by the name of "endemic Seresponsive muscular dystrophy".

This paper reports the results of 15 cases of chronic hepatitis B treated with alpha-inter -feron (?-IFN) combined with silybum marinanum compound at a small dose (?-IFN 5000 units/day)in a long treatment course(3-6 months). The cure rate was 66.6% in the treatment group and 33.3% in the placebo group. The amount of ?-IFN in the serum of the patients was determined before and after treatment, but no ?-IFN in the serum was detected before treatment, ?-IFN elevated to 5-40 units/ml after treatment for 3-7 months. However, it would drop when the treatment was stopped. The therapeutic effect was remarkable when the amount of ?-IFN in the serum was increased. The results indicate that the anti-viral drug(IFN)combined with silyoum marinanum compound may be used to cure chronic hepatitis B.

In this paper, the author firstly introduces the developments of "hospice" in our country,and then analyzes the problems that it still faces.After that,the author also summarizes the practical experiences and theoretical outcomes that we have obtained in the past more than ten years,and lastly puts forward some suggestions on this foundation.

Objective To investigate the clinical efficacy of trimetazidine combined with atorvastatin in the treatment of patients with coronary heart disease and its influence on serum lipid and hemorheology.Methods 140 cases with coronary heart disease were randomly divided into two groups.The control group was given trimetazidine on the basis of the conventional treatment,while the observation group was given atorvastatin on the basis of the control group.The clinical efficacy,changes of hemorheology and blood lipid were compared between the two groups.Results The effective rate of the observation group was 95.71％,which was significantly higher than 77.14％ of the control group (x2 =10.620,P ＜ 0.05).After treatment,TC,TG,LDL-C and HDL-C of the observation group were (2.54 ± 0.32) mmol/L,(1.41 ± 0.32) mmol/L,(1.53 ± 0.33) mmol/L and (0.73 ± 0.35) mmol/L,which were significantly lower than those of the control group (t =7.42,8.09,8.11,9.03,all P ＜ 0.05).After treatment,the nbh,nbl,np and HCT of the observation group were (5.11 ± 0.51) mPa/s,(8.03 ± 1.12) mPa/s,(1.02 ± 0.21) mPa/s and (40.34 ± 3.28) ％,which were significantly lower than those of the control group(t =8.33,9.14,8.08,9.44,all P ＜ 0.05).Conclusion Trimetazidine combined with atorvastatin in the treatment of coronary heart disease can significantly improve patients' lipid and hemorheology,and it can be promoted and applied in clinical practice.

The lack and absence of knowledge in humanities and medical ethics among nursing staff have a negative influence on the mutual communication between nurses and patients and the implementation of humanistic nursing.The implementation of humanistic nursing can improve the general quality of nursing staff,especially their knowledge in humanities and medical ethics,and increase patients' satisfaction towards medical service.Meanwhile,it can also upgrade hospital's overall image,and increase social and economic profits of hospitals as well.

This paper analyzes ethical issues to be concerned in the nursing process for patients' with severe degree burn,and comes up with countermeasures including building up comfortable environment of medical service,showing enough respect for patients,improving mutual communication between doctors and patients,applying flexible regulations of visiting and accompany,strengthening health propaganda,and making best efforts to maintain the original appearance and function of the burnt organs.

In the present study, comparison have been made between two kinds of double labelling methods; single side labelling and both sides labelling. To make the different groups be comparable, all sections were continuously cut from the same specimen and all labelling processes were carried out at the same ti- me. The groups were divided as follows: A, one side anti-?-amylase labelling and one side antitrypsin labelling; B, single side labelling of anti-?-amylase and antitrypsin; C, one incubation with anti-?-amylase, followed by two incu- bations with protein A-gold (PAG) complexes of varied size, C1: 7nm and 20nm, C2: 10nm and 20nm, C3: 20nm and 7nm; D, single side labelling of anti- ?-amylase and an unrelated antiserum (antichathepsin D), applying free protein A between two labellings; E, as C, but with free protein A between two PAG incubations; F, as control. Group A and B showed that the two labelling me- thods had almost the same sensitivity.Group C indicated that the interaction of single side labelling were resulted from the combination of the second PAG with the free Fc region of the first antiserum. To decrease the interaction, it was necessary for the second PAG to be much larger than the first one.Group D de- monstrated that the interaction between the second antiserum and the first PAG was very feeble. Group E proved that free protein A could completely prevent the interaction of single side labelling method. The both sides labelling method avoids interaction, but mistakes resulting from the ultrastructural differences on two sides of the sections may happen. Which method to be selected is dependent upon what to be labelled.