The inverted terminal repeat (ITR) is the only cis element of AAV genome essential for rAAV rescue, replication and packaging. It is prone to mutation or loss when it is latent in host cell or in plasmid. Plasmids with different ITR types were cloned to compare the influence of ITR types on the AAV packaging and infectivity. The vector plasmids were transformed the competent SURE cells to get different colonies. The ITR types of plasmids were screened by digestion with SmaⅠ. AAV vector plasmid pScGFPud has two ITRs at both ends of AAV genome and plasmid pScGFPu has only one ITR at upstream end of AAV genome. When the two plasmids were co-transfected 293 cells to prepare rAAVs, 1.08?1013 viral particles (AAV1-GFPud) were produced from 20-dishes of 293 cells cotransfected with plasmid pScGFPud, 4.28?1012 viral particles (AAV1-GFPu) were produced from 20-dishes of 293 cells cotransfected with plasmid pScGFPu. Virus AAV1-GFPud infected 293, HeLa and NCI H446 cells more efficiently than did virus AAV1-GFPu. This suggests that defect ITRs in AAV genome is deleterious to AAV packaging and AAV infectivity and vector with complete ITRs is favorable to the yield and activity of rAAV.

Objective To investigate whether the combination of maintenance hemodialysis (MHD) with hemoperfusion (HP) could improve the left ventricular structure and function of MHD patients with resistant hypertension. Methods This study was a prospective, randomized, controlled clinical trial. 65 MHD patients with resistant hypertension of our hospital were enrolled and then randomly divided into Group HD+HP and Group HD. The follow-up study was 12 months. The major outcome measures were related to the cardiac structure and function, and the serum inflammatory factors. Results No statistical differences of related factors such as blood pressure and cardiac function was observed in HD group. The serum concentration of inflammatory factors was higher after one-year HD treatment alone. But the serum concentration of inflammatory related factors , blood pressure, cardiac function and related indexes were lower in HD + HP than in Group HD (P < 0.05);HD + HP group had higher ejection fraction (EF) and SV, which demonstrated the improved cardiac function (P < 0.05). Conclusions HD + HP was superior to HD in the treatment in alleviating cardiac after-load and improving the left ventricular structure and function of MHD patients with resistance hypertension.

Objective To investigate immunoprophylactic potential of genetic engineering vaccines of enterohaemorrhagie Escherichia coli O157:H7 in BALB/c mice after immunization with these vaccines. Methods Sixty BALB/c mice (3 weeks old) were randomized averagely into 5 groups. Group 1-3 were im-munized respectively with IntiminC300, Stx2B and HIyAN436, group 4 with a combination of these three vaccines, and group 5 with PBS. Each mouse was immunized with vaccine(100 μg)and Al(OH)3 adjuvant (100 μg) for 3 times. After 7 d of the second and third immunization, serum of each mouse was collected and the different antibodies were detected. After 10 d of the last immunization, all mice were given drinking water containing streptomycin for 3 d before and following oral challenge with O157:H7 (109 CFU), and treated with clinical, microbiological and pathological examination. Results The three vaccines elicited high titer antiserum, and some mice were died after infection with O157. The livability of group 1-4 was re-spectively 73%, 64%, 36% and 91%. And these vaccines depressed fecal and colon shedding with O157. Condusion IntiminC300, Stx2B and HIyAN436 have certain protective efficacy for infection of O157, and combined immunization was more effective than single vaccine.

OBJECTIVE:To prepare a three-pulse drug release system of nimodipine and study its drug release.METHODS:Solid dispersion technique and dry-coating method were respectively applied to prepare immediate-release mini-tablets and Pulsatile mini-tablets with lag-time of 4 h or 8 h.Then those mini-tablets were filled into capsule to obtain a three-pulse drug release system and subjected to in vitro dissolution test.DSC was employed to determine drug status in solid dispersion.RESULTS:Immediate-release mini-tablets were released more than 95% in 30 min.Pulsatile mini-tablets were released less than 10% in 4 h or 8 h of lag-time period.After lag-time period,pulsatile mini-tablets were released completely in 3 h.The whole pulsatile drug release system achieved three times of drug release at 5 min,4 h,8 h,respectively.Nimodipine kept amorphous form and were delivered into carrier evenly.CONCLUSION:A three-pulse drug release system of nimodipine has been prepared successfully.

Air ; analysis ; Chromatography, Gas ; methods ; Styrenes ; analysis ; Workplace

Objective To explore the circadian rhythm of neurally mediated syncope (NMS)in children. Methods There were 21 6 children with NMS included in the study,including 91 male and 1 25 female,aged from 4 to 1 7 years old with a mean age of (1 1 .34 ±2.65)years,who came from the Specialist Syncope Outpatient Department or Inpatient Department of the Second Xiangya Hospital of Central South University from December 201 3 to October 201 5. The patients were divided into vasovagal syncope (VVS)group and postural tachycardia syndrome (POTS)group ac-cording to head -up tilt test (HUTT)results,including 1 78 VVS patients and 38 POTS patients.Ninety -four patients with NMS were in the <1 2 years old group[(8.88 ±1 .88)years old]and 1 22 subjects with NMS were in the ≥1 2 years old group[(1 3.24 ±1 .1 8)years old].All patients or guardians were carefully asked about the number of synco-pal attacks and the periods in which episodes occurred in before HUTT [24 hours of a day were divided into 4 periods:morning (0600 AM-1 200 AM),afternoon (1 200 AM-1 800 PM),evening (1 800 PM-2400 PM), night (0000 AM-0600 AM)].Results (1 )General data:the total syncopal episodes of 21 6 children with NMS were 61 4 episodes,including 1 78 VVS patients with 471 syncopal episodes in total and 1 43 attacks of 38 children with POTS.There were 273 episodes of 94 patients in the <1 2 years old group and 341 episodes of 1 22 subjects in the ≥1 2 years old group.There were no significant differences in the diurnal variation of syncopal episodes between the VVS group and POTS group regarding age and gender (P >0.05).(2)The number of syncopal episodes in patients with VVS which occurred in the morning hours was strikingly higher than that of afternoon,evening or nighttime (P <0.05).But there was no significant difference in the frequency of episodes in different periods through the day in the POTS group (P >0.05).Patients with VVS had a higher proportion of episodes in the morning but a lower proportion in the evening when compared with the POTS group (P <0.05).(3)The male children with NMS tended to have a higher proportion of episodes in the morning than the female patients(χ2 =1 1 .001 ,P =0.01 2).(4)There seemed to be no difference in the frequency of syncopal episodes through the day between the <1 2 years old group and the ≥1 2 years old group(χ2 =1 .995,P =0.573).Conclusions The frequency of syncopal episodes in children with VVS displayed a clear circadian rhythm,with a peak in the morning,but the POTS patients did not show a circadian variation.The male children with NMS tended to have a higher proportion of episodes in the morning than the female patients.

Objective To study the intervention effectiveness of intensive training in medical interns’occupational exposure protection.Methods Medical interns in seven colleges and universities in Wuhan between 2011 and 2015 were chosen and divided into control group (675 medical interns in 2011 - 2013,comprehensive training about healthcare-associated infection control was conducted for once before internship )and intervention group (682 medical interns in 2014 - 2015,hospital-department-tutor three steps of intensive training was conducted),the occurrence of occupational exposure and exposure knowledge awareness between two groups were compared. Results The awareness rate of knowledge about occupational exposure protection and the implementation rate of protective measures in intervention group were both higher than control group (all P <0.05).Incidence of occupa-tional exposure in intervention group was lower than control group (21 .70%[n=148]vs 65.33%[n=441 ],χ2 =262.91 ,P <0.01);percentage of interns who conducted active serological virus detection and intensified vaccination of hepatitis B virus were both higher than control group (14.66%[n=100]vs 2.96%[n=20];11 .73%[n=80]vs 2.67%[n=18],respectively).Occupational exposure before and after intervention occurred most frequently when interns were preparing medicine,occupational exposure reporting rate in intervention group was higher than control group (72.97%[108/148]vs 50.11 % [221/441 ],χ2 =52.78,P <0.01 ).Conclusion The intensive training of occupational exposure protection among medical interns can improve the awareness and skills of occupational protec-tion,and reduce the occurrence of occupational exposure.

Objective To explore the changes in serum and urine electrolytes of children with neurally media-ted syncope (NMS)after oral rehydration salts (ORS)[Ⅰ]treatment.Methods The study group included 135 patients [60 male and 75 female,aged 4 -16 years,average of (10.20 ±2.68)years old]with unexplained syncope and prodro-mal symptoms of syncope in our hospital between May 2014 and April 2015.The patients underwent head -up tilt test (HUTT),and completed serum electrolytes and 24 -hour urine electrolytes,and the serum electrolytes and 24 -hour u-rine electrolytes in different hemodynamic type of HUTT were compared.Positive HUTT patients were treated with health education and ORS[Ⅰ],while negative HUTT patients were received health education.Then 21 -154(42.63 ±27.71) days later,the patients returned to hospital,for the inquiry of symptom improvement,and review of HUTT,24 -hour urine and serum electrolytes.Results (1)The total effective rate of ORS[Ⅰ]treatment was 62.96% (17 /27 cases),while negative conversion rate of HUTT was 48.15% (13 /27 cases).(2)There was no significant difference in serum electro-lytes,24 -hour urine electrolytes or 24 -hour urine volume between HUTT positive group and negative group during the first visit (all P >0.05).(3)In return visit,serum calcium [(2.30 ±0.10)mmol/L vs (2.20 ±0.09)mmol/L,t =2.72,P <0.05]and serum phosphorus [(1.73 ±0.22)mmol/L vs (1.51 ±0.23)mmol/L,t =2.671,P <0.05]in HUTT positive group were significantly higher than those in negative group.The serum sodium,potassium,chloride,mag-nesium and 24 -hour urine electrolytes,24 -hour urine volume had no statistical difference(all P >0.05).(4)24 -hour urine sodium [(159.06 ±72.76)mmol/24 h vs (118.97 ±52.75)mmol/24 h,t =2.712,P <0.05],24 -hour urine chloride [(139.08 ±66.53)mmol/24 h vs (111.34 ±47.33)mmol/24 h,t =2.116,P <0.05]and 24 -hour urine volume [(1 564.21 ±829.39)mL vs (1 058.95 ±509.92)mL,t =3.371,P <0.01]after ORS[Ⅰ]treatment were sig-nificantly higher than those before ORS[Ⅰ]treatment.The serum electrolytes and 24 -hour urine potassium,calcium, phosphorus,magnesium had no statistical difference (all P >0.05).(5)There was no significant difference in serum elec-trolytes,24 -hour urine electrolytes or 24 -hour urine volume between vasovagal syncope group and postural orthostatic tachycardia syndrome group during the first visit(all P >0.05).Conclusions ORS[Ⅰ]treatment can obviously increase the 24 -hour urine sodium,24 -hour urine chloride in children with NMS.ORS[Ⅰ]is an effective therapy for NMS.

Objective:To investigate the clinical characteristics and outcome of novel coronavirus pneumonia (COVID-19)patients with different body mass index (BMI), and to provide the basis for disease assessment and prognosis.Methods:The clinical data of 541 patients with COVID-19 diagnosed in Xiaogan Hospital Affiliated to Wuhan University of Science and Technology from January 16 to March 28, 2020 were collected. The patients were divided into normal weight group, overweight group, and obesity group according to BMI. The clinical characteristics and outcomes of the three groups were compared. The correlation between BMI and clinical classification was analyzed by ordinal logistic regression.Results:There were 288 cases (53.23%) in normal weight group, 193 cases (35.67%) in overweight group, and 60 cases (11.09%) in obesity group. Compared with normal weight group, overweight and obesity groups displayed higher proportion of hypertension, with increased levels of white blood cells, neutrophils, C reactive protein, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and triglyceride in laboratory test results, and higher proportion of severe and critical illness ( P<0.05 or P<0.01). After adjusting for age, gender, and underlying diseases, regression analysis showed that higher BMI predicted more severe clinical classification ( OR=1.079, 95% CI 1.010-1.154). Conclusion:In COVID-19 patients, overweight and obese patients are more likely to develop into severe and critical illness, suggesting that obesity may be an important risk factor affecting the clinical outcome of COVID-19.

Objective:To analyze the association between the types of response to head-up tilt test (HUTT) and levels of various vitamins in children and adolescents with orthostatic intolerance.Methods:Sixty-five cases of children and adolescents 35 males and 30 females aged 5-14 (10.5±2.0) years old who complained orthostatic intolerance symptoms, including syncope, dizziness, headache, and chest tightness, in Children′s Medical Center, the Se-cond Xiangya Hospital, Central South University between November 2018 and April 2019, were enrolled.By comprehensive history, physical examination, and necessary laboratory testing, heart, cerebrovascular and other system diseases were excluded.According to HUTT result, these patients were divided into 2 groups, namely negative HUTT group and positive HUTT group.The serum levels of vitamin A 1, B 1, B 2, B 6, B 9, B 12, C, and E were compared between the 2 groups.Continuous data were analyzed by conducting t-test and categorical data were analyzed by performing χ2 test. Results:(1) There were 32 cases (18 males and 14 females) in negative HUTT group and 33 cases (17 males and 16 females) in positive HUTT groups, with 28 cases of vaso vagal syncope and 5 cases of postural tachycardia syndrome.There were no significant differences in age, gender ratio, and body mass index between the negative HUTT group and positive HUTT group.(2) The serum level of vitamin B 1 of positive HUTT group was lower than that of negative HUTT group [(77.43±16.70) nmol/L vs.(88.56±18.98) nmol/L, t=-2.513, P<0.05], while the levels of vitamin A 1, B 2, B 6, B 9, B 12, C, and E had no significant differences between the two groups (all P>0.05). Conclusions:Patients with positive responses to HUTT relatively lacked vitamin B 1, suggesting the role of vitamin B 1 in the pathophysiological process of neurally mediated syncope.