Objective To explore a method for detecting the integrity of target nucleic acid in recombinant adeno-associated virus(rAAV),and establish a preliminary analysis algorithm.Methods The free DNA fragment of rAAV was digested,and the virus genome was extracted.Five pairs of overlapping primers were designed,using the orthogonal array method,which were detected by digital PCR respectively,with conventional conditions:20 μL reaction system with EveGreen and4 μL template,and digital PCR conditions:95 ℃ 5 min;95 ℃ 15 s,55 ℃ 30 s,72 ℃ 90 s for 45 cycles.The enhancement condition of ultra-long nucleic acid fragment was 25 μL reaction system with Mix B-2 000 bp and 2 μL template,and the quantitative analysis was performed by using the software attached to the instrument.Using the least square method,the number of full-length fragments was fitted and analyzed,and then the integrity distribution of target nucleic acid was interpreted.Results The fragments with the distance between primers of no more than 1 200 nt were amplified effectively,and a series of effective copies of fragments with a length of about 1 000 nt were obtained by systematic analysis.The copy number of common fragments was fitted and analyzed by the least square method.It was estimated that the full-length fragments in the sample were no more than 1 234 copies/μL,and there was a signficant difference(P < 0.05)between this value and the maximum measured value of 1 443 copies/μL,with the difference of approximately 16.9%.Conclusion A preliminary detection method for the integrity of target nucleic acid in rAAV has been developed,and a certain amount of incomplete target nucleic acids were analyzed in the test sample,laying a foundation for further in-depth research.

Objective: To study the effects of variou s absorption enhancers and enzyme inhibitors on in sulin permeation in vitro and in vivo . Methods: The pene tration of insulin throu gh hamster and rabbit buccal membrane was investigated by measuring in vitro transbuccal flux. Buccal insulin absorption was estimated in vivo from th e cumula tive response of serum glucose concentrations and comparison was made with the results of subcutaneous experiments. Results: There was a statistically significant permeabil ity increas e of insulin over controls after co-administration with the SDCh, Brij78, SLS or lecithin, aprotinin, bacitracin, whereas 1-menthol or poloxamer were less e ffective. Buccal insulin efficacy in the absence of co-administration adjuvants was very low relative t o subcutaneous administration of insulin. When co-administered buccally with SD Ch, SLS, l ecithin or Brij78, Fr (relative pharmacological bioavailability) values were al l increased significantly. Conclusion: The present studies showed that with the most effective absorption adjuvants, buccal insulin was one-fifth to one-fourth as effective as subcutaneous insulin. Results of in vitro experiments were in agreement with the in vivo re sults with respect to the enhancement of these adjuvants.

Circular RNA(circRNA) is a type of RNA with a circular coding structure, characterized by a circular topological structure formed during the RNA splicing process. Compared to traditional linear mRNA, circRNA exhibits enhanced stability and immunogenicity, leading to its wide-ranging potential applications in areas such as vaccines, biomarkers, and targeted therapies. This paper summarizes the research progress of circRNA, including its synthesis and degradation in vivo and in vitro, components, biological functions, and applications, aiming to provide novel insights for their utilization in vaccine design and development.

Objective To observe the relationship of blood serum bilirubin level with coronary heart disease and the degree of coronary artery narrowing. Methods A total of 126 patients were divided into the coronary disease group (83) and the control group (43). According to the coronary artery narrowing integral, the 83 patients in the coronary disease group were divided into three subgroups: mild narrowing group (15 people), moderate narrowing group (35 people), and severe narrowing group (33 people). The coronary arteriography of the patients in the control group was normal. 5mL venous blood was drawn on empty stomach, and the enzyme method was used to determine glycerin, the total cholesterol, the low density lipoprotein cholesterol, the high density lipoprotein cholesterol, and the blood serum bilirubin. Results The blood serum total bilirubin in the coronary disease group was (12.30?3.84)?mol/L, direct bilirubin was (4.07?1.45)?mol/L, and indirect bilirubin was (8.23 ? 2.82 )?mol/L. The total bilirubin in the normal group was (14.59?4.37)?mol/L, direct bilirubin was (4.66 ? 1.55 )?mol/L, and indirect bilirubin was (9.93?3.33)?mol/L. The total bilirubin, indirect bilirubin and direct bilirubin were lower in coronary disease group than in normal group (P0.05). Conclusion The blood serum bilirubin level and coronary disease have remarkable negative correlation, and the blood serum bilirubin level of patients with coronary disease is lower than that of healthy people. ② The blood serum bilirubin level and the degree of coronary artery narrowing do not have remarkable correlation.

AIM: To prepare a novel long-circulating paclitaxel-containing liposomes, evaluate its quality and study its toxicity to gastric cancer cell line BGC823. METHODS: From May to November in 2006, the experiment was carried out in the Laboratory of Chemistry, Kaili College and the Laboratory of Physics and Chemistry Department, Southeast Guizhou Center for Disease Control and Prevention.①The novel long-circulating paclitaxel-containing liposomes was prepared by ultrasonic film method. Encapsulation efficiency and stability of the paclitaxel liposomes were measured by RP-HPLC.②The paclitaxel liposomes were dispersed in phosphate buffer at the pH value of 6.5, at 4 ℃ for 30 days, and the change of drug loads was observed.③The proliferation of BGC823 cell at logarithmic phase was tested by MTT method under the paclitaxel liposomes concentrations of 5, 10, 15, 20 mg/L. Meanwhile the paclitaxel free drug was taken as controls. RESULTS: ①Encapsulation efficiency of this paclitaxel liposomes was high to 97.6% detected by RP-HPLC.②When the paclitaxel liposomes were stored in refrigerator of 4 ℃, there was no flocculent precipitation during one month. Drug loaded in liposome after 3, 8, 15, 22, 30 days of preservation was respectively 97.3%, 96.2%, 95.6%, 94.9%, and 94.2%. The drug leakage of the paclitaxel liposomes was only 6% one month after dispersing in this buffer.③The result of in vitro cell toxicity showed that, under the drug concentration of 5, 10, 15, 20 mg/L, the inhibition rate to BGC823 cell of paclitaxel liposomes was 33.46%, 44.15%, 51.94%, 68.64% respectively, where those of free paclitaxel was 35.24%, 48.37%, 57.49%, 74.84%. CONCLUSION: ①This high-encapsulated and stable long-circulating paclitaxel-containing liposomes is expected to provide a more effective paclitaxel preparation.②The inhibition rate to BGC823 cell of the paclitaxel liposomes is low to that of free paclitaxel at the same drug concentration, indicating that it can present the slow release of liposomes.

Objective To describe the importance of anesthesia management in performing interventional procedures. Methods A total of 24 382 patients, who were admitted to authors’ hospital during the period from April 2011 to April 2015 to receive level Ⅲ or level Ⅳ interventional procedures, were enrolled in this study. According to the anesthesia method, the patients were divided into (1) mechanical ventilation group (group A), i.e. intravenous general anesthesia combined with laryngeal mask or endotracheal intubation, (2) intravenous general anesthesia and autonomous respiratory group (group B), (3) conscious sedation group (group C) and (4) local anesthesia group (group D). The heart rate (HR), mean arterial pressure (ABP), blood oxygen saturation (SpO2) and anesthesia-related complications of the patients of all four groups were kept under close observation before, during and after the interventional procedures, the results were statistically analyzed. Results The anesthesia was successfully implemented according to the operation plan in all 24 382 patients. Interventional procedure of level Ⅲ was performed in 16 702 patients(68.5%) and interventional procedure of levelⅣwas adopted in 7 680 patients (31.5%). The patients receiving interventional procedure of level Ⅲof group A, B, C and D were 6 797 (40.7%), 3 608 (21.6%), 5 095(30.5%) and 1 202(7.2%) respectively;while the patients receiving interventional procedure of level Ⅳ of group A, B, C and D were 4 193 (54.6%), 2 527 (32.9%), 699 (9.1%) and 261 (3.4%)respectively. No statistically significant differences in preoperative HR, ABP and SpO2 existed between each other among the four groups (P>0.05). In group A, B and C the HR and ABP values determined in operation were not statistically different from the preoperative ones(P>0.05), and the differences in HR and ABP values among the three groups were also not statistically different (P>0.05);SpO2 levels showed no obvious changes (P>0.05). In group D, the HR and ABP values determined in operation were significantly higher than the preoperative ones (P<0.05), and also these values were statistically different from those of other three groups (P<0.05);SpO2 levels showed no obvious changes (P>0.05). In 22 patients of group D the operation had to be stopped as they were unable to tolerate the procedure. Conclusion In performing different levels of interventional procedures, level Ⅲ and level Ⅳ intervention surgeries in particular, careful selection of individualized anesthesia plan on the base of patient’s condition and operation requirement is an important guarantee for ensuring a safe operation with no interference, and it is also a good way to reduce the pain severity of patient. Therefore, individualized anesthesia plan is worth to be widely used in interventional procedures.

With the continuous innovation and development in the field of interventional therapy, the complexity of surgery has been gradually increased, meanwhile, the psychological expectation from patients for a minimally invasive surgery has also become higher and higher. Patients expect not only to be safe to complete the surgery, but also to maintain the comfort during operation process. For ensuring these demands, anesthesia plays a key role. This paper aims to discuss the current situation, problems and development direction of anesthesia application in interventional surgeries in order to help promote the anesthesia management concepts in performing interventional procedures.

Objectives To evaluate mini/VITAL and BacT/Alert, two fully automated blood culture systems. Methods Standard mini/VITAL AER and ANA bottles (bioM′erieux, Marcy 1′Etoile, France) and BacT/Alert FAN aerobic and FAN anerobic bottles (organon Netherlands) were used. The bottles were analysed over a period of 7 days. A total of 600 samples of blood, 108 samples of body fluid, and 100 strains of imitation were detected by mini/VITAL and BacT/Alert parallelly. The bottles of flagged positive were smeared and Gram stained before being examined under a microscope. Subcultures were performed in parallel on fresh blood agar and chocolate agar incubated in microaerophilic Conditions with 5% CO 2. All the bottles flagged negative were examined in the same conditions as the positive bottles. Results mini/VITAL: The positive rate for aerobic and anaerobic blood cultures was 10% and 5% respectively, for aerobic body fluid culture was 26 9%. No contaminants were detected. 0.5% false positive and 0.8% false negative were detected. 90% of positive sample were detected in 48 h. The shortest time of report of positive bottles was 1.5 h for blood culture. BacT/Alert: The rate of positive of aerobic and anaerobic blood culture was 9.8% and 5% respectively. The positive rate of body fluid culturs was 26.9%. There were used 3 bottles contaminated. The false positive and false negative rate were 0.5% and 0 98% respective. 95.5% of positive bottles were detected in 48 h. The shortest time of flagged positive was 2.5 h. Conclusion mini/VITAL and BacT/Alert were accurate automated blood culture systems for the detection of bacteremia. There was no significant difference on the speed of detection between BacT/Alert FAN bottles and mini/VITAL standard bottles.

Objective: To provide references for the locating the third ventriculos tomy by measuring normal brain MRI data. Methods: Three hundreds normal brain MRI data were tagged and measured. The relevant reference data about third ventricle fistulization were obtained after statistics processing. Rosults: There were no statistical differences in the ratio of the distance before and after into the skull (BC / AC) in all age groups (P＞0.05). There were statistical differences in parameters of surgical approach-the distance of the middle line beside the incision (L),the depth inside the scalp (H) and the angle between the long axle of endoscope and the center sagittal plane (a) between ≤ 9-year--old and other age groups. The 95 percent confidence interval about parameters of surgical approach was obtained among all age groups. Conclusion: The results provide the reference of surgical treatment for the individual, enjoying the advantage of accuracy location and wide application.