OBJECTIVE: To construct and obtain ideal protein delivery vectors by researching the delivery efficiency and cytotoxicity to Hela cells using mPEG-PLGA-BSA-FITC-NPs. METHOD: The mPEG-PLGA nanoparticle was obtained through surface modification of PLGA with PEG, and deliver BSA-FITC into Hela cells in vitro. The positive cells were counted by Laser scanning confocal microscopy and the survival rate of Hela cells was calculated by MTT assay at different time points. RESULT: mPEG-PLGA-BSA-FITC-NPs shows the classic nanometer size, and the encapsulation efficiency reached 51. 2%. At the same time, the nanoparticles possess characteristics of slow release. By optimizing the delivery conditions, the highest efficiency of mPEG-PLGA-BSA-FITC-NPs was above 65.2%, and the cellular viability was about 85.7%. CONCLUSION mPEG-PLGA-BSA-FITC-NPs nanoparticles can successfully carry the target protein into cells as safe and effective as novel delivery materials of protein in vitro, and has shown slow release characteristics. The mPEG-PLGA-BSA-FITC-NPs provide ideal delivery vector for future application in clinical treatment of disease using nano-materials.
Drug Carriers ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; HeLa Cells ; Humans ; Nanoparticles ; Particle Size ; Polyesters ; Polyethylene Glycols ; Serum Albumin, Bovine

Objective To investigate the risk factors of anastomotic leakage after radical gastrectomy for gastric cancer and establish a risk prediction scoring model for postoperative anastomotic leakage.Methods The retrospective case-control study was conducted.The clinicopathological data of 1 757 patients who underwent radical gastrectomy for gastric cancer in the First People's Hospital of Yibin from February 2000 to December 2017 were collected.There were 1 207 males and 550 females,aged (59± 11) years,with a range from 48 to 70 years.Observation indicators:(1) follow-up situations;(2) risk factors analysis of anastomotic leakage after radical gastrectomy for gastric cancer;(3) establishment of risk prediction scoring model and verification;(4) risk prediction scores and probability of anastomotic leakage in patients with different scores.Follow-up using outpatient examination and telephone interview to detect anastomotic leakage after radical gastrectomy and reoperation or death caused by anastomotic leakage up to June 2018.Measurement data with normal distribution were represented as Mean±SD.Count data were described as absolute number.The univariate analysis and multivariate analysis were performed using the chi-square test and Logistic regression model respectively.The discrimination and fitting degree of the model were detected by the Hosmer-Lemeshow test and receiver operating characteristic (ROC) curve.Results (1) Follow-up situations:800 of 1 757 patients were followed up at 1 week,1 month,3 months and 6 months after discharge.During the follow-up,75 had anastomotic leakage,60 of which recovered after conservative treatment,9 recovered after reoperation,6 died of complications including septic shock and cardiac and respiratory failure.(2) Risk factors analysis of anastomotic leakage after radical gastrectomy for gastric cancer:results of univariate analysis showed that patients with diabetes,preoperative pulmonary insufficiency,preoperative level of albumin and volume of intraoperative blood loss were related factors affecting anastomotic leakage after radical gastrectomy for gastric cancer (x2=5.604,4.975,18.563,35.688,P＜0.05).Results of multivariate analysis showed that patients with diabetes,preoperative pulmonary insufficiency,preoperative level of albumin ＜ 30 g/L and volume of intraoperative blood loss ＞ 400 mL were independent risk factors affecting anastomotic leakage after radical gastrectomy for gastric cancer (odds ratio=2.337,1.946,3.478,4.357,95％ confidence interval:1.136-4.804,1.022-3.705,1.871-6.464,2.678-7.090,P＜0.05).(3) Establishment of risk prediction scoring model and verification.Risk prediction equation was established according to the multivariate Logistic regression results:P =1/1 +exp (4.092-0.666 * X1-0.849 * X2-1.246 * X3-1.472 * X4).The fitting degree of the model was detected by the Hosmer-Lemeshow test (P=0.287).The discrimination of the model was detected by the ROC curve,with the area under curve as 0.734 (95％ confidence interval:0.689-0.834,P=0.002).(4) Risk prediction scores and probability of anastomotic leakage in patients with different scores:the risk prediction scores of anastomotic leakage after radical gastrectomy for gastric cancer were 1,1,2,2 in patients with diabetes,preoperative pulmonary insufficiency,preoperative level of albumin ＜ 30 g/L and volume of intraoperative blood loss ＞400 mL,respectively.The incidence of anastomotic leakage of patients with risk prediction scores of 0,1,2,3,4,5,6 was 1.6％,3.2％,5.9％,10.1％,19.3％,31.8％ and 47.6％,respectively.The incidence of anastomotic leakage was 13.7％ of patients with score ≥3 and 3.5％ of patients with score ＜ 3.Conclusions Patients with diabetes,preoperative pulmonary insufficiency,preoperative level of albumin ＜ 30 g/L and volume of intraoperative blood loss ＞ 400 mL are independent risk factors affecting anastomotic leakage after radical gastrectomy for gastric cancer.Establishment of a risk prediction scoring model for anastomotic leakage after radical gastrectomy for gastric cancer can effectively identify high-risk patients with anastomotic leakage after radical gastrectomy.

Objective:To explore the predictive effect of European treatment and outcome study long term survival (ELTS) score on survival outcomes in chronic myeloid leukemia of chronic phase (CML-CP) children.Methods:A single-center retrospective cohort study was conducted. Clinical data of 216 children with CML-CP in Peking University People′s Hospital from January 2010 to December 2023 were analyzed. Children were divided into low, intermediate and high-risk groups according to ELTS score. The survival outcomes and prognostic factors were analyzed. Kaplan-Meier method and Log-Rank test were used for survival analysis.Cox regression model was applied for analysis of prognostic factors.Results:Among the 216 children with CML-CP, there were 122 males and 94 females, with the diagnosis age of 11.0 (8.0, 14.7) years. The follow-up time was 77 (57, 99) months. According to ELTS score, 145, 52, and 19 children were classified as low, intermediate and high-risk group. For the low-risk and intermediate/high-risk groups, the 6-year failure-free survival (FFS) rates were (83.0±3.1)% and (64.6±5.7)%, the 6-year progression-free survival (PFS) rates were (91.4±2.3)% and (78.7±4.8)%, and the 6-year event-free survival (EFS) rates were (80.8±3.3)% and (64.2±5.7)%, with statistically significant difference ( χ2=9.45, 7.16, 7.40, P=0.002, 0.007, 0.007), respectively.The 6-year overall survival (OS) rates were (98.5±1.0)% and (95.6±2.4)%, without statistically significant difference ( χ2=0.35, P=0.550). Multivariate analysis showed that ELTS score was an independent prognostic factor or tendency for FFS ( HR=1.97, 95% CI 1.11-3.49), PFS ( HR=2.95, 95% CI 1.18-7.39), and no independent prognostic factor for EFS and OS were found. Conclusions:ELTS score at diagnosis can help stratify the risk of children with CML-CP. The children in intermediate/high-risk group are more likely to have treatment failure, disease progression than those in low-risk group, but the predictive ability of ELTS score for OS is limited.

Objective: To understand the epidemiological and etiological characteristics of enterovirus (EV)-associated diseases among children in Beijing from 2010 to 2016. Methods: This was a repeated cross-sectional study. The throat swabs were collected from children with probable EV-associated diseases at the Children' s Hospital Affiliated to Capital Institute of Pediatrics from 2010 to 2016. The samples were sent for pan-EV, enterovirus 71 (EV-A71) and coxsackievirus A16 (CV-A16) detection by real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR) . The viral types of non-EV-A71 and non-CV-A16 EV-positive samples were identified using modified RT-PCR and sequencing with CV-A6, EV-A/B group and 5 'UTR universal primers. The constituent ratios of the prevalence of different EV types in different age and gender groups were compared. Results: Of the 2 703 throat swabs, 1 992 (73.7%) samples were positive for EV, including EV-A71 (19.1%, 516/2 703), CV-A16 (24.3%, 658/2 703), CV-A6 (22.2%, 600/2 703), CV-A10 (4.5%, 122/2 703) and other types of EV (3.5%, 95/2 703). There was 1 case of EV-A71 and CV-A16 co-infection. The positive detection rate of EV-A group (excluding EV-A71, CV-A16, CV-A6 and CV-A10) increased from 11.3% (7/62) to 95.2% (59/62) after using the modified VP1-specific primers and PCR amplification conditions. During the period between 2010 and 2012, CV-A16 and EV-A71 predominated in EV-positive samples. However, CV-A6 accounted for 60.7% (68/112) in 2013, much higher than CV-A16 (23.2%, 26/112) and EV-A71 (12.5%, 14/112). In 2014, EVs were mainly of CV-A16 and EV-A71, but CV-A6 was the predominant type in 2015 (68.2%, 232/340) and in 2016 (38.6%, 151/391). The epidemic season of EVs was mostly from April to August, but CV-A6 showed a small epidemic peak from October to November. The male-to-female ratio of EV-positive patients was 1.50∶1, and EV-associated diseases mostly occurred in children under 5 years of age. Younger children were more susceptible to CV-A6 than to EV-A71 and CV-A16. Conclusions From 2010 to 2016, there was a significant change in the spectrum of EVs in children with EV-associated diseases in Beijing. Since 2013, non-EV-A71 and non-CV-A16 increased, and CV-A6 gradually became one of the major pathogens of EV-associated diseases. The modified PCR primers and amplification conditions can effectively improve the reliability of test results.

Objective: To observe the root and root canal morphology of mandibular second molars in Western Guangxi by CBCT, to provide a reference for clinical diagnosis and treatment. Methods: In total, 564 patients′ 1 128 mandibular second molars that satisfy the inclusion criteria were analyzed with a planmecaromexis CBCT machine and its own image analysis software. The patients′ gender, age and ethnic differences in the root and canal morphology and the symmetry of the bilateral root and canal were statistically analyzed. Results: Among the 1 128 mandibular second molars, 662 were the Zhuang ethnic group and 384 were the Han ethnic group, and 82 were other ethnic groups; the double root type and C-shaped root type accounted for a relatively high proportion: 73.94% and 24.47%, respectively. The detection rates of the double root type were higher in males than in females (P < 0.05); the detection rates of the C-shaped root type were higher in females than in males (P <0.05); the root type of the teeth was mainly double-rooted in the Zhuang ethnic group (P<0.01). The incidence of type IV in the mesial root of the double root type mandibular second molar was the highest (P < 0.01), and the incidence of type I in the distal root was the highest (P < 0.01). The C-shaped root canal is more continuous at the mouth of the root canal, more downward corresponds to a worse continuity: in three different levels of root canal orifice, root middle and root apex, the root canal orifice is dominated by the C1 type, and both root middle and root apex are mainly C3-type (P < 0.01). The difference in symmetry of bilateral roots and root canals was statistically significant among different gender groups, age groups, and ethnic groups (P < 0.05): there were more males than females, the results in the 18-35-year-old group and the Zhuang ethnic group were higher. Conclusion The root and root canal morphology of mandibular second molars in western Guangxi people are complex and changeable. The roots are mainly double root type in the Han ethnic group and the Zhuang ethnic group. C-shaped roots are also common. The detection rate of C-shaped roots in the Zhuang ethnic group was higher, and the symmetry rate of bilateral roots and that of bilateral root canals was higher in the Zhuang ethnic group than in the Han ethnic group.

Objective: To analyze the clinical characteristics of hand, foot and mouth disease (HFMD) caused by coxsackievirus A6 (CV-A6) in different age groups. Methods: From January 2015 to December 2017, throat swabs were collected from children with or suspected of having HFMD then quantitative real-time PCR was performed to detect enterovirus nucleic acid. HFMD cases caused by CV-A6 were divided into different groups according to age for comparison. Results: In total, there were 467 cases of HFMD caused by CV-A6 with the age ranging from 3 months to 16 years. There were 273 cases in the infants and young children group (< 3 years old), 131 cases in the pre-school group (3-6 years old), and 63 cases in the school-age group (> 6 years old). The peak incidence was found between May and November.Fever was the common symptom, and the rate of fever in infant group was the highest (220/273, 80.5%); The proportion of cases with leucocyte elevation in the infant group was the highest (127/273, 46.5%) than that in the school-age group (17/63, 27.0%) with a statistical significance. The skin erythra of the HFMD caused by CV-A6 were diverse in forms. Over two forms of skin erythra accounted for 53.9% (257/476) of all cases, and the cases in the infant group showing more forms of skin erythra (163/273, 59.7%). The oral herpes were mainly distributed in the upper palate and pharyngeal isthmus, but the school age group had the least number of distribution sites (0.89±0.86). The cases in the infant group showed higher incidence of skin rash at the elbow joint (109/273, 39.9%), knee (88/273, 32.6%), thigh (112/273, 41%), buttock (122/273, 44.7%) than the other two groups, However, the school age group showed lower incidence of skin rash in the lower leg (0/63, 0%) and thigh (6/63, 9.5%) than the other two groups. The differences between groups were statistically significant. All cases were cured clinically, no severe cases occurred. Among the 288 cases followed up for 6 months, 33 (33/288, 11.5%) suffered from nail exfoliation. Conclusions Different age groups of HFMD caused by CV-A6 had different clinical manifestations. In the infant group, more cases had fever and the erythra were more diverse in forms and wider in distribution. In addition, the increased leukocytes in routine blood test was also more common in the infant group.

Objective: To investigate the positive rate for 2019-nCoV tests and co-infections in Wuhan district. Methods: A total of 8 274 cases in Wuhan were enrolled in this cross-sectional study during January 20 to February 9, 2020, and were tested for 2019-nCoV using fluorescence quantitative PCR. Both respiratory tract samples (nasopharynx, oropharynx, sputum and alveolar lavage fluid) and non-respiratory tract samples (urine, feces, anal swabs, blood and conjunctival sac swabs) were collected. If both orf1ab and N genes are positive, they are classified as nucleic acid test positive group; if both orf1ab and N genes are negative, they are classified as negative group; if single gene target is positive, they are classified as suspicious group. Individuals were divided into male group and female group according to sex. At the same time, 316 patients were tested for 13 respiratory pathogens by multiplex PCR. Results: Among the 8 274 subjects, 2 745 (33.2%) were 2019-nCoV infected; 5 277 (63.8%) subjects showed negative results in the 2019-nCoV nucleic acid test; and 252 cases (3.05%) was not definitive (inconclusive result). The age of cases with COVID-19 patients and inconclusive cases was significantly higher than that of cases without 2019-nCoV infection (40 vs 56, t=27.569, P<0.001; 52 vs 56, t=6.774, P<0.001). The positive rate of 13 respiratory pathogens multiple tests was significantly lower in 104 subjects who were positive for 2019-nCoV compared with those in subjects who were negative for 2019-nCoV test (5.77% vs 18.39%, χ²=24.105, P=0.003). Four types of respiratory tract samples and five types of non-respiratory tract samples were found to be positive for 2019-nCoV nucleic acid test. Conclusion The 2019-nCoV nucleic acid positive rate in male is higher than in female. Co-infections should be pay close attention in COVID-19 patients. 2019-nCoV nucleic acid can be detected in non-respiratory tract samples.