BACKGROUND:Current osteoarthritis modeling methods include anterior cruciate ligament transection(ACLT)and ACLT combined with medial meniscal anterior horn resection.ACLT requires excessive postoperative exercise,which is time and labor-intensive.Complete removal of anterior horn of the medial meniscus can cause collateral damage and increase variability in modeling outcomes,requiring higher surgical skills from the surgeon. OBJECTIVE:To modify and simplify the traditional method to create animal osteoarthritis model and compare osteoarthritis symptoms of different modeling methods under a low-load exercise environment. METHODS:Forty-eight Sprague-Dawley rats were randomly assigned in four groups(n=12 per group):sham operation(complete exposure of the knee cavity of the left hind limb followed by suturing the joint cavity and skin),ACLT,ACLT+anterior horn resection(removal of the anterior horn of the medial meniscus)and ACLT+anterior horn tear(anterior horn tear of the medial meniscus).At 4 weeks after modeling,the rats were euthanized and their knee specimens were collected for gross observation,X-ray and CT scans,pathological observation,and PCR detection. RESULTS AND CONCLUSION:Gross observation:Mild meniscal wear was observed in the ACLT group.In the ACLT+anterior horn tear group,severe wear of the lateral condyle articular surface,mild wear of the medial condyle articular surface,severe meniscal wear,and full wear of the medial meniscus were observed.The ACLT+resection group showed severe wear of the lateral condyle articular surface,mild wear of the medial condyle articular surface,absence of the anterior horn of the medial meniscus,and meniscus wear area＞50%.Imaging examinations showed no significant difference among the four groups.However,the anterior tibial translocation sign was observed in the three operation groups and the anterior horn of the medial meniscus was missing in the ACLT+anterior horn resection group.Histopathological section observation:Hematoxylin-eosin,toluidine blue,and Sirius red staining showed smooth joint surfaces in the sham operation group and ACLT group;cartilage damage and matrix degradation were evident in the ACLT+anterior horn tear and ACLT+anterior horn transection groups,with less cartilage damage and matrix degradation in the ACLT+anterior horn tear group.PCR results showed higher mRNA expressions of interleukin 1β,interleukin 6,interleukin 8,tumor necrosis factor α,matrix metalloproteinase 1 and matrix metalloproteinase 3 and lower mRNA expressions of aggrecan in the ACLT+anterior horn tear group and ACLT+anterior horn resection group than in the sham operation group and ACLT group(P＜0.05).The mRNA expressions of interleukin 6,matrix metalloproteinase 1,and matrix metalloproteinase 3 were higher in the ACLT + anterior horn resection group than in the ACLT +anterior horn tear group(P＜0.05).To conclude,ACLT alone is less likely to induce osteoarthritis with obvious cartilage wear.ACLT combined with anterior horn resection or tear of the medial meniscus can induce obvious symptoms of osteoarthritis and achieve similar modeling effects.

Objective To evaluate the molluscicidal effects and costs of spraying 20% suspension concentrate of pyricloben-zuron sulphate (SCPS) with drones against Pomacea canaliculata in paddy environments, so as to provide insights into the extensive applications of pyriclobenzuron against P. canaliculata. Methods On July 2022, a paddy field was selected from Nanchang City, Jiangxi Province as the study area, and 72 independent rectangular plots measuring 2 m × 1 m were allocated in the study area, with 1 m interval between each plot, and 20 P. canaliculata snails gently placed in each plot. The activity of 25% wettable powder of pyriclobenzuron sulphate (WPPS) by manual spraying at doses of 0.50, 1.00, 2.00 g/m² and 4.00 g/m² against P. canaliculata was tested in 54 plots, and manual spraying of 50% wettable powder of niclosamide ethanolamine salt (WPNES) at a dose of 0.10 g/m² served as a chemical control, while manual spraying of the same volume of clean water served as a blank control, with 9 plots in each group. The activity of SCPS against P. canaliculata was tested in the remaining 18 plots. Based on the molluscicidal tests of WPPS, the molluscicidal effect of SCPS by manual spraying at doses of 0.20, 0.30, 0.40 g/m² and 0.50 g/m² against P. canaliculata was evaluated, and manual spraying of WPNES at a dose of 0.10 g/m² served as a chemical control, while manual spraying of the same volume of clean water served as a blank control, with three plots in each group. On July 2023, 14 paddy fields with a mean living P. canaliculata density of > 5 snails/m² were selected from Yujiang District, Yingtan City, Jiangxi Province for molluscicidal tests. Based on the molluscicidal effect of pyriclobenzuron against P. canaliculata in plots, the molluscicidal effects of WPPS by manual spraying at doses of 0.25, 0.50 g/m² and 1.00 g/m² and manual applications of WPPS at dose of 0.25, 0.50, 1.00 g/m² and 2.00 g/m² mixed with soil were tested, and manual spraying of 0.10 g/m² WPNES served as a chemical control group, while manual spraying of the same volume of clean water served as a blank control, with one paddy field in each group. Based on the effect of pyriclobenzuron against P. canaliculata in plots, the activity of SCPS sprayed with drones at doses of 0.25 g/m² and 0.50 g/m² mixed in water at 2 kg/667 m² and 4 kg/667 m² was tested against P. canaliculata, and spraying of the same volume of clean water with drones served as a blank control. All P. canaliculata snails were captured 3 days and 7 days following chemical treatment in plots and paddy fields and identified for survival, and the mortality and corrected mortality of P. canaliculata snails were estimated. In addition, the areas of chemical treatment, amount of molluscicide use and labor costs of chemical treatment were estimated in molluscicidal tests in paddy fields, and the costs of chemical treatment for an area covering 667 m² by drones and manual applications were calculated. Results The mortality of P. canaliculata snails was all 100% in plots 3 days and 7 days following spraying WPPS at doses of 0.50, 1.00, 2.00 g/m² and 4.00 g/m², and the mortality rates of P. canaliculata snails were 66.67% to 100.00% 3 days post-treatment with SCPS at various doses (χ² = 277.897, P < 0.05) and 76.67% to 100.00% 7 days post-treatment (χ² = 274.206, P < 0.05). The mortality rates of P. canaliculata snails were 98.19% to 100.00% 3 days post-treatment with WPPS at various doses in paddy fields. There was a significant difference in the mortality of P. canaliculata snails among WPPS treatment groups and controls (χ² = 270.778, P < 0.05), and there were no significant differences between WPPS treatment groups and the chemical control group (all P values > 0.05), while there were significant differences in the mortality of P. canaliculata snails between WPPS treatment groups and the blank control group (all P values < 0.05). The mortality rates of P. canaliculata snails were 89.83% to 95.31% 3 days post-treatment with SCPS at various doses sprayed with drones, and there was a significant difference in the mortality of P. canaliculata snails among SCPS treatment groups and the blank control group (χ² = 1 132.892, P < 0.05). There were no significant differences in the mortality of P. canaliculata snails among SCPS treatment groups or water mixture groups (all P values > 0.05), and there were significant differences in the mortality of P. canaliculata snails between SCPS treatment groups and the blank control group (all P values < 0.05). The mortality rates of P. canaliculata snails were 94.62% to 100.00% 7 days post-treatment with SCPS at various doses sprayed with drones, and there was a significant difference in the mortality of P. canaliculata snails among SCPS treatment groups and the blank control group (χ² = 1 266.932, P < 0.05), with the highest mortality found following spraying 0.50 g/m² SCPS mixed in 2 kg/667 m² water with drones (P < 0.05). The costs of P. canaliculata snail control by drones and manually were 35.85 Yuan/667 m² and 43.33 Yuan/667 m²; however, the snail control efficiency was 6.67 times higher by drones than by manual applications. Conclusions SCPS sprayed with drones is highly active against P. canaliculata snails in paddy fields. SCPS sprayed with drones is highly efficient and low in cost for P. canaliculata snail control in paddy fields, beaches and river courses.

Objective:To investigate the value of systemic inflammatory response syndrome (SIRS), pediatric sequential organ failure assessment (pSOFA) and pediatric critical illness score (PCIS) in predicting mortality of pediatric sepsis in pediatric intensive care units (PICU) from Southwest China.Methods:This was a prospective multicenter observational study. A total of 447 children with sepsis admitted to 12 PICU in Southwest China from April 2022 to March 2023 were enrolled. Based on the prognosis, the patients were divided into survival group and non-survival group. The physiological parameters of SIRS, pSOFA and PCIS were recorded and scored within 24 h after PICU admission. The general clinical data and some laboratory results were recorded. The area under the curve (AUC) of the receiver operating characteristic curve was used to compare the predictive value of SIRS, pSOFA and PCIS in mortality of pediatric sepsis.Results:Amongst 447 children with sepsis, 260 patients were male and 187 patients were female, aged 2.5 (0.8, 7.0) years, 405 patients were in the survival group and 42 patients were in the non-survival group. 418 patients (93.5%) met the criteria of SIRS, and 440 patients (98.4%) met the criteria of pSOFA≥2. There was no significant difference in the number of items meeting the SIRS criteria between the survival group and the non-survival group (3(2, 4) vs. 3(3, 4) points, Z=1.30, P=0.192). The pSOFA score of the non-survival group was significantly higher than that of the survival group (9(6, 12) vs. 4(3, 7) points, Z=6.56, P<0.001), and the PCIS score was significantly lower than that of the survival group (72(68, 81) vs. 82(76, 88) points, Z=5.90, P<0.001). The predictive value of pSOFA (AUC=0.82) and PCIS (AUC=0.78) for sepsis mortality was significantly higher than that of SIRS (AUC=0.56) ( Z=6.59, 4.23, both P<0.001). There was no significant difference between pSOFA and PCIS ( Z=1.35, P=0.176). Platelet count, procalcitonin, lactic acid, albumin, creatinine, total bilirubin, activated partial thromboplastin time, prothrombin time and international normalized ratio were all able to predict mortality of sepsis to a certain degree (AUC=0.64, 0.68, 0.80, 0.64, 0.68, 0.60, 0.77, 0.75, 0.76, all P<0.05). Conclusion:Compared with SIRS, both pSOFA and PCIS had better predictive value in the mortality of pediatric sepsis in PICU.

Objective To compare the size discrepancy between ultrasonic and pathological measurement of solitary cN0M0 papillary thyroid microcarcinoma(PTMC),and to explore their correlation with lymph node metastasis.Methods From April 2021 to January 2022,234 patients with solitary cN0M0 PTMC who received thyroid lobectomy or total thyroidectomy in the Department of Thyroid and Breast Surgery of Nanjing University of Chinese Medicine,Affiliated Hospital of Integrated Traditional Chinese and Western Medicine were analyzed retrospectively.The size discrepancy between ultrasonic and pathological measurement were compared,and the risk factors of central lymph node metastasis were analyzed.Results The mean of maximum diameter of PTMC measured by ultrasound was 6.8(range 5.6 to 8.4)mm,which was significantly bigger than that measured by pathology 5.0(range 4.0 to 7.0)mm(P=0.000).Of them,37.2%of the tumor size measured by ultrasound is consistent with pathology,61.1%of the tumor size measured by ultrasound is bigger than that measured by pathology,and only 1.7%of the tumor size measured by ultrasound is smaller than that measured by pathology.There was a linear correlation between the diameter measured by ultrasound and pathology.And the regression equation can be expressed as:pathological diameter=0.799×ultrasonic diameter-0.221.In addition,28.6%patients had central lymph node metastasis.Multivariate Logistic regression analysis showed that the diameter measured by pathology is a risk factor for central lymph node metastasis in patients(OR=17.845,95%CI:2.507-127.025,P=0.004),and the cutoff value is 5.5 mm which corresponded to the diameter measured by ultrasound as 7.2 mm.Conclusions The sizes of solitary cN0M0 PTMC measured by ultrasound and pathology are different but also correlated.PMTC with pathological diameter>5.5 mm with its corresponding ultrasonic diameter as 7.2 mm indicated an increased risk of central lymph node metastasis.

Complicated crown root fracture is a serious combined fracture of the enamel,dentin,and cementum in dental trauma.The treatment method is complicated.During the procedure,the condition of pulp,periodontal,and tooth body should be thoroughly evaluated,and a multidisciplinary approach combined with sequential treatment is recom-mended.This case reported the different treatment and repair processes of one case of two affected teeth after complicat-ed crown root fracture of upper anterior teeth,including regrafting of broken crown after flap surgery at the first visit,di-rect resin repair to remove broken fragments,and pulp treatment and post-crown repair at the second visit.After 18 months of follow-up,the preservation treatment of the affected teeth with complicated crown root fracture was achieved.Therefore,fragment reattachment and post-crown restoration are feasible treatment options for children with complicated crown root fracture.

Objective:To provide references for the clinical popularization of stainless steel crowns by analyzing the clinical application and affecting factors of stainless steel crowns used in primary molars based on a questionnaire survey.Methods:Pediatric dentists from all over the country in June to December 2013 and June to December 2022 were selected to answer the survey questionnaire through the online or offline methods for analyzing the clinical application and influencing factors of stainless steel crowns used in primary molars.Results:A total of 712 and 580 questionnaires were distributed, and 608 and 512 valid questionnaires were recovered respectively in 2013 and 2022. The effective rates of the questionnaires were 85.4% and 88.3%. Among them, the usage rates of stainless steel crowns in 2013 and 2022 were 52.6% (320/608) and 78.1% (400/512) respectively, with a statistically significant difference (χ 2=78.68, P<0.001). The main reasons for not selecting stainless steel crowns in 2013 were high technical sensitivity [52.8% (152/288)], followed by the impact of crown edge on gingiva [50.0% (144/288)] and poor aesthetics [27.8% (80/288)], while in 2022 were the inadequate equipage in hospitals [53.6% (60/112)] and poor aesthetics [21.4% (24/112)]. The main reasons for choosing stainless steel crowns restoration were good protection effect, less tooth fracture [78.1% (250/320) in 2013 and 82.5% (330/400) in 2022], followed by low dropout rate [62.5% (200/320) in 2013 and 68.0% (272/400) in 2022]. The proportions of stainless steel crowns restoration for primary molars after root canal therapy were 51.3% (312/608) and 76.6% (392/512) respectively with statistically significant (χ 2=75.88, P<0.001). The proportions after pulp capping treatment for deep caries were 13.2% (80/608) and 53.1% (272/512) respectively, and the difference was statistically significant (χ 2=206.01, P<0.001). Conclusions:During the 10 years from 2013 to 2022, the technical sensitivity of stainless steel crowns had gradually decreased, and the application in China had gradually increased. However, some hospitals have not yet equipped, which limits the application of stainless steel crowns. Therefore, training efforts should be derived to promote the clinical popularization of stainless steel crowns.

A cross-sectional study was conducted on 1 052 patients with coronary heart disease (CHD) who visited 10 community health service centers in Chaoyang and Haidian districts of Beijing from March 2019 to September 2019. Basic information, medical history, cardiovascular risk factors, β-blocker use, height, weight, blood pressure level and resting heart rate were collected by face-to-face interview;biochemical tests and on-site physical examination were performed;and the influencing factors of heart rate control were analyzed by logistic regression. There were 563 males (53.5%) and 489 females(46.5%),with a mean age of (66.7±9.5) years. The rate of β-blockers use was 43.6% (459/1 052). The average resting heart rate was (70±8)/min, and only 14.5% (153/ 1 052) patients had the heart rate under control. The resting heart rate control rate in patients with β-blockers use was 17.9% (82/459), higher than that of those without β-blockers use [12.0% (71/593), χ 2=7.23, P=0.007]. Moderate leisure activities were beneficial to the control of resting heart rate ( OR=2.14, 95% CI: 1.26-3.62). The study shows that β-blockers use and resting heart rate control in CHD patients in Beijing community health institutions were both at low levels, and the resting heart rate was not well controlled even in patients taking beta-blockers.

Objective:To investigate the effects of different chimerism strategies and different immune ways on the two antigen-dominant regions of Xinjiang hemorrhagic fever virus (XHFV) glycoprotein.Methods:The 5' end was added or not added with interleukin-2 (IL-2) signal peptide and the general-purpose auxiliary T cell epitopes as different design strategies. GcⅠ and GcⅡ and the epitopes previously identified on GcⅠ (Gc 233-248, Gc 241-256 and Gc 281-296) were fused and constructed into the eukaryotic expression vector pVAX1 and the prokaryotic expression vector pET-28a. The recombinant prokaryotic plasmid transformed into E.coli BL21 was induced and purified, and the recombinant eukaryotes were extracted by indirect immunofluorescent assay. BALB/c mice were immunized by protein immunity, gene immunity, and DNA prime-protein boost immunity. The IgG antibody level was measured by ELISA. The immune effect was evaluated by the proliferation of T-lymphocytes and the content of cytokines in the spleen. Results:The results of double enzyme digestion and sequencing showed that eight recombinant plasmids were successfully constructed, and the recombinant eukaryotes were successfully expressed in vitro by fluorescence microscopy. After three times of immunization, the IgG level and the proliferation of T-lymphocytes in the spleen of mice in the experimental group were significantly higher than those in the control group ( P<0.01). The mass concentration test results of Th2 cytokines IL-4 and Th1 cytokines interferon-gamma (IFN-γ) revealed that the response of the DNA prime-protein boost immunity was biased to Th1. Conclusions:The multi-epitope chimeric vaccine of XHFV glycoprotein was successfully constructed, and the target antigen could be expressed effectively in vivo. The immune groups stimulated stronger humoral and cellular immune responses compared with the control group. Among them, the immune effect of pVAX1-ST(GcⅠe+GcⅡ) combined with recombinant protein r(GcⅠe+GcⅡ) was the best, and it is expected to be a new candidate vaccine for XHFV.

With the emergence of DNA nanotechnology in the 1980s, self-assembled DNA nanostructures have attracted considerable attention worldwide due to their inherent biocompatibility, unsurpassed programmability, and versatile functions. Especially promising nanostructures are tetrahedral framework nucleic acids (tFNAs), first proposed by Turberfield with the use of a one-step annealing approach. Benefiting from their various merits, such as simple synthesis, high reproducibility, structural stability, cellular internalization, tissue permeability, and editable functionality, tFNAs have been widely applied in the biomedical field as three-dimensional DNA nanomaterials. Surprisingly, tFNAs exhibit positive effects on cellular biological behaviors and tissue regeneration, which may be used to treat inflammatory and degenerative diseases. According to their intended application and carrying capacity, tFNAs could carry functional nucleic acids or therapeutic molecules through extended sequences, sticky-end hybridization, intercalation, and encapsulation based on the Watson and Crick principle. Additionally, dynamic tFNAs also have potential applications in controlled and targeted therapies. This review summarized the latest progress in pure/modified/dynamic tFNAs and demonstrated their regenerative medicine applications. These applications include promoting the regeneration of the bone, cartilage, nerve, skin, vasculature, or muscle and treating diseases such as bone defects, neurological disorders, joint-related inflammatory diseases, periodontitis, and immune diseases.
Nucleic Acids/chemistry* ; Regenerative Medicine ; Consensus ; Reproducibility of Results ; DNA/chemistry*

Objective:To obtain purified protein antigen of guertu virus (GTV) nucleoprotein (NP) and establish a rapid and accurate enzyme-linked immunosorbent assay (ELISA) method for detection of GTV antibody.Methods:Codon optimized GTV NP encoding genes were synthesized, cloned into the pet32a (+) vector, and recombinant expression plasmids were constructed and transformed into BL21 (DE3). Recombinant protein (rNP) obtained from the optimized expression were purified over a Ni column and identified by SDS-PAGE and Western blot. The purified protein was used as the antigen to optimize the reaction conditions, and an indirect ELISA assay for GTV IgG antibody was developed and optimized, which was evaluated and initially applied.Results:The prokaryotic expression plasmid pet32a-NP was successfully constructed, the recombinant protein was highly expressed in E. coli in the form of inclusion bodies, the size was about 44 kD, and the results of Western blot indicated that the recombinant protein had good antigenicity with GTV positive serum. The optimized ELISA (GTV-rNP-iELISA) established in this study showed strong specificity, high sensitivity, and the coefficient of variation within and between batches is less than 10%, and has good repeatability; the detection results are consistent with the IFA detection results. Using the established ELISA method to detect 162 sheep sera from some regions of Xinjiang in 2017-2019, the total positive rate of antibodies was 39.8%.Conclusions:The GTV NP antibody detection ELISA method has good sensitivity, reproducibility, and specificity and has the potential to be a powerful tool for the diagnosis and serological investigation of GTV infection.