Objective:To compare the efficacy between Kirschner wire and clavicular hook plate in the fixation of distal clavicle fractures in older children.Methods:A retrospective analysis was conducted of the 28 pediatric patients with distal clavicle fracture who had undergone surgical treatment at Department of Pediatric Orthopedics, The Second General Hospital of Fuzhou from December 2014 to December 2023. There were 20 boys and 8 girls. Age: 10.0 (9.0, 12.8) years old; 22 left sides and 6 right sides; by the Craig modified Neer classification: 22 cases of type Ⅱ, 5 cases of type Ⅳ, and 1 case of type Ⅴ. The patients were divided into 2 groups based on their internal fixation methods. The Kirschner wire group of 12 cases were treated with open reduction and Kirschner wire fixation; the hook plate group of 16 cases were treated with open reduction and clavicular hook plate fixation. Operation time, hospital stay, fracture healing time, incidence of complications, and scores at the final follow-up (the Constant-Murley shoulder function score, quick disabilities of the arm, shoulder, and hand (quickDASH) score, and visual analog scale (VAS) for aesthetic satisfaction) were recorded and compared between the 2 groups.Results:There were no statistically significant differences in the baseline data between the 2 groups, indicating comparability ( P>0.05). The postoperative follow-up period for the 28 patients was (14.8±6.5) months. The operation time in the Kirschner wire group [(34.3±12.1) min] was significantly shorter than that in the hook plate group [(49.4±16.8) min] ( P<0.05), and the VAS score for aesthetic satisfaction at the final follow-up in the Kirschner wire group [(8.2±1.1) points] was significantly higher than that in the hook plate group [(6.2±2.6) points] ( P<0.05). There were no statistically significant differences between the 2 groups in terms of fracture healing time, hospital stay, incidence of complications, or Constant-Murley score and quickDASH score at the final follow-up ( P>0.05). Conclusions:For pediatric patients with distal clavicle fracture, both Kirschner wire fixation and clavicular hook plate fixation can achieve good functional outcomes. However, Kirschner wire fixation has advantages of shorter operation time, higher postoperative aesthetic satisfaction, and no need of a secondary surgery for implant removal.

Objective: To investigate the osteogenic effect of β-tricalcium phosphate (β-TCP) and bone morphogenetic protein-2 (BMP-2) in the repair of the alveolar cleft. Methods : Fifty-nine patients with unilateral alveolar cleft who visited Capital Medical University School of Stomatology from January 2016 to May 2021 were included. They were divided into three groups according to the different bone repair materials: autologous bone, β-TCP and BMP-2 +β-TCP. The preoperative and postoperative CBCT data of the patients were imported into Mimics 21.0 software. The preoperative volume of the bone defect and the new volume of bone formation were calculated by the three-dimensional reconstruction method. The osteogenesis rate was calculated to evaluate the osteogenesis effect Results: The wounds in the three groups healed well after the operation, without implant material discharge, infection, dehiscence, rejection or other symptoms. Twelve months after the operation, CBCT scanning and three⁃dimensional reconstruction images of the three groups of patients showed the formation of new bone bridges in the alveolar ridge fissure area. The image density of the new bone tissue was not significantly different from that of normal bone tissue, and the continuity of the maxilla was re⁃ stored to varying degrees. The bone rate of autogenous bone was 65.00% ± 16.66%, β⁃ TCP group and BMP⁃2+ β⁃ The bone composition rate of TCP was 69.82% ± 17.60%, 71.35% ± 17.51%, respectively, and there was no significant dif⁃ ference compared with the autogenous bone group (P = 0.382, P = 0.244). The β⁃TCP and BMP⁃2+ β⁃TCP groups had no significant differences in bone rate (P = 0.789). Conclusion β⁃TCP could be used to replace autologous bone for alveolar cleft repair. The addition of BMP⁃2 to β⁃TCP did not significantly improve the osteogenesis rate.

OBJECTIVE: To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology. METHODS: We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated. RESULTS: This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively. CONCLUSION By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.
Humans ; COVID-19 ; CRISPR-Cas Systems ; Genotype ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; SARS-CoV-2/genetics* ; RNA ; COVID-19 Testing

Objective:To explore the diagnosis and treatment of humeral medial epicondyle fracture combined with radial neck fracture in children and adolescents.Methods:The clinical data were retro-spectively analyzed of the 12 pediatric patients with fractures of the humeral medial epicondyle plus the radial neck who had been admitted from February 2015 to August 2021 to Department of Pediatric Orthopedics, The Second Hospital of Fuzhou. There were 6 males and 6 females, with an age of (11.1 ± 2.5) years. According to the Papavasiliou classification, the humeral medial epicondyle fractures were type Ⅱ in 7 cases, type Ⅲ in 2 cases and type Ⅳ in 3 cases; according to the Judet classification, the radial neck fractures were type Ⅰ in 3 cases, type Ⅱ in 4 cases and type Ⅲ in 5 cases. Two Judet-Ⅰ radial neck fractures were missed by X-ray exam-ination but diagnosed by CT examination. Of the humeral medial epicondylar fractures, 9 were treated by open reduction and hollow screwing and 3 by closed reduction and Kirschner wiring. Of the radial neck fractures, 8 were treated by closed reduction and elastic intramedullary nailing and 4 conservatively. Fracture healing was followed up by postoperative radiographs. At the last follow-up, the carrying angles were measured, Kim Elbow Function Score (KEPS) was used to evaluate the functional recovery of the injured limb, and related complications were recorded.Results:All the 12 patients were followed up for (40.0±25.6) months. Fractures headed after (6.3±1.2) weeks. At the last follow-up, the carrying angle was 15.5°±2.6° on the injured side and 14.7°±2.0° on the healthy side, showing no significant difference ( P>0.05); KEPS was (96.3±5.3) points on the injured side and (98.8±2.3) points on the healthy side, showing no significant difference either ( P>0.05). No incision infection, bone nonunion, elbow valgus, joint stiffness or other complications were found; the postoperative elbow stability recovered well. Conclusions:As the fracture of the humeral medial epicondyle combined with the radial neck fracture is a special type of injury of straightened elbow during valgus stress in children and adolescents, it is likely to be missed in diagnosis. The goal of treatment is good functional recovery by restoring the articular match and elbow stability.

Objective:To investigate the clinical results of Bouquet technique [multiple elastic stable intramedullary nails (ESIN)] in the treatment of unstable pediatric femoral shaft fractures.Methods:From August 2016 to October 2019, 25 pediatric patients (26 sides) with unstable femoral shaft fracture were treated with Bouquet technique at Department of Pediatric Orthopedics, The Second Fuzhou Hospital Affiliated to Xiamen University. They were 17 boys and 8 girls, with a mean age of 7.8 years at injury (from 5 to 11 years). Their body weight ranged from 23 kg to 62 kg, with an average of 36.3 kg. Long oblique fractures happened in 6 cases (6 sides) and comminuted fractures in 19 cases (20 sides). Ten cases (11 sides) were fixated with 3 ESINs (type 3A) and 15 cases (15 sides) with 4 ESINs (type 4A in 6 cases and type 2A+2B in 9 cases). The multiple ESINs were inserted from the medial and lateral cortex of the distal femur through the fracture sides until the head ends of the nails were distributed evenly at the proximal femur. Recorded were the fracture healing time, complications, fracture reduction quality and hip function at the last follow-up.Results:All the 25 patients were followed up for 18 to 27 months (mean, 22.4 months). Their fractures got united after 4 to 8 weeks (mean, 6.3 weeks). One case presented with femoral overgrowth of 1.1 cm and another case with nail tail irritation. Follow-ups revealed that none of the patients had such complications as incision infection, limited joint movement, nonunion or delayed fracture union. The limb alignment recovered well with no angulation, shortening or rotational deformity. At the last follow-up, according to the efficacy evaluation system for intramedullary nails by Flynn et al., the quality of reduction of femoral shaft fractures was assessed as excellent in 24 sides and as good in 2 sides, giving an excellent and good rate of 100% (26/26). According to the hip Harris scoring system, the hip function was assessed as excellent in all the 26 sides, giving an excellent rate of 100% (26/26).Conclusion:Bouquet technique is safe and effective in the treatment of unstable pediatric femoral shaft fractures, leading to fine clinical results.

Aim: To investigate the effects of Scutellarin (Scu) on microvascular endothelial cells (MEC) with ischemia/reperfusion injury (I/R) and its mechanisms involved. Methods Microvascular endothelial cells were used to build model of I/R, and the protective effects of Scu were observed. Cell viability was determined by MTT assay; SOD. MDA and LDH levels were determined by biochemical method; ICAM-1, VCAM-1, caspase-3 and LC3 mRNA expressions were detected by RT-PCR; LC3, CREB and TLR4 expressions were detected by Western blot. Results MTT assay showed that the cell viability was rescued by 50 μmol • L

OBJECTIVE：To e stablish the method for simultaneous determination of 10 kinds of active components in Tibetan medicine Siwei jianghuang prescription ，and to optimize its decoction technology. METHODS ：HPLC method was adopted. Using soaking time ，the amount of added water ，decoction time and decoction times as factors ，comprehensive score of the contents of 10 kinds of components and solid extracts rate as response values ，one the basis of single factor test ，Box-Behnken response surface method was used to optimize its decoction technology. RESULTS ：The linear range of gallic acid ，corilagin，magnoflorine， ellagic acid ， hydrochloric jatrorrhizine ， hydrochloride palmatine ， hydrochloride berberine ， bisdemethoxycurcumin， demethoxycurcumin and curcumin were 0.280 6-1.683 6，0.289 6-1.737 6，0.320 8-1.924 8，0.116 0-0.696 0，0.018 9-0.113 5， 0.013 3-0.079 9，0.092 3-0.553 8，0.025 5-0.153 0，0.036 1-0.216 3，0.041 0-0.245 7 µg（all r were 0.999 9），respectively. The limits of quantitation were 0.28，14.48，3.21，11.60，1.89，4.44，0.46，0.26，0.36，0.41 ng，respectively. The limits of detection were 0.11，4.14，1.24，3.32，0.58，1.33，0.13，0.09，0.14，0.12 ng，respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％. The recoveries were 92.56％-103.69％（RSDs were 0.90％-3.81％，n＝6）. The optimal decoction technology included soaking 60 min，adding 8-fold（mL/g）water，decoction for twice ，lasting for 65 min each time. In 6 validation tests ，comprehensive scores were 3.323 2-3.422 4，and the absolute value of the relative error with the predicted value （3.437 4）was less than 2％.CONCLUSIONS：Established method is simple and repeatable ，and can be used for simultaneous determination of 10 kinds of active components in Siwei jianghuang prescription. Optimized decoction technology is stable and feasible.

Objective To analyze the effect of Noggin silencing on the BMP and Wnt signaling pathways in hair follicle development.Methods The expression of BMP-2, BMP-4, BMPR-IA, BMP-6, BMP-7, LEF-1 andβ-catenin in Noggin silencing MC3T3-E1 stable cell line was detected by RT-PCR and western blot.Results RT-PCR results showed that the expressions of five genes in BMP signaling pathway were all significantly influenced by Noggin silencing, the ex-pressions of BMP-2 (P<0.001), BMP-4 (P<0.01), BMP-6 (P<0.001) and BMP-7 (P<0.001) were all increased and the expression of BMPR-IA (P<0.01) was decreased.While the expressions of the two genes LEF-1 (P<0.001) and β-catenin ( P<0.001) in Wnt signaling pathway were significantly decreased.Western blot results showed that the ex-pressions of these proteins in the two signaling pathways were also affected.The expressions of BMP-2 (P<0.05), BMP-4 (P<0.05), BMP-6 (P<0.05) and BMP-7 (P<0.05) were all increased, while the expressions of BMPR-IA (P<0.05), LEF-1 (P<0.01) andβ-catenin (P<0.001) were decreased.Conclusions There may be a negative feedback regulation of Noggin on the BMP signaling pathway in vitro, but a positive feedback regulation on the Wnt signaling pathway in vitro.It provides certain evidence for studies on the effect of Noggin gene on BMP and Wnt signaling pathways in vivo. There may be an interaction between hair follicle development-related signaling pathways, which still needs further experi-ments to prove.

Objective To establish a stable transfection cell line of iRhom2 and its mutant through recombinant lentivirus infection.Methods The full-length gene of iRhom2 and its mutant were cloned into the lentivirus vector Lenti-OE-Flag, and got recombinant lentiviral vector of Lenti-OE-iRhom2 and Lenti-OE-iRhom2mut.The constructed recombi-nant lentivirus vectors were transfected into HEK-293T packaging cells to obtain the recombinant virus.Vero cells were in-fected with recombinant virus.The stable expressing cell lines were obtained by pressure screening with puromycin. Results The recombinant lentivirus vectors were constructed and the recombinant virus was obtained.The stable express-ing cell lines were obtained using virus infection and the protein expression was testified with Western blotting.Conclu-sions Stable iRhom2-expressing Vero cell line and its mutant are achieved by recombinant lentivirus infection.It paves the way for future study on biological functions and mechanism of iRhom2.

Objective To explore the effects of electroacupuncture at Shenting (DU24) and Baihui (DU20) on learning and memory ability in the cerebral ischemia-reperfusion rats and its possible mechanism. Methods 45 male Sprague-Dawley rats were randomly divided into sham group (n=15), ischemia group (n=15) and electroacupuncture group (n=15). The latter 2 groups were modeled as focal cerebral ischemia-reperfusion injury, and the electroacupuncture group received electroacupuncture at Shenting (DU24) and Baihui (DU20) for 7 days. They were tested with Morris Water Maze, observed with Nissl's staining. The protein expression of RhoA was detected with Western blotting. Results The learning and memory ability improved in the electroacupuncture group (P<0.05), the injury of the neurons reduced (P<0.05) and the expression of RhoA in hippocampus decreased compared with the ischemia group (P<0.05). Conclusion Electroacupuncture could ameliorate the learning and memory ability in ischemia- reperfusion rats, which may relate with the inhibition of the expression of RhoA in hippocampus.