ABSTRACT
Objective To investigate antigen optimization of Shisa like protein 1 (SHISAL1) for preparing mouse anti-human SHISAL1 polyclonal antibody and to identify the specificity of the prepared antibody. Methods Bioinformatics was employed to predict the antigenic epitope region of SHISAL1 protein, and then a polypeptide composed of amino acid residues from the site of 28 to 97 of SHISAL1, termed SHISAL1-N, was selected as the antigen. The coding region of SHISAL1-N was cloned by molecular cloning technique, and then it was inserted into pET-28a to generate pET28a-SHISAL1-N recombinant plasmid. The two recombinant plasmids pET28a-SHISAL1-N and pET28a-SHISAL1 were transformed into BL21 (DE3) bacteria and induced to express by IPTG. The two proteins were purified and immunized to female Kunming mice, respectively. The specificities and sensitivities of the acquired antibodies were detected by Western blot analysis, immunoprecipitation and immunofluorescent cytochemical staining. Results pET28a-SHISAL1-N recombinant plasmid was successfully constructed, and the two fused proteins, SHISAL1 and SHISAL1-N, were induced to express. Moreover, two types of SHISAL1 mouse polyclonal antibodies, derived from SHISAL1-N and SHISAL1 antigens, were obtained. Western blot results showed that the antibody prepared from SHISAL1 antigen was less specific and sensitive compared with the antibody prepared from SHISAL1-N antigen which could specifically identify different endogenous SHISAL1 protein. Immunoprecipitation results showed that SHISAL1-N antibody could specifically pull down SHIISAL1 protein in hepatocellular carcinoma cells and immunofluorescence results demonstrated that SHISAL1-N antibody could specifically bind to SHISAL1 protein in the cytoplasm. Conclusion We have optimized the SHISAL1 antigen and prepared the mouse anti-human SHISAL1 polyclonal antibodies successfully, which can be used for Western blot analysis, immunoprecipitation and immunofluorescence cytochemical staining.
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Animals , Female , Humans , Mice , Antibodies , Antibody Specificity , Blotting, Western , Cloning, Molecular , Epitopes/geneticsABSTRACT
Objective:To explore the curative effect of modified double Kirschner wire compression fixation in the treatment of skeletal mallet fingers, and analyze the related influencing factors of curative effect.Methods:The clinical data of 100 patients with skeletal mallet fingers from January 2018 to December 2020 in Yanzhou Branch of Affiliated Hospital of Jining Medical University were retrospectively analyzed, and the patients were treated with modified double Kirschner wire compression fixation. Before surgery and 8 weeks after surgery, the Fugl-Meyer score, simple test for evaluating hand function (STEF) score and visual analogue score (VAS) were assessed, and the flexion range of knuckles was measured. The curative effect was evaluated by Crawford standard, excellent and good means was good prognosis, medium and poor was poor prognosis. Binary Logistic regression was used to analyze the influencing factors of curative effect in patients with skeletal mallet fingers, Spearman method was used to analyze correlation.Results:All incisions healed in the first stage 14 d after surgery, the function and structure of the hand recovered well, and related incision skin necrosis and needle tract infection did not occur. Compared with before surgery, the Fugl-Meyer score, STEF score and flexion range of knuckles 8 weeks after surgery were significantly higher: (62.58 ± 4.56) scores vs. (32.33 ± 2.84) scores, (91.31 ± 3.19) scores vs. (62.51 ± 3.42) scores and (66.72 ± 3.65)° vs. (45.56 ± 2.31)°, the VAS was significantly lower: (2.65 ± 1.19) scores vs. (5.68 ± 1.43) scores, and there were statistical differences ( P<0.01). According to Crawford standard, good prognosis was in 79 cases, and poor prognosis was in 21 cases. There were no statistical differences in gender composition, injury location, cause of injury and type of injury between good prognosis patients and poor prognosis patients ( P>0.05); compared with the poor prognosis patients, the good prognosis patients were younger, the time from injury to surgery was shorter, the injured parts were mainly the little finger and ring finger, and there were statistical differences ( P<0.05 or <0.01). Binary Logistic regression analysis result showed that age, time from injury to operation and injury site were independent risk factors of curative effect in patients with skeletal mallet fingers ( OR = 4.62, 5.94 and 2.33; 95% CI 1.06 to 20.14, 2.23 to 15.81 and 1.12 to 4.82; P<0.05 or <0.01). Spearman correlation analysis result showed that the curative effect was positively correlated with age, time from injury to operation and injury location ( r = 0.25, 0.62 and 0.43; P<0.05 or <0.01). Conclusions:Modified double Kirschner wire compression fixation in the treatment of skeletal mallet fingers can promote the recovery of finger function after surgery, but age, damaged location and time of visit are independent risk factors of curative effect.
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Objective:To explore the method and effect in repairing the defect of fingertip with lateral V-Y advancement flap with one side palmar proper digital artery.Methods:From October 2014 to May 2019, Department of the Hand and Foot Surgery, the Third People's Hospital of Jining(Yanzhou District People's Hospital of Jining City) treated 34 digits of 27 cases with a defect area of 0.5 cm×0.5 cm-1.5 cm×2.0 cm. A lateral V-Y advancement flap with one side palmar proper artery was used to repair the fingertip defect, and the flap size was 1.7 cm×1.0 cm-4.5 cm×1.5 cm. Twenty cases entered long-time follow-up after operation, with 7 cases lost in follow-up, 16 cases were reviewed at outpatient and 4 by WeChat.Results:All the flaps of 34 digits of 27 cases survived. The color of the flaps were close to or completely normal to the surrounding tissue, the texture was soft and the appearance was good. The TPD of the flap was 2.0-6.0 mm. The follow-up time ranged from 22 to 77 months, with an average of 31.45 months. The flexion and extension function of the digits were good with total range of motion(ROM) of the thumb was > 90 °; total active motion (TAM) of the fingers was 260 °-200 °. The fingers of 1 case had hook nail or hook finger deformity. According to the Evaluation Trial Standard of Upper Limb Partial Function of Hand Surgery of Chinese Medical Association, 18 cases were excellent and 2 cases were good.Conclusion:The lateral V-Y advancement flap with one side palmar proper digital artery is easy to operate. The blood supply of the flap is reliable, with good sensation. The flexion and extension of the digits are good, and the appearance and texture of the flap are good.